消囊调经汤调控NF-κB/IRS-1/PI3K/AKT通路治疗肥胖型多囊卵巢综合征的作用机制OA

Objective:To investigate the mechanism of Xiaonang Tiaojing Decoction in treating obese polycystic ovary syndrome(PCOS)by regulating the nuclear factor kappa-B(NF-κB)/insulin receptor substrate-1(IRS-1)/phosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods:An obese PCOS mouse model was established using a high-fat diet combined with dehydroepiandrosterone.Mice were divided into a normal control group(Control),a model group(Model),a metformin group(200 mg·kg-1,Met),a low-dose Xiaonang Tiaojing Decoction group(5 g·kg-1,XNTJT-L),and a high-dose Xiaonang Tiaojing Decoctiion group(20 g·kg-1,XNTJT-H),with10 mice per group,administered orally for8 weeks.Ovarian granulosa cells were di-vided into a blank group(Control),a model group(Model),a drug-containing serum group(XNTJT),a drug-containing serum si-NC group(XNTJT si-NC),and a drug-containing serum si-IRS-1 group(XNTJT si-IRS-1).Mouse insulin resistance in-dex(HOMA-IR)and serum free fatty acids(FFA)levels were measured;HE staining was used to assess mouse estrous cycle and o-varian tissue pathology;ELISA was used to detect serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),tes-tosterone(T),and LH/FSH ratio;Western blot analyzed key protein expression levels of the NF-κB/IRS-1/PI3K/AKT pathway in mice and cells;ELISA measured tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and NF-κB levels in mice and cells;CCK-8 assay assessed cell proliferation;and flow cytometry analyzed cell apoptosis.Results:Animal experiments:Compared with the Control group,the levels of HOMA-IR and FFA in the Model group were increased(P<0.01).Compared with the Model group,the levels of HOMA-IR and FFA in the Met group and XNTJT-H group were decreased(P<0.05).The estrous cycle of the mice in the Control group was regular,and follicles at different stages of development could be seen.The mice in the model group had an estrotic cy-cle disorder,with a decrease in primary and secondary follicles,and an increase in atresia follicles.The estrous cycle of mice in the Met group and XNTJT-H group was significantly improved,with an increase in primary and secondary follicles,and a decrease in atresia follicles.Compared with the Control group,the serum levels of LH,T,LH/FSH in the Model group were increased,and the FSH level decreased(P<0.01).Compared with the Model group,the serum levels of LH,T,LH/FSH,and FSH in the Met group and XNTJT-H group were decreased,and the FSH level was increased(P<0.01).Compared with the Control group,the p-NF-κB p65/NF-κB p65 ratio of mice in the Model group was increased,and the ratio of p-IRS-1/IRS-1,p-PI3K/PI3K and p-AKT/AKT decreased(P<0.01).Compared with the model group,the ratio of p-NF-κB p65/NF-κB p65 decreased in the XNTJT-H group and in-creased the ratio of p-IRS-1/IRS-1,p-PI3K/PI3K and p-AKT/AKT(P<0.05).Compared with the Control group,the serum levels of TNF-α,IL-6 and NF-κB in the Model group were increased(P<0.05).Compared with the Model group,the serum lev-els of TNF-α,IL-6 and NF-κB in the Met group and XNTJT-H group were decreased(P<0.05).Cell experiments:Compared with the Control group,the p-NF-κB p65/NF-κB p65 ratio of the cells in the Model group was increased,and the p-IRS-1/IRS-1,p-PI3K/PI3K,and p-AKT/AKT ratios were decreased(P<0.01).Compared with the Model group,the p-NF-κB p65/NF-κB p65 ratio of the cells in the XNTJT group decreased(P<0.01),and the ratio of p-IRS-1/IRS-1,p-PI3K/PI3K and p-AKT/AKT increased(P<0.05).Compared with the XNTJT group,the p-PI3K/PI3K and p-AKT/AKT ratios of the cells in the XNTJT si-IRS-1 group decreased,and the p-NF-κB p65/NF-κB p65 ratio increased(P<0.01).Compared with the Control group,the cell proliferation rate of the Model group was reduced(P<0.01).Compared with the Model group,the XNTJT group showed an increased cell proliferation rate(P<0.01).Compared with the XNTJT group,the XNTJT si-IRS-1 group had a decreased cell proliferation rate(P<0.01).The apoptosis rate of cells in the Model group was higher than that in the Control group(P<0.01);the apoptosis rate in the XNTJT group was lower than that in the Model group(P<0.01);the apoptosis rate in the XNTJT si-IRS-1 group was higher than that in the XNTJT group(P<0.01).Levels of TNF-α,IL-6,and NF-κB in cells of the Model group were higher than those in the Control group(P<0.01);levels in the XNTJT group were lower than those in the Model group(P<0.01);levels in the XNTJT si-IRS-1 group were higher than those in the XNTJT group(P<0.01).Conclusion:Xiaonang Tiaojing Decoc-tion may improve metabolic and reproductive abnormalities in obese PCOS mice and modulate ovarian granulosa cell function by regula-ting the NF-κB/IRS-1/PI3K/AKT pathway.