Regulation of endometrial cell proliferation and migration in endometriosis by total flavonoids of astragalus through the ERα/STAT3 signaling pathway
CHEN Xi
OUYANG Zi-ting
LUO Lan
SHEN Yan
NING Dong-hong
LIANG Xu
Abstract：Objective To investigate the effects of Total Flavonoids of Astragalus(TFA)on the proliferation and migration of endometrial cells in Endometriosis(EMs)and explore the potential molecular mechanisms based on the ERα/STAT3 signaling pathway.Methods Endometrial cells were isolated from human EMs patients.In order to preliminative-ly explore the effects of different doses of TFA on the proliferation and migration of EMs endometrial cells,the experiment was divided into 5 groups,Control(NC)group,ectopic(EMs)group,Astragalus flavone high-dose group(TFA-H,2 mg/mLTFA intervention in ectopic endometrial cells),Astragalus flavone medium-dose group(TFA-M,1 mg/mLT-FA intervention in ectopic endometrial cells),and Astragalus flavone low-dose group(TFA-L,0.5 mg/mLTFA for ec-topic endometrial cells).Plate clone formation assay and CCK-8 assay were used to detect cell proliferation ability,cell scratch assay was used to detect cell migration ability,Western blotting and RT-qPCR were used to detect ERα,STAT3 protein and mRNA expression levels of cells.In order to investigate the effects of ERα/STAT3 signaling pathway on EMs endometrial cells,ERα inducer Ferutinin was used to intervene EMs endometrial cells.They were divided into two groups,NC group and ERα inducer(Ferutinin)group.Western blotting and RT-qPCR were used to detect the expres-sion levels of ERα,STAT3 protein and mRNA,CCK-8 was used to detect cell proliferation ability,and Transwell assay was used to detect cell migration ability.In order to further explore the mechanism of TFA and ERα/STAT3 signaling pathway in EMs endometrial cells,the experiments were divided into four groups:NC group,EMs group,TFA-M group,and TFA-M+Ferutinin group.Western blotting and RT-qPCR were used to detect the expression levels of ERα,STAT3 protein and mRNA,CCK-8 was used to detect cell proliferation ability,and Transwell assay was used to detect cell migration ability Human endometrial cells from EMs patients were isolated.In the preliminary exploration of the im-pact of different doses of TFA on EMs endometrial cell proliferation and migration,the study was divided into five groups,Control(NC)group,Endometriosis(EMs)group,High-dose TFA group(TFA-H,2 mg/mL TFA intervention in EMs endometrial cells),Medium-dose TFA group(TFA-M,1 mg/mL TFA intervention in EMs endometrial cells),and Low-dose TFA group(TFA-L,0.5 mg/mL TFA intervention in EMs endometrial cells).Clonogenic formation as-say,CCK-8 assay,scratch assay,Western blotting,and RT-qPCR were employed to evaluate cell proliferation,migra-tion,and the expression levels of ERα and STAT3 proteins and mRNA.Results(1)Compared to the NC group,the EMs group showed increased clonogenic proliferative ability(P＜0.01).Compared to the EMs group,the TFA-L group(P＜0.05),TFA-M group(P＜0.05),and TFA-H group(P＜0.01)exhibited decreased clonogenic proliferative a-bility.Compared to the NC group,the EMs group demonstrated increased cell viability(P＜0.01).Compared to the EMs group,the TFA-L group,TFA-M group,and TFA-H group showed decreased cell viability(P＜0.05).Compared to the NC group,the EMs group exhibited an increase in relative migration distance(P＜0.01).Compared to the EMs group,the TFA-L group(P＜0.05),TFA-M group(P＜0.05),and TFA-H group(P＜0.01)showed a decrease in relative migration distance.Compared to the NC group,the EMs group showed increased expression levels of ERα and STAT3 proteins and mRNA(P＜0.01).Compared to the EMs group,the TFA-L group(P＜0.05),TFA-M group(P＜0.01),and TFA-H group(P＜0.01)exhibited decreased expression levels of ERα and STAT3 proteins and mR-NA.Based on these results,TFA-M was selected for subsequent experiments.(2)Compared to the NC group,the Ferutinin group showed increased expression levels of ERα and STAT3 proteins and mRNA(P＜0.01).Compared to the NC group,the Ferutinin group demonstrated increased cell viability(P＜0.01).Compared to the NC group,the Feruti-nin group exhibited increased clonogenic proliferative ability(P＜0.01).(3)Compared to the NC group,the EMs group showed increased expression levels of ERα and STAT3 proteins and mRNA(P＜0.05).Compared to the EMs group,the TFA-M group exhibited decreased expression levels of ERα and STAT3 proteins and mRNA(P＜0.05).Compared to the TFA-M group,the TFA-M+Ferutinin group showed increased expression levels of ERα and STAT3 proteins and mRNA(P＜0.05).Compared to the NC group,the EMs group demonstrated increased cell viability(P＜0.05).Con-clusion TFA has the potential to downregulate the proliferation and migration capabilities of endometrial cells in EMs,which may be associated with the ERα/STAT3 signaling pathway.
Keywords：endometriosisendometrial cellstotal flavonoids of astragalusERαSTAT3proliferation and migration
Publication Date：2024-08-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 951-958 )