miR-96-5p prevents hepatic stellate cell activation by inhibiting autophagy via ATG7
miR-96-5p prevents hepatic stellate cell activation by inhibiting autophagy via ATG7
复制标题
miR-96-5p 通过 ATG7 抑制自噬来防止肝星状细胞活化
DOI：
10.1007/s00109-017-1593-6
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发表时间：
2018-01-01
影响因子：
4.7
通讯作者：
Shi, Guangfeng
中科院分区：
文献类型：
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作者：
Yu, Kangkang;Li, Ning;Shi, Guangfeng
肝星状细胞（HSC）是细胞外基质的主要来源，其活化在肝纤维化的发生中起着关键作用。肝星状细胞自噬与肝纤维化有密切关系，但其在肝纤维化过程中的调控机制尚不清楚。在此，我们首先鉴定了miR-96- 5 p作为纤维化肝组织中异常表达的miRNA。接下来，我们将miR-96- 5 p模拟物转染到人肝星状细胞系LX-2中，观察到α-SMA和Col 1A 1的蛋白和mRNA水平降低。此外，转染miR-96- 5 p模拟物可显著降低LX-2细胞的自噬活性，而转染miR-96- 5 p抑制剂可促进LX-2细胞的自噬。自噬相关蛋白7（autophagy-related protein 7，ATG 7）是miR-96- 5 p的潜在作用靶点，荧光素酶检测证实ATG 7与miR-96- 5 p直接相互作用。最后，将ATG 7重新引入LX-2细胞逆转了miR-96- 5 p介导的自噬抑制以及α-SMA和Col 1A 1表达。总之，我们证明了miR-96- 5 p可以通过ATG 7阻断自噬来抑制肝星状细胞活化。这些发现为开发基于miRNA的抗肝纤维化策略提供了新的见解。关键信息·在肝纤维化组织中观察到改变的miRNA表达谱。·miR-96- 5 p可抑制HSC活化。HSC的自噬在活化过程中被miR-96- 5 p抑制。ATG 7是miR-96- 5 p的直接靶点。ATG 7可以挽救miR-96- 5 p介导的自噬和HSC活化的抑制。
AbstractActivation of hepatic stellate cell (HSC), which is the main source of extracellular matrix, plays a pivotal role in liver fibrogenesis. Autophagy of hepatic stellate cell has been recently implicated in liver fibrosis, but the regulation of hepatic stellate cell autophagy during this process remains poorly understood. Here, we first identified miR-96-5p as an aberrantly expressed miRNA in fibrotic liver tissues. Next, we transfected miR-96-5p mimic into human hepatic stellate cell line LX-2 and observed decreased protein and mRNA levels of α-SMA and Col1A1. In addition, transfection of miR-96-5p mimic significantly reduced autophagy activity of LX-2 cells, while transfection of miR-96-5p inhibitor promoted LX-2 cell autophagy. Moreover, autophagy-related protein 7 (ATG7) was predicted as a potential target of miR-96-5p and luciferase assay confirmed its direct interaction with miR-96-5p. Finally, reintroduction of ATG7 into LX-2 cells reversed miR-96-5p-mediated inhibition of autophagy as well as α-SMA and Col1A1 expression. In conclusion, we demonstrated that miR-96-5p can inhibit hepatic stellate cell activation by blocking autophagy via ATG7. These findings provide new insight into the development of miRNA-based anti-fibrotic strategies.Key messages• Altered miRNA expression profile is observed in fibrotic liver tissues.• miR-96-5p can inhibit HSC activation.• Autophagy of HSC is repressed by miR-96-5p during activation.• ATG7 is a direct target of miR-96-5p.• ATG7 can rescue miR-96-5p-mediated inhibition of autophagy and HSC activation.