Study On Effect Of Partial Oxidation Products Of LPS-induced KGN Cell Lines By DHEA

Objective:To study the effect of dehydroepiandrosterone(DHEA)on oxidative stress model induced by LPS(lipopolysaccharide)in KGN cell line(ovarian granuloidal cell tumor cell line),and to explore the ameliorative effect of DHEA on the effect of oxidative stress on female reproductive function.Method:The total antioxidant capacity(T-AOC),superoxide dismutase(SOD)active,reducing glutathione(GSH)content and lipid peroxide malondialdehyde(MDA)content of KGN cell line were observed by the intervention of LPS on KGN cell line,And the expression of the uncoupled protein 2(UCP2)content,glutathione peroxidase4(GPX4)were observed under different concentrations of DHEA.1.According to the conclusion drawn from previous studies,the optimal LPS concentration for LPS-induced oxidation of KGN cell lines is 2ug/mL,and the optimal time for intervention is 24 h.2.Study on oxidation products of KGN cell lines induced by 2ug/mL LPS during different concentrations of DHEA: After the KGN cell lines were cultured for48 hours,2ug/mL LPS was added for 24 hours,and then different concentrations of DHEA were given for the intervention of KGN cell lines under oxidative stress at concentrations of 0.05umol/ L,0.5umol/ L,5umol/ L,10umol/ L,1/1000 DMSO in blank group was set for intervention for 24 hours.Western blot was used to detect the expression of GPX4 and UCP2 proteins,and the supernatant was used to detect the expression of SOD activity,MDA content,T-AOC and GSH content with related kit.Result:Effects of different concentrations of DHEA on oxidation products of 2ug/mL LPS-induced KGN cell lines:1.Compared with blank group(KGN cell line+2ug/mL LPS),T-AOC SOD activity,,GSH content secretion and GPX4,UCP2 content expression showed an upward trend with the increase of DHEA concentration.0.05umol/ L DHEA group,0.5umol/ L DHEA group,5umol/ L DHEA group,10umol/ L DHEA group compared with the blank group had statistically significant differences(P<0.05).2.Compared with blank group(KGN cell line+2ug/mL LPS),with the increase of DHEA concentration,the expression of MDA oxide decreased with the increase of DHEA concentration,0.05umol/ L DHEA group,0.5umol/ L DHEA group,5umol/ L DHEA group,10umol/ L DHEA group compared with the blank grouphad statistically significant differences(P<0.05).Conclusion:1 In this study,the antioxidant T-AOC,SOD activity,GSH content and the expression of UCP2,GPX4 protein content of KGN cell lines under oxidative stress induced by LPS were treated with different concentrations of DHEA was increased,suggesting that DHEA could achieve the purpose of antioxidant effect by increasing the production of antioxidant products and related proteins.2.After different concentrations of DHEA were used to interfere with LPS-induced oxidative stress,the content of MDA in KGN cells decreased,suggesting that DHEA could reduce the content of oxidation products to achieve the purpose of antioxidant effect.3.In this study,the antioxidant effect of DHEA on KGN cell lines under oxidized state was initially explored,which laid the experimental foundation for the subsequent exploration of DHEA on patients with premature ovarian failure.This study provides the experimental basis for the application of DHEA antioxidant in clinical practice...