冬虫夏草通过调控AMPK/mTOR通路保护高糖诱导的足细胞损伤

doi:10.11958/20240816

天津医药 ›› 2025, Vol. 53 ›› Issue (3): 225-229.doi: 10.11958/20240816

• 细胞与分子生物学 • 下一篇

冬虫夏草通过调控AMPK/mTOR通路保护高糖诱导的足细胞损伤

李冰心¹(), 许军英¹, 张雅茹¹, 周小兵²^,^△()

1 湖南省第二人民医院（湖南省脑科医院）肾内科（邮编410021）
2 湖南省第二人民医院（湖南省脑科医院）微创外科（邮编410021）

收稿日期:2024-06-24 修回日期:2024-08-22 出版日期:2025-03-15 发布日期:2025-03-31
通讯作者: △E-mail：150634509@qq.com
作者简介:李冰心（1982），女，副主任医师，主要从事慢性肾脏病方面研究。E-mail：libignxin_9@163.com
基金资助:
湖南省卫生健康委科研计划项目(D202303057259);湖南省自然科学基金项目(2025JJ90079)

Effect of Cordyceps sinensis on podocyte damage induced by high glucose by regulating the AMPK/mTOR pathway

LI Bingxin¹(), XU Junying¹, ZHANG Yaru¹, ZHOU Xiaobing²^,^△()

1 Department of Nephrology, the Second People’s Hospital of Hunan Province (Brain Hospital of Hunan Province), Changsha 410021, China
2 Department of Minimally Invasive Center for General Surgery, the Second People’s Hospital of Hunan Province (Brain Hospital of Hunan Province), Changsha 410021, China

Received:2024-06-24 Revised:2024-08-22 Published:2025-03-15 Online:2025-03-31
Contact: △E-mail：150634509@qq.com

李冰心, 许军英, 张雅茹, 周小兵. 冬虫夏草通过调控AMPK/mTOR通路保护高糖诱导的足细胞损伤[J]. 天津医药, 2025, 53(3): 225-229.

LI Bingxin, XU Junying, ZHANG Yaru, ZHOU Xiaobing. Effect of Cordyceps sinensis on podocyte damage induced by high glucose by regulating the AMPK/mTOR pathway[J]. Tianjin Medical Journal, 2025, 53(3): 225-229.

摘要/Abstract

摘要：

目的探讨冬虫夏草通过调节腺苷酸活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路对高糖诱导的足细胞损伤的影响。方法体外培养小鼠足细胞，分为正常葡萄糖浓度（NG）组、高糖（HG）组、高糖+冬虫夏草（HG+CS）组、高糖+冬虫夏草+自噬抑制剂（HG+CS+3MA）组、高糖+冬虫夏草+AMPK抑制剂（HG+CS+Compound C）组。CCK-8法检测足细胞活力；蛋白免疫印迹法检测足细胞标志蛋白podocin、nephrin，自噬相关蛋白Beclin-1、P62以及通路相关蛋白磷酸化AMPK（p-AMPK）、磷酸化mTOR（p-mTOR）的表达。结果与NG组比较，HG组足细胞活力降低，podocin、nephrin、Beclin-1、p-AMPK蛋白表达水平降低，P62、p-mTOR蛋白表达水平升高（P<0.05）。与HG组相比，HG+CS组足细胞活力升高，podocin、nephrin、Beclin-1、p-AMPK蛋白表达水平升高，P62、p-mTOR蛋白表达水平降低（P<0.05）。与HG+CS组相比，HG+CS+3MA组和HG+CS+Compound C组足细胞活力均降低，podocin、nephrin、Beclin-1蛋白表达水平均降低，P62蛋白表达水平均升高（P<0.05）；HG+CS+Compound C组p-AMPK蛋白表达水平降低，p-mTOR蛋白表达水平升高（P<0.05）。结论冬虫夏草可能通过上调AMPK/mTOR信号传导通路来激活自噬、改善高糖诱导的足细胞损伤，从而发挥糖尿病肾病足细胞保护作用。

关键词: 冬虫夏草, 糖尿病肾病, 自噬, 足细胞, AMPK/mTOR信号通路

Abstract:

Objective To investigate effects of Cordyceps sinensis (CS) on high glucose (HG) induced podocyte injury by regulating the adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Methods Mouse podocytes were cultured in vitro and divided into the normal glucose (NG) group, the HG group, the HG+CS group, the HG+CS+autophagy inhibitor (HG+CS+3MA) group and the HG+CS+AMPK inhibitor (HG+CS+Compound C) group. Podocyte viability was detected by CCK-8 method. Western blot assay was used to detect the expression levels of podocyte marker proteins podocin and nephrin, autophagy-related proteins Beclin-1 and P62, and pathway related proteins p-AMPK and p-mTOR. Results Compared with the NG group, the cell viability of podocytes decreased, the expression levels of podocin, nephrin，Beclin-1 and p-AMPK protein were decreased, and the expression levels of P62 and p-mTOR protein were increased in the HG group (P<0.05). Compared with the HG group, the cell viability of podocytes was increased, the expression levels of podocin, nephrin, Beclin-1 and p-AMPK protein were significantly increased (P<0.05), and the expression levels of P62 and p-mTOR protein were decreased in the HG+CS group (P<0.05). Compared with the HG+CS group, the cell viability decreased in the HG+CS+3MA group and the HG+CS+Compound C group (P<0.05). Compared with the HG+CS group, the HG+CS+3MA group and the HG+CS+Compound C group showed decreased expression levels of podocin, nephrin and Beclin-1 protein, and increased expression of P62 protein (P<0.05). Compared with the HG+CS group, the expression of p-AMPK protein decreased and the expression of p-mTOR protein increased in the HG+CS+Compound C group (P<0.05). Conclusion Cordyceps sinensis may play a protective role in diabetic nephropathy by up-regulating AMPK/mTOR signaling pathway to induce podocyte autophagy, alleviate high glucose induced podocyte injury and apoptosis.

Key words: cordyceps, diabetic nephropathies, autophagy, podocytes, AMPK/mTOR signaling pathway

中图分类号:

R285.5

图/表 7

图1 各组足细胞活力比较 A：NG组；B：HG组；C：HG+CS组；D：HG+CS+3MA组；E：HG+CS+Compound C组。a与NG组比较，b与HG组比较，c与HG+CS组比较，P<0.05。

Fig.1 Comparison of cell viability levels of podocytes between the five groups

图2 自噬抑制剂干预下各组细胞podocin、nephrin、Beclin-1、P62蛋白的表达 A：NG组；B：HG组；C：HG+CS组；D：HG+CS+3MA组。

Fig.2 The expression levels of podocin, nephrin, Beclin-1 and P62 in cells of each group under autophagy inhibitor intervention

表1 自噬抑制剂干预下各组细胞podocin、nephrin、Beclin-1、P62蛋白表达水平比较（n=5，$\bar{x} \pm s$）

Tab.1 Comparison of protein expression levels of podocin, nephrin, Beclin-1 and P62 under autophagy inhibitor intervention between the four groups of cells

组别	podocin	nephrin	Beclin-1	P62
NG组	1.00±0.00	1.00±0.01	1.00±0.00	1.00±0.00
HG组	0.27±0.03^a	0.32±0.05^a	0.33±0.04^a	4.62±0.57^a
HG+CS组	0.63±0.07^b	0.81±0.08^b	0.61±0.06^b	1.87±0.23^b
HG+CS+3MA组	0.20±0.02^c	0.20±0.05^c	0.25±0.05^c	3.61±0.23^c
F	454.211^＊	425.352^＊	322.695^＊	124.156^＊

图3 AMPK抑制剂干预下各组细胞p-AMPK和p-mTOR蛋白的表达 A：NG组；B：HG组；C：HG+CS组；D：HG+CS+Compound C组。

Fig.3 The expression levels of p-AMPK and p-mTOR protein in each group affected by AMPK inhibitor intervention

表2 AMPK抑制剂干预的各组细胞的p-AMPK和p-mTOR蛋白表达水平比较（n=5，$\bar{x} \pm s$）

Tab.2 Comparison of protein expression levels of p-AMPK and p-mTOR under AMPK inhibitor intervention between the four groups of cells

组别	p-AMPK/AMPK	p-mTOR/mTOR
NG组	1.00±0.00	1.00±0.01
HG组	0.35±0.03^a	1.40±0.05^a
HG+CS组	0.96±0.02^b	1.02±0.05^b
HG+CS+Compound C组	0.27±0.02^c	1.66±0.03^c
F	496.373^＊	305.433^＊

图4 AMPK抑制剂干预下各组细胞podocin、nephrin、Beclin-1、P62蛋白的表达 A：NG组；B：HG组；C：HG+CS组；D：HG+CS+Compound C组。

Fig.4 The expression levels of podocin, nephrin, Beclin-1 and P62 protein in each group affected by AMPK inhibitor intervention

表3 AMPK抑制剂干预下各组细胞podocin、nephrin、Beclin-1、P62蛋白水平比较（n=5，$\bar{x} \pm s$）

Tab.3 Comparison of protein expression levels of podocin, nephrin, Beclin-1 and P62 under AMPK inhibitor intervention between the four groups of cells

组别	podocin	nephrin	Beclin-1	P62
NG组	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00
HG组	0.20±0.03^a	0.43±0.05^a	0.23±0.04^a	4.40±0.67^a
HG+CS组	0.88±0.13^b	0.74±0.05^b	0.86±0.10^b	2.53±0.36^b
HG+CS+ Compound C组	0.30±0.05^c	0.31±0.04^c	0.40±0.07^c	4.26±0.47^c
F	151.175^＊	309.741^＊	160.780^＊	64.570^＊

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冬虫夏草通过调控AMPK/mTOR通路保护高糖诱导的足细胞损伤

Effect of Cordyceps sinensis on podocyte damage induced by high glucose by regulating the AMPK/mTOR pathway

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