miR-146b-5p activation of hepatic stellate cells contributes to the progression of fibrosis by directly targeting HIPK1.
miR-146b-5p activation of hepatic stellate cells contributes to the progression of fibrosis by directly targeting HIPK1.
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肝星状细胞的miR-146b-5p激活通过直接靶向HIPK1促进纤维化进展
DOI：
10.3892/etm.2022.11474
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发表时间：
2022-08
影响因子：
2.7
通讯作者：
中科院分区：
文献类型：
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作者：
本研究旨在探讨microRNA (miR)-146b-5p和同源结构域相互作用蛋白激酶1 （HIPK1）在肝纤维化（HF）进展中的生物学功能，并确定其潜在机制。采用皮下注射四氯化碳（CCl4）建立大鼠心衰模型。采用定量反转录PCR、western blotting和免疫组织化学检测纤维化大鼠肝组织和大鼠肝星状细胞（HSC）系HSC- t6中miR-146b-5p和HIPK1的相对水平。脂多糖（LPS）诱导HSC-T6细胞活化后，采用MTT法检测细胞活力。将miR-146b-5p模拟物或抑制剂转染到HSC-T6细胞中，目的是鉴定miR-146b-5p对HSC-T6细胞行为的影响。通过TargetScan 7.2和StarBase 3.0预测miR-146b-5p和HIPK1之间的靶向关系，随后通过双荧光素酶报告基因试验验证。通过慢病毒转染，研究HIPK1调控HF进展的生物学功能及其机制。结果显示，miR-146b-5p在HF及活化的HSC-T6细胞大鼠肝组织中表达上调，HIPK1在HF及活化的HSC-T6细胞大鼠肝组织中表达下调。miR-146b-5p能够上调lps诱导的HSC-T6细胞的激活标志物，上调COL1A1和TGF-β，提高细胞活力，促进纤维化进展。HIPK1被证实是miR-146b-5p的直接靶点，其过表达可以有效降低miR-146b-5p在HF进展中的作用。总之，miR-146b-5p在HF的进展过程中显著上调。通过靶向和下调HIPK1， miR-146b-5p可显著激活hsc，上调COL1A1和TGF-β，促进纤维化进展。miR-146b-5p是HF的潜在生物标志物和治疗靶点。
The present study aimed to explore the biological functions of microRNA (miR)-146b-5p and homeodomain interacting protein kinase 1 (HIPK1) in the progression of hepatic fibrosis (HF) and to identify the underlying mechanism. A rat HF model was established by administering a subcutaneous injection of carbon tetrachloride (CCl4). Relative levels of miR-146b-5p and HIPK1 in fibrotic rat liver tissues and the rat hepatic stellate cell (HSC) line HSC-T6 were measured by quantitative reverse transcription PCR, western blotting and immunohistochemistry. Following activation of HSC-T6 cells by lipopolysaccharide (LPS) induction, cell viability was examined by MTT assay. Transfection of miR-146b-5p mimic or inhibitor into HSC-T6 cells was performed, with the aim to identify the influence of miR-146b-5p on HSC-T6 cell behavior. The targeting relationship between miR-146b-5p and HIPK1 was predicted by TargetScan 7.2 and StarBase 3.0 and it was later verified by a dual-luciferase reporter assay. Through lentivirus transfection, the biological function of HIPK1 in regulating the progression of HF and the underlying mechanism were investigated. The results showed that miR-146b-5p was upregulated in liver tissues of rats with HF and activated HSC-T6 cells, while HIPK1 was downregulated in liver tissues of rats with HF and activated HSC-T6 cells. miR-146b-5p was able to upregulate the activation markers of LPS-induced HSC-T6 cells, upregulate COL1A1 and TGF-β, increase cell viability and contribute to fibrosis progression. HIPK1 was validated as the direct target of miR-146b-5p and its overexpression could effectively reduce the effect of miR-146b-5p in contribution to the progression of HF. In conclusion, miR-146b-5p was significantly upregulated during the progression of HF. By targeting and downregulating HIPK1, miR-146b-5p could significantly activate HSCs, upregulate COL1A1 and TGF-β and contribute to fibrosis progression. miR-146b-5p is a potential biomarker and therapeutic target for HF.
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影响因子：
4.2
作者：
Ouyang, Bin;Pan, Ningfeng;Ji, Wu
通讯作者：
Ji, Wu
影响因子：
3.7
作者：
Nakamura M;Kanda T;Sasaki R;Haga Y;Jiang X;Wu S;Nakamoto S;Yokosuka O
通讯作者：
Yokosuka O
影响因子：
3.2
作者：
Lee, Ursula E.;Friedman, Scott L.
通讯作者：
Friedman, Scott L.
影响因子：
3.8
作者：
Ge, Shanfei;Zhang, Lunli;Xiang, Tianxing
通讯作者：
Xiang, Tianxing
影响因子：
5.6
作者：
Fodor A;Lazar AL;Buchman C;Tiperciuc B;Orasan OH;Cozma A
通讯作者：
Cozma A