Exploring the Mechanism of Action of Astragaloside on Oxidative Damage in Rat Hepatic Stellate Cells Based on SIRT1/NRF2/HO-1 Signaling Pathway
Objective To investigate the mechanism of astragaloside in treating hepatic fibrosis(HF).Methods HSC-T6 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 100 µmol·L-1 H2O2 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 Collagen 1 levels in cell supernatants;biochemical kit assay to determine oxidative stress-related 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 of α-SMA,Collagen1 and cellular MDA and ROS in the supernatants of HSC-T6 treated with H2O2 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 the α-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 of α-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 of α-SMA and Collagen1 were decreased(P＜0.05).Conclusion Astragaloside can attenuate HSC-T6 oxidative stress caused by H2O2 stimulation,reduce oxidative stress product production and collagen fiber deposition,thus achieving anti-HF,and its mechanism of action may be related to the regulation of the Sirt1/Nrf2/HO-1 signaling pathway.
万方数据
