甘氨酸[14] -人胰岛素对环磷酰胺诱导的卵巢早衰的改善作用及潜在机制

Ameliorative effects of Gly[14]-humanin on cyclophosphamide-induced premature ovarian insufficiency and underlying mechanisms.

作者信息

Huang Jin, Liu Yumeng, Zou Liping, Zhu Changhong, Xia Wei

机构信息

Department of Obstetrics and Gynaecology, Wuhan Third Hospital (Tongren Hospital of Wuhan University), Wuhan, Hubei, People's Republic of China.

Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People's Republic of China.

出版信息

Reprod Biomed Online. 2025 Sep;51(3):104901. doi: 10.1016/j.rbmo.2025.104901. Epub 2025 Mar 7.

DOI:10.1016/j.rbmo.2025.104901

PMID:40639309

Abstract

RESEARCH QUESTION

Can Gly[14]-humanin (HNG) improve cyclophosphamide-induced premature ovarian insufficiency (POI)?

DESIGN

A POI model was induced in female rats by intraperitoneal injection of cyclophosphamide, followed by treatment with an intraperitoneal injection of HNG. Ovarian weight, ovarian index, regularity of the oestrous cycle, numbers of various types of follicle, hormone concentrations, and expression of the proliferation marker Ki67 were assessed in rats, and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) staining was performed. The human ovarian granulosa cell line COV434 was exposed to the cyclophosphamide metabolite 4-hydroperoxy cyclophosphamide (4-HC), followed by HNG treatment. Cell viability and single-cell clone formation after HNG treatment were also evaluated. Reactive oxygen species (ROS), mitochondrial ROS (mtROS), 5-ethynyl-2'-deoxyuridine (EdU) and Ki67 expression, and Western blot analysis were performed to evaluate the expression of apoptosis-related proteins (Bcl-2 and Bax) and phosphorylated STAT3 protein.

RESULTS

HNG mitigated cyclophosphamide-induced ovarian damage significantly in rats by restoring the disrupted oestrous cycle and sex hormone concentrations while reducing follicular depletion (P < 0.05) and apoptosis (P < 0.001). HNG treatment enhanced cell viability (P < 0.01) without causing abnormal cell proliferation. Furthermore, HNG ameliorated the 4-HC-induced damage in COV434 cells by enhancing cell viability, increasing the expression of proliferation markers (EdU and Ki67) and reducing ROS and mtROS concentrations (P < 0.05). HNG also decreased the 4-HC-induced elevation in Bax expression, and increased the expression of Bcl-2 and phosphorylated STAT3 (P < 0.05).

CONCLUSIONS

The mitochondrial-derived peptide analogue HNG confers protection against cyclophosphamide-induced POI in rats by reducing oxidative stress and apoptosis. These findings suggest the potential of HNG as a therapeutic agent for preventing cyclophosphamide-induced POI in women.

摘要

研究问题

甘氨酸[14] -人胰岛素（HNG）能否改善环磷酰胺诱导的卵巢早衰（POI）？

设计

通过腹腔注射环磷酰胺诱导雌性大鼠建立POI模型，随后腹腔注射HNG进行治疗。评估大鼠的卵巢重量、卵巢指数、发情周期规律、各类卵泡数量、激素浓度以及增殖标志物Ki67的表达，并进行末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色。将人卵巢颗粒细胞系COV434暴露于环磷酰胺代谢产物4 -氢过氧环磷酰胺（4 - HC），随后进行HNG处理。还评估了HNG处理后的细胞活力和单细胞克隆形成情况。进行活性氧（ROS）、线粒体ROS（mtROS）、5 -乙炔基-2'-脱氧尿苷（EdU）和Ki67表达以及蛋白质印迹分析，以评估凋亡相关蛋白（Bcl - 2和Bax）和磷酸化STAT3蛋白的表达。

结果

HNG通过恢复紊乱的发情周期和性激素浓度，同时减少卵泡耗竭（P < 0.05）和细胞凋亡（P < 0.001），显著减轻了环磷酰胺诱导的大鼠卵巢损伤。HNG处理增强了细胞活力（P < 0.01），且未导致细胞异常增殖。此外，HNG通过增强细胞活力、增加增殖标志物（EdU和Ki67）的表达以及降低ROS和mtROS浓度（P < 0.05），改善了4 - HC诱导的COV434细胞损伤。HNG还降低了4 - HC诱导的Bax表达升高，并增加了Bcl - 2和磷酸化STAT3的表达（P < 0.05）。