{"id":2197,"date":"2026-05-20T22:29:58","date_gmt":"2026-05-20T14:29:58","guid":{"rendered":"https:\/\/www.fanyamin.com\/wordpress\/?p=2197"},"modified":"2026-05-20T22:29:58","modified_gmt":"2026-05-20T14:29:58","slug":"%e9%80%9a%e8%bf%87%e4%bf%83%e8%bf%9b%e8%83%86%e5%9b%ba%e9%86%87%e6%8e%92%e6%b3%84%ef%bc%8c%e6%8a%91%e5%88%b6-asgr1-%e5%8f%af%e9%99%8d%e4%bd%8e%e8%84%82%e8%b4%a8%e6%b0%b4%e5%b9%b3-full","status":"publish","type":"post","link":"https:\/\/www.fanyamin.com\/wordpress\/?p=2197","title":{"rendered":"\u901a\u8fc7\u4fc3\u8fdb\u80c6\u56fa\u9187\u6392\u6cc4\uff0c\u6291\u5236 ASGR1 \u53ef\u964d\u4f4e\u8102\u8d28\u6c34\u5e73 – full"},"content":{"rendered":"

\u901a\u8fc7\u4fc3\u8fdb\u80c6\u56fa\u9187\u6392\u6cc4\uff0c\u6291\u5236 ASGR1 \u53ef\u964d\u4f4e\u8102\u8d28\u6c34\u5e73<\/h1>\n

\u539f\u6587\uff1aJu-Qiong Wang et al., Nature<\/em> 608, 413-420 (2022)
\nDOI: https:\/\/doi.org\/10.1038\/s41586-022-05006-3<\/a>
\n\u6e90\u6587\u4ef6\uff1a~\/Downloads\/thesis_0520_xx.pdf<\/code><\/p>\n

\n

\u8bf4\u660e\uff1a\u8fd9\u662f\u5b8c\u6574\u7248\u8bd1\u6587\u3002\u76f8\u6bd4 thesis_0520_xx_zh.md<\/code> \u9605\u8bfb\u7248\uff0c\u672c\u6587\u4ef6\u8865\u5145\u4e86 Methods\u3001\u4e3b\u8981\u56fe\u6ce8\u3001\u6269\u5c55\u6570\u636e\u56fe\u6ce8\u3001Reporting Summary\u3001\u53c2\u8003\u6587\u732e\u548c\u6297\u4f53\/\u6750\u6599\u6e05\u5355\u3002PDF \u62bd\u53d6\u51fa\u7684\u56fe\u5185\u5750\u6807\u8f74\u3001\u6cf3\u9053\u7f16\u53f7\u3001P \u503c\u7b49\u788e\u7247\u5316\u6807\u7b7e\u4e0d\u9010\u9879\u91cd\u6392\uff0c\u4e3b\u8981\u5b9e\u9a8c\u542b\u4e49\u4ee5\u56fe\u6ce8\u8bd1\u6587\u4e3a\u51c6\u3002<\/p>\n<\/blockquote>\n

\u6587\u7ae0\u4fe1\u606f<\/h2>\n

\u9898\u76ee<\/strong>\uff1a\u901a\u8fc7\u4fc3\u8fdb\u80c6\u56fa\u9187\u6392\u6cc4\uff0c\u6291\u5236 ASGR1 \u53ef\u964d\u4f4e\u8102\u8d28\u6c34\u5e73
\n\u4f5c\u8005<\/strong>\uff1a\u738b\u83ca\u743c\u3001\u674e\u4eae\u4eae\u3001\u80e1\u5965\u3001\u9093\u521a\u3001\u9b4f\u5065\u3001\u674e\u4e91\u5cf0\u3001\u5218\u8fdc\u658c\u3001\u5362\u6653\u827a\u3001\u90b1\u5fd7\u5e73\u3001\u65bd\u96c4\u6770\u3001\u8d75\u6653\u9732\u3001\u7f57\u6770\u3001\u5b8b\u4fdd\u4eae
\n\u901a\u8baf\u4f5c\u8005<\/strong>\uff1a\u5b8b\u4fdd\u4eae\uff0c\u6b66\u6c49\u5927\u5b66\u751f\u547d\u79d1\u5b66\u5b66\u9662\u3001\u6cf0\u5eb7\u751f\u547d\u533b\u5b66\u4e2d\u5fc3\u3001\u6cf0\u5eb7\u533b\u5b66\u9662
\n\u63a5\u6536\u65e5\u671f<\/strong>\uff1a2021 \u5e74 8 \u6708 22 \u65e5
\n\u63a5\u53d7\u65e5\u671f<\/strong>\uff1a2022 \u5e74 6 \u6708 22 \u65e5
\n\u5728\u7ebf\u53d1\u8868<\/strong>\uff1a2022 \u5e74 8 \u6708 3 \u65e5<\/p>\n

\u6458\u8981<\/h2>\n

\u9ad8\u80c6\u56fa\u9187\u662f\u5fc3\u8840\u7ba1\u75be\u75c5\u7684\u4e3b\u8981\u5371\u9669\u56e0\u7d20\u3002\u76ee\u524d\u5c1a\u65e0\u836f\u7269\u80fd\u591f\u901a\u8fc7\u76f4\u63a5\u4fc3\u8fdb\u80c6\u56fa\u9187\u6392\u6cc4\u6765\u964d\u4f4e\u80c6\u56fa\u9187\u3002\u4eba\u7c7b\u9057\u4f20\u5b66\u7814\u7a76\u53d1\u73b0\uff0c\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u53d7\u4f53 1\uff08asialoglycoprotein receptor 1, ASGR1\uff09\u7684\u529f\u80fd\u7f3a\u5931\u53d8\u5f02\u4e0e\u8f83\u4f4e\u80c6\u56fa\u9187\u6c34\u5e73\u4ee5\u53ca\u8f83\u4f4e\u5fc3\u8840\u7ba1\u75be\u75c5\u98ce\u9669\u76f8\u5173\u3002<\/p>\n

ASGR1 \u53ea\u5728\u809d\u810f\u4e2d\u8868\u8fbe\uff0c\u8d1f\u8d23\u4ecb\u5bfc\u8840\u6db2\u4e2d\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7684\u5185\u541e\u548c\u6eb6\u9176\u4f53\u964d\u89e3\u3002ASGR1 \u5f71\u54cd\u80c6\u56fa\u9187\u4ee3\u8c22\u7684\u673a\u5236\u6b64\u524d\u5e76\u4e0d\u6e05\u695a\u3002\u672c\u6587\u53d1\u73b0\uff0cAsgr1 \u7f3a\u5931\u901a\u8fc7\u7a33\u5b9a LXR\u03b1\uff0c\u964d\u4f4e\u8840\u6e05\u548c\u809d\u810f\u4e2d\u7684\u8102\u8d28\u6c34\u5e73\u3002LXR\u03b1 \u4e0a\u8c03 ABCA1 \u548c ABCG5\/G8\uff0c\u5206\u522b\u4fc3\u8fdb\u80c6\u56fa\u9187\u8f6c\u8fd0\u5230\u9ad8\u5bc6\u5ea6\u8102\u86cb\u767d\uff08HDL\uff09\uff0c\u4ee5\u53ca\u6392\u6cc4\u5230\u80c6\u6c41\u548c\u7caa\u4fbf\u4e2d\u3002<\/p>\n

ASGR1 \u7f3a\u5931\u4f1a\u963b\u65ad\u7cd6\u86cb\u767d\u7684\u5185\u541e\u548c\u6eb6\u9176\u4f53\u964d\u89e3\uff0c\u964d\u4f4e\u6eb6\u9176\u4f53\u4e2d\u7684\u6c28\u57fa\u9178\u6c34\u5e73\uff0c\u7531\u6b64\u6291\u5236 mTORC1 \u5e76\u6fc0\u6d3b AMPK\u3002\u4e00\u65b9\u9762\uff0cAMPK \u901a\u8fc7\u964d\u4f4e LXR\u03b1 \u7684\u6cdb\u7d20\u8fde\u63a5\u9176 BRCA1\/BARD1 \u6765\u589e\u52a0 LXR\u03b1\uff1b\u53e6\u4e00\u65b9\u9762\uff0cAMPK \u6291\u5236\u63a7\u5236\u8102\u80aa\u751f\u6210\u7684 SREBP1\u3002\u6297 ASGR1 \u4e2d\u548c\u6297\u4f53\u53ef\u901a\u8fc7\u589e\u52a0\u80c6\u56fa\u9187\u6392\u6cc4\u6765\u964d\u4f4e\u8102\u8d28\u6c34\u5e73\uff0c\u5e76\u4e0e\u4e24\u79cd\u5e38\u7528\u964d\u80c6\u56fa\u9187\u836f\u7269\u963f\u6258\u4f10\u4ed6\u6c40\u6216\u4f9d\u6298\u9ea6\u5e03\u4ea7\u751f\u534f\u540c\u6709\u76ca\u4f5c\u7528\u3002<\/p>\n

\u603b\u4e4b\uff0c\u672c\u7814\u7a76\u8868\u660e\uff0c\u9776\u5411 ASGR1 \u53ef\u4e0a\u8c03 LXR\u03b1\u3001ABCA1 \u548c ABCG5\/G8\uff0c\u6291\u5236 SREBP1 \u548c\u8102\u80aa\u751f\u6210\uff0c\u4ece\u800c\u4fc3\u8fdb\u80c6\u56fa\u9187\u6392\u6cc4\u5e76\u964d\u4f4e\u8102\u8d28\u6c34\u5e73\u3002<\/p>\n

\u5f15\u8a00<\/h2>\n

\u80c6\u56fa\u9187\u7a33\u6001\u4f9d\u8d56\u80a0\u9053\u80c6\u56fa\u9187\u5438\u6536\u3001\u8840\u6d46\u8102\u86cb\u767d\u6444\u53d6\u3001\u4ece\u5934\u5408\u6210\u3001\u80c6\u56fa\u9187\u5206\u89e3\u4ee3\u8c22\u548c\u6392\u6cc4\u4e4b\u95f4\u590d\u6742\u7684\u76f8\u4e92\u4f5c\u7528\u3002\u8fc4\u4eca\u4e3a\u6b62\uff0c\u4e3b\u8981\u6709\u4e09\u7c7b\u964d\u80c6\u56fa\u9187\u836f\u7269\u3002<\/p>\n

\u7b2c\u4e00\u7c7b\u662f\u4ed6\u6c40\u7c7b\u836f\u7269\uff0c\u5373 3-\u7f9f\u57fa-3-\u7532\u57fa\u620a\u4e8c\u9170\u8f85\u9176 A \u8fd8\u539f\u9176\uff08HMGCR\uff09\u7684\u7ade\u4e89\u6027\u6291\u5236\u5242\u3002\u5b83\u901a\u8fc7\u964d\u4f4e\u80c6\u56fa\u9187\u751f\u7269\u5408\u6210\u3001\u4e0a\u8c03 LDL \u53d7\u4f53\uff08LDLR\uff09\u5e76\u589e\u5f3a\u4f4e\u5bc6\u5ea6\u8102\u86cb\u767d\uff08LDL\uff09\u6444\u53d6\u6765\u964d\u4f4e\u8840\u6d46\u80c6\u56fa\u9187\u3002\u7b2c\u4e8c\u7c7b\u662f\u4f9d\u6298\u9ea6\u5e03\uff0c\u5b83\u662f\u80a0\u9053\u80c6\u56fa\u9187\u5438\u6536\u6291\u5236\u5242\uff0c\u901a\u8fc7\u6291\u5236 Niemann-Pick C1-like 1\uff08NPC1L1\uff09\u7684\u5185\u541e\u6765\u963b\u65ad\u80c6\u56fa\u9187\u6444\u5165\u3002\u7b2c\u4e09\u7c7b\u662f PCSK9 \u6291\u5236\u5242\uff0c\u5b83\u901a\u8fc7\u7a33\u5b9a LDLR \u6765\u589e\u52a0\u809d\u810f\u5bf9 LDL \u7684\u6444\u53d6\u3002<\/p>\n

\u867d\u7136\u8fd9\u4e9b\u836f\u7269\u5df2\u7ecf\u88ab\u5e7f\u6cdb\u4f7f\u7528\uff0c\u4ecd\u6709\u76f8\u5f53\u4e00\u90e8\u5206\u60a3\u8005\u51fa\u73b0\u590d\u53d1\u6027\u5fc3\u8840\u7ba1\u75be\u75c5\uff0c\u800c\u4e14\u5176 LDL \u80c6\u56fa\u9187\u6c34\u5e73\u4e5f\u672a\u80fd\u8fbe\u5230\u6307\u5357\u63a8\u8350\u76ee\u6807\u3002\u66f4\u91cd\u8981\u7684\u662f\uff0c\u73b0\u6709\u8fd9\u4e9b\u964d\u80c6\u56fa\u9187\u836f\u7269\u90fd\u4e0d\u662f\u901a\u8fc7\u76f4\u63a5\u4fc3\u8fdb\u80c6\u56fa\u9187\u5206\u89e3\u4ee3\u8c22\u6216\u6392\u6cc4\u6765\u964d\u4f4e\u80c6\u56fa\u9187\u3002<\/p>\n

\u80c6\u56fa\u9187\u7531 ABCG5 \u548c ABCG8 \u5f02\u4e8c\u805a\u4f53\u6392\u6cc4\u5230\u80c6\u6c41\u548c\u80a0\u8154\u3002ABCG5 \u6216 ABCG8 \u7a81\u53d8\u4f1a\u5bfc\u81f4\u8c37\u56fa\u9187\u8840\u75c7\uff0c\u8fd9\u662f\u4e00\u79cd\u4ee5\u753e\u9187\u7d2f\u79ef\u548c\u65e9\u53d1\u6027\u52a8\u8109\u7ca5\u6837\u786c\u5316\u4e3a\u7279\u5f81\u7684\u7f55\u89c1\u75be\u75c5\u3002\u5728\u5c0f\u9f20\u809d\u810f\u4e2d\u8fc7\u8868\u8fbe ABCG5\/G8 \u53ef\u589e\u52a0\u809d\u80c6\u80c6\u56fa\u9187\u5206\u6ccc\u5e76\u964d\u4f4e\u8840\u6d46\u80c6\u56fa\u9187\u3002<\/p>\n

ABCG5\/G8 \u7684\u8868\u8fbe\u4e3b\u8981\u5728\u8f6c\u5f55\u6c34\u5e73\u53d7 LXR \u8c03\u63a7\u3002\u836f\u7406\u6027\u6fc0\u6d3b LXR \u53ef\u901a\u8fc7\u4e0a\u8c03 ABCG5\/G8 \u589e\u52a0\u80c6\u56fa\u9187\u5916\u6392\u3002\u7136\u800c\uff0cLXR \u4e5f\u4f1a\u589e\u52a0 SREBP1\uff08\u53c8\u79f0 ADD1\uff09\uff0c\u540e\u8005\u9a71\u52a8\u8102\u80aa\u9178\u751f\u7269\u5408\u6210\u57fa\u56e0\u8868\u8fbe\uff0c\u5bfc\u81f4\u6709\u5bb3\u7684\u809d\u8102\u80aa\u53d8\u6027\u548c\u9ad8\u7518\u6cb9\u4e09\u916f\u8840\u75c7\u3002\u56e0\u6b64\uff0c\u76f4\u63a5 LXR \u6fc0\u52a8\u5242\u5e76\u4e0d\u9002\u5408\u4e34\u5e8a\u6cbb\u7597\u9ad8\u80c6\u56fa\u9187\u8840\u75c7\u3002<\/p>\n

\u4e00\u9879\u9488\u5bf9\u51b0\u5c9b\u4eba\u7684\u5927\u89c4\u6a21\u9057\u4f20\u5b66\u7814\u7a76\u53d1\u73b0\uff0cASGR1 \u529f\u80fd\u7f3a\u5931\u53d8\u5f02\u4e0e\u8f83\u4f4e\u80c6\u56fa\u9187\u6c34\u5e73\u548c\u8f83\u4f4e\u5fc3\u8840\u7ba1\u75be\u75c5\u98ce\u9669\u76f8\u5173\u3002ASGR1 \u662f\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u53d7\u4f53\u7684\u4e3b\u8981\u4e9a\u578b\uff0c\u662f\u4e00\u79cd\u809d\u810f\u7279\u5f02\u6027\u53d7\u4f53\uff0c\u4ecb\u5bfc\u591a\u79cd\u8131\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7684\u5185\u541e\u548c\u6eb6\u9176\u4f53\u964d\u89e3\u3002ASGR1 \u4f18\u5148\u8bc6\u522b\u5176\u7cd6\u94fe\u672b\u7aef\u7684\u534a\u4e73\u7cd6\u6216 N-\u4e59\u9170\u534a\u4e73\u7cd6\u80fa\u3002\u7f3a\u5931 Asgr1 \u7684\u5c0f\u9f20\u8868\u578b\u603b\u4f53\u6b63\u5e38\uff0cAsgr1 \u7f3a\u5931\u6240\u5e26\u6765\u7684\u964d\u80c6\u56fa\u9187\u4f5c\u7528\u4e5f\u5728\u5c0f\u9f20\u548c\u732a\u4e2d\u5f97\u5230\u590d\u73b0\u3002\u7136\u800c\uff0cASGR1 \u7a76\u7adf\u5982\u4f55\u8c03\u63a7\u80c6\u56fa\u9187\u4ee3\u8c22\uff0c\u4ecd\u4e0d\u6e05\u695a\u3002<\/p>\n

Asgr1\u2212\/\u2212 \u589e\u52a0\u80c6\u56fa\u9187\u5916\u6392<\/h2>\n

\u4e3a\u63ed\u793a ASGR1 \u529f\u80fd\u7f3a\u5931\u53d8\u5f02\u5f71\u54cd\u80c6\u56fa\u9187\u4ee3\u8c22\u7684\u673a\u5236\uff0c\u7814\u7a76\u8005\u5728\u4eba\u809d\u764c\u7ec6\u80de\u7cfb Huh7 \u4e2d\u6572\u4f4e ASGR1\uff0c\u5e76\u8fdb\u884c\u4e86 RNA \u6d4b\u5e8f\u3002\u57fa\u56e0\u96c6\u5bcc\u96c6\u5206\u6790\uff08GSEA\uff09\u663e\u793a\uff0cLXR \u9776\u57fa\u56e0\u663e\u8457\u5bcc\u96c6\u3002RT-qPCR \u8bc1\u5b9e\uff0c\u5305\u62ec ABCG5\u3001ABCG8 \u548c ABCA1 \u5728\u5185\u7684\u7ecf\u5178 LXR \u9776\u57fa\u56e0\u8868\u8fbe\u5347\u9ad8\u3002<\/p>\n

\u6709\u8da3\u7684\u662f\uff0c\u5728 ASGR1 \u6572\u4f4e\u6216\u6572\u9664\u7ec6\u80de\u4e2d\uff0cLXR\u03b1 \u548c LXR\u03b2 \u7684\u86cb\u767d\u6c34\u5e73\u5347\u9ad8\u3002\u76f8\u53cd\uff0c\u8fc7\u8868\u8fbe ASGR1 \u5219\u964d\u4f4e LXR\u03b1\u3001LXR\u03b2 \u86cb\u767d\u6c34\u5e73\uff0c\u4ee5\u53ca ABCG5\u3001ABCG8 \u548c ABCA1 \u7684 mRNA \u6c34\u5e73\u3002<\/p>\n

ASGR1 \u5728\u809d\u810f\u4e2d\u9ad8\u5ea6\u8868\u8fbe\u3002\u7814\u7a76\u8005\u6784\u5efa\u4e86 Asgr1 \u7f3a\u5931\u5c0f\u9f20\u3002\u6742\u5408\uff08Asgr1+\/\u2212\uff09\u548c\u7eaf\u5408\uff08Asgr1\u2212\/\u2212\uff09\u5c0f\u9f20\u5916\u89c2\u5747\u57fa\u672c\u6b63\u5e38\u3002\u4e0e\u91ce\u751f\u578b\u5c0f\u9f20\u76f8\u6bd4\uff0cAsgr1+\/\u2212 \u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u8840\u6e05\u603b\u80c6\u56fa\u9187\uff08TC\uff09\u548c\u7518\u6cb9\u4e09\u916f\uff08TG\uff09\u6c34\u5e73\u663e\u8457\u964d\u4f4e\uff0c\u5e76\u5448\u57fa\u56e0\u5242\u91cf\u4f9d\u8d56\u5173\u7cfb\u3002Asgr1+\/\u2212 \u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u809d\u810f TC \u548c TG \u6c34\u5e73\u4e5f\u4f4e\u4e8e\u91ce\u751f\u578b\u5c0f\u9f20\u3002<\/p>\n

\u4e0e\u6b64\u4e00\u81f4\uff0cAsgr1 \u7f3a\u5931\u663e\u8457\u51cf\u5c11\u809d\u810f\u8102\u6ef4\u7d2f\u79ef\uff0c\u4e14\u672a\u9020\u6210\u809d\u635f\u4f24\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0cAsgr1 \u7f3a\u5931\u5c0f\u9f20\u80c6\u56ca\u4e2d\u7684\u80c6\u6c41\u4f53\u79ef\u589e\u52a0\uff0c\u80c6\u6c41\u80c6\u56fa\u9187\u548c\u80c6\u6c41\u9178\u6d53\u5ea6\u4e5f\u5347\u9ad8\u3002\u4e0e\u91ce\u751f\u578b\u5c0f\u9f20\u76f8\u6bd4\uff0cAsgr1+\/\u2212 \u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u5206\u522b\u589e\u52a0 71% \u548c 114%\uff0c\u603b\u80c6\u6c41\u9178\u4e5f\u5347\u9ad8\u3002Asgr1\u2212\/\u2212 \u5c0f\u9f20\u80c6\u6c41\u5448\u6d51\u6d4a\u72b6\uff0c\u63d0\u793a\u80c6\u6c41\u80c6\u56fa\u9187\u589e\u52a0\u3002<\/p>\n

\u6b64\u5916\uff0cAsgr1 \u7f3a\u5931\u964d\u4f4e\u80c6\u9178\u4e0e\u9f20\u80c6\u9178\u7684\u6bd4\u4f8b\uff0c\u5e76\u4f7f\u80c6\u6c41\u78f7\u8102\u6d53\u5ea6\u7565\u6709\u5347\u9ad8\u3002\u7531\u4e8e\u80c6\u9178\u6bd4\u9f20\u80c6\u9178\u66f4\u80fd\u4fc3\u8fdb\u80a0\u9053\u80c6\u56fa\u9187\u5438\u6536\uff0cAsgr1 \u7f3a\u5931\u5c0f\u9f20\u80c6\u6c41\u9178\u7ec4\u6210\u7684\u53d8\u5316\u4e5f\u53ef\u80fd\u6709\u52a9\u4e8e\u5176\u8f83\u4f4e\u80c6\u56fa\u9187\u6c34\u5e73\u3002<\/p>\n

LXR\u03b1 \u662f\u809d\u810f\u4e2d\u5360\u4e3b\u5bfc\u5730\u4f4d\u7684 LXR \u4e9a\u578b\uff0c\u5728 Asgr1\u2212\/\u2212 \u809d\u810f\u4e2d\u5927\u5e45\u5347\u9ad8\u3002\u4e0e\u6b64\u4e00\u81f4\uff0cAbcg5\u3001ABCG8 \u548c CYP7A1 \u7684\u8868\u8fbe\u589e\u52a0\uff0c\u8fd9\u89e3\u91ca\u4e86\u80c6\u6c41\u4e2d\u80c6\u56fa\u9187\u548c\u80c6\u6c41\u9178\u6c34\u5e73\u5347\u9ad8\uff0c\u4ee5\u53ca\u8840\u6e05\u548c\u809d\u810f\u8102\u8d28\u6c34\u5e73\u964d\u4f4e\u3002<\/p>\n

\u5c3d\u7ba1 LXR\u03b1 \u5347\u9ad8\uff0cAsgr1\u2212\/\u2212 \u809d\u810f\u4e2d SREBP1 \u524d\u4f53\uff08pre-SREBP1\uff09\u3001\u5176\u6838\u578b\u5f62\u5f0f\uff08n-SREBP1\uff09\u548c\u8102\u80aa\u9178\u5408\u9176\uff08FASN\uff09\u7684\u86cb\u767d\u6c34\u5e73\u5747\u660e\u663e\u4e0b\u964d\u3002\u8102\u80aa\u9178\u751f\u7269\u5408\u6210\u57fa\u56e0\u7684 mRNA \u6c34\u5e73\u4e5f\u4e00\u81f4\u4e0b\u8c03\u3002\u8fdb\u4e00\u6b65\u7814\u7a76\u6392\u9664\u4e86 LDLR \u548c HMGCR \u86cb\u767d\u6c34\u5e73\u3001Dil-LDL \u6444\u53d6\u6216 PCSK9 \u4fc3\u8fdb LDLR \u964d\u89e3\u7684\u6539\u53d8\u3002Asgr1 \u7f3a\u5931\u5c0f\u9f20 LDL \u6e05\u9664\u672a\u53d7\u635f\u3002TC \u548c TG \u5206\u6ccc\u4e0b\u964d\uff0c\u53ef\u80fd\u662f Asgr1\u2212\/\u2212 \u5c0f\u9f20\u80c6\u56fa\u9187\u6392\u6cc4\u589e\u52a0\u540e\u7684\u7ee7\u53d1\u6548\u5e94\u3002Asgr1 \u7f3a\u5931\u5e76\u672a\u6539\u53d8\u5185\u6e90\u6027 LXR \u914d\u4f53\u6c34\u5e73\u3002\u96cc\u6027 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\u4e5f\u89c2\u5bdf\u5230\u7c7b\u4f3c\u7684\u57fa\u56e0\u8868\u8fbe\u53d8\u5316\u548c\u4ee3\u8c22\u8868\u578b\u3002<\/p>\n

Asgr1\u2212\/\u2212 \u7684\u4ee3\u8c22\u83b7\u76ca\u9700\u8981 LXR\u03b1<\/h2>\n

\u4e3a\u786e\u5b9a Asgr1\u2212\/\u2212 \u5c0f\u9f20\u7684\u4ee3\u8c22\u6539\u5584\u662f\u5426\u6e90\u4e8e LXR\u03b1 \u5347\u9ad8\uff0c\u7814\u7a76\u8005\u83b7\u5f97\u4e86 Asgr1\u2212\/\u2212Lxr\u03b1\u2212\/\u2212 \u5c0f\u9f20\u3002Asgr1\u2212\/\u2212 \u809d\u810f\u4e2d ABCG8\u3001ABCA1 \u548c CYP7A1 \u86cb\u767d\u6c34\u5e73\u7684\u5347\u9ad8\uff0c\u5728\u7f3a\u5931 Lxr\u03b1 \u540e\u5b8c\u5168\u6d88\u5931\u3002<\/p>\n

\u7531\u4e8e LXR\u03b1 \u9a71\u52a8 Srebp1c \u8f6c\u5f55\uff0cLxr\u03b1\u2212\/\u2212 \u5c0f\u9f20\u4e2d SREBP1c\u3001FASN\u3001LDLR \u548c\u8102\u80aa\u9178\u751f\u7269\u5408\u6210\u57fa\u56e0\u5747\u4e0b\u8c03\u3002Lxr\u03b1 \u7f3a\u5931\u5c0f\u9f20\u4e2d Abcg5\u3001Abcg8\u3001Abca1 \u548c Cyp7a1 \u7684 mRNA \u6c34\u5e73\u4e0b\u964d\uff1b\u5728\u7f3a\u4e4f LXR\u03b1 \u7684\u60c5\u51b5\u4e0b\uff0cAsgr1 \u7f3a\u5931\u65e0\u6cd5\u589e\u52a0\u8fd9\u4e9b\u57fa\u56e0\u7684\u8868\u8fbe\u3002<\/p>\n

\u867d\u7136\u5404\u7ec4\u5c0f\u9f20\u4f53\u91cd\u3001\u98df\u7269\u6444\u5165\u548c\u8840\u7cd6\u6c34\u5e73\u76f8\u4f3c\uff0c\u6572\u9664 Lxr\u03b1 \u5bfc\u81f4\u809d\u810f\u8102\u6ef4\u5927\u91cf\u7d2f\u79ef\u3001\u809d\u7ec6\u80de\u6c14\u7403\u6837\u53d8\uff0c\u4ee5\u53ca ALT \u548c AST \u5347\u9ad8\u3002Asgr1 \u7f3a\u5931\u663e\u8457\u964d\u4f4e\u809d\u810f\u8102\u8d28\u542b\u91cf\u548c\u8840\u6e05 TC\u3001TG\uff0c\u4f46\u8fd9\u4e9b\u6548\u679c\u5728 Lxr\u03b1 \u6d88\u878d\u540e\u5b8c\u5168\u6d88\u5931\u3002<\/p>\n

\u4e0e\u91ce\u751f\u578b\u5c0f\u9f20\u76f8\u6bd4\uff0cAsgr1\u2212\/\u2212 \u5c0f\u9f20 VLDL \u548c LDL \u4e2d\u7684\u80c6\u56fa\u9187\u542b\u91cf\u660e\u663e\u4e0b\u964d\uff0c\u800c HDL \u80c6\u56fa\u9187\u6c34\u5e73\u5347\u9ad8\u3002Lxr\u03b1 \u6d88\u878d\u663e\u8457\u589e\u52a0 VLDL \u548c LDL \u7ec4\u5206\u4e2d\u7684\u80c6\u56fa\u9187\uff0c\u5e76\u62b5\u6d88 Asgr1 \u7f3a\u5931\u9020\u6210\u7684\u8102\u8d28\u53d8\u5316\u3002\u4e0e\u91ce\u751f\u578b\u76f8\u6bd4\uff0cAsgr1\u2212\/\u2212 \u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u589e\u52a0\u7ea6\u4e24\u500d\uff0c\u4f46\u5728 Asgr1\u2212\/\u2212Lxr\u03b1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\u964d\u81f3\u57fa\u7840\u6c34\u5e73\u3002\u7caa\u4fbf\u80c6\u56fa\u9187\u4e5f\u5728 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\u589e\u52a0 48%\uff0c\u8fd9\u4e00\u73b0\u8c61\u88ab Lxr\u03b1 \u7f3a\u5931\u524a\u5f31\u3002\u8fd9\u4e9b\u7ed3\u679c\u4e0e Abcg5\u3001Abcg8 \u548c Abca1 \u7684\u8868\u8fbe\u53d8\u5316\u4e00\u81f4\uff1aABCG5\/G8 \u5c06\u80c6\u56fa\u9187\u6392\u5165\u80c6\u6c41\uff0c\u968f\u540e\u901a\u8fc7\u7caa\u4fbf\u6392\u51fa\u4f53\u5916\uff1bABCA1 \u5219\u5c06\u80c6\u56fa\u9187\u8f6c\u8fd0\u81f3 HDL\u3002<\/p>\n

LXR\u03b1 \u964d\u89e3\u9700\u8981 BRCA1 \u548c BARD1<\/h2>\n

\u7814\u7a76\u8005\u968f\u540e\u8003\u5bdf ASGR1 \u8c03\u63a7 LXR\u03b1 \u7684\u673a\u5236\u3002MG132 \u53ef\u963b\u65ad ASGR1 \u8bf1\u5bfc\u7684 LXR\u03b1 \u964d\u89e3\u3002\u8fc7\u8868\u8fbe ASGR1 \u589e\u52a0 LXR\u03b1 \u6cdb\u7d20\u5316\uff0c\u800c\u6572\u9664 ASGR1 \u964d\u4f4e\u5176\u6cdb\u7d20\u5316\u3002<\/p>\n

\u7531 BRCA1 \u548c BARD1 \u7ec4\u6210\u7684 E3 \u6cdb\u7d20\u8fde\u63a5\u9176\u590d\u5408\u7269\u53ef\u4ecb\u5bfc LXR\u03b1 \u6cdb\u7d20\u5316\u548c\u964d\u89e3\u3002\u6c89\u9ed8 BRCA1 \u548c BARD1 \u663e\u8457\u524a\u5f31 ASGR1 \u4fc3\u8fdb\u7684 LXR\u03b1 \u6cdb\u7d20\u5316\u548c\u964d\u89e3\u3002ASGR1 \u6572\u9664\u6216\u6572\u4f4e\u964d\u4f4e BRCA1 \u548c BARD1\uff0c\u5e76\u589e\u52a0 LXR\u03b1\u3002\u76f8\u53cd\uff0cASGR1 \u4ee5\u5242\u91cf\u4f9d\u8d56\u65b9\u5f0f\u589e\u52a0 BRCA1 \u548c BARD1\u3002<\/p>\n

ASGR1 \u6572\u9664\u6216\u8fc7\u8868\u8fbe\u5e76\u4e0d\u6539\u53d8\u5176\u4ed6 BRCA1\/BARD1 \u5e95\u7269\uff0c\u5982 cyclin B1 \u548c Cdc25C\u3002Ki-67 \u548c PCNA \u67d3\u8272\u663e\u793a Asgr1 \u7f3a\u5931\u809d\u810f\u4e2d\u6ca1\u6709\u5f02\u5e38\u7ec6\u80de\u589e\u6b96\uff0c\u86cb\u767d\u9176\u4f53\u4e9a\u57fa\u4e5f\u672a\u6539\u53d8\u3002\u7efc\u4e0a\uff0c\u8fd9\u4e9b\u7ed3\u679c\u8868\u660e\uff0cAsgr1 \u7f3a\u5931\u901a\u8fc7\u964d\u4f4e BRCA1 \u548c BARD1\uff0c\u7279\u5f02\u6027\u589e\u52a0 LXR\u03b1\u3002<\/p>\n

\u914d\u4f53-ASGR1 \u901a\u8fc7 AMPK \u8c03\u63a7 LXR\u03b1<\/h2>\n

ASGR1 \u662f\u4e00\u79cd\u5355\u8de8\u819c\u86cb\u767d\uff0c\u53ef\u7ed3\u5408\u8840\u6db2\u4e2d\u7684\u591a\u79cd\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u3002\u4e3a\u68c0\u9a8c\u914d\u4f53-ASGR1 \u76f8\u4e92\u4f5c\u7528\u662f\u5426\u53c2\u4e0e LXR\u03b1 \u8c03\u63a7\uff0c\u7814\u7a76\u8005\u6bd4\u8f83\u4e86\u6709\u65e0\u80ce\u725b\u8840\u6e05\u65f6 ASGR1 \u4fc3\u8fdb LXR\u03b1 \u964d\u89e3\u7684\u60c5\u51b5\u3002\u5728\u65e0\u8840\u6e05\u57f9\u517b\u57fa\u4e2d\uff0cASGR1 \u4e0d\u80fd\u8bf1\u5bfc LXR\u03b1 \u964d\u89e3\u3002<\/p>\n

\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u6216\u53bb\u553e\u6db2\u9178\u8840\u6e05\u7c7b\u9ecf\u86cb\u767d\u662f ASGR1 \u7684\u9ad8\u4eb2\u548c\u529b\u5929\u7136\u914d\u4f53\u3002\u5728\u65e0\u8840\u6e05\u57f9\u517b\u57fa\u4e14\u5b58\u5728 ASGR1 \u65f6\uff0c\u5b83\u4eec\u53ef\u4ee5\u5242\u91cf\u4f9d\u8d56\u6027\u523a\u6fc0 LXR\u03b1 \u964d\u89e3\u3002\u540c\u65f6\uff0c\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u4e0e ASGR1 \u534f\u540c\u589e\u5f3a LXR\u03b1 \u6cdb\u7d20\u5316\u3002\u914d\u4f53\u7ed3\u5408\u7f3a\u9677\u7a81\u53d8\u4f53 ASGR1(6A) \u4e0d\u80fd\u4fc3\u8fdb LXR\u03b1 \u964d\u89e3\u3002<\/p>\n

\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u5904\u7406\u589e\u52a0\u5185\u6e90\u6027 BRCA1 \u548c BARD1\uff0c\u5e76\u964d\u4f4e LXR\u03b1\u3002\u6c89\u9ed8 ASGR1 \u6216 BRCA1\/BARD1 \u4f7f LXR\u03b1 \u7ef4\u6301\u5728\u8f83\u9ad8\u6c34\u5e73\uff0c\u5e76\u6d88\u9664\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u8bf1\u5bfc\u7684 LXR\u03b1 \u964d\u89e3\u3002\u8fd9\u4e9b\u7ed3\u679c\u8bf4\u660e\uff0c\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7ed3\u5408 ASGR1 \u540e\u589e\u52a0 BRCA1\/BARD1\uff0c\u4ece\u800c\u4fc3\u8fdb LXR\u03b1 \u964d\u89e3\u3002<\/p>\n

\u8840\u6db2\u4e2d\u7684\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u901a\u8fc7\u809d\u810f ASGR1 \u7ecf\u7f51\u683c\u86cb\u767d\u4ecb\u5bfc\u7684\u5185\u541e\u8fdb\u5165\u7ec6\u80de\u3002\u5185\u4f53\u4e2d\u7684\u9178\u6027\u73af\u5883\u8bf1\u5bfc ASGR1 \u4e0e\u7cd6\u86cb\u767d\u89e3\u79bb\uff0cASGR1 \u56de\u6536\u5230\u8d28\u819c\uff0c\u800c\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u88ab\u9012\u9001\u81f3\u6eb6\u9176\u4f53\u964d\u89e3\u3002\u7814\u7a76\u8005\u8fdb\u4e00\u6b65\u5206\u6790\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7684\u5185\u541e\u548c\u6eb6\u9176\u4f53\u964d\u89e3\u662f\u5426\u662f ASGR1 \u8c03\u63a7 LXR\u03b1 \u6240\u5fc5\u9700\u3002<\/p>\n

\u6c89\u9ed8\u7f51\u683c\u86cb\u767d\u91cd\u94fe\uff08CHC\uff09\u6d88\u9664 ASGR1 \u6216\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u8bf1\u5bfc\u7684 LXR\u03b1 \u964d\u89e3\u3002\u65e0\u8bba\u662f\u5426\u63a5\u53d7\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u5904\u7406\uff0cCHC \u6c89\u9ed8\u7ec6\u80de\u4e2d\u7684 BRCA1 \u548c BARD1 \u6c34\u5e73\u5747\u4f4e\u4e8e\u5bf9\u7167\u7ec6\u80de\u3002<\/p>\n

\u6eb6\u9176\u4f53\u964d\u89e3\u91ca\u653e\u7684\u7cd6\u548c\u6c28\u57fa\u9178\u7b49\u8425\u517b\u7269\u8d28\u53ef\u4ee5\u6fc0\u6d3b mTORC1 \u5e76\u6291\u5236 AMPK\u3002mTORC1 \u6fc0\u6d3b SREBP \u548c USP20\uff0c\u4fc3\u8fdb\u8102\u8d28\u751f\u7269\u5408\u6210\u3002\u76f8\u53cd\uff0cAMPK \u6291\u5236 SREBP \u5e76\u4e0b\u8c03 BRCA1\u3002\u4e0e\u8fd9\u4e9b\u53d1\u73b0\u4e00\u81f4\uff0c\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u5904\u7406\u5728\u5bf9\u7167\u7ec6\u80de\u4e2d\u6fc0\u6d3b mTORC1 \u5e76\u6291\u5236 AMPK\u3002\u540c\u65f6\uff0c\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u589e\u52a0 BRCA1 \u548c BARD1\uff0c\u5e76\u964d\u4f4e LXR\u03b1\u3002\u8fd9\u4e9b\u4f5c\u7528\u5728 CHC \u6c89\u9ed8\u7ec6\u80de\u4e2d\u6d88\u5931\u3002<\/p>\n

Bafilomycin A1\uff08BFA\uff09\u901a\u8fc7\u5347\u9ad8\u6eb6\u9176\u4f53 pH \u7834\u574f\u6eb6\u9176\u4f53\u964d\u89e3\uff0c\u4ece\u800c\u6291\u5236 mTORC1 \u5e76\u6fc0\u6d3b AMPK\u3002\u5728 BFA \u5904\u7406\u7684\u91ce\u751f\u578b\u7ec6\u80de\u4e2d\uff0cBRCA1 \u548c BARD1 \u4e0b\u964d\uff0cLXR\u03b1 \u4e0a\u5347\u3002BFA \u4e5f\u963b\u65ad ASGR1 \u8bf1\u5bfc\u7684 LXR\u03b1 \u964d\u89e3\u3002ASGR1 \u7f3a\u5931\u6291\u5236 mTORC1\u3001\u6fc0\u6d3b AMPK\uff0c\u8fdb\u4e00\u6b65\u5bfc\u81f4 BRCA1 \u548c BARD1 \u964d\u4f4e\u4ee5\u53ca LXR\u03b1 \u5347\u9ad8\uff1b\u800c BFA \u5904\u7406\u963b\u65ad ASGR1 \u7f3a\u5931\u5bf9 mTORC1\u3001AMPK\u3001BRCA1\/BARD1 \u6216 LXR \u7684\u5f71\u54cd\u3002<\/p>\n

\u968f\u540e\u7814\u7a76\u8005\u68c0\u6d4b ASGR1 \u7684\u4f5c\u7528\u662f\u5426\u4f9d\u8d56 AMPK\u3002AMPK \u6fc0\u52a8\u5242 A769662 \u589e\u52a0 LXR\u03b1 \u548c p-ACC\uff0c\u5e76\u964d\u4f4e n-SREBP1\u3001n-SREBP2\u3001BRCA1 \u548c BARD1\u3002AMPK \u6291\u5236\u5242 dorsomorphin \u5219\u4ea7\u751f\u76f8\u53cd\u6548\u679c\u3002A769662 \u51cf\u5f31 ASGR1 \u8bf1\u5bfc\u7684 LXR\u03b1 \u964d\u89e3\uff1b\u76f8\u53cd\uff0cdorsomorphin \u963b\u65ad ASGR1 \u7f3a\u5931\u8bf1\u5bfc\u7684 LXR\u03b1 \u589e\u52a0\u3002<\/p>\n

\u4e3a\u660e\u786e\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u6d88\u5316\u4ea7\u751f\u54ea\u4e9b\u8425\u517b\u7269\u8d28\u8c03\u63a7 mTORC1\uff0c\u7814\u7a76\u8005\u5feb\u901f\u5206\u79bb\u6eb6\u9176\u4f53\u5e76\u76f4\u63a5\u6d4b\u91cf\u6c28\u57fa\u9178\u6c34\u5e73\u3002\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u5904\u7406\u589e\u52a0\u91ce\u751f\u578b\u7ec6\u80de\u6eb6\u9176\u4f53\u4e2d\u5927\u591a\u6570\u6c28\u57fa\u9178\u6c34\u5e73\uff0c\u4f46\u5728 ASGR1-KO \u7ec6\u80de\u4e2d\u65e0\u6b64\u4f5c\u7528\u3002\u8f6c\u8fd0\u86cb\u767d SLC38A9 \u5bf9\u4eae\u6c28\u9178\u548c\u5176\u4ed6\u6c28\u57fa\u9178\u6fc0\u6d3b mTORC1 \u662f\u5fc5\u9700\u7684\u3002\u786e\u5b9e\uff0c\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u4ee5 SLC38A9 \u4f9d\u8d56\u65b9\u5f0f\u589e\u52a0 p-S6K\u3002\u91ce\u751f\u578b SLC38A9 \u53ef\u5728 SLC38A9 \u6572\u9664\u7ec6\u80de\u4e2d\u6062\u590d S6K \u78f7\u9178\u5316\uff0c\u800c\u5f71\u54cd SLC38A9-Ragulator \u76f8\u4e92\u4f5c\u7528\u6216 SLC38A9 \u8f6c\u8fd0\u6d3b\u6027\u7684\u7a81\u53d8\u4e0d\u80fd\u6062\u590d S6K \u78f7\u9178\u5316\u3002<\/p>\n

\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u53ef\u4f7f mTOR \u91cd\u65b0\u5b9a\u4f4d\u81f3\u6eb6\u9176\u4f53\u3002\u767d\u86cb\u767d\u6fc0\u6d3b mTORC1 \u7684\u80fd\u529b\u5f31\u4e8e\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A\uff0c\u63d0\u793a ASGR1 \u4ecb\u5bfc\u7684\u7cd6\u86cb\u767d\u6444\u53d6\u975e\u5e38\u9ad8\u6548\u3002<\/p>\n

\u8fd9\u4e9b\u7ed3\u679c\u5171\u540c\u652f\u6301\u56fe 3h \u6240\u793a\u7684\u5de5\u4f5c\u6a21\u578b\uff1aASGR1 \u7ed3\u5408\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\uff0c\u5e76\u901a\u8fc7\u7f51\u683c\u86cb\u767d\u4ecb\u5bfc\u7684\u5185\u541e\u5c06\u5176\u9012\u9001\u81f3\u6eb6\u9176\u4f53\u3002\u7cd6\u86cb\u767d\u88ab\u6d88\u5316\u540e\u91ca\u653e\u6c28\u57fa\u9178\uff0c\u6fc0\u6d3b\u6eb6\u9176\u4f53 mTORC1 \u5e76\u6291\u5236 AMPK\u3002\u5176\u7ed3\u679c\u662f\uff0c\u4e00\u65b9\u9762 SREBP \u88ab\u6fc0\u6d3b\u5e76\u589e\u52a0\u8102\u80aa\u751f\u6210\uff1b\u53e6\u4e00\u65b9\u9762 BRCA1 \u548c BARD1 \u8868\u8fbe\u589e\u52a0\uff0c\u4fc3\u8fdb LXR\u03b1 \u964d\u89e3\u5e76\u964d\u4f4e\u80c6\u56fa\u9187\u5916\u6392\u3002\u901a\u8fc7\u8fd9\u4e00\u8fc7\u7a0b\uff0c\u6444\u5165\u7684\u8425\u517b\u7269\u8d28\u652f\u6301\u7ec6\u80de\u751f\u957f\u3002<\/p>\n

\u5f53 ASGR1 \u7a81\u53d8\u6216\u88ab\u6291\u5236\u65f6\uff0c\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7684\u5185\u541e\u548c\u6eb6\u9176\u4f53\u964d\u89e3\u88ab\u963b\u65ad\uff0c\u5bfc\u81f4 mTORC1 \u6291\u5236\u548c AMPK \u6fc0\u6d3b\u3002AMPK \u4f7f BRCA1\/BARD1 \u4e0d\u7a33\u5b9a\uff0c\u4ece\u800c\u589e\u52a0 LXR\u03b1\uff0c\u5e76\u4e0a\u8c03\u5305\u62ec ABCA1 \u548c ABCG5\/G8 \u5728\u5185\u7684\u80c6\u56fa\u9187\u8f6c\u8fd0\u86cb\u767d\u3002ABCA1 \u5c06\u80c6\u56fa\u9187\u9012\u9001\u81f3 HDL\uff0c\u4fc3\u8fdb\u9006\u5411\u80c6\u56fa\u9187\u8f6c\u8fd0\uff1bABCG5\/G8 \u5c06\u80c6\u56fa\u9187\u6cf5\u5165\u80c6\u6c41\uff0c\u4f7f\u80c6\u56fa\u9187\u901a\u8fc7\u7caa\u4fbf\u6392\u51fa\u3002\u540c\u65f6\uff0c\u6fc0\u6d3b\u7684 AMPK \u901a\u8fc7\u6291\u5236 SREBP \u6291\u5236\u8102\u80aa\u751f\u6210\u3002\u7efc\u4e0a\uff0cASGR1 \u7f3a\u5931\u53ef\u589e\u52a0\u80c6\u56fa\u9187\u5916\u6392\uff0c\u800c\u4e0d\u50cf\u6cdb LXR \u6fc0\u52a8\u5242\u90a3\u6837\u8bf1\u5bfc\u8102\u80aa\u751f\u6210\u3002<\/p>\n

Asgr1\u2212\/\u2212 \u901a\u8fc7 AMPK \u4ea7\u751f\u4ee3\u8c22\u83b7\u76ca<\/h2>\n

\u63a5\u4e0b\u6765\uff0c\u7814\u7a76\u8005\u8bbe\u8ba1\u4e86\u4e00\u7cfb\u5217\u52a8\u7269\u5b9e\u9a8c\u8fdb\u884c\u4f53\u5185\u9a8c\u8bc1\u3002\u9996\u5148\uff0c\u5411\u91ce\u751f\u578b\u5c0f\u9f20\u9759\u8109\u6ce8\u5c04\u4e00\u6b21\u6027\u5927\u5242\u91cf\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A\uff0c\u5bfc\u81f4\u809d\u810f\u4e2d p-S6K \u548c p-4E-BP1 \u5347\u9ad8\uff0c\u63d0\u793a mTORC1 \u6fc0\u6d3b\uff1b\u540c\u65f6 p-AMPK\u3001p-Raptor\u3001p-ACC \u548c p-ULK1 \u964d\u4f4e\uff0c\u63d0\u793a AMPK \u88ab\u6291\u5236\u3002BARD1 \u589e\u52a0\uff0cLXR\u03b1 \u964d\u4f4e\u3002\u4e0e\u6b64\u4e00\u81f4\uff0cAbcg5\u3001Abcg8\u3001Abca1 \u548c Cyp7a1 \u8868\u8fbe\u4e0b\u964d\u3002Srebp1c\u3001Fasn \u548c Scd1 mRNA \u4e0a\u8c03\u3002\u809d\u810f\u548c\u8840\u6e05 TC\u3001TG \u660e\u663e\u589e\u52a0\uff0c\u80c6\u56fa\u9187\u548c\u80c6\u6c41\u9178\u5411\u80c6\u6c41\u7684\u6392\u6cc4\u5927\u5e45\u51cf\u5c11\u3002<\/p>\n

\u4e0e\u6b64\u5f62\u6210\u9c9c\u660e\u5bf9\u6bd4\u7684\u662f\uff0cAsgr1\u2212\/\u2212 \u5c0f\u9f20\u8868\u73b0\u4e3a mTORC1 \u53d7\u6291\u3001AMPK \u6fc0\u6d3b\u3001LXR\u03b1 \u589e\u52a0\u3001\u8102\u80aa\u9178\u751f\u7269\u5408\u6210\u57fa\u56e0\u8868\u8fbe\u4e0b\u964d\u4ee5\u53ca\u4ee3\u8c22\u8c31\u6539\u5584\uff0c\u4e14\u8fd9\u4e9b\u8868\u578b\u4e0d\u53d7\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A \u6ce8\u5c04\u5f71\u54cd\u3002<\/p>\n

\u5176\u6b21\uff0c\u7814\u7a76\u8005\u4f7f\u7528 mTOR \u6291\u5236\u5242\u96f7\u5e15\u9709\u7d20\u6a21\u62df Asgr1 \u7f3a\u5931\u8bf1\u5bfc\u7684 mTORC1 \u6291\u5236\u3002\u96f7\u5e15\u9709\u7d20\u663e\u8457\u589e\u52a0 p-AMPK\u3001p-Raptor\u3001p-ULK1 \u548c p-ACC\uff0c\u8bc1\u5b9e\u6291\u5236 mTORC1 \u53ef\u6fc0\u6d3b AMPK\u3002\u540c\u65f6\uff0cLXR\u03b1\u3001ABCG5\u3001ABCG8\u3001ABCA1 \u548c CYP7A1 \u6c34\u5e73\u5347\u9ad8\uff0c\u800c FASN \u6c34\u5e73\u4e0b\u964d\uff0c\u8fd9\u4e0e AMPK \u6291\u5236 SREBP \u901a\u8def\u4e00\u81f4\u3002\u96f7\u5e15\u9709\u7d20\u964d\u4f4e\u8840\u6e05\u548c\u809d\u810f\u4e2d\u7684 TC\u3001TG\uff0c\u5e76\u589e\u52a0\u80c6\u6c41\u80c6\u56fa\u9187\u3001\u80c6\u6c41\u9178\u548c\u7caa\u4fbf\u80c6\u56fa\u9187\u3002<\/p>\n

\u7b2c\u4e09\uff0c\u7814\u7a76\u8005\u901a\u8fc7\u9776\u5411 AMPK \u50ac\u5316\u4e9a\u57fa\u4e9a\u578b Prkaa1 \u548c Prkaa2 \u7684 AAV-shRNA\uff0c\u8003\u5bdf AMPK \u5bf9 Asgr1 \u7f3a\u5931\u8bf1\u5bfc\u7684\u4ee3\u8c22\u6539\u5584\u662f\u5426\u5fc5\u9700\u3002AMPK \u8017\u7aed\u4e00\u81f4\u6fc0\u6d3b mTORC1\uff0c\u5e76\u5728\u5f88\u5927\u7a0b\u5ea6\u4e0a\u9006\u8f6c Asgr1 \u7f3a\u5931\u809d\u810f\u4e2d LXR\u03b1\u3001ABCA1\u3001ABCG5\u3001ABCG8 \u548c CYP7A1 \u7684\u5347\u9ad8\u3002\u809d\u810f AMPK \u8017\u7aed\u589e\u52a0\u8840\u6e05\u548c\u809d\u810f TC\u3001TG \u6c34\u5e73\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0cAMPK shRNA \u5728\u91ce\u751f\u578b\u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\u5747\u964d\u4f4e\u80c6\u6c41\u80c6\u56fa\u9187\u3001\u80c6\u6c41\u9178\u4ee5\u53ca\u7caa\u4fbf\u80c6\u56fa\u9187\u3002<\/p>\n

\u6700\u540e\uff0c\u7814\u7a76\u8005\u4f7f\u7528 LXR \u6fc0\u52a8\u5242 GW3965 \u548c AMPK \u6fc0\u52a8\u5242 A769662\uff0c\u68c0\u9a8c AMPK \u6fc0\u6d3b\u662f\u5426\u53ef\u6291\u5236\u8102\u8d28\u751f\u7269\u5408\u6210\uff0c\u5e76\u533a\u5206 ASGR1 \u7f3a\u5931\u4e0e\u76f4\u63a5 LXR \u6fc0\u6d3b\u3002A769662 \u786e\u5b9e\u6fc0\u6d3b AMPK \u5e76\u968f\u540e\u6291\u5236 mTORC1\u3002\u5b83\u8fd8\u964d\u4f4e BARD1 \u5e76\u589e\u52a0 LXR\u03b1\uff0c\u8bf4\u660e AMPK \u6fc0\u6d3b\u786e\u5b9e\u4f1a\u5347\u9ad8 LXR\u03b1\u3002\u6b64\u5916\uff0cA769662 \u589e\u52a0 ABCG8\u3001ABCA1 \u548c CYP7A1\u3002<\/p>\n

\u5355\u72ec\u6216\u8054\u5408\u7ed9\u4e88 GW3965 \u548c A769662 \u5747\u964d\u4f4e\u8840\u6e05\u548c\u809d\u810f\u80c6\u56fa\u9187\u6c34\u5e73\uff0c\u5e76\u589e\u52a0\u80c6\u56fa\u9187\u5411\u80c6\u6c41\u548c\u7caa\u4fbf\u7684\u6392\u6cc4\uff0c\u56e0\u4e3a\u4e8c\u8005\u5747\u53ef\u6fc0\u6d3b LXR\u3002\u7531\u4e8e AMPK \u6291\u5236 SREBP\uff0cA769662 \u663e\u8457\u964d\u4f4e Srebp2\u3001Hmgcr\u3001Srebp1c\u3001Fasn \u548c Scd1 mRNA\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0cGW3965 \u589e\u52a0 LXR \u9776\u57fa\u56e0\u8868\u8fbe\uff0c\u800c\u8054\u5408 A769662 \u53ef\u964d\u4f4e Srebp1c\u3001Fasn \u548c Scd1 mRNA\uff0c\u4f46\u4e0d\u964d\u4f4e Abcg5\u3001Abcg8\u3001Abca1 \u548c Cyp7a1 \u7b49\u5176\u4ed6 LXR \u9776\u57fa\u56e0\u3002GW3965 \u589e\u52a0\u809d\u810f TG\uff0c\u8fd9\u662f\u6cdb LXR \u6fc0\u52a8\u5242\u7684\u4e3b\u8981\u526f\u4f5c\u7528\uff1b\u7136\u800c\uff0c\u4e0e A769662 \u8054\u5408\u7ed9\u836f\u540e\uff0c\u809d\u810f TG \u8f83 GW3965 \u7ec4\u4e0b\u964d 47.5%\u3002\u603b\u4e4b\uff0cGW3965 \u4e0e A769662 \u8054\u5408\u7ed9\u836f\u53ef\u590d\u73b0 Asgr1 \u7f3a\u5931\u7684\u57fa\u56e0\u8868\u8fbe\u548c\u4ee3\u8c22\u8868\u578b\u3002<\/p>\n

\u6291\u5236 ASGR1 \u7684\u6cbb\u7597\u6f5c\u529b<\/h2>\n

RNA \u6291\u5236\uff08RNAi\uff09\u6b63\u6210\u4e3a\u4e00\u79cd\u6709\u529b\u7684\u6cbb\u7597\u65b9\u6cd5\u3002\u7814\u7a76\u8005\u4f7f\u7528 AAV-shRNA \u5728\u809d\u810f\u4e2d\u6c89\u9ed8 Asgr1\u3002LXR\u03b1\u3001ABCG8\u3001ABCA1 \u548c CYP7A1 \u86cb\u767d\u6c34\u5e73\u663e\u8457\u5347\u9ad8\uff0c\u800c Srebp1c\u3001Fasn \u548c Scd1 mRNA \u964d\u4f4e\u3002Asgr1 \u6572\u4f4e\u540e\uff0c\u8840\u6e05\u548c\u809d\u810f\u4e2d TC\u3001TG \u6c34\u5e73\u663e\u8457\u4e0b\u964d\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0c\u6ce8\u5c04 AAV-shAsgr1 \u7684\u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u589e\u52a0\u8d85\u8fc7 1.4 \u500d\uff0c\u603b\u80c6\u6c41\u9178\u4e5f\u540c\u6b65\u589e\u52a0\u3002AAV-shAsgr1 \u672a\u6539\u53d8\u4f53\u91cd\uff0c\u4e5f\u672a\u9020\u6210\u809d\u635f\u4f24\u3002\u5728 Ldlr\u2212\/\u2212 \u5c0f\u9f20\u4e2d\uff0c\u6572\u4f4e Asgr1 \u663e\u8457\u9884\u9632\u9ad8\u8102\/\u9ad8\u80c6\u56fa\u9187\/\u80c6\u76d0\u996e\u98df\u8bf1\u5bfc\u7684\u52a8\u8109\u7ca5\u6837\u786c\u5316\u6591\u5757\u5f62\u6210\u548c\u809d\u810f\u8102\u8d28\u6c89\u79ef\u3002<\/p>\n

\u4e3a\u8fdb\u4e00\u6b65\u7406\u89e3 ASGR1 \u6291\u5236\u7684\u6cbb\u7597\u6548\u679c\uff0c\u7814\u7a76\u8005\u5f00\u53d1\u4e86\u6297 ASGR1 \u5355\u514b\u9686\u4e2d\u548c\u6297\u4f53\u30024B9-IgG \u514b\u9686\u4ee5\u5242\u91cf\u4f9d\u8d56\u65b9\u5f0f\u589e\u52a0 LXR\u03b1\uff0c\u5e76\u4e0a\u8c03 Abcg5 \u548c Abcg8 \u8868\u8fbe\u30024B9-IgG \u8bc6\u522b ASGR1 \u4e2d\u4f4d\u4e8e\u7cd6\u8bc6\u522b\u7ed3\u6784\u57df\u7684\u533a\u57df\uff08\u6c28\u57fa\u9178 182-274\uff09\u3002<\/p>\n

\u91ce\u751f\u578b\u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u63a5\u53d7\u5bf9\u7167 IgG \u6216 4B9-IgG \u8179\u8154\u6ce8\u5c04\u30024B9-IgG \u5904\u7406\u7684\u91ce\u751f\u578b\u5c0f\u9f20\u4e2d LXR\u03b1 \u663e\u8457\u589e\u52a0\uff0cLXR\u03b2 \u7565\u6709\u589e\u52a0\uff0cABCG8\u3001CYP7A1 \u548c ABCA1 \u8868\u8fbe\u660e\u663e\u5347\u9ad8\u30024B9-IgG \u6fc0\u6d3b AMPK\uff0c\u5e76\u964d\u4f4e SREBP1\u3001FASN \u548c BARD1\u3002\u4e0e\u6b64\u4e00\u81f4\uff0c4B9-IgG \u5728\u91ce\u751f\u578b\u5c0f\u9f20\u809d\u810f\u4e2d\u4e0a\u8c03 Abcg5\u3001Abcg8\u3001Abca1 \u548c Cyp7a1 \u8f6c\u5f55\uff0c\u5e76\u964d\u4f4e Srebp1c\u3001Fasn \u548c Scd1 mRNA\u3002\u5728 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\uff0c4B9-IgG \u4e0d\u6539\u53d8 AMPK \u6fc0\u6d3b\u6216\u4e0a\u8ff0\u57fa\u56e0\u8868\u8fbe\u3002<\/p>\n

\u5728\u91ce\u751f\u578b\u5c0f\u9f20\u4e2d\uff0c4B9-IgG \u5206\u522b\u4f7f\u8840\u6e05 TC \u548c TG \u964d\u4f4e 50% \u548c 22%\uff0c\u809d\u810f TC \u548c TG \u4e5f\u663e\u8457\u4e0b\u964d\u30024B9-IgG \u5904\u7406\u8fd8\u4f7f\u80c6\u56ca\u80c6\u6c41\u4f53\u79ef\u589e\u52a0 62%\uff0c\u80c6\u6c41\u80c6\u56fa\u9187\u6d53\u5ea6\u589e\u52a0 67%\uff0c\u5bfc\u81f4\u91ce\u751f\u578b\u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u589e\u52a0\u8d85\u8fc7\u4e00\u500d\u3002\u5bf9\u7167 IgG \u5904\u7406\u7684\u91ce\u751f\u578b\u5c0f\u9f20\u80c6\u6c41\u6e05\u6f88\uff0c\u800c 4B9-IgG \u6ce8\u5c04\u5c0f\u9f20\u6216 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u80c6\u6c41\u4e0d\u900f\u660e\u30024B9-IgG \u5904\u7406\u7684\u91ce\u751f\u578b\u5c0f\u9f20\u7caa\u4fbf\u80c6\u56fa\u9187\u589e\u52a0\u7ea6 87%\u3002\u80c6\u6c41\u80c6\u6c41\u9178\u6d53\u5ea6\u548c\u603b\u80c6\u6c41\u9178\u4e5f\u56e0 4B9-IgG \u800c\u589e\u52a0\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0c4B9-IgG \u4e0d\u6539\u5584 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4ee3\u8c22\u8c31\uff0c\u652f\u6301 4B9-IgG \u7684\u7279\u5f02\u6027\u3002\u672a\u68c0\u6d4b\u5230\u5176\u4ed6\u4ee3\u8c22\u6539\u53d8\u6216\u809d\u635f\u4f24\u3002<\/p>\n

\u6297 ASGR1 \u4e0e\u4ed6\u6c40\/\u4f9d\u6298\u9ea6\u5e03\u534f\u540c<\/h2>\n

\u7814\u7a76\u8005\u968f\u540e\u8003\u5bdf 4B9-IgG \u662f\u5426\u4e0e\u73b0\u6709\u836f\u7269\u963f\u6258\u4f10\u4ed6\u6c40\u548c\u4f9d\u6298\u9ea6\u5e03\u4ea7\u751f\u534f\u540c\u964d\u8102\u4f5c\u7528\u3002\u4e0d\u8bba\u662f\u5426\u4f7f\u7528\u963f\u6258\u4f10\u4ed6\u6c40\uff0c4B9-IgG \u90fd\u5728\u91ce\u751f\u578b\u5c0f\u9f20\u4e2d\u6fc0\u6d3b AMPK\u3001\u589e\u52a0 LXR\u03b1\u3001\u4e0a\u8c03 Abcg5\u3001Abcg8\u3001Abca1 \u548c Cyp7a1 \u8868\u8fbe\uff0c\u5e76\u964d\u4f4e Srebp1c\u3001Fasn \u548c Scd1 \u8868\u8fbe\u3002<\/p>\n

\u5355\u72ec\u4f7f\u7528 4B9-IgG \u6216\u963f\u6258\u4f10\u4ed6\u6c40\u5747\u53ef\u76f8\u4f3c\u5730\u964d\u4f4e\u91ce\u751f\u578b\u5c0f\u9f20 TC \u548c TG\u3002\u4f53\u91cd\u548c\u5176\u4ed6\u4ee3\u8c22\u53c2\u6570\u76f8\u4f3c\u3002\u503c\u5f97\u6ce8\u610f\u7684\u662f\uff0c4B9-IgG \u4e0e\u963f\u6258\u4f10\u4ed6\u6c40\u8054\u5408\u7ed9\u836f\u663e\u8457\u964d\u4f4e\u91ce\u751f\u578b\u5c0f\u9f20\u8840\u6e05\u548c\u809d\u810f\u8102\u8d28\u6c34\u5e73\u3002Asgr1\u2212\/\u2212 \u5c0f\u9f20 TC \u548c TG \u6c34\u5e73\u8fdc\u4f4e\u4e8e\u91ce\u751f\u578b\u5c0f\u9f20\u3002\u963f\u6258\u4f10\u4ed6\u6c40\u53ef\u8fdb\u4e00\u6b65\u964d\u4f4e\u8840\u6e05\u8102\u8d28\u6c34\u5e73\uff0c\u800c 4B9-IgG \u5728 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\u65e0\u6548\u3002\u4e0e\u56fe 4 \u7ed3\u679c\u4e00\u81f4\uff0c\u65e0\u8bba\u662f\u5426\u4f7f\u7528\u963f\u6258\u4f10\u4ed6\u6c40\uff0c4B9-IgG \u5747\u663e\u8457\u589e\u52a0\u91ce\u751f\u578b\u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u3001\u603b\u80c6\u6c41\u9178\u548c\u7caa\u4fbf\u80c6\u56fa\u9187\u3002\u5355\u72ec\u963f\u6258\u4f10\u4ed6\u6c40\u5bf9\u80c6\u6c41\u80c6\u56fa\u9187\u3001\u80c6\u6c41\u9178\u6216\u7caa\u4fbf\u80c6\u56fa\u9187\u6c34\u5e73\u65e0\u5f71\u54cd\u3002<\/p>\n

\u6700\u540e\uff0c\u7814\u7a76\u8005\u7528 4B9-IgG \u548c\u4f9d\u6298\u9ea6\u5e03\u5904\u7406\u5c0f\u9f20\u3002\u5728\u91ce\u751f\u578b\u5c0f\u9f20\u4e2d\uff0c\u65e0\u8bba\u662f\u5426\u5b58\u5728\u4f9d\u6298\u9ea6\u5e03\uff0c4B9-IgG \u5747\u4ee5\u963b\u65ad ASGR1 \u7684\u65b9\u5f0f\u53d1\u6325\u4f5c\u7528\u30024B9-IgG \u4e0e\u4f9d\u6298\u9ea6\u5e03\u534f\u540c\u964d\u4f4e\u91ce\u751f\u578b\u5c0f\u9f20\u8840\u6e05\u548c\u809d\u810f\u4e2d\u7684 TC\u3001TG\uff0c\u5c3d\u7ba1\u5355\u72ec\u5904\u7406\u4e5f\u80fd\u6539\u5584\u8102\u8d28\u8c31\u3002\u5728 Asgr1\u2212\/\u2212 \u5c0f\u9f20\u4e2d\uff0c\u4f9d\u6298\u9ea6\u5e03\u4ecd\u53ef\u964d\u4f4e\u8840\u6e05\u548c\u809d\u810f\u4e2d\u7684\u8102\u8d28\u542b\u91cf\uff0c\u800c 4B9-IgG \u65e0\u6548\u3002\u4f9d\u6298\u9ea6\u5e03\u663e\u8457\u964d\u4f4e\u91ce\u751f\u578b\u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\uff0c\u56e0\u4e3a\u5b83\u6291\u5236\u81b3\u98df\u80c6\u56fa\u9187\u5438\u6536\u3002\u5373\u4f7f\u5408\u7528\u4f9d\u6298\u9ea6\u5e03\uff0c4B9-IgG \u4ecd\u53ef\u589e\u52a0\u91ce\u751f\u578b\u5c0f\u9f20\u603b\u80c6\u6c41\u80c6\u56fa\u9187\u548c\u80c6\u6c41\u9178\u3002<\/p>\n

\u8ba8\u8bba<\/h2>\n

mTORC1 \u548c AMPK \u662f\u4e24\u4e2a\u611f\u77e5\u7ec6\u80de\u8425\u517b\u72b6\u6001\u5e76\u63a7\u5236\u4ee3\u8c22\u7684\u4e3b\u8c03\u63a7\u56e0\u5b50\u3002\u5b83\u4eec\u901a\u8fc7\u591a\u79cd\u673a\u5236\u53cd\u5411\u8c03\u63a7\u3002\u867d\u7136 AMPK \u88ab\u8ba4\u4e3a\u53ef\u80fd\u6210\u4e3a\u4ee3\u8c22\u75be\u75c5\u6cbb\u7597\u9776\u70b9\uff0c\u4f46\u6cdb AMPK \u6fc0\u52a8\u5242\u4f1a\u5bfc\u81f4\u5fc3\u810f\u80a5\u5927\uff0c\u9650\u5236\u5176\u4e34\u5e8a\u5e94\u7528\u3002\u9664 AMPK \u6fc0\u6d3b\u7684\u7ec4\u7ec7\u7279\u5f02\u6027\u4f5c\u7528\u5916\uff0c\u7ec6\u80de\u5185 AMPK \u8fd8\u5b58\u5728\u533a\u5ba4\u5316\uff0c\u5e76\u53d7\u836f\u7269\u3001\u8425\u517b\u53ef\u7528\u6027\u548c AMP \u7684\u5dee\u5f02\u8c03\u63a7\uff0c\u4ece\u800c\u5bfc\u81f4\u4e0d\u540c\u9776\u6807\u88ab\u78f7\u9178\u5316\u3002\u56e0\u6b64\uff0c\u5728\u836f\u7269\u5f00\u53d1\u4e2d\uff0c\u9009\u62e9\u6027 AMPK \u6fc0\u6d3b\u5bf9\u83b7\u5f97\u4ee3\u8c22\u83b7\u76ca\u5e76\u907f\u514d\u4e0d\u826f\u53cd\u5e94\u81f3\u5173\u91cd\u8981\u3002<\/p>\n

\u672c\u6587\u63ed\u793a mTORC1 \u548c AMPK \u53ef\u53d7 ASGR1 \u8c03\u8282\u3002\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u7ecf ASGR1 \u4ecb\u5bfc\u7684\u5185\u541e\u8fdb\u5165\u809d\u7ec6\u80de\uff0c\u5176\u6eb6\u9176\u4f53\u6d88\u5316\u91ca\u653e\u7684\u6c28\u57fa\u9178\u6fc0\u6d3b mTORC1\u3002\u4ece\u6982\u5ff5\u4e0a\u770b\uff0c\u8fd9\u4e00\u53d1\u73b0\u4e0e\u6b64\u524d\u5173\u4e8e\u81ea\u566c\u6216\u5de8\u80de\u996e\u4ea7\u751f\u7684\u6eb6\u9176\u4f53\u6c28\u57fa\u9178\u53ef\u6fc0\u6d3b mTORC1 \u7684\u8ba4\u8bc6\u4e00\u81f4\u3002\u6291\u5236 ASGR1 \u53ef\u963b\u65ad\u53d7\u4f53\u4ecb\u5bfc\u7684\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u5185\u5316\u53ca\u968f\u540e\u7684\u6eb6\u9176\u4f53\u6d88\u5316\uff0c\u4ece\u800c\u6fc0\u6d3b AMPK \u5e76\u6291\u5236 mTORC1\u3002\u8fd9\u4e00\u673a\u5236\u63d0\u4f9b\u4e86\u4e00\u79cd\u9ad8\u5ea6\u5c40\u90e8\u5316\u7684\u4fe1\u53f7\uff0c\u53ef\u5b9e\u73b0\u9009\u62e9\u6027 AMPK \u6fc0\u6d3b\u3002<\/p>\n

\u80c6\u56fa\u9187\u5916\u6392\u901a\u8fc7 LXR\u03b1 \u548c ABCG5\/G8 \u589e\u52a0\u3002\u7531\u4e8e ABCA1 \u88ab LXR\u03b1 \u4e0a\u8c03\uff0c\u8102\u86cb\u767d\u8c31\u4e5f\u5f97\u5230\u6539\u5584\uff0c\u8868\u73b0\u4e3a HDL \u80c6\u56fa\u9187\u5347\u9ad8\u3001LDL \u80c6\u56fa\u9187\u964d\u4f4e\u3002\u4f5c\u4e3a mTORC1 \u6291\u5236\u548c AMPK \u6fc0\u6d3b\u7684\u7ed3\u679c\uff0cSREBP1 \u88ab\u6291\u5236\uff0c\u4ece\u800c\u9632\u6b62\u8102\u80aa\u751f\u6210\u3002\u6b64\u5916\uff0cTtc39b \u6d88\u878d\u53ef\u589e\u52a0 LXR\u03b1 \u548c ABCG5\/G8 \u800c\u4e0d\u6fc0\u6d3b SREBP1\uff0c\u8fd9\u4e5f\u8bc1\u5b9e ABCG5\/G8 \u8868\u8fbe\u53ef\u4e0e SREBP1 \u8868\u8fbe\u8131\u8026\u3002\u7531\u4e8e ASGR1 \u51e0\u4e4e\u53ea\u5728\u809d\u7ec6\u80de\u4e2d\u8868\u8fbe\uff0c\u9776\u5411 ASGR1 \u4e3a\u809d\u810f\u7279\u5f02\u6027\u6fc0\u6d3b AMPK \u548c\u6291\u5236 mTORC1 \u5f00\u8f9f\u4e86\u8def\u5f84\uff0c\u53ef\u7ed5\u5f00\u6cdb AMPK \u6fc0\u52a8\u5242\u7684\u4e0d\u826f\u526f\u4f5c\u7528\u3002<\/p>\n

\u5728\u6cbb\u7597\u9ad8\u8840\u7cd6\u7684\u836f\u7269\u4e2d\uff0c\u94a0-\u8461\u8404\u7cd6\u534f\u540c\u8f6c\u8fd0\u86cb\u767d 2\uff08SGLT2\uff09\u6291\u5236\u5242\u901a\u8fc7\u963b\u65ad\u80be\u810f\u8461\u8404\u7cd6\u91cd\u5438\u6536\uff0c\u4fc3\u8fdb\u8461\u8404\u7cd6\u7ecf\u5c3f\u6db2\u6392\u6cc4\u3002\u8fd9\u4e9b\u6291\u5236\u5242\u4ea7\u751f\u591a\u91cd\u6709\u76ca\u4f5c\u7528\uff0c\u4e3b\u8981\u662f\u56e0\u4e3a\u5b83\u4eec\u4fc3\u4f7f\u80be\u810f\u5c06\u8461\u8404\u7cd6\u79fb\u51fa\u4f53\u5916\u3002\u80c6\u56fa\u9187\u662f\u4e00\u79cd\u5e26\u6709\u56db\u73af\u7ed3\u6784\u7684\u8102\u8d28\uff0c\u96be\u4ee5\u5206\u89e3\uff0c\u5e76\u4e14\u662f\u5f3a\u4fc3\u708e\u5206\u5b50\uff0c\u53ef\u52a0\u901f\u52a8\u8109\u7ca5\u6837\u786c\u5316\u548c\u975e\u9152\u7cbe\u6027\u8102\u80aa\u6027\u809d\u708e\u3002\u6291\u5236 ASGR1 \u53ef\u589e\u52a0\u80c6\u56fa\u9187\u5411\u80c6\u6c41\u548c\u968f\u540e\u5411\u7caa\u4fbf\u7684\u6392\u6cc4\uff0c\u4e3a\u964d\u4f4e\u80c6\u56fa\u9187\u63d0\u4f9b\u4e86\u4e00\u79cd\u72ec\u7279\u65b9\u5f0f\u3002\u4e8b\u5b9e\u4e0a\uff0cASGR1 \u529f\u80fd\u7f3a\u5931\u53d8\u5f02\u5df2\u88ab\u53d1\u73b0\u4e0e\u8f83\u4f4e\u975e HDL \u80c6\u56fa\u9187\u548c\u8f83\u4f4e\u5fc3\u8840\u7ba1\u75be\u75c5\u98ce\u9669\u76f8\u5173\uff0c\u8fd9\u63d0\u793a ASGR1 \u6291\u5236\u662f\u4e00\u79cd\u5b89\u5168\u6709\u6548\u7684\u5fc3\u8840\u7ba1\u75be\u75c5\u6cbb\u7597\u65b9\u5f0f\u3002<\/p>\n

\u5728\u7ebf\u5185\u5bb9<\/h2>\n

\u4efb\u4f55\u65b9\u6cd5\u3001\u989d\u5916\u53c2\u8003\u6587\u732e\u3001Nature Research \u62a5\u544a\u6458\u8981\u3001\u6e90\u6570\u636e\u3001\u6269\u5c55\u6570\u636e\u3001\u8865\u5145\u4fe1\u606f\u3001\u81f4\u8c22\u3001\u540c\u884c\u8bc4\u8bae\u4fe1\u606f\uff1b\u4f5c\u8005\u8d21\u732e\u548c\u5229\u76ca\u51b2\u7a81\u58f0\u660e\uff1b\u6570\u636e\u548c\u4ee3\u7801\u53ef\u7528\u6027\u58f0\u660e\uff0c\u5747\u53ef\u5728 https:\/\/doi.org\/10.1038\/s41586-022-05006-3<\/a> \u83b7\u53d6\u3002<\/p>\n

\u65b9\u6cd5<\/h2>\n

\u5c0f\u9f20<\/h3>\n

Asgr1 \u6572\u9664\u5c0f\u9f20\u7531 CRISPR-Cas9 \u7cfb\u7edf\u751f\u6210\u3002\u5355\u5bfc RNA\uff08sgRNA\uff09\u548c\u4f9b\u4f53\u5171\u540c\u6ce8\u5c04\u5165 C57BL\/6J \u5408\u5b50\u3002sgRNA \u5f15\u5bfc Cas9 \u6838\u9178\u5185\u5207\u9176\u5728\u5185\u542b\u5b50 2-3 \u548c\u5185\u542b\u5b50 8-9 \u4e2d\u5207\u5272\uff0c\u5982\u6269\u5c55\u6570\u636e\u56fe 1 \u6240\u793a\u3002LXR\u03b1 \u6572\u9664\u5c0f\u9f20\u7531\u4e2d\u56fd\u79d1\u5b66\u9662\u4e0a\u6d77\u8425\u517b\u4e0e\u5065\u5eb7\u7814\u7a76\u6240\u5c39\u6167\u52c7\u9988\u8d60\u3002<\/p>\n

C57\/B6J \u5c0f\u9f20\uff086-8 \u5468\u9f84\uff09\u8d2d\u81ea GemPharmatech \u548c\u6e56\u5317\u7701\u75be\u63a7\u4e2d\u5fc3\u3002\u56fe\u6ce8\u6240\u8ff0\u5b9e\u9a8c\u4f7f\u7528\u4e86\u96cc\u96c4\u5c0f\u9f20\u3002C57\/B6J Ldlr\u2212\/\u2212 \u5c0f\u9f20\u8d2d\u81ea GemPharmatech\u3002Asgr1+\/\u2212 \u6742\u4ea4\u4ea7\u751f\u7684\u91ce\u751f\u578b\u540c\u7a9d\u5c0f\u9f20\u4f5c\u4e3a\u52a8\u7269\u5b9e\u9a8c\u5bf9\u7167\u3002\u4e3a\u751f\u6210 Asgr1\u2212\/\u2212Lxr\u03b1\u2212\/\u2212 \u53cc\u6572\u9664\u5c0f\u9f20\uff0c\u5c06 Asgr1+\/\u2212 \u4e0e Lxr\u03b1+\/\u2212 \u5c0f\u9f20\u6742\u4ea4\u3002\u6240\u6709\u5c0f\u9f20\u9972\u517b\u4e8e SPF \u73af\u5883\uff0c\u6e29\u5ea6 22 \u00b1 1 \u00b0C\uff0c\u76f8\u5bf9\u6e7f\u5ea6 50-60%\uff0c\u5149\u7167\u5468\u671f\u4e3a 06:00-18:00\u3002\u5904\u7406\u524d\u6240\u6709\u52a8\u7269\u5747\u7ef4\u6301\u666e\u901a\u9972\u6599\u3002\u6309\u56fe\u6ce8\u6240\u8ff0\uff0c\u5e74\u9f84\u5339\u914d\u5c0f\u9f20\u7ed9\u4e88\u9ad8\u8102\/\u9ad8\u80c6\u56fa\u9187\/\u80c6\u76d0\u996e\u98df\u3002\u5904\u6b7b\u524d\u6240\u6709\u5c0f\u9f20\u7981\u98df 4 \u5c0f\u65f6\u3002\u7531\u4e8e\u7814\u7a76\u8005\u5fc5\u987b\u6267\u884c\u76f8\u5e94\u64cd\u4f5c\uff0c\u65e0\u6cd5\u5bf9\u5b9e\u9a8c\u8005\u5b9e\u65bd\u76f2\u6cd5\u3002\u5c0f\u9f20\u968f\u673a\u5206\u914d\u81f3\u4e0d\u540c\u7ec4\uff0c\u5e76\u5c3d\u53ef\u80fd\u63a5\u53d7\u7b49\u540c\u5904\u7406\u3002\u6ca1\u6709\u6570\u636e\u88ab\u6392\u9664\u3002\u6240\u6709\u52a8\u7269\u64cd\u4f5c\u5747\u7ecf\u6b66\u6c49\u5927\u5b66\u52a8\u7269\u62a4\u7406\u4e0e\u4f7f\u7528\u59d4\u5458\u4f1a\u6279\u51c6\u3002<\/p>\n

\u6750\u6599\u548c\u8d28\u7c92<\/h3>\n

MG132 \u8d2d\u81ea Boston Biochem\uff1bdorsomorphin 2HCl\uff08AMPK \u6291\u5236\u5242\uff09\u548c A769662\uff08AMPK \u6fc0\u6d3b\u5242\uff09\u8d2d\u81ea Target Molecule\uff1bbafilomycin A1 \u8d2d\u81ea MCE\uff1b\u6d1b\u4f10\u4ed6\u6c40\u8d2d\u81ea\u4e0a\u6d77 Pharm Valley\uff1b\u7532\u7f9f\u620a\u9178\u548c\u80ce\u7403\u86cb\u767d A \u8d2d\u81ea Sigma\uff1bEZ18D03841 \u8d2d\u81ea Steroloids\uff1b\u963f\u6258\u4f10\u4ed6\u6c40\u8d2d\u81ea Energy Chemical\uff1bDil-LDL \u8d2d\u81ea Yeasen\uff1b\u8840\u6e05\u7c7b\u9ecf\u86cb\u767d\u8d2d\u81ea Abcam\uff1bPMSF \u8d2d\u81ea Thermo Scientific\uff1b\u96f7\u5e15\u9709\u7d20\u548c GW3965 \u8d2d\u81ea Topscience\uff1bpoloxamer 407 \u8d2d\u81ea Sigma\uff1b\u653e\u5c04\u6027\u80c6\u56fa\u9187\u6cb9\u9178\u916f\u8d2d\u81ea Perkin Elmer\uff1bDOPC \u548c DAB \u5e95\u7269\u8bd5\u5242\u76d2\u5206\u522b\u8d2d\u81ea Sigma \u548c Abcam\u3002\u8102\u86cb\u767d\u7f3a\u4e4f\u8840\u6e05\uff08LPDS\uff09\u7531\u65b0\u751f\u725b\u8840\u6e05\u5728\u672c\u5b9e\u9a8c\u5ba4\u7ecf\u8d85\u901f\u79bb\u5fc3\u5236\u5907\u3002<\/p>\n

\u7814\u7a76\u8005\u4f7f\u7528\u6807\u51c6\u5206\u5b50\u514b\u9686\u65b9\u6cd5\u6784\u5efa\u4e86\u591a\u79cd\u8d28\u7c92\u3002pCMV-ASGR1-3\u00d7FLAG \u7f16\u7801\u5e26\u8868\u4f4d\u6807\u7b7e\u7684\u5168\u957f\u5c0f\u9f20 Asgr1\u3002pCMV-EGFP-LXR\u03b1 \u548c pCMV-5\u00d7MYC-LXR\u03b1 \u5206\u522b\u7f16\u7801\u5e26 EGFP \u6216 5\u00d7MYC \u8868\u4f4d\u6807\u7b7e\u7684\u5c0f\u9f20 LXR\u03b1\u3002pEF-HA-Ubi \u7f16\u7801\u5e26 HA \u8868\u4f4d\u6807\u7b7e\u7684\u4eba Ubi\u3002pCMV-BARD1-5\u00d7MYC \u548c pCMV-LDLR-5\u00d7MYC \u5206\u522b\u7f16\u7801\u5e26 5\u00d7MYC \u8868\u4f4d\u6807\u7b7e\u7684\u4eba BARD1 \u548c\u5c0f\u9f20 LDLR\u3002ASGR1 \u622a\u77ed\u8d28\u7c92\u548c pCMV-ASGR1(6A)-3\u00d7FLAG \u5747\u901a\u8fc7 QuikChange \u6784\u5efa\u3002pCMV-EGFP-BRCA1 \u6765\u81ea Addgene\uff08\u7f16\u53f7 71116\uff09\u3002TMEM192 \u7f16\u7801\u533a\u4ece Huh7 \u7ec6\u80de cDNA \u6269\u589e\u5e76\u8fde\u63a5\u81f3 pCMV14-3\u00d7FLAG \u8f7d\u4f53\uff0c\u968f\u540e\u4e9a\u514b\u9686\u81f3 pLVX-IRES-puro\u3002\u91ce\u751f\u578b\u548c\u7a81\u53d8\u578b SLC38A9 cDNA \u7531\u540c\u6d4e\u5927\u5b66\u738b\u9e4f\u63d0\u4f9b\u5e76\u4e9a\u514b\u9686\u81f3 pLVX-IRES-puro\u3002<\/p>\n

\u7ec6\u80de\u57f9\u517b<\/h3>\n

Huh7 \u548c HEK293T \u7ec6\u80de\u5728 37 \u00b0C\u30015% CO2 \u6761\u4ef6\u4e0b\u57f9\u517b\u3002Huh7 \u548c HEK293T \u7ec6\u80de\u4f7f\u7528 Dulbecco's Modified Eagle Medium\uff08DMEM\uff09\u57f9\u517b\uff0c\u57f9\u517b\u57fa\u542b 100 units\/ml \u9752\u9709\u7d20\u3001100 \u03bcg\/ml \u94fe\u9709\u7d20\u548c 10% FBS\u3002\u539f\u4ee3\u5c0f\u9f20\u809d\u7ec6\u80de\u4ece\u666e\u901a\u9972\u6599\u5582\u517b\u7684 8 \u5468\u9f84\u96c4\u6027 C57BL\/6J \u5c0f\u9f20\u65b0\u9c9c\u5206\u79bb\uff0c\u5e76\u5728\u542b 5% FBS\u3001100 units\/ml \u9752\u9709\u7d20\u548c 100 \u03bcg\/ml \u94fe\u9709\u7d20\u7684 Medium 199 \u4e2d\u57f9\u517b\u3002\u80c6\u56fa\u9187\u8017\u7aed\u57f9\u517b\u57fa\u4e3a\u542b 5% LPDS\u30011 \u03bcM \u6d1b\u4f10\u4ed6\u6c40\u548c 10 \u03bcM \u7532\u7f9f\u620a\u9178\u94a0\u7684 DMEM\u3002<\/p>\n

\u7ec6\u80de\u7cfb\u6784\u5efa<\/h3>\n

ASGR1 KO \u7ec6\u80de\uff08ASGR1 KO-A\u3001KO-B\uff09\u4f7f\u7528 CRISPR-Cas9 \u57fa\u56e0\u7ec4\u7f16\u8f91\u7cfb\u7edf\u6784\u5efa\u3002\u7b80\u8a00\u4e4b\uff0c\u4eba ASGR1 \u9776\u5411 sgRNA\uff08\u5e8f\u5217\uff1a5'-CTGAAGGTCTTGATACTCCT-3'\uff09\u514b\u9686\u5165 pX330-U6-Chimeric_BB-CBh-hSpCas9 \u8f7d\u4f53\uff08#42230\uff0cAddgene\uff09\uff0c\u968f\u540e\u4e0e pLKO.1 \u5171\u8f6c\u67d3 Huh7 \u7ec6\u80de\u3002\u8f6c\u67d3 8 \u5c0f\u65f6\u540e\uff0c\u7ec6\u80de\u5728\u542b 10% FBS \u548c 2 \u03bcg\/ml \u560c\u5464\u9709\u7d20\u7684 DMEM \u4e2d\u57f9\u517b\uff0c\u76f4\u81f3\u5f62\u6210\u5355\u514b\u9686\u3002<\/p>\n

\u7a33\u5b9a\u8868\u8fbe ASGR1 \u7684 Huh7 \u7ec6\u80de\uff08ASGR1 OE-A\u3001OE-B\uff09\u6309\u65e2\u5f80\u65b9\u6cd5\u751f\u6210\u3002\u7b80\u8a00\u4e4b\uff0c\u7528 FuGENE HD \u77ac\u65f6\u8f6c\u67d3 pCMV-ASGR1-3\u00d7Flag\u30028 \u5c0f\u65f6\u540e\u66f4\u6362\u4e3a\u542b 10% FBS \u548c 400 \u03bcg\/ml G418 \u7684 DMEM\u3002\u6bcf 2 \u5929\u66f4\u6362\u57f9\u517b\u57fa\uff0c\u76f4\u81f3\u5f62\u6210\u5355\u514b\u9686\u3002\u9884\u9a8c\u8bc1 guide RNA \u5e8f\u5217\u5408\u6210\u540e\u8fde\u63a5\u5165 pX330 \u8f7d\u4f53\uff08GGCTCAAACTGGATATTCATAGG\uff09\u3002Huh7 \u7ec6\u80de\u4e0e pX330 \u548c\u63d0\u4f9b\u560c\u5464\u9709\u7d20\u6297\u6027\u7684 pLKO.1 \u5171\u8f6c\u67d3\u3002\u7ec6\u80de\u7528 5 \u03bcg\/ml \u560c\u5464\u9709\u7d20\u7b5b\u9009 48 \u5c0f\u65f6\uff0c\u7136\u540e\u4ee5\u6bcf\u5b54 0.8 \u4e2a\u7ec6\u80de\u7684\u5bc6\u5ea6\u63a5\u79cd\u5230 96 \u5b54\u677f\u4e2d\uff0c\u57f9\u517b 2 \u5468\u3002SLC38A9 \u6572\u9664\u901a\u8fc7\u514d\u75ab\u5370\u8ff9\u786e\u8ba4\u3002<\/p>\n

\u6cdb\u7d20\u5316\u68c0\u6d4b<\/h3>\n

Huh7 \u7ec6\u80de\u5728\u7b2c 0 \u5929\u4ee5 6 \u00d7 10^5 \u4e2a\u7ec6\u80de\u5bc6\u5ea6\u63a5\u79cd\u4e8e 6 cm \u57f9\u517b\u76bf\u4e2d\uff0c\u6309\u56fe\u6ce8\u8fdb\u884c\u8d28\u7c92\u8f6c\u67d3\u548c\u5904\u7406\u300248 \u5c0f\u65f6\u540e\uff0c\u7ec6\u80de\u7528\u9884\u51b7 PBS \u6e05\u6d17\uff0c\u5e76\u7528\u514d\u75ab\u6c89\u6dc0\u7f13\u51b2\u6db2\u88c2\u89e3\u3002\u88c2\u89e3\u6db2\u5728 4 \u00b0C\u300114,000g \u79bb\u5fc3 20 \u5206\u949f\u3002\u968f\u540e\u4e0a\u6e05\u4e0e anti-FLAG M2 \u743c\u8102\u7cd6\u73e0\u5728 4 \u00b0C \u5b75\u80b2 4 \u5c0f\u65f6\u3002<\/p>\n

\u5bf9\u4e8e\u5185\u6e90\u6027 LXR \u7684\u6cdb\u7d20\u5316\u68c0\u6d4b\uff0c\u88c2\u89e3\u6db2\u5148\u5728 4 \u00b0C \u4e0b\u4e0e\u6297 LXR \u6297\u4f53\u548c protein A\/G \u743c\u8102\u7cd6\u73e0\u514d\u75ab\u6c89\u6dc0 2 \u5c0f\u65f6\u3002\u73e0\u5b50\u968f\u540e\u7528\u542b\u86cb\u767d\u9176\u6291\u5236\u5242\u7684\u514d\u75ab\u6c89\u6dc0\u7f13\u51b2\u6db2\u6e05\u6d17\u4e09\u6b21\uff08\u6392\u9664 PMSF \u548c DTT\uff09\uff0c\u518d\u7528\u4e0d\u542b \u03b2-\u5def\u57fa\u4e59\u9187\u7684\u4e0a\u6837\u7f13\u51b2\u6db2\u5728 95 \u00b0C \u716e\u6cb8 10 \u5206\u949f\u3002\u6536\u96c6\u6837\u54c1\u5e76\u8fdb\u884c western blot \u5206\u6790\u3002<\/p>\n

Dil-LDL \u5185\u541e<\/h3>\n

siRNA \u4ecb\u5bfc\u7684 ASGR1 \u6572\u4f4e\u7ec6\u80de\uff08siASGR1\uff09\u548c\u5bf9\u7167\u7ec6\u80de\uff08siCtrl\uff09\u5728\u7b2c 0 \u5929\u63a5\u79cd\u4e8e\u73bb\u7247\u4e0a\u3002\u6536\u83b7\u524d\uff0c\u7ec6\u80de\u5728 4 \u00b0C \u4e0b\u7528\u80c6\u56fa\u9187\u8017\u7aed\u57f9\u517b\u57fa\u5b75\u80b2 30 \u5206\u949f\uff0c\u518d\u8f6c\u5165\u542b 10 \u03bcg\/ml Dil-LDL \u7684\u65b0\u9c9c\u80c6\u56fa\u9187\u8017\u7aed\u57f9\u517b\u57fa\u4e2d\uff0c\u4e8e 4 \u00b0C \u5b75\u80b2 1 \u5c0f\u65f6\u3002PBS \u6e05\u6d17\u4e24\u6b21\u540e\uff0c\u5207\u6362\u81f3 37 \u00b0C\uff0c\u52a0\u5165\u65b0\u9c9c\u80c6\u56fa\u9187\u8017\u7aed\u57f9\u517b\u57fa\uff0c\u5e76\u5728\u6307\u5b9a\u65f6\u95f4\u70b9\u6536\u83b7\u3002\u6700\u540e\uff0c\u7ec6\u80de\u7528 4% \u591a\u805a\u7532\u919b\u56fa\u5b9a\u3001PBS \u6e05\u6d17\uff0c\u5e76\u5728 Leica SP8 \u5171\u805a\u7126\u663e\u5fae\u955c\u4e0b\u6210\u50cf\u3002\u5185\u541e Dil-LDL \u7684\u8367\u5149\u5f3a\u5ea6\u6309\u65e2\u5f80\u65b9\u6cd5\u7528 ImageJ \u5b9a\u91cf\u3002<\/p>\n

RNA-seq \u548c GSEA<\/h3>\n

Huh7 \u7ec6\u80de\u8f6c\u67d3\u4e71\u5e8f siRNA\uff08siCtrl\uff09\u6216 ASGR1 \u9776\u5411 siRNA\uff08siASGR1\uff09\uff0c\u6bcf\u7ec4 3 \u4e2a\u751f\u7269\u5b66\u72ec\u7acb\u91cd\u590d\u300272 \u5c0f\u65f6\u540e\u6536\u83b7\u7ec6\u80de\u63d0\u53d6 RNA\uff0c\u5e76\u5c06\u6837\u54c1\u5206\u522b\u547d\u540d\u4e3a WT \u548c AS2\u3002\u968f\u540e\u5bf9\u8fd9\u4e9b\u6837\u54c1\u8fdb\u884c RNA-seq\uff0c\u5e76\u6309\u8f6f\u4ef6\u8bf4\u660e\u4f7f\u7528 GSEA \u8fdb\u884c\u5206\u6790\u3002\u7b80\u8a00\u4e4b\uff0c\u7814\u7a76\u8005\u81ea\u5b9a\u4e49\u4e86 GO pathway \u7684 C5 \u5b50\u96c6\uff0c\u5e76\u4ece\u5df2\u53d1\u8868\u6570\u636e\u4e2d\u9009\u62e9 LXR \u9776\u57fa\u56e0\u4f5c\u4e3a\u65b0\u5b50\u96c6\u3002\u6700\u7ec8\u83b7\u5f97\u5bcc\u96c6\u5206\u6570\uff08ES\uff09\u3001|NOM|\u3001P \u503c\u7b49\u6570\u636e\uff0c\u5e76\u7528 Prism v9.0 \u5bfc\u51fa\u6298\u7ebf\u56fe\u3002<\/p>\n

AAV2\/8 \u4ecb\u5bfc\u7684\u57fa\u56e0\u8868\u8fbe\u548c\u6572\u4f4e<\/h3>\n

AAV \u514b\u9686\u8f7d\u4f53 AAV-shRNA \u548c pAKD-CMV-bGlobin-EGFP-H1-shRNA \u5206\u522b\u6765\u81ea Obio Technology \u548c PackGene Biotech\uff08\u5e7f\u5dde\uff09\u3002AAV2\/8-shControl\u3001AAV2\/8-shAsgr1\u3001AAV2\/8-shAMPK\u03b11 \u548c AAV2\/8-shAMPK\u03b12 \u901a\u8fc7\u5c06\u76f8\u5e94 shRNA \u514b\u9686\u5165 pAKD-CMV-bGlobin-EGFP-H1-shRNA \u5236\u5907\u3002\u6240\u7528\u5e8f\u5217\u5305\u62ec\u5bf9\u7167 shRNA\u3001ASGR1 shRNA\u3001AMPK\u03b11 shRNA \u548c AMPK\u03b12 shRNA\u3002\u5c0f\u9f20\u4f7f\u7528 29G \u80f0\u5c9b\u7d20\u6ce8\u5c04\u5668\u7ecf\u5c3e\u9759\u8109\u6ce8\u5c04 AAV8\u3002<\/p>\n

\u53bb\u553e\u6db2\u9178\u7cd6\u86cb\u767d\u5236\u5907<\/h3>\n

\u80ce\u7403\u86cb\u767d A \u548c\u8840\u6e05\u7c7b\u9ecf\u86cb\u767d\u6309\u65e2\u5f80\u65b9\u6cd5\u8131\u553e\u6db2\u9178\u3002\u7b80\u8a00\u4e4b\uff0c\u5c06\u7cd6\u86cb\u767d\u7f6e\u4e8e 0.1 mol\/l HCl \u4e09\u6c1f\u4e59\u9178\u4e2d\uff0c80 \u00b0C \u5904\u7406 2 \u5c0f\u65f6\u3002<\/p>\n

\u6eb6\u9176\u4f53\u5feb\u901f\u7eaf\u5316<\/h3>\n

\u8868\u8fbe TMEM192-3\u00d7FLAG \u6216\u5bf9\u7167\u6162\u75c5\u6bd2\u7684\u9897\u7c92\u5728 HEK293T \u7ec6\u80de\u4e2d\u4ea7\u751f\u3002\u75c5\u6bd2\u4e0a\u6e05\u7528\u4e8e\u8f6c\u5bfc Huh7 \u7ec6\u80de\uff0c\u52a0\u5165 8 \u03bcg\/ml polybrene \u5904\u7406 24 \u5c0f\u65f6\u3002\u7ec6\u80de\u7528 5 \u03bcg\/ml \u560c\u5464\u9709\u7d20\u7b5b\u9009 48 \u5c0f\u65f6\uff0c\u5e76\u5728 2 \u03bcg\/ml \u560c\u5464\u9709\u7d20\u5b58\u5728\u4e0b\u7ef4\u6301\u57f9\u517b\u3002\u901a\u8fc7\u6297 FLAG \u67d3\u8272\u786e\u8ba4\u8f6c\u5bfc\u6548\u7387\u8d85\u8fc7 95%\u3002<\/p>\n

\u5bf9\u7167\u6216 TMEM192-3\u00d7FLAG \u8fc7\u8868\u8fbe Huh7 \u7ec6\u80de\u63a5\u79cd\u4e8e 10 \u00d7 10 cm\u00b2 \u57f9\u517b\u76bf\u4e2d\uff0c\u5e76\u751f\u957f\u81f3\u63a5\u8fd1\u6c47\u5408\u3002\u7ec6\u80de\u7528 EBSS \u6e05\u6d17\u4e24\u6b21\uff0c\u5728\u542b 5 mM \u8461\u8404\u7cd6\u7684 EBSS \u4e2d\u5b75\u80b2 2 \u5c0f\u65f6\uff0c\u968f\u540e\u52a0\u5165\u53bb\u553e\u6db2\u9178\u80ce\u7403\u86cb\u767d A\uff0830 \u03bcg\/ml\uff0c2 \u5c0f\u65f6\uff09\u3002\u7ec6\u80de\u7528\u51b0\u51b7 KPBS \u6e05\u6d17\u4e24\u6b21\uff0c\u522e\u53d6\u5e76\u6536\u96c6\u3002\u7ec6\u80de\u6c89\u6dc0\u91cd\u60ac\u4e8e 1 ml KPBS \u4e2d\uff0c\u5e76\u7528 Dounce \u5300\u6d46\u5668\u88c2\u89e3\u3002\u7ec6\u80de\u88c2\u89e3\u6db2\u4e0e 30 \u03bcl \u6297 FLAG \u78c1\u73e0\u5728\u51b0\u4e0a\u65cb\u8f6c\u5b75\u80b2 3 \u5206\u949f\u3002\u78c1\u73e0\u7528 KPBS \u6e05\u6d17\u4e94\u6b21\uff0c\u5e76\u7528 0.5% SDS\uff08\u7528\u4e8e\u514d\u75ab\u5370\u8ff9\uff09\u6216 80% \u7532\u9187\uff08\u7528\u4e8e\u6c28\u57fa\u9178\u5b9a\u91cf\uff09\u6d17\u8131\u3002<\/p>\n

\u4ee3\u8c22\u7ec4\u5b66\u5206\u6790<\/h3>\n

\u4ece\u6eb6\u9176\u4f53\u63d0\u53d6\u4ee3\u8c22\u7269\u65f6\uff0c\u5c06\u7ed3\u5408\u6eb6\u9176\u4f53\u7684\u78c1\u73e0\u91cd\u60ac\u4e8e 200 \u03bcl \u51b0\u51b7\u4ee3\u8c22\u7269\u63d0\u53d6\u7f13\u51b2\u6db2\uff0880% \u7532\u9187\u300120% \u6c34\uff0c\u542b\u5185\u6807\uff09\u4e2d\uff0c\u6da1\u65cb 30 \u79d2\u3002\u6df7\u5408\u7269\u5728\u6db2\u6c2e\u4e2d\u51b7\u51bb\u3001\u5ba4\u6e29\u89e3\u51bb\uff0c\u91cd\u590d\u4e09\u6b21\u3002\u6837\u54c1\u8d85\u58f0 10 \u5206\u949f\uff0c\u5e76\u5728 4 \u00b0C\u300113,000 r.p.m. \u79bb\u5fc3 15 \u5206\u949f\u3002\u4e0a\u6e05\u8f6c\u79fb\u5230\u5e72\u51c0\u7ba1\u4e2d\uff0c\u53d6 50 \u03bcl \u4e0a\u6e05\u7528\u4e8e Amplex Red \u6cd5\u80c6\u56fa\u9187\u5b9a\u91cf\u3002\u5269\u4f59\u4e0a\u6e05\u7528 speedvac \u5b8c\u5168\u6d53\u7f29\u5e72\u71e5\u3002\u4ee3\u8c22\u7269\u6b8b\u6e23\u91cd\u65b0\u6eb6\u89e3\u4e8e 50% \u4e59\u8148\u4e2d\uff0c\u5e76\u7528 Waters Acquity UPLC \u7cfb\u7edf\u8054\u7528 SCIEX 5500 QTRAP \u7cfb\u7edf\u8fdb\u884c\u8d85\u9ad8\u6548\u6db2\u76f8\u8272\u8c31-\u4e32\u8054\u8d28\u8c31\u5206\u6790\u3002<\/p>\n

\u8272\u8c31\u5206\u79bb\u4f7f\u7528 Waters Acquity UPLC BEH Amide \u67f1\uff082.1 mm \u00d7 100 mm\uff0c1.7 \u03bcm\uff09\uff0c\u6d41\u901f 0.4 ml\/min\uff0c\u6e29\u5ea6 40 \u00b0C\uff0c15 \u5206\u949f\u68af\u5ea6\u6d17\u8131\u3002\u7f13\u51b2\u6db2 A \u4e3a\u6c34\u300110 mM \u7532\u9178\u94f5\u548c 0.15% \u7532\u9178\uff1b\u7f13\u51b2\u6db2 B \u4e3a 99% \u4e59\u8148\u30011% \u6c34\u30012 mM \u7532\u9178\u94f5\u548c 0.15% \u7532\u9178\u3002\u8d28\u8c31\u4ee5\u6b63\u79bb\u5b50\u6a21\u5f0f\u548c\u7535\u55b7\u96fe\u79bb\u5b50\u6e90\u8fd0\u884c\u3002\u5206\u6790\u7269\u68c0\u6d4b\u91c7\u7528\u591a\u53cd\u5e94\u76d1\u6d4b\u3002\u5cf0\u8bc6\u522b\u548c\u9762\u79ef\u79ef\u5206\u4f7f\u7528 Analyst v1.7.1 \u548c SCIEX OS 1.4.0\u3002\u80c6\u56fa\u9187\u5b9a\u91cf\u7528\u4e8e\u5f52\u4e00\u5316\u3002<\/p>\n

VLDL \u5206\u6ccc\u5b9e\u9a8c<\/h3>\n

10 \u5468\u9f84\u96c4\u6027\u91ce\u751f\u578b\u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u7ed9\u4e88\u9ad8\u8102\/\u9ad8\u80c6\u56fa\u9187\/\u80c6\u76d0\u996e\u98df 2 \u5929\u3002\u968f\u540e\u5c0f\u9f20\u7981\u98df 4 \u5c0f\u65f6\uff0c\u5e76\u8179\u8154\u6ce8\u5c04 1 g\/kg \u4f53\u91cd\u7684 poloxamer 407\uff08PBS \u4e2d\uff09\u3002\u6309\u6307\u5b9a\u65f6\u95f4\u70b9 0\u30010.5\u30011\u30012\u30013\u30014 \u548c 5 \u5c0f\u65f6\u91c7\u8840\uff0c\u5e76\u7528\u4e0a\u6d77\u79d1\u534e\u751f\u7269\u5de5\u7a0b\u516c\u53f8 TC \u548c TG \u8bd5\u5242\u76d2\u6d4b\u5b9a\u3002<\/p>\n

\u8102\u86cb\u767d\u6e05\u9664\u5b9e\u9a8c<\/h3>\n

\u4eba LDL \u4ece\u5065\u5eb7\u4f9b\u8005\u8840\u6d46\u4e2d\u901a\u8fc7\u8fde\u7eed\u8d85\u901f\u79bb\u5fc3\u5206\u79bb\u3002\u7814\u7a76\u8005\u5236\u5907\u4e86 ^3H-\u80c6\u56fa\u9187\u6cb9\u9178\u916f\u6807\u8bb0\u8102\u8d28\u4f53\u3002\u7ea6 200 \u03bcCi ^3H-\u80c6\u56fa\u9187\u6cb9\u9178\u916f\u52a0\u5165\u6eb6\u89e3\u4e8e\u6c2f\u4eff\u7684 15 mg DOPC \u4e2d\u5e76\u84b8\u5e72\u3002\u52a0\u5165 1 ml PBS \u91cd\u60ac\u8102\u8d28\uff0c\u968f\u540e\u5728\u6db2\u6c2e\u548c 37 \u00b0C \u6c34\u6d74\u4e4b\u95f4\u8fdb\u884c 5 \u6b21\u51bb\u878d\u5faa\u73af\u3002\u968f\u540e\u4f7f\u7528 mini-extruder \u6324\u51fa\u6280\u672f\u5236\u5907\u8102\u8d28\u4f53\u3002^3H-\u80c6\u56fa\u9187\u6cb9\u9178\u916f\u88ab\u63ba\u5165 LDL\u3002\u7b80\u8a00\u4e4b\uff0c\u5c06 LDL\uff082.5 mg\/ml\uff0c10 ml\uff09\u5728 37 \u00b0C \u4e0b\u6301\u7eed\u632f\u8361 24 \u5c0f\u65f6\u4ee5\u4fc3\u8fdb\u4ea4\u6362\u3002\u968f\u540e\u7ecf\u5bc6\u5ea6\u68af\u5ea6\u79bb\u5fc3\u91cd\u65b0\u5206\u79bb ^3H-\u80c6\u56fa\u9187\u6cb9\u9178\u916f\u6807\u8bb0 LDL\uff0c\u5e76\u5728\u542b 150 mmol\/l NaCl \u7684\u7f13\u51b2\u6db2\u4e2d\u4e8e 4 \u00b0C \u900f\u6790\uff0c\u518d\u5b9a\u91cf\u6bcf\u5206\u949f\u8870\u53d8\u6570\uff08DPM\uff09\u3002<\/p>\n

10 \u5468\u9f84\u91ce\u751f\u578b\u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u7ed9\u4e88\u9ad8\u8102\/\u9ad8\u80c6\u56fa\u9187\/\u80c6\u76d0\u996e\u98df 2 \u5929\u3002\u968f\u540e\u7981\u98df 17 \u5c0f\u65f6\u5e76\u590d\u5582 6 \u5c0f\u65f6\u3002\u653e\u5c04\u6027\u6807\u8bb0 LDL \u7528 PBS \u7a00\u91ca\u540e\u9759\u8109\u6ce8\u5c04\u5165\u5c0f\u9f20\uff08\u6bcf\u53ea\u5c0f\u9f20 5 \u00d7 10^5 dpm\uff0c\u603b\u4f53\u79ef\u7ea6 200 \u03bcl\uff09\u3002\u5728 0 \u548c 30 \u79d2\uff0c\u4ee5\u53ca 20\u300140\u300160\u300180\u3001100 \u548c 120 \u5206\u949f\u91c7\u6837\u3002\u53d6 5 \u03bcl \u8840\u6d46\u7528\u95ea\u70c1\u8ba1\u6570\u4eea\u8ba1\u6570\u3002<\/p>\n

\u514d\u75ab\u7ec4\u7ec7\u5316\u5b66<\/h3>\n

\u5236\u5907 p53\u2212\/\u2212 \u548c Ptch1+\/\u2212 \u9ad3\u6bcd\u7ec6\u80de\u7624\u3001\u91ce\u751f\u578b\u5c0f\u9f20\u548c Asgr1\u2212\/\u2212 \u5c0f\u9f20\u7684\u77f3\u8721\u5207\u7247\u3002\u5207\u7247\u5728 37 \u00b0C \u590d\u6e29 15 \u5206\u949f\uff0c\u968f\u540e\u7528\u65b0\u9c9c\u4e8c\u7532\u82ef\u6d17\u6da4 15 \u5206\u949f\uff0c\u91cd\u590d\u4e09\u6b21\u3002\u968f\u540e\u7528 50% \u4e8c\u7532\u82ef\u548c 50% \u4e59\u9187\u5237\u65b0\u7f13\u51b2\u6db2 15 \u5206\u949f\uff0c\u5e76\u7528\u4e59\u9187\u6e05\u6d17\u4e24\u6b21\u3002\u5207\u7247\u5728\u6d78\u5165\u53cc\u84b8\u6c34\u524d\uff0c\u5206\u522b\u7528 95%\u300180%\u300170%\u300150% \u548c 30% \u4e59\u9187\u68af\u5ea6\u6d17\u8131\uff0c\u6bcf\u6b65 5 \u5206\u949f\u3002\u5207\u7247\u7f6e\u4e8e pH 6.0 \u67e0\u6aac\u9178\u94a0\u7f13\u51b2\u6db2\u4e2d\u716e\u6cb8 15 \u5206\u949f\uff0c\u5e76\u7528\u53cc\u84b8\u6c34\u6e05\u6d17\u4e09\u6b21\uff0c\u6bcf\u6b21 5 \u5206\u949f\u3002\u968f\u540e\u7528 3% H2O2\uff08\u7532\u9187\u7a00\u91ca\uff09\u706d\u6d3b\u8fc7\u6c27\u5316\u7269\u9176 10 \u5206\u949f\uff0c\u7528\u53cc\u84b8\u6c34\u6e05\u6d17\u4e24\u6b21\uff0c\u6bcf\u6b21 5 \u5206\u949f\uff0c\u518d\u7f6e\u4e8e PBS \u7f13\u51b2\u6db2 5 \u5206\u949f\u3002\u6837\u54c1\u7528 10% \u5c71\u7f8a\u8840\u6e05\uff08PBS \u7a00\u91ca\uff09\u5ba4\u6e29\u5c01\u95ed 30 \u5206\u949f\uff0c\u968f\u540e\u8986\u76d6\u6307\u5b9a\u6d53\u5ea6\u7684\u4e00\u6297\u8fc7\u591c\u3002\u7b2c\u4e8c\u5929\uff0c\u5207\u7247\u7528 PBS \u6e05\u6d17\u4e09\u6b21\uff0c\u6bcf\u6b21 5 \u5206\u949f\uff0c\u5e76\u5728\u5ba4\u6e29\u4e0b\u4e0e\u4e8c\u6297\u5b75\u80b2 1 \u5c0f\u65f6\u3002\u968f\u540e\u7528 DAB \u5e95\u7269\u8bd5\u5242\u76d2\u663e\u8272\uff0c\u7528\u82cf\u6728\u7cbe\u6838\u67d3 4 \u5206\u949f\u3002\u6700\u540e\uff0c\u6837\u54c1\u7528\u6d41\u6c34\u51b2\u6d17\uff0c\u5e76\u4f9d\u6b21\u7f6e\u4e8e 75%\u300185%\u300195% \u548c 100% \u4e59\u9187\u4e2d 2-3 \u79d2\uff0c\u968f\u540e\u7528 100% \u4e59\u9187\u8131\u6c34\uff0c\u51b2\u6d17\u4e24\u6b21\uff0c\u6bcf\u6b21 1 \u5206\u949f\uff0c\u5c01\u7247\u540e\u5728\u8367\u5149\u663e\u5fae\u955c\u4e0b\u89c2\u5bdf\u3002<\/p>\n

\u52a8\u8109\u7ca5\u6837\u786c\u5316\u75c5\u53d8\u5206\u6790<\/h3>\n

\u5206\u79bb\u5c0f\u9f20\u4e3b\u52a8\u8109\u6811\uff0c\u5e76\u7528 4% \u591a\u805a\u7532\u919b\u56fa\u5b9a\u3002Sudan IV \u67d3\u8272\u524d\u53bb\u9664\u591a\u4f59\u8840\u7ba1\u5468\u56f4\u8102\u80aa\u7ec4\u7ec7\u3002\u968f\u540e\u7528 Sudan IV \u67d3\u8272 10 \u5206\u949f\uff0c\u7528 70% \u4e59\u9187\u6d17\u6da4 3 \u5206\u949f\uff0c\u518d\u7528 70% \u4e59\u9187\u6bcf\u6b21 1.5 \u5206\u949f\u6e05\u6d17\u4e24\u6b21\uff0c\u7136\u540e\u6210\u50cf\u3002<\/p>\n

siRNA \u6572\u4f4e<\/h3>\n

\u57fa\u56e0\u7279\u5f02\u6027 siRNA \u7531\u5e7f\u5dde\u9510\u535a\u751f\u7269\u8bbe\u8ba1\u5408\u6210\u3002siRNA \u6309\u5382\u5bb6\u8bf4\u660e\u4f7f\u7528 Lipofectamine RNAiMAX \u8f6c\u67d3\u8bd5\u5242\u5206\u522b\u8f6c\u67d3\u5165\u7ec6\u80de\u3002\u5e8f\u5217\u5982\u4e0b\uff1a<\/p>\n