Investigation on mechanisms of Danggui Shaoyao Powder in treating premature ovarian insufficiency

WANG Jing; ZHAO Rui; CHEN Yang; ZHANG Xiaojuan; XIA Xiaoyan

doi:10.3969/j.issn.1674-3806.2025.08.07

Abstract：Objective To investigate mechanisms of Danggui Shaoyao Powder in treating premature ovarian insufficiency(POI).Methods The rat model of POI was constructed by intraperitoneal injection of cyclophosphamide(CTX).The blank group,model group,gonadotropin-releasing hormone analogue(GnRHa)group(with intraperitoneal injection of GnRHa at 0.1 mg/kg on the 10th and 5th days before constructing the model)and Danggui Shaoyao Powder group(with gavage administration of Danggui Shaoyao Powder at 10 mL/kg after constructing the model)were set up,with 6 rats in each group,and continuous intervention for 14 days.The morphological structure of rat ovarian tissues was observed by using hematoxylin-eosin(HE)staining and transmission electron microscopy.The levels of serum follicle-stimulating hormone(FSH),estradiol(E2),and anti-Müllerian hormone(AMH)were detected by using enzyme-linked immunosorbent assay(ELISA).The expression levels of key factors in the transforming growth factor β1(TGFβ1)-Smad2 signaling pathway in ovarian tissues were detected by using Western blot assay and reverse transcription-polymerase chain reaction(RT-PCR)assay.Results In the rats of the model group,the levels of serum E2 and AMH were decreased significantly,while the level of FSH was increased significantly,with an increase in atretic follicles,granulosa cells aligned loosely,severe edema of organelles,disordered tissue spaces,cell membrane damage,and irregular depression of cell nuclei in ovarian tissues,indicating that the ovarian function was severely impaired.After the interventions with Danggui Shaoyao Powder and GnRHa,the levels of serum E2 and AMH in the POI model rats were rebounded,while the level of FSH was decreased,with reduced vacuoles in ovarian tissues,increased ovarian follicles at all levels,damages to cell membrane and nuclear membrane similar to those in the model group,and alleviated swelling of organelles.The expression levels of phosphorylated transforming growth factor β receptor 1(p-TGFβR1)and phosphorylated Smad2(p-Smad2)protein in the ovarian tissues of the model group were significantly increased(P＜0.05),and the expression levels of TGFβ1,transforming growth factor β receptor 1(TGFβR1),transforming growth factor β receptor 2(TGFβR2),Smad2 protein and mRNA were significantly increased(P＜0.05),and the expression levels of Smad6 and Smad7 proteins and mRNA were significantly decreased(P＜0.05),indicating that the TGFβ1-Smad2 signaling pathway was overactivated.After the intervention with Danggui Shaoyao Powder,the expression levels of Smad2 and p-Smad2 proteins were significantly decreased(P＜0.05),and the expression levels of TGFβ1,TGFβR1,TGFβR2 proteins and mRNA were significantly decreased(P＜0.05).The expression levels of Smad6 and Smad7 proteins and mRNA were significantly increased(P＜0.05).The indicator changes after the intervention of Danggui Shaoyao Powder were similar to those after the intervention of GnRHa.Conclusion Danggui Shaoyao Powder may exert its therapeutic effect on POI by inhibiting the abnormal activation of the TGFβ1-Smad2 signaling pathway.