Choline theophyllinate promotes activation of primordial follicles in mice through PI3K/Akt signaling pathway
10.3969/j.issn.1000-4718.2025.06.011
- VernacularTitle:胆茶碱通过PI3K/Akt信号通路促进小鼠原始卵泡激活
- Author:
Tiantian HAO
1
;
Wenbo ZHANG
1
;
Weiyong WANG
1
;
Meijia ZHANG
1
Author Information
1. 华南理工大学医学院,广东 广州 510006
- Publication Type:Journal Article
- Keywords:
choline theophyllinate;
mouse;
primordial follicle activation;
PI3K/Akt signaling pathway
- From:
Chinese Journal of Pathophysiology
2025;41(6):1145-1152
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the function and mechanism of the smooth muscle relaxant choline theophylli-nate(CT)in activating primordial follicles in mice.METHODS:Experiments were conducted using in vitro culture of 3-day postpartum(dpp)neonatal SPF-grade female mice,intraperitoneal injection in 3-dpp neonatal mice,and oral adminis-tration in 21-dpp adolescent female mice.The mice were divided into control and CT groups.The ovaries were isolated from 3-dpp neonatal mice for the in vitro culture.Hematoxylin staining was used to count the number of primordial and growing follicles,with 10 mice in each group.qPCR was performed to analyze the expression levels of genes related to fol-licle growth,proliferation,and apoptosis,including growth differentiation factor 9(Gdf9),zona pellucida glycoprotein 3(Zp3),proliferating cell nuclear antigen(PCNA),Ki-67,B-cell lymphoma 2(BCL2),BCL2-associated X protein(BAX),and caspase-3,with 9 mice in each group.Immunofluorescence staining was used to detect the expression of pro-liferation-and apoptosis-related proteins,including PCNA,Ki-67,5-bromo-2'-deoxyuridine(BrdU),cleaved caspase-3,and forkhead box O3a(FOXO3a),with 15 mice in each group.Western blot was performed to measure the expression of DEAD-box helicase 4(DDX4),phosphorylated mammalian target of rapamycin(p-mTOR),and phosphorylated protein kinase B(p-Akt),with 9 mice in each group.Intraperitoneal injection of CT was administered to 3-dpp mice,and follicle counting was performed with 10 mice in each group.Western blotting was used to detect p-mTOR and p-Akt expression,with 9 mice in each group.Immunofluorescence was employed to assess FOXO3a nuclear export,with 15 mice in each group.For the oral administration of CT in drinking water to 21-dpp mice,immunofluorescence and hematoxylin staining was used to count follicles,with 9 mice in each group.RESULTS:Compared with control group,CT treatment signifi-cantly increased the number of growing follicles in mice.The mRNA and/or protein levels of Gdf9,Zp3,Ki-67,PCNA and DDX4 were markedly elevated.Further studies revealed that CT treatment significantly increased p-Akt levels in the ovaries but had no significant effect on p-mTOR levels.The PI3K/Akt inhibitor LY294002 also reversed the choline the-ophyllinate-induced increase in growing follicles.CONCLUSION:Choline theophyllinate promotes the activation of pri-mordial follicles in mice via the PI3K/Akt signaling pathway in oocytes.
