Zhang Qian, Liu Yan, Yuan Yujie, Yao Feifei, Zhang Hongmei, Zhao Caiyan, Luo Yanli
Department of Geriatrics, the third Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Neurology, The Gucheng County Hospital of Hebei Province, Hebei, China.
Toxicol Mech Methods. 2022 Jun;32(5):325-332. doi: 10.1080/15376516.2021.2003919. Epub 2021 Nov 25.
miR-26a-5p is a short noncoding RNA that is abnormally expressed in drug-induced liver injury (DILI), but its pathophysiologic role in the mechanism of disease in DILI is still vague.
The expression of miR-26a-5p, viability of hepatic stellate cells (HSCs) proliferation, and apoptosis were explored via real-time PCR, CCK-8 assay, Tunel fluorescence, and flow cytometry. The expression of Bid was detected via Western blot assays, real-time PCR, and immunofluorescence. The apoptosis-associated proteins were determined through Western blot. The interaction between miR-26a-5p and Bid was measured via Dual luciferase reporter assay.
miR-26a-5p expression was greatly decreased in HSCs and serum treated with azithromycin, simvastatin and diclofenac sodium, respectively. Hepatocyte viability was largely suppressed while hepatocyte apoptosis was markedly increased in DILI. Correspondingly, the apoptosis-associated proteins including Bid, caspase-8 and cytochrome C in HSCs were significantly upregulated when treated with either of these drugs. Moreover, miR-26a-5p interacted with Bid, and hepatocyte proliferation and apoptosis influenced by miR-26a-5p mimics were obviously reversed when co-treated with overexpressed Bid plasmids.
miR-26a-5p played a protective role against DILI via targeting Bid.
miR-26a-5p是一种短链非编码RNA，在药物性肝损伤（DILI）中异常表达，但其在DILI疾病机制中的病理生理作用仍不明确。
通过实时PCR、CCK-8检测、Tunel荧光和流式细胞术探讨miR-26a-5p的表达、肝星状细胞（HSCs）增殖活力及凋亡情况。通过蛋白质免疫印迹分析、实时PCR和免疫荧光检测Bid的表达。通过蛋白质免疫印迹法测定凋亡相关蛋白。通过双荧光素酶报告基因检测法检测miR-26a-5p与Bid之间的相互作用。
在用阿奇霉素、辛伐他汀和双氯芬酸钠处理的HSCs和血清中，miR-26a-5p的表达分别大幅降低。在DILI中，肝细胞活力受到很大抑制，而肝细胞凋亡明显增加。相应地，当用这些药物中的任何一种处理时，HSCs中包括Bid、半胱天冬酶-8和细胞色素C在内的凋亡相关蛋白显著上调。此外，miR-26a-5p与Bid相互作用，当与过表达的Bid质粒共同处理时，受miR-26a-5p模拟物影响的肝细胞增殖和凋亡明显逆转。
miR-26a-5p通过靶向Bid对DILI起保护作用。
