Study On The Antioxidation Of Astragaloside On MRC-5 By Nrf2 Signaling Pathway And Discussion On Clinical Application

Objective :Pulmonary fibrosis(PF)is a disease of the lungs caused by multiple etiologies whose pathogenesis is incomplete.It has been confirmed that oxidative stress-induced cellular oxidation/antioxidation imbalance is the key to promoting the formation and progression of pulmonary fibrosis,and Nrf2/ARE pathway is an important endogenous antioxidant signaling pathway.Previous studies have found that Yiqi type herbs have a significant protective effect on Oxidative damage of pulmonary fibrosis,and at the same time,the clinical diagnosis and treatment of tutor Zhang Wei found that anti-fibrosis prescriptions are compatible with Chinese herbal medicines such as Astragalus(Fang Ji Huang Qi Tang,etc)can significantly improve the prognosis of patients with pulmonary fibrosis,and clinical efficacy is significant.This experiment was based on the Nrf2 signaling pathway to investigate the anti-oxidation effect of astragaloside on human embryonic lung fibroblasts MRC-5.Through observing the expression of related genes and downstream proteins in this pathway,the mechanism of antioxidative stress was identified,which providing effective theoretical for Clinical treatment of pulmonary fibrosis.Methods:First,the oxidative stress model of MRC-5 cells was constructed and the optimal concentration of astragaloside was selected.Secondly,experimental groups: blank group,H2O2 model group,astragaloside group,H2O2 model group,astragaloside group,and astragaloside group were given to H2O2 group after pre-intervention,with 6,12 and 24 hours as monitoring points.The expressions of SOD,CAT,MDA and GSH were detected.The expression of Nrf2,keap1,NQO1 and HO-1 m RNA in each group were detected by RT-PCR and compared.The best time point immunofluorescence chemical method was used to observe the distribution of Nrf2,and the above values were compared and analysis.Results:1.When the concentration of H2O2 was 600?mol/L for 24 h,the inhibition rate of the cells reached 33.96%.The stimulation concentration,the action time,and the extent of damage inhibition for the cells were in accordance with the best modeling requirements.Therefore,600umol/L was used as the model concentration.2.When the drug concentration of astragaloside is 0.3125 mg/ml,the inhibition rate of the cell is 11.04%.Under the drug dosage,the cell inhibition rate is low,the damage to MRC-5 cell is small,the drug concentration is relatively safe,and the concentration is determined.The maximum non-toxic concentration of the drug is chosed for this experiment.3.Astragaloside can significantly increase the expression of SOD,CAT,and GSH in the anti-oxidation parameters of MRC-5 cells.At the same time,it can significantly reduce the expression of MDA,which is an indicator of oxidative damage,and has a time-dependent,statistically significant(P< 0.05).4.Astragaloside can significantly increase the expression levels of HO-1,NQO1 and Nrf2,and decrease the expression of Keap1 in MRC-5 cells,which has significant statistical significance(P<0.05).5.Astragaloside can significantly increase the expression of Nrf2 in the nucleus,and it can promote the transfer of Nrf2 from the cytoplasm to the nucleus and increase the antioxidant capacity of the body.Conclusion:Astragaloside can up-regulate the expression of Nrf2 in MRC-5 cells,enhance the transcriptional activation of Nrf2 and up-regulate the expression of antioxidases and related detoxification enzymes in the lungs to exert a protective effect.It is suspected that astragaloside may interfere with Nrf2/ARE signaling.Transduction pathways exert antifibrotic effects and have a certain degree of preprotection.Clinical anti-fibrosis prescriptions in combination with Chinese herbal medicines such as Astragalus membranaceus may have therapeutic effects on pulmonary fibrosis in terms of anti-oxidative stress.