Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian, 116044, China.
Drug Clinical Trial Institution, The Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, 214023, China.
Eur J Pharmacol. 2021 Jan 5;890:173653. doi: 10.1016/j.ejphar.2020.173653. Epub 2020 Oct 15.
Chronic alcohol assumption has been recognized as a major cause of alcoholic liver disease (ALD), which ranges from alcoholic steatohepatitis to fibrosis and hepatocellular carcinoma. Alcoholic liver disease has become the leading cause of liver-related health problem in the world. Herewith, effective therapeutic strategy for alcoholic liver disease is necessary. Yangonin (Yan), a bioactive compound extract from Kava, has been reported to exert hepatoprotective effects via Farnesoid X receptor (FXR) activation. The present study aims to investigate whether Yan ameliorated the ethanol-stimulated liver injury and further to elucidate the mechanisms in vivo and in vitro. Yan improved cell viabilities via cell count kit-8 (CCK-8) methods and obviously reduced aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC) and total triglyceride (TG) levels. We detected miR-194 levels in ethanol-induced LO cells and male C57BL/6 mice by quantitative real-time PCR. Also, the effects of miR-194 on modulating cellular senescence via targeting FXR were further verified. The cellular senescence markers p16, p21, telomerase activity and senescence-related β-galactosidase (SA-β-gal) were evaluated by quantitative real-time PCR and Western blot. Also, LO cells or liver tissues were stained with special primary antibodies and 4',6'-Diamidino-2-phenylindole (DAPI). The cell cycle was detected by flow cytometry. We observed that Yan significantly inhibited ethanol-induced cellular senescence via FXR activation (P < 0.05). Our results demonstrate that Yan significantly reduced the cellular markers p16, p21 and Hmga1 expression and inhibited the cell cycle arrest (P < 0.05). MiR-194 was upregulated in the alcoholic liver disease, which was significantly suppressed by Yan (P < 0.05). Moreover, miR-194 mimic inhibited FXR expression in vitro. In summary, these aggregated data demonstrate that Yan alleviates chronic ethanol-induced liver injury through inhibition of cellular senescence via regulating miR-194/FXR axis.
慢性酒精摄入已被认为是酒精性肝病（ALD）的主要原因，其范围从酒精性肝炎发展到纤维化和肝细胞癌。酒精性肝病已成为世界范围内与肝脏相关的健康问题的主要原因。因此，有必要开发有效的酒精性肝病治疗策略。Yangonin（Yan）是从卡瓦植物中提取的一种生物活性化合物，据报道，它通过激活法尼醇 X 受体（FXR）发挥肝脏保护作用。本研究旨在探讨 Yan 是否改善了乙醇刺激的肝损伤，并进一步在体内和体外阐明其机制。Yan 通过细胞计数试剂盒-8（CCK-8）方法提高细胞活力，明显降低天冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）、总胆固醇（TC）和总甘油三酯（TG）水平。我们通过定量实时 PCR 检测了乙醇诱导的 LO 细胞和雄性 C57BL/6 小鼠中的 miR-194 水平。此外，还通过靶向 FXR 验证了 miR-194 调节细胞衰老的作用。通过定量实时 PCR 和 Western blot 评估了细胞衰老标志物 p16、p21、端粒酶活性和衰老相关的β-半乳糖苷酶（SA-β-gal）。此外，用特殊的一抗和 4'，6'-二脒基-2-苯基吲哚（DAPI）对 LO 细胞或肝组织进行染色。通过流式细胞术检测细胞周期。我们观察到 Yan 通过 FXR 激活显著抑制了乙醇诱导的细胞衰老（P<0.05）。我们的结果表明，Yan 显著降低了细胞标志物 p16、p21 和 Hmga1 的表达并抑制了细胞周期停滞（P<0.05）。miR-194 在酒精性肝病中上调，Yan 显著抑制了 miR-194 的表达（P<0.05）。此外，miR-194 模拟物在体外抑制了 FXR 的表达。总之，这些综合数据表明，Yan 通过调节 miR-194/FXR 轴抑制细胞衰老来缓解慢性乙醇诱导的肝损伤。
