基于SIRT1/NRF2/HO-1信号通路探讨黄芪甲苷对大鼠肝星状细胞氧化损伤的作用机制  

Exploring the Mechanism of Action of Astragaloside on Oxidative Damage in Rat Hepatic Stellate Cells Based on SIRT1/NRF2/HO-1 Signaling Pathway

作　　者：田萌媛 张铭 张秋菊[1] 陈光顺[1] TIAN Mengyuan;ZHANG Ming;ZHANG Qiuju;CHEN Guangshun(Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,兰州730000

出　　处：《世界科学技术-中医药现代化》2024年第9期2492-2499,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：甘肃省优秀研究生“创新之星”基金项目(2023CXZX-744):基于Nrf2/HO-1信号通路探讨黄芪甲苷抗肝纤维化的作用机制,负责人:田萌媛。

摘　　要：目的探讨黄芪甲苷治疗肝纤维化(HF)的作用机制。方法HSC-T6分为空白组、模型组、黄芪甲苷组、抑制剂组(采用Sirt1抑制剂EX527)、黄芪甲苷加抑制剂组,除空白组外均采用100μmol·L^(-1)的H_(2)O_(2)制造HSC-T6氧化应激的模型,模型组干预4 h,除模型组外干预24 h。ELISA法测定细胞上清中α-SMA、Collagen1水平;生化试剂盒法测定细胞中氧化应激相关指标;流式细胞术检测细胞中ROS的含量;免疫荧光法检测细胞中α-SMA的含量;实时荧光定量聚合酶链式反应(Real-time qPCR)和蛋白免疫印迹法(Western blot)检测各组细胞中Sirt1、Nrf2、HO-1、α-SMA、Collagen1 mRNA和蛋白表达水平。结果与空白组比较,经H_(2)O_(2)处理的HSC-T6上清中α-SMA、Collagen1及细胞中MDA、ROS的含量明显增加,而细胞中CAT含量及SOD活性明显降低,细胞中Sirt1、Nrf2、HO-1 mRNA和蛋白表达水平降低,α-SMA、Collagen1mRNA表达水平升高(P<0.05);与模型组比较,各给药组细胞中MDA、ROS的含量明显降低,CAT含量及SOD活性明显增加,黄芪甲苷组、黄芪甲苷加抑制剂组上清中α-SMA、Collagen1表达量降低,Sirt1、Nrf2、HO-1 mRNA和蛋白表达水平增加,α-SMA、Collagen1 mRNA表达水平降低(P<0.05)。结论黄芪甲苷可以减轻因H_(2)O_(2)刺激造成的HSC-T6氧化应激反应,减少氧化应激产物产生及胶原纤维沉积,从而达到抗HF的目的,其作用机制可能与调节Sirt1/Nrf2/HO-1信号通路有关。Objective To investigate the mechanism of astragaloside in treating hepatic fibrosis(HF).Methods HSCT6 was divided into a blank group,astragaloside group,inhibitor group(using the Sirt1 inhibitor EX527),and astragaloside plus inhibitor group,Except for the blank group using 100μmol·L^(-1) H_(2)O_(2) to create a model of oxidative stress in HSC-T6,with 4 h of intervention in the modeling group,and 24 h of intervention in the modeling group,except for the modeling group.and Collagen1 levels in cell supernatants;biochemical kit assay to determine oxidative stressrelated indexes in cells;flow cytometry to detect the content of ROS in cells;immunofluorescence to detect the content ofα-SMA in cells;and real-time fluorescence quantitative polymerase chain reaction(Real-time qPCR)and protein immunoblotting(Western blot)to detect SIRT1,NRF2,HO-1,α-SMA,Collagen1 mRNA and protein expression levels in each group of cells.Results Compared with the blank control group,the contents ofα-SMA,Collagen1 and cellular MDA and ROS in the supernatants of HSC-T6 treated with H_(2)O_(2) were significantly increased,whereas the contents of CAT and the activity of SOD in the cells were significantly decreased,the expression levels of mRNA and protein of Sirt1,Nrf2 and HO-1 in the cells were decreased,and theα-SMA,Collagen1 mRNA expression levels increased(P<0.05);Compared with the model group,the contents of MDA and ROS in cells were significantly decreased,the CAT content and SOD activity were significantly increased,the expression levels ofα-SMA and Collagen1 in the supernatant were decreased in the astragaloside IV group and astragaloside IV plus inhibitor group,the mRNA and protein expression levels of Sirt1,Nrf2 and HO-1 were increased,and the mRNA expression levels ofα-SMA and Collagen1 were decreased(P<0.05).Conclusion Astragaloside can attenuate HSC-T6 oxidative stress caused by H_(2)O_(2) stimulation,reduce oxidative stress product production and collagen fiber deposition,thus achieving anti-HF,and its mechanism of action

关 键 词：肝纤维化 黄芪甲苷 大鼠肝星状细胞 氧化损伤 Sirtuin1/核因子红系2相关因子2(Nrf2)/血红素氧合酶1(HO-1) 

分 类 号：R285.5[医药卫生—中药学]