Mechanism of Luoshi Neiyi Formula in treatment of SF-1 on endometriosis by improving hypoxia
This study investigated the effect of Luoshi Neiyi Formula(LSNYF)on hypoxia-inducible factor-1α(HIF-1α)and ste-roidogenic factor 1(SF-1)in endometriosis(EMs),aiming to explore the mechanism of Luoshi Neiyi Formula in treating EMs.Immu-nohistochemistry and quantitative real-time PCR(qPCR)were employed to determine the expression of HIF-1α and SF-1 in the endo-metriotic tissue.Human primary endometrial stromal cells(ESCs)were extracted and identified,in which the expression levels of HIF-1α and SF-1 were measured by immunofluorescence and qPCR.Cobalt chloride(CoCl2)was used to simulate a low-oxygen envi-ronment in vitro,and the lentivirus with down-regulated expression of HIF-1α was constructed.The expression of HIF-1α or SF-1 was then determined by qPCR or Western blot.The effects of Luoshi Neiyi Formula-containing serum on the proliferation,migration,estra-diol(E2),HIF-1α,and SF-1 of ESCs in the low-oxygen environment were observed.Finally,the rat model of EMs with the syndrome of Qi stagnation and blood stasis was established by multi-factor intervention combined with autologous endometrium transplantation,and the effect of Luoshi Neiyi Formula on HIF-1α and SF-1 in the endometriotic tissue were studied.Compared with the normal endo-metrial tissue and ESCs,the endometriotic tissue and ectopic ESCs presented up-regulated mRNA and protein levels of HIF-1α and SF-1.Compared with the control group,treating eutopic ESCs with 50 pimol·L-1 CoCl2 for 24 h promoted cell proliferation and up-regulated the mRNA level of HIF-1α and protein level of SF-1,while knocking down HIF-1α down-regulated the expression of SF-1 and E2.Compared with the model group,the Luoshi Neiyi Formula-containing serum inhibited the proliferation and migration of ESCs and down-regulated the expression of E2,HIF-1α,and SF-1.Similarly,the treatment with Luoshi Neiyi Formula attenuated the pathologi-cal injury of the endometriotic tissue and down-regulated the mRNA and protein levels of HIF-1α and SF-1 in the rat model of EMs.In summary,LSNYF may treat EMs by alleviating hypoxia and inhibiting the HIF-1α/SF-1 axis.
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