环孢霉素A对葡萄糖氧化酶诱导的HepG2线粒体功能损伤和细胞凋亡的保护作用

发布时间：2018-01-10 11:27

本文关键词：环孢霉素A对葡萄糖氧化酶诱导的HepG2线粒体功能损伤和细胞凋亡的保护作用　出处：《癌变.畸变.突变》2014年01期 　论文类型：期刊论文

【摘要】：目的:探讨环孢霉素A(CsA)对葡萄糖氧化酶(GOX)诱导的肝细胞线粒体功能损伤和细胞凋亡的影响。方法:胞分别给予不同浓度的GOX和CsA处理,MTT法检测细胞活力,确定所要选用的剂量和时间点。以10μmol/L的CsA预处理2 h,并给予25 U/L GOX处理12 h作为CsA+GOX组,同时设立空白对照组、GOX阳性对照组、CsA阴性对照组。分别采用Annexin V/PI双染色结合流式细胞术检测细胞凋亡率;GENMED活体细胞线粒体膜通道孔荧光试剂盒检测线粒体膜通透性转换;JC-1染色激光共聚焦检测线粒体膜电位变化;ATP检测试剂盒检测细胞内ATP水平;液相氧电极法检测线粒体耗氧量。结果:MTT结果显示GOX能诱导HepG2细胞活力下降,且呈现时间和剂量效应关系(P均0.05);与空白对照组比较,25 U/L GOX作用12 h可诱导HepG2细胞凋亡,同时伴有线粒体膜通道开放和膜电位下降,细胞内ATP水平和线粒体耗氧量下降(P均0.05),而10μmol/L的CsA预处理2 h有一定的保护作用,可以有效阻断线粒体结构功能的损伤,降低凋亡率,CsA+GOX组与GOX阳性对照组比较差异均具有统计学意义(P均0.05)。结论:CsA对氧化应激诱导的肝细胞线粒体功能损伤和凋亡具有保护作用。
[Abstract]:Objective: to study the effect of cyclosporine (CsA) on glucose oxidase (GOX). Methods: cells were treated with different concentrations of GOX and CsA respectively. The cell viability was detected by MTT, and the dosage and time point were determined. The cells were pretreated with 10 渭 mol/L CsA for 2 h. The CsA GOX group was treated with 25 U / L GOX for 12 h, and the blank control group was established as the positive control group. CsA negative control group. Annexin V / Pi double staining and flow cytometry were used to detect the apoptosis rate. Mitochondrial membrane permeability was detected by fluorescence kit of mitochondrial membrane channel in GENMED living cells. The changes of mitochondrial membrane potential were detected by confocal laser with JC-1 staining. ATP assay kit was used to detect intracellular ATP level. Results the results showed that GOX could induce the decrease of HepG2 cell viability, and the relationship between time and dose-effect was 0.05%. Compared with the control group, 25 U / L GOX could induce apoptosis of HepG2 cells for 12 h, accompanied by the opening of mitochondrial membrane channel and the decrease of membrane potential. Both intracellular ATP level and mitochondrial oxygen consumption decreased by 0.05%, while 10 渭 mol/L CsA pretreatment for 2 h had a protective effect to some extent. It can effectively block the damage of mitochondrial structure and function and reduce the apoptosis rate. The difference between CsA GOX group and GOX positive control group was statistically significant (P < 0.05). Conclusion: CSA has protective effect on oxidative stress induced mitochondrial dysfunction and apoptosis.
【作者单位】：第四军医大学军事预防医学系毒理学教研室;
【分类号】：R329
【正文快照】： 研究表明,线粒体结构功能改变与细胞凋亡密切相关,大量肝细胞急性死亡是临床急性肝损伤、肝衰[1]竭的病理基础。机体氧化还原失衡引发的氧化应激是多种肝病发病的共同病理生理基础,活性氧主要通过肝细胞线粒体的膜通透性和流动性降低,细胞钙内流,与细胞因子(如NF-κB、TNF-α

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