Chikusetsusaponin IVa通过Nrf2/HO-1信号通路改善帕罗西汀诱导的间质细胞（TM3细胞）损伤

IF 2.8 4区医学 Q2 REPRODUCTIVE BIOLOGY

Reproductive toxicology Pub Date : 2025-08-28 DOI:10.1016/j.reprotox.2025.109039

Qianqian Huang , Haiying Wu , Xiangxin Xiao , Zhen Lu , Xiuping Fan , Wenhong Cao , Suqing Liu , Xiaoming Qin

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引用次数: 0

摘要

帕罗西汀（PRX）对男性生殖系统有明显的毒性作用。以往的动物研究表明，茯苓提取物可以改善prx诱导的雄性小鼠性功能障碍，但其有效成分和机制尚不清楚。菊叶皂苷IVa （CHS-IVa）是番麻的主要皂苷成分，具有抗氧化和抗凋亡活性，已成为研究热点。本研究旨在探讨CHS-IVa是否通过调节氧化应激、凋亡和雄激素合成途径，减轻prx诱导的小鼠间质细胞（TM3细胞）损伤。为此，我们建立了prx诱导的TM3细胞损伤模型，并通过CCK-8细胞活力测定、ELISA检测性激素水平、DCFH-DA荧光探针检测活性氧（ROS）、RT-qPCR/Western blot分析mRNA和蛋白表达进行综合评价。结果显示，与PRX组相比，CHS-IVa（6.25 μg/mL）显著提高细胞活力12.9 % (p <； 0.05)；激活Nrf2/HO1信号通路，使细胞内ROS水平降低至少21.9% % (p <； 0.05)，从而减轻氧化应激损伤；雄激素合成相关基因（StAR、CYP11a1、CYP17a1、LHr） mRNA表达上调2倍以上（p <； 0.05），其中睾酮、二氢睾酮和黄体生成素水平最高分别升高8.4 %、50.4 %和13.0 % (p <； 0.05)；增强抗凋亡因子bcl - 2 mRNA和蛋白表达了3.4倍和1.6倍(p & lt; 0.05),同时减少pro-apoptotic因子Bax mRNA和蛋白表达40.5 % 44.6 % (p & lt; 0.05),和减少Caspase-3 mRNA表达61.5 % (p & lt; 0.05),最终减少PRX-induced TM3细胞异常凋亡。综上所述，CHS-IVa是肾小球法菲的关键活性成分，通过多种机制改善TM3细胞的损伤，从而保护肾小球法菲免受prx诱导的生殖毒性。

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Chikusetsusaponin IVa ameliorates paroxetine-induced Leydig cells (TM3 cells) injury via the Nrf2/HO-1 signaling pathway

Paroxetine (PRX) exhibits significant toxic effects on the male reproductive system. Previous animal studies have demonstrated that Pfaffia glomerata extract can ameliorate PRX-induced sexual dysfunction in male mice, but its active components and underlying mechanisms remain unclear. Chikusetsusaponin IVa (CHS-IVa), a major saponin component of Pfaffia glomerata with well-documented antioxidant and anti-apoptotic activities, has become a research focus. This study aimed to investigate whether CHS-IVa could mitigate PRX-induced injury in mouse Leydig cells (TM3 cells) by regulating oxidative stress, apoptosis, and androgen synthesis pathways. To achieve this, a PRX-induced injury model in TM3 cells was established and subjected to comprehensive evaluation using CCK-8 assay for cell viability, ELISA for sex hormone levels, DCFH-DA fluorescent probe for reactive oxygen species (ROS) detection, and RT-qPCR/Western blot for mRNA and protein expression analysis. Results showed that compared to PRX group, CHS-IVa (6.25 μg/mL) significantly increased cell viability by 12.9 % (p < 0.05); activated Nrf2/HO1 signaling pathway, reducing intracellular ROS levels by at least 21.9 % (p < 0.05), thereby alleviating oxidative stress injury; upregulated mRNA expression of androgen synthesis-related genes (StAR, CYP11a1, CYP17a1, LHr) by over 2-fold (p < 0.05), with maximal increases in testosterone, dihydrotestosterone and luteinizing hormone levels by 8.4 %, 50.4 % and 13.0 % respectively (p < 0.05); enhanced anti-apoptotic factor Bcl-2 mRNA and protein expression by up to 3.4-fold and 1.6-fold (p < 0.05), while reducing pro-apoptotic factor Bax mRNA and protein expression by 40.5 % and 44.6 % (p < 0.05), and decreasing Caspase-3 mRNA expression by 61.5 % (p < 0.05), ultimately reducing PRX-induced abnormal apoptosis in TM3 cells. In conclusion, CHS-IVa serves as the key active component in Pfaffia glomerata that protects against PRX-induced reproductive toxicity by ameliorating injury in TM3 cells through multiple mechanisms.

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来源期刊

Reproductive toxicology 生物-毒理学

CiteScore

6.50

自引率

3.00%

发文量

131

审稿时长

45 days

期刊介绍： Drawing from a large number of disciplines, Reproductive Toxicology publishes timely, original research on the influence of chemical and physical agents on reproduction. Written by and for obstetricians, pediatricians, embryologists, teratologists, geneticists, toxicologists, andrologists, and others interested in detecting potential reproductive hazards, the journal is a forum for communication among researchers and practitioners. Articles focus on the application of in vitro, animal and clinical research to the practice of clinical medicine. All aspects of reproduction are within the scope of Reproductive Toxicology, including the formation and maturation of male and female gametes, sexual function, the events surrounding the fusion of gametes and the development of the fertilized ovum, nourishment and transport of the conceptus within the genital tract, implantation, embryogenesis, intrauterine growth, placentation and placental function, parturition, lactation and neonatal survival. Adverse reproductive effects in males will be considered as significant as adverse effects occurring in females. To provide a balanced presentation of approaches, equal emphasis will be given to clinical and animal or in vitro work. Typical end points that will be studied by contributors include infertility, sexual dysfunction, spontaneous abortion, malformations, abnormal histogenesis, stillbirth, intrauterine growth retardation, prematurity, behavioral abnormalities, and perinatal mortality.