补肾还精方调节Klotho表达改善卵巢早衰的机制研究OA

Objective To investigate the underlying mechanism of Bushen Huanjing Formula(补肾还精方,BSHJF)in ameliorating cyclophosphamide(CTX)-induced premature ovarian failure(POF)in mice.Methods Twenty-four 9-week-old female C57BL/6 mice were randomly divided into four groups(n=6 per group):blank control,model,BSHJF treatment,and growth hormone(GH)treat-ment.Except for the control group,POF was induced by intraperitoneal injection of CTX.The BSHJF group received intragastric admin-istration of BSHJF,and the GH group received GH injections.Ovarian morphology was observed via hematoxylin-eosin(HE)staining,and atretic follicles were counted.Serum levels of anti-Müllerian hormone(AMH),follicle-stimulating hormone(FSH),and estradiol(E2)were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of Klotho,receptor for advanced glycation end products(RAGE),and cellular senescence-associated proteins(p53,p21,p16)were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and immunofluorescence(IF).Protein expression of Klotho,RAGE,p53,p21,and p16 was assessed by IF.Results Compared with the control group,the model group exhibited significant ovarian atrophy,reduced ovarian weight,and an increased number of atretic follicles(P<0.01).Serum AMH and E2 levels were decreased(P<0.05),while FSH levels were increased(P<0.05).Compared with the model group,both the BSHJF and GH treatment groups showed an increased number and proportion of growing follicles(P<0.05),elevated serum AMH and E2 levels(P<0.05),and reduced FSH levels(P<0.05).Furthermore,these treatments significantly up-regulated the expression of Klotho(gene and protein)and down-regulated the expression of RAGE,p53,p21,and p16(gene and protein)(P<0.05).Conclusion BSHJF ameliorates CTX-induced premature ovar-ian failure by upregulating the expression of the anti-aging protein Klotho,which subsequently negatively regulates RAGE expression and downregulates senescence-associated proteins,suggesting a potential therapeutic target for POF.