摘要
目的:探讨重组人MG53蛋白对人脐带间充质干细胞(hUC-MSCs)氧化损伤的保护作用及分子机制。方法:实验分为对照组(CON组,不处理)、LPS组(加入200mg/LLPS)、MG53组(加入30mg/L重组人MG53蛋白)和MG53+LPS组(同时加入重组人MG53蛋白和LPS),48h后,采用CCK-8法检测细胞活力,AO-EB染色检测细胞凋亡,JC-1染色评价线粒体膜电位变化,DCFH-DA荧光探针法检测细胞内活性氧(ROS)水平,酶标法检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和过氧化氢酶(CAT)活性,Westernblot法检测磷酸化Akt和GSK-3β蛋白的表达。结果:与LPS组相比,MG53+LPS组细胞存活率升高,凋亡率和线粒体膜电位降低,细胞内ROS和MDA含量降低,SOD和CAT活性升高,同时磷酸化Akt和磷酸化GSK-3β的表达升高(P<0.05)。结论:重组人MG53蛋白可能通过激活Akt/GSK-3β信号通路,保护hUC-MSCs免受LPS诱导的氧化损伤。
Aim :To investigate the protective effects of rhMG53 protein on LPS-induced oxidative damage to human umbilical cord mesenchymal stem cells(hUC-MSCs). Methods :hUC-MSCs were divided into control group(CON group), LPS group(200 mg/L LPS), MG53 group(30 mg/L rhMG53),and MG53+LPS group.After 48 hours,cell viability was detected by CCK-8 assay, apoptosis was detected by AO-EB staining, mitochondrial membrane potential was measured by JC-1 staining, intracellular reactive oxygen species(ROS) level was detected by DCFH-DA fluorescence probing method, malondialdehyde(MDA) content, and superoxide dismutase(SOD) and catalase(CAT) activity were detected by enzyme labeling method,and Western blot was used to analyze the expressions of phosphorylated Akt and GSK-3β. Results :Compared with LPS group, the viability rate in MG53+LPS group increased, apoptosis rate and mitochondrial membrane potential decreased,ROS production and MDA content were lower, and SOD and CAT activities were higher( P <0.05). In addition, the expressions of LPS-induced phosphorylated Akt and phosphorylated GSK-3β also increased in MG53+LPS group( P <0.05). Conclusion :rhMG53 could protect hUC-MSCs from LPS-induced oxidative damage by activating the Akt/GSK-3β signaling pathway.
作者
王亚苹
张振坤
李喆
刘腾飞
黄团结
周馨魁
麻建杰
关方霞
马珊珊
WANG Yaping;ZHANG Zhenkun;LI Zhe;LIU Tengfei;HUANG Tuanjie;ZHOU Xinkui;MA Jianjie;GUAN Fangxia;MA Shanshan(College of Life Sciences,Zhengzhou University,Zhengzhou 450001;Davis Heart and Lung Research Institute,the Ohio State University,Columbus,OH,43210 USA)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第4期522-526,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
国家自然科学基金资助项目(81601078,81471306)
河南省高校科技创新团队项目(15IRTSTHN022)
河南省重点科技攻关项目(152102310272)
