Exploration of Protective Effect of Zhuluan Decoction on Ovarian Function in Rats with Premature Ovarian Insufficiency via AMPK/mTOR/ULK1 Signaling Pathway
10.13422/j.cnki.syfjx.20260209
- VernacularTitle:基于AMPK/mTOR/ULK1信号通路探讨助卵汤对早发性卵巢功能不全大鼠卵巢功能的保护作用
- Author:
Li TANG
1
;
Yao CHEN
1
;
Sainan TIAN
1
;
Jing ZENG
1
;
Lei LEI
1
;
Wen'e LIU
1
Author Information
1. The First Hospital of Hunan University of Chinese Medicine， Changsha 410007，China
- Publication Type:Journal Article
- Keywords:
Zhuluan decoction;
premature ovarian insufficiency;
adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(21):110-119
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism by which Zhuluan decoction improves ovarian function in a cyclophosphamide （CTX）-induced premature ovarian insufficiency （POI） rat model through regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway. MethodsA POI rat model was established via intraperitoneal injection of CTX. Ten rats were randomly selected as the normal control group. The remaining 50 successfully modeled rats were randomly divided into the model group， the estradiol valerate group （0.1 mg·kg-1）， low-dose （7 g·kg-1）， medium-dose （14 g·kg-1）， and high-dose （28 g·kg-1） Zhuluan decoction groups， with 10 rats per group. All groups received intragastric administration for 21 consecutive days. Changes in body weight， uterine， and ovarian indices were observed across groups. Hematoxylin and eosin （HE） staining was used to examine ovarian pathological alterations and count follicles at various stages in cross-sections. Enzyme-linked immunosorbent assay （ELISA） measured serum estradiol （E2）， follicle-stimulating hormone （FSH）， and anti-Müllerian hormone （AMH） levels. Western blot （WB） and immunohistochemistry （IHC） were used to detect the expression levels of AMPK， mTOR， ULK1 and their phosphorylated proteins， Beclin1， Bcl-2-associated X protein （Bax）， and Bcl-2 in ovarian tissues of each group. ResultsCompared with the normal group， model rats exhibited slower body weight gain （P<0.01）， significantly reduced uterine and ovarian organ indices （P<0.01）， reduced ovarian volume， irregular follicular morphology， unclear cumulus and oocyte structures， and a significant reduction in the number of primordial follicles （P<0.01）. Conversely，the numbers of secondary and atretic follicles significantly increased （P<0.01）. Serum E2 and AMH levels significantly decreased， while FSH levels significantly elevated（P<0.01）. The expression of AMPK， ULK1， Bax， and Beclin1 was increased （P<0.01）， while that of mTOR and Bcl-2 was decreased （P<0.01）. Compared with the model group， the estradiol valerate group and the medium- and high-dose Zhuluan decoction groups showed significantly increased body weight， uterine organ index， and ovarian organ index （P<0.01）， while the low-dose Zhuluan decoction group showed no statistically significant difference. Ovarian volume increased across all treatment groups， with improved follicular morphology and structure. The number of primordial follicles significantly increased （P<0.01）， while the number of atretic follicles significantly decreased （P<0.01）. The number of secondary follicles decreased in the estradiol valerate group and the high-dose Zhuluan decoction group （P<0.01）， with no significant difference in the low- and medium-dose Zhuluan decoction groups. E2 and AMH levels significantly increased， while FSH levels significantly decreased in all treatment groups （P<0.05， P<0.01）. The estradiol valerate group and medium- and high-dose Zhuluan decoction groups showed significantly reduced expression of AMPK， ULK1， Bax， and Beclin1 （P<0.05， P<0.01）， while mTOR and Bcl-2 expression levels were significantly elevated （P<0.01）. The low-dose Zhuluan decoction group exhibited significantly reduced expression of ULK1， Beclin1， and Bax （P<0.05）， while Bcl-2 expression was significantly increased （P<0.05）. ConclusionZhuluan decoction improves serum hormone levels in POI rats and mitigates chemotherapy （CTX）-induced damage to ovarian structure and function， exerting ovarian protective effect. This mechanism may involve regulating the AMPK/mTOR/ULK1 signaling pathway to inhibit excessive autophagy and apoptosis.
