The Mechanism And Clinical Study Of Ginsenoside Rg1 On Human Lung Fibroblast MRC-5 Based On PI3k/Akt Pathway

Objective:PI-3k/Akt pathway based on the research of ginsenoside Rg1 on MRC-5 hum an embryonic lung fibroblast intervention,so as to clarify its mechanism of anti f ibrosis,provide effective drugs for the treatment of interstitial pneumonia,so as t o reduce the mortality rate,help reduce patient suffering and economic burden,to provide new ideas for clinical treatment.Methods:according to the design requirements of the study,the human embryonic lung fibroblast MRC-5 was taken as the research object,ginsenoside Rg1 as an experi mental drug,and hydrogen peroxide induced MRC-5 cell oxidative damage model was established.The experiment was conducted in 6 groups.1.to construct the oxidative stress model of MRC-5 cells and to select the o ptimal concentration of ginsenoside Rg1.The oxidative damage model of hydroge n peroxide as a reagent to establish MRC-5 cells,with different concentrations of H2O2 and different time to MRC-5 cells,detect cell inhibition rate of change,R OS index,combining the data,select the best concentration,using H2O2 modeling time.The best concentration of ginsenoside Rg1 on MRC-5 cells was selected.I C10 was the largest non-toxic concentration.Four concentrations of drugs were se t to treat MRC-5 cells,the cell inhibition rate was detected,and the best concentr ation of ginsenoside Rg1 was selected.2.the protective effect of ginsenoside Rg1 on the oxidative damage of MRC-5 cells induced by H2O2 and the role of PI-3k/AKT pathway were observed.Human embryo lung fibroblast MRC-5 was cultured in vitro and successfully resuscitated and passaged.Then human embryo lung fibroblasts were intervened by hydrogen peroxide and ginsenoside Rg1 respectively.Cells were divided into model group(control group),hydrogen peroxide treated group,ginsenoside Rg1 gr oup and ginsenoside Rg1 intervention after 1h intervention group treated with H2O2 and hydrogen peroxide treatment after 1h given ginsenoside Rg1 treatment gro up and AKT inhibitor 1H intervention after ginsenoside Rg1 to hydrogen peroxide treatment group,a total of 6 groups.Time monitoring points were 6,12 and 24 h.CCK-8 assay was used to detect cell proliferation,and DCFH-DA fluorescence probe was used to detect the content of reactive oxygen species(flow type)in c ells.The expressions of SOD,CAT,MDA and GSH in each time point were co mpared and compared.Western blotting was used to detect and compare the expr ession of Akt,p-Akt,PI-3k protein at each time point of each group.RT-PCR w as used to detect the expression of Akt,p-Akt and PI-3k at each time point of e ach group.And the best time point immunofluorescence chemical method was use d to observe the changes of collagen Ⅰ and collagen Ⅲ.Results:ginsenoside Rg1 can inhibit the proliferation of MRC-5 in human embryonic lung fibroblasts,reduce extracellular collagen secretion,increase the levels of antio xidants such as SOD,GSH and CAT,reduce MDA level,and regulate the protei n expression of Akt,p-Akt and PI-3k.After the use of the AKT pathway inhibit or,the effect changes.Conclusion:ginsenoside Rg1 has obvious inhibitory effect on human embryonic lung fibro blast MRC-5 cell proliferation,decrease in collagen secretion,improve the level o f anti oxidative damage plays an important role,has a clear effect of anti fibrosis,and play the role of the AKT pathway and close relation.