摘要
目的探讨丙酮醛(MGO)介导人脑微血管内皮细胞系hCMEC/D3的损伤作用及机制。方法不同浓度MGO(0.25、0.50、0.75、1.00和1.25 mmol/L)作用hCMEC/D3细胞24 h,用CCK-8法检测hCMEC/D3细胞活力;用试剂盒检测胞内乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)活性。线粒体膜电位检测试剂盒(JC-1)荧光探针检测线粒体膜电位水平,MitoSOX荧光探针观察线粒体活性氧(mROS)产生。结果MGO组与对照组相比,hCMEC/D3细胞活力呈浓度依赖性下降(P<0.01),细胞内LDH活性显著升高(P<0.01),同时SOD活性降低(P<0.01),差异有统计学意义。与对照组相比,MGO处理12 h后的hCMEC/D3细胞经JC-1染色红绿荧光比值降低,提示MGO诱导细胞线粒体膜电位下降。MitoSOX染色后,与对照组相比,MGO处理过的hCMEC/D3细胞的红色荧光表达数量增加,提示细胞内mROS过量生成。结论MGO降低hCMEC/D3细胞活力及线粒体膜电位,并诱导胞内mROS过量生成。
Objective To explore the effects and mechanism of methylglyoxal(MGO)in human brain microvascular endothelial cell line(hCMEC/D3).Methods The hCMEC/D3 cells were incubated with MGO(0.25,0.50,0.75,1.00 and 1.25 mmol/L)for 24 h.Cell viability was detected by CCK-8 assay.Cellular LDH and SOD activities were determined by commercially available kits.The change of mitochondrion membrane potential was assessed by JC-1 probe.MitoSOX probe was used to observe mitochondrial reactive oxygen species(mROS)release.Results Compared with control,cell viability of hCMEC/D3 treated with MGO decreased in a concentration-dependent manner(P<0.01).Increased LDH activity and decreased SOD activity were observed with MGO exposure(P<0.01).The decrease of mitochondrion membrane potential and the excessive increase of mROS were induced after MGO exposure for 12 h,which were detected by JC-1 or MitoSOX probe.Conclusions MGO reduces cell viability and mitochondria membrane potential in hCMEC/D3 cells with excessive mROS generation.
作者
李婷婷
韩臻平
高立
王思懿
钟恺
黄萍
严洁萍
LI Tingting;HAN Zhenping;GAO Li;WANG Siyi;ZHONG Kai;HUANG Ping;YAN Jieping(Center for Clinical Pharmacy,Department of Pharmacy,Zhejiang Provincial People′s Hospital,People′s Hospital Affiliated to Hangzhou Medical College,Hangzhou 310014;Department of Pharmacology,College of Pharmaceutical Sciences,Zhejiang University of Technology,Hangzhou 310014;Key Laboratory of Endocrine Gland Diseases of Zhejiang Province,Hangzhou 310014,China)
出处
《基础医学与临床》
2023年第11期1631-1635,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81903597)
浙江省自然科学基金(LQ16H310003)
浙江省医药卫生科技项目(2021KY016,2022KY061)
浙江省中医药科技计划(2023ZL015)。
关键词
糖尿病脑血管并发症
丙酮醛
人脑微血管内皮细胞系
膜电位
线粒体活性氧
diabetic cerebral vascular complication
methylglyoxal
human brain microvascular endothelial cells line
membrane potential
mitochondrial reactive oxygen species