补肾中药通过AMPK-mTOR通路提高肾精亏虚大鼠睾丸间质细胞自噬能力

doi:10.3969/j.issn.1000-1565.2024.06.009

摘要/Abstract

摘要： 为了探究补肾中药提高肾精亏虚大鼠睾丸间质细胞自噬能力的机制,采用单因素电刺激法建立肾精亏虚大鼠模型,免疫组化及Western blot检测各组睾丸细胞自噬标志蛋白Beclin1、p62、LC3B表达水平以及AMPK和mTOR表达变化.基于肾精亏虚与衰老的密切联系,通过自由基氧化损伤法建立睾丸间质细胞(Leydig细胞)衰老模型,联合AMPK抑制剂,探究何首乌饮激活细胞自噬的分子机制.结果显示:何首乌饮可上调睾丸组织Beclin1蛋白、LC3B表达,下调p62蛋白表达,促进AMPK磷酸化并抑制mTOR激活(P<0.05);细胞实验表明,何首乌饮可通过激活AMPK-mTOR信号通路提高Leydig细胞自噬能力(P<0.05),促进肾精亏虚大鼠睾丸Leydig细胞自噬进程.

关键词: 何首乌饮, 肾精亏虚, 睾丸组织, 自噬, AMPK-mTOR

Abstract: In order to explore the mechanism of kidney-tonifying Chinese medicine in improving autophagy of testicular mesenchymal cells in rats with kidney-essence deficiency, a rat model of kidney-essence deficiency was established by single-factor electrical stimulation. Immunohistochemistry and Western blot were used to detect the expression levels of autophagy marker proteins Beclin1, p62, LC3B, AMPK and mTOR in testicular cells of each group. Based on the close relationship between kidney essence deficiency and aging, Leydig cell aging model of testicular mesenchymal cells was established by free radical oxidative damage method, and AMPK inhibitor was combined to explore the molecular mechanism of Heshouwuyin activating autophagy. The results showed that Heshouwuyin could up-regulate the expression of Beclin1 protein and LC3B in testicular tissue, down-regulate the expression of p62 protein,- DOI:10.3969/j.issn.1000-1565.2024.06.009补肾中药通过AMPK-mTOR通路提高肾精亏虚大鼠睾丸间质细胞自噬能力李朝英¹,李思睿²,张伟³,马昊飞²,牛双²,段豫磊²,牛嗣云² (1.河北大学附属医院新生儿科, 河北保定 071000;2.河北大学基础医学院, 河北保定 071000;3.河北大学附属医院医学工程中心, 河北保定 071000)摘要:为了探究补肾中药提高肾精亏虚大鼠睾丸间质细胞自噬能力的机制,采用单因素电刺激法建立肾精亏虚大鼠模型,免疫组化及Western blot检测各组睾丸细胞自噬标志蛋白Beclin1、p62、LC3B表达水平以及AMPK和mTOR表达变化.基于肾精亏虚与衰老的密切联系,通过自由基氧化损伤法建立睾丸间质细胞(Leydig细胞)衰老模型,联合AMPK抑制剂,探究何首乌饮激活细胞自噬的分子机制.结果显示:何首乌饮可上调睾丸组织Beclin1蛋白、LC3B表达,下调p62蛋白表达,促进AMPK磷酸化并抑制mTOR激活(P<0.05);细胞实验表明,何首乌饮可通过激活AMPK-mTOR信号通路提高Leydig细胞自噬能力(P<0.05),促进肾精亏虚大鼠睾丸Leydig细胞自噬进程.关键词:何首乌饮;肾精亏虚;睾丸组织;自噬;AMPK-mTOR中图分类号:Q291 文献标志码:A 文章编号:1000-1565(2024)06-0633-10Kidney-tonifying Chinese medicine improves autophagy of Leydig cells in rats with kidney essence deficiency through AMPK-mTOR pathwayLI Chaoying¹, LI Sirui², ZHANG Wei³, MA Haofei², NIU Shuang², DUAN Yulei², NIU Siyun²(1. Department of Neonatology, Affiliated Hospital of Hebei University, Baoding 071000, China;2. School of Basic Medical Sciences, Hebei University, Baoding 071000, China;3. Medical Engineering Center, Affiliated Hospital of Hebei University, Baoding 071000, China)Abstract: In order to explore the mechanism of kidney-tonifying Chinese medicine in improving autophagy of testicular mesenchymal cells in rats with kidney-essence deficiency, a rat model of kidney-essence deficiency was established by single-factor electrical stimulation. Immunohistochemistry and Western blot were used to detect the expression levels of autophagy marker proteins Beclin1, p62, LC3B, AMPK and mTOR in testicular cells of each group. Based on the close relationship between kidney essence deficiency and aging, Leydig cell aging model of testicular mesenchymal cells was established by free radical oxidative damage method, and AMPK inhibitor was combined to explore the molecular mechanism of Heshouwuyin activating autophagy. The results showed that Heshouwuyin could up-regulate the expression of Beclin1 protein and LC3B in testicular tissue, down-regulate the expression of p62 protein,- 收稿日期:2024 -02-19;修回日期:2024-05-08 基金项目:国家自然科学基金资助项目(82374180);河北省中医药管理局科研项目(2020234);河北省重点研发计划项目-中医药创新专项(223777134D);河北大学医学学科培育项目(2023B10);河北省高等学校科学技术研究项目(ZD2022060) 第一作者: 李朝英(1980—),女,河北大学附属医院主治医师,主要从事早产儿营养评估.E-mail:lizhaoying725@126.com 通信作者:段豫磊(1994—),男,河北大学实验师,主要从事中药药理方向研究.E-mail:dyl15771@163.com牛嗣云(1967—),女,满族,河北大学教授,博士,博士生导师,主要从事生殖衰老方向研究.E-mail:nsy1688@163.com 第6期李朝英等:补肾中药通过AMPK-mTOR通路提高肾精亏虚大鼠睾丸间质细胞自噬能力河北大学学报(自然科学版) 第44卷promote AMPK phosphorylation and inhibit mTOR activation(P<0.05). Cell experiments showed that Heshouwuyin could affect the autophagy ability of Leydig cells by activating AMPK-mTOR signaling pathway(P<0.05).Therefore, Heshouwuyin may promote the autophagy process of Leydig cells in rats with kidney essence deficiency by activating AMPK-mTOR signaling pathway.

Key words: Heshouwuyin, kidney essence deficiency, testicular tissue, autophagy, AMPK-mTOR

中图分类号:

Q291

李朝英,李思睿,张伟,马昊飞,牛双,段豫磊,牛嗣云. 补肾中药通过AMPK-mTOR通路提高肾精亏虚大鼠睾丸间质细胞自噬能力[J]. 河北大学学报(自然科学版), 2024, 44(6): 633-642.

LI Chaoying, LI Sirui, ZHANG Wei, MA Haofei, NIU Shuang, DUAN Yulei, NIU Siyun. Kidney-tonifying Chinese medicine improves autophagy of Leydig cells in rats with kidney essence deficiency through AMPK-mTOR pathway[J]. Journal of Hebei University(Natural Science Edition), 2024, 44(6): 633-642.

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