谷胱甘肽对镉致猪肾PK-15细胞氧化损伤及凋亡的修复作用

doi:10.11843/j.issn.0366-6964.2025.05.037

摘要/Abstract

摘要：

本研究探索了谷胱甘肽(glutathione，GSH)对镉(cadmium，Cd)所引起的PK-15细胞损伤的修复功能及其背后的作用机制。本研究主要针对猪肾PK-15细胞，利用MTT检测、光学显微镜观察、透射电镜观察、JC-1染色、彗星试验和流式细胞术等多种研究方法，检测GSH对Cd所造成的细胞氧化损伤及细胞凋亡的修复作用。结果显示，GSH显著缓解了Cd引起的PK-15细胞活性下降，维持ROS的稳态，从而缓解DNA和线粒体损伤及细胞凋亡等现象。Western blot结果表明，GSH可能通过上调抑制凋亡蛋白Bcl-2的表达、下调促凋亡蛋白Bax的表达，进而对Cd诱导的PK-15细胞凋亡产生抑制作用。综上所述，GSH可以通过抑制氧化应激反应和减少细胞凋亡来缓解Cd诱导的PK-15细胞损伤。

关键词: 谷胱甘肽, 镉, PK-15细胞, 氧化损伤, 细胞凋亡

Abstract:

This study explored the reparative effect and mechanism of glutathione (GSH) on cadmium (Cd)-induced PK-15 cell injury. In the present study, we used MTT assay, cellular microscopy observation, JC-1 staining, comet assay, and flow cytometry to investigate the effects of GSH on Cd induced oxidative damage and apoptosis in PK-15 cells. Results showed that GSH significantly alleviated the decrease in cell viability and increase in ROS, attenuated DNA damage, mitochondrial damage, and apoptosis in Cd-treated PK-15 cell, and western blot analysis showed that GSH may inhibit Cd induced PK-15 cell apoptosis by upregulating the expression of anti-apoptotic protein Bcl-2 and downregulating the expression of pro-apoptotic protein Bax. Taken together, these data suggest that GSH can alleviate Cd induced PK-15 cell damage by inhibiting oxidative stress and cell apoptosis.

Key words: glutathione, cadmium, PK-15 cells, oxidative damage, apoptosis

中图分类号:

S856.9

董志芳, 张丽, 朱向博. 谷胱甘肽对镉致猪肾PK-15细胞氧化损伤及凋亡的修复作用[J]. 畜牧兽医学报, 2025, 56(5): 2403-2412.

DONG Zhifang, ZHANG Li, ZHU Xiangbo. Repair Effects of Glutathione on Cadmium Induced Oxidative Damage in Pig Kidney PK-15 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2403-2412.

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主要材料/试剂/仪器 Main materials/reagents/instruments	名称 Name	来源 Origin
主要材料与试剂 Main materials and reagents	PK-15细胞	ATCC细胞库
	DMEM高糖培养基	GIBCO公司
	胎牛血清	生工生物工程有限公司
	氯化镉(CdCl₂)	Sigma公司
	谷胱甘肽	阿拉丁生化科技股份有限公司
	Hoechst 33342	北京索莱宝科技有限公司
	MTT、活性氧检测试剂盒、线粒体膜电位检测试剂盒(JC-1)、Annexin V-FITC/PI细胞凋亡检测试剂盒、BCA蛋白浓度测定试剂盒(增强型)	碧云天生物技术有限公司
	Bax、Bcl-2、β-actin抗体 Cleaved-caspase-3兔单抗	Santa Cruz公司 CTSC(Cell Signaling Technology)
	辣根过氧化物酶/FITC标记的二抗	北京中杉金桥生物技术有限公司
	PVDF膜、超敏化学发光试剂ECL	美国Millipore公司
主要仪器 Main instruments	超净工作台	北京亚泰科隆仪器技术有限公司
	CO₂细胞培养箱	日本三洋
	移液器、Centrifuge 5415R型离心机	德国Eppendorf
	荧光显微镜	日本Olympus
	全波长自动酶标仪	美国Bio-Tek
	DYY-7C型电泳仪	北京六一仪器厂
	凝胶成像系统	美国Bio-Rad
	-80 ℃冰箱	日本三洋
	蛋白电泳槽、转移电泳槽	美国Bio-Rad
	流式细胞仪	美国Beckman
	透射电镜	日本JEOL

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