Effects of medicated serum of Siwutang on autophagy of ovarian granulosa cells in polycystic ovarian syndrome
10.6039/j.issn.1001-0408.2025.02.09
- VernacularTitle:四物汤含药血清对多囊卵巢综合征卵巢颗粒细胞自噬的影响
- Author:
Yanshe SHAO
1
;
Xuemei XU
;
Baoqin YANG
;
Huijuan LI
;
Xia JI
Author Information
1. 河南省中医院/河南中医药大学第二附属医院妇产科,郑州 450011
- Publication Type:Journal Article
- Keywords:
Siwutang;
medicated serum;
polycystic ovarian syndrome;
ovarian granulosa cells;
FBP1/mTOR signaling pathway
- From:
China Pharmacy
2025;36(2):185-190
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the effects of medicated serum of Siwutang on autophagy of ovarian granulosa cells(KGN cells)in polycystic ovarian syndrome(PCOS)and its underlying mechanism.METHODS Blank serum and different-concentration medicated serum of Siwutang were prepared by intragastric administration of normal saline and different doses of Siwutang[0.52,1.04,2.08 g/(kg·d)]in 3-month-old female SD rats.After screening the intervention concentration of Siwutang medicated serum,KGN cells were divided into control group(without any treatment),dehydroepiandrosterone(DHEA)group(treated with 50 μmol/L DHEA for 48 h),blank serum group(treated with 50 μmol/L DHEA for 48 h and with 10％blank serum for 72 h)and medium-concentration of Siwutang medicated serum group(treated with 50 μmol/L DHEA for 48 h and with 10％medium-concentration Siwutang medicated serum for 72 h).The number of autophagosomes was observed in each group,and protein expressions of pathway-related proteins[fructose-1,6-bisphosphatase 1(FBP1),mammalian target of rapamycin(mTOR),phosphorylated mTOR(p-mTOR)],autophagy-related proteins[p62,microtubule-associated protein 1 light chain 3(LC3)]and mRNA expression of FBP1 were also detected.The(transfected)cells were further divided into Siwutang group(treated with 10％medium dose of Siwutang medicated serum for 72 h after 48 h intervention with 50 μmol/L DHEA),Siwutang+si-NC group[negative control small interfering RNA(siRNA)transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration of Siwutang medicated serum for 72 h]and Siwutang+si-FBP1 group(FBP1 siRNA transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration Siwutang medicated serum for 72 h).The effects of knocking down FBP1 on the above-mentioned effects of Siwutang were detected.RESULTS Compared with control group,DHEA group exhibited an increase in the number of autophagosomes,an elevated LC3-Ⅱ/LC3-Ⅰ and p-mTOR/mTOR,as well as increases in protein and mRNA expressions of FBP1,and decreased protein expression of p62(P＜0.05).Compared to both DHEA group and blank serum group,the medium-concentration of Siwutang medicated serum group showed a decrease in the number of autophagosomes,a decrease in LC3-Ⅱ/LC3-Ⅰ,and increases in p-mTOR/mTOR,protein expression of p62,protein and mRNA expressions of FBP1(P＜0.05).After knocking down FBP1,compared with Siwutang+si-NC group,Siwutang+si-FBP1 group showed a significant decrease in cell viability,protein expression of p62,protein and mRNA expressions of FBP1 as well as p-mTOR/mTOR,and an increase in LC3-Ⅱ/LC3-Ⅰ(P＜0.05).CONCLUSIONS Siwutang can promote the phosphorylation of mTOR protein by up-regulating the protein and mRNA expressions of FBP1 in KGN cells,thus inhibiting autophagy of KGN cells.
