Zuogui Wan preserves Leydig cell steroidogenesis via G6PD/NADPH/GSH-dependent ferroptosis inhibition in D-galactose-induced hypogonadism

IF 6.8 2区医学 Q1 CHEMISTRY, MEDICINAL

Journal of ethnopharmacology Pub Date : 2027-02-10 Epub Date: 2026-10-17 DOI:10.1016/j.jep.2026.122507

Ruonan Li, Shenghan Xu, Yuhao Liu, Shuo Sun, Han Wu, Zhitao Wei, Yong Yang

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引用次数: 0

Abstract

Ethnopharmacological relevance

Zuogui Wan (ZGW), a classical formula for aging-related disorders, may delay reproductive decline, but its effects on late-onset hypogonadism (LOH) remain unclear.

Aim of the study

To investigate the effects and underlying mechanisms of ZGW in LOH using rat and cellular models by integrating serum pharmacochemistry, network pharmacology, and transcriptomic analyses.

Materials and methods

A D-galactose-induced accelerated aging rat model was established to evaluate the ameliorative effects of ZGW. Blood-absorbed constituents were identified by UHPLC–MS/MS, and candidate pathways and targets were screened by integrating network pharmacology, testicular transcriptomics, molecular docking, and molecular dynamics simulations. Mechanistic validation was performed in rat testicular tissues and TM3 Leydig cells, and the functional role of G6PD was further evaluated using G6PDi-1-mediated pharmacological inhibition and G6pdx knockdown/overexpression rescue experiments.

Results

ZGW ameliorated D-galactose-induced reproductive endocrine dysfunction and testicular aging-related injury in rats and attenuated injury in TM3 Leydig cells. Based on the identification of 36 blood-absorbed constituents, integrated analyses prioritized G6PD and suggested that it may be involved in redox regulation and ferroptosis-related mechanisms. In vivo and in vitro experiments showed that ZGW restored G6PD-related NADPH/GSH redox homeostasis, strengthened GPX4-dependent defense against lipid peroxidation, attenuated ferroptosis-related injury, and preserved mitochondrial function and StAR/CYP11A1-associated steroidogenesis. Both G6PDi-1 and G6pdx knockdown partially attenuated the effects of ZGW, whereas G6pdx rescue partially restored these changes.

Conclusions

ZGW may alleviate D-gal-induced LOH-like dysfunction by restoring G6PD-related NADPH/GSH homeostasis and GPX4-dependent lipid-peroxide defense. These effects may limit ferroptosis-related injury and preserve the mitochondrial conditions required for StAR/CYP11A1-associated steroidogenesis.

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左桂丸通过G6PD/NADPH/ gsh依赖性铁下垂抑制d -半乳糖诱导的性腺功能减退，保存间质细胞的甾体生成。

民族药理学相关性：左归丸（ZGW）是治疗衰老相关疾病的经典配方，可能延缓生殖功能下降，但其对迟发性性腺功能减退（LOH）的影响尚不清楚。目的：结合血清药物化学、网络药理学和转录组学分析，利用大鼠和细胞模型研究ZGW对LOH的作用及其机制。材料与方法：建立d -半乳糖诱导的加速衰老大鼠模型，评价ZGW的改善作用。通过UHPLC-MS/MS鉴定血吸收成分，结合网络药理学、睾丸转录组学、分子对接和分子动力学模拟筛选候选通路和靶点。在大鼠睾丸组织和TM3间质细胞中进行机制验证，并通过g6pdi -1介导的药理抑制和G6pdx敲低/过表达拯救实验进一步评价G6PD的功能作用。结果：ZGW可改善d -半乳糖诱导的大鼠生殖内分泌功能障碍和睾丸衰老相关损伤，减轻TM3间质细胞损伤。在鉴定了36种血吸收成分的基础上，综合分析优先考虑了G6PD，并认为其可能参与氧化还原调节和凋亡相关机制。体内和体外实验表明，ZGW恢复了g6pd相关的NADPH/GSH氧化还原稳态，增强了gpx4依赖的脂质过氧化防御，减轻了铁中毒相关的损伤，并保持了线粒体功能和StAR/ cyp11a1相关的甾体生成。G6PDi-1和G6pdx的敲除都部分减弱了ZGW的作用，而G6pdx的恢复部分恢复了这些变化。结论：ZGW可能通过恢复g6pd相关的NADPH/GSH稳态和gpx4依赖的脂质过氧化物防御来缓解d- gal诱导的loh样功能障碍。这些作用可能限制了铁中毒相关的损伤，并保留了StAR/ cyp11a1相关类固醇生成所需的线粒体条件。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of ethnopharmacology 医学-全科医学与补充医学

CiteScore

10.30

自引率

5.60%

发文量

967

审稿时长

77 days

期刊介绍： The Journal of Ethnopharmacology is dedicated to the exchange of information and understandings about people''s use of plants, fungi, animals, microorganisms and minerals and their biological and pharmacological effects based on the principles established through international conventions. Early people confronted with illness and disease, discovered a wealth of useful therapeutic agents in the plant and animal kingdoms. The empirical knowledge of these medicinal substances and their toxic potential was passed on by oral tradition and sometimes recorded in herbals and other texts on materia medica. Many valuable drugs of today (e.g., atropine, ephedrine, tubocurarine, digoxin, reserpine) came into use through the study of indigenous remedies. Chemists continue to use plant-derived drugs (e.g., morphine, taxol, physostigmine, quinidine, emetine) as prototypes in their attempts to develop more effective and less toxic medicinals.