Jiang Zhi-Bin, Gao Jie, Chai Yi-Hui, Li Wen, Luo Yun-Feng, Chen Yun-Zhi
College of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, China.
Department of Preclinical Medicine, Guizhou University of Traditional Chinese Medicine, Guiyang, China.
Kaohsiung J Med Sci. 2021 Aug;37(8):718-729. doi: 10.1002/kjm2.12390. Epub 2021 May 11.
Alcoholic fatty liver disease (AFLD) is the most common liver disease and can progress to fatal liver cirrhosis and carcinoma, affecting millions of patients worldwide. The functions of astragaloside on the cardiovascular system have been elucidated. However, its role in AFLD is unclear. Ethanol-treated AML-12 cells were used as a cell model of alcoholic fatty liver. Real-time quantitative reverse transcription-PCR and Western blotting detected genes and proteins expressions. Reactive oxygen species (ROS), triglyceride, total cholesterol, low-density lipoprotein, albumin, ferritin, bilirubin, superoxide dismutase, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were examined using commercial kits. Lipid accumulation was assessed by Oil red O staining. MTT and flow cytometry measured cell viability and apoptosis. JC-1 was used to analyze mitochondrial membrane potential. A rat model of AFLD was established by treating rats with ethanol. Astragaloside suppressed ethanol-induced lipid accumulation, oxidative stress, and the production of AST and ALT in AML-12 cells. Ethanol induced TNF-α and reduced IL-10 expression, which were reversed by astragaloside. Ethanol promoted Bax expression and cytochrome C release and inhibited Bcl-2 and ATP expression. Astragaloside hampered these apoptosis effects in AML-12 cells. Impaired mitochondrial membrane potential was recovered by astragaloside. However, all these astragaloside-mediated beneficial effects were abolished by the ROS inducer pyocyanin. Ethanol-induced activation of NF-κB signaling was suppressed by astragaloside in vitro and in vivo, suggesting that astragaloside inhibited oxidative stress by suppressing the activation of NF-κB signaling, thus improving liver function and alleviating AFLD in rats. Our study elucidates the pharmacological mechanism of astragaloside and provides potential therapeutic strategies for AFLD.
酒精性脂肪性肝病(AFLD)是最常见的肝脏疾病,可进展为致命的肝硬化和肝癌,影响着全球数百万患者。黄芪甲苷对心血管系统的作用已得到阐明。然而,其在AFLD中的作用尚不清楚。用乙醇处理的AML-12细胞作为酒精性脂肪肝的细胞模型。通过实时定量逆转录PCR和蛋白质免疫印迹法检测基因和蛋白质表达。使用商业试剂盒检测活性氧(ROS)、甘油三酯、总胆固醇、低密度脂蛋白、白蛋白、铁蛋白、胆红素、超氧化物歧化酶、天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)。通过油红O染色评估脂质蓄积。MTT法和流式细胞术检测细胞活力和凋亡。使用JC-1分析线粒体膜电位。通过用乙醇处理大鼠建立AFLD大鼠模型。黄芪甲苷抑制乙醇诱导的AML-12细胞脂质蓄积、氧化应激以及AST和ALT的产生。乙醇诱导肿瘤坏死因子-α(TNF-α)表达并降低白细胞介素-10(IL-10)表达,而黄芪甲苷可逆转这些变化。乙醇促进Bax表达和细胞色素C释放,并抑制Bcl-2和ATP表达。黄芪甲苷阻碍AML-12细胞中的这些凋亡效应。黄芪甲苷可恢复受损的线粒体膜电位。然而,活性氧诱导剂绿脓菌素消除了黄芪甲苷介导的所有这些有益作用。在体外和体内,黄芪甲苷均抑制乙醇诱导的核因子-κB(NF-κB)信号通路激活,这表明黄芪甲苷通过抑制NF-κB信号通路激活来抑制氧化应激,从而改善肝功能并减轻大鼠的AFLD。我们的研究阐明了黄芪甲苷的药理机制,并为AFLD提供了潜在的治疗策略。