医学研究与教育 ›› 2024, Vol. 41 ›› Issue (4): 1-7.DOI: 10.3969/j.issn.1674-490X.2024.04.001

• 基础医学 •    下一篇

齐墩果酸对衰老睾丸间质细胞(Leydig细胞)的作用以及EGF/EGFR表达的影响

段豫磊1,牛嗣云1,梁思2,刘子铭2,朱娟3,王红杰1,2   

  1. 1.河北大学基础医学院, 河北 保定 071000;
    2.河北大学附属医院麻醉科, 河北 保定 071000;
    3.郑州市第一人民医院药学部, 河南 郑州 450004
  • 收稿日期:2024-06-25 出版日期:2024-08-25 发布日期:2024-08-25
  • 通讯作者: 王红杰(1968—),女,河北保定人,主任医师,教授,博士,博士生导师,主要从事麻醉与复苏的临床及基础研究。E-mail: hongjiew68@163.com
  • 作者简介:段豫磊(1994—),男,河南焦作人,实验师,硕士,主要从事男性生殖衰老研究。 E-mail: 497305795@qq.com
  • 基金资助:
    国家自然科学基金资助项目(82374180);河北省中医药管理局项目(2020234);河北大学医学学科培育项目(2023B10)

  • Received:2024-06-25 Online:2024-08-25 Published:2024-08-25

摘要: 目的 观察齐墩果酸对衰老睾丸间质细胞(Leydig细胞)的影响以及对表皮生长因子(epidermal growth factor, EGF)和表皮生长因子受体(epidermal growth factor receptor, EGFR)的调控作用。方法 原代培养大鼠Leydig细胞,将其分为正常组、模型组和齐墩果酸组。模型组和齐墩果酸组采用H2O2(300 μmol/L)和FeSO4(100 μmol/L)每日处理2 h,持续4 d,造细胞衰老模型。之后正常组和模型组在DMEM/F12 培养液培养3 d,齐墩果酸组在齐墩果酸终浓度为20 μmol/L的DMEM/F12 培养液中培养3 d。ELISA法检测细胞睾酮分泌情况;BrdU免疫荧光染色检测细胞增殖率;DAPI荧光染色检测细胞凋亡率;qRT-PCR检测细胞EGF和EGFR mRNA表达。结果 模型组睾酮分泌水平、细胞增殖率、EGF和EGFR mRNA表达水平均低于正常组(P<0.05),凋亡率均高于正常组(P<0.05)。齐墩果酸可抑制Leydig细胞凋亡(P<0.05),并提高细胞睾酮分泌水平、细胞增殖水平、EGF和EGFR mRNA表达水平(P<0.05)。结论 齐墩果酸可促进衰老Leydig细胞睾酮分泌及细胞增殖、抑制细胞凋亡,其机制与调控EGF和EGFR mRNA表达有关。

关键词: Leydig细胞, 齐墩果酸, 衰老, 表皮生长因子, 表皮生长因子受体

Abstract: Objective To observe the effect of oleanolic acid on senescent Leydig cells and the regulation of EGF and EGFR. Methods Primary cultured rat Leydig cells were divided into normal group, model group and oleanolic acid group. The model group and the neat fruit acid group were treated with H2O2(300 μmol/L)and FeSO4(100 μmol/L)daily for 2 h for 4 d. Thereafter, the normal and model groups were cultured in DMEM/F12 medium for 3 d, and the oleanolic acid group was cultured in DMEM/F12 medium with a final concentration of 20 μmol/L. Testosterone secretion was measured by ELISA, proliferation by immunofluorescence staining by BrdU, apoptosis by DAPI staining, and EGF and EGFR mRNA expression by qRT-PCR. Results Compared with the normal group, the testosterone secretion level, cell proliferation rate, EGF and EGFR mRNA expression level were significantly decreased(P<0.05), and the apoptosis rate was significantly increased(P<0.05). Oleanolic acid significantly inhibited apoptosis in the model group(P<0.05)and significantly increased testosterone secretion, cell proliferation levels, and EGF and EGFR mRNA expression levels(P<0.05). Conclusion Oleanolic acid can promote testosterone secretion and cell proliferation and inhibit cell apoptosis, and its mechanism may be related to the regulation of EGF and EGFR mRNA expression.

Key words: Leydig cells, oleanolic acid, senescence, epidermal growth factor, epidermal growth factor receptor

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