摘要
目的探讨NADPH氧化酶在高脂诱导的MIN6胰岛β细胞损伤中的作用。方法不同浓度(0.1、0.3、0.5、0.8mmol·L^(-1))高脂刺激MIN6胰岛β细胞(24、48、72、96 h),MTT法检测细胞增殖情况;免疫印迹法检测细胞内p22^(phox)、p47^(phox)、p67^(phox)以及gp91^(phox)等NADPH氧化酶亚基的表达水平及凋亡相关蛋白(Bcl-2及Bax)的表达水平。结果 0.5mmol·L^(-1)高脂刺激MIN6胰岛细胞48 h时,细胞增殖率出现明显下降;p22^(phox)、p47^(phox)、p67^(phox)以及gp91^(phox)等NADPH氧化酶亚基和Bax的表达水平明显升高(P<0.05),而Bcl-2表达水平明显降低(P<0.05);NADPH氧化酶抑制剂diphenyliodonium(DPI,10μmol·L^(-1))预处理后并高脂刺激MIN6胰岛β细胞48 h,与高脂刺激组相比,DPI处理组细胞Bcl-2表达明显上升(P<0.05),Bax表达明显下调(P<0.05)。结论 NADPH氧化酶活性对高脂引起的MIN6胰岛β细胞损伤的防治有重要意义。
Aim To explore the role of nicotinamide adenine dinucleotide phosphate（ NADPH） oxidase in high fat-induced injury in MIN6 islet β cells. Methods MIN6 islet β cells were exposed to different concentrations of palmitic acid（ 0. 1,0. 3,0. 5,0. 8 mmol ·L^（-1）） for 48 h and different time points of 0. 5 mmol·L^（-1）palmitic acid（ 24,48,72,96 h）. Cell viability was measured by MTT,the protein expression of NADPH oxidase subunits such as p22 ^（phox）,p47^（phox）,p67 ^（phox）and gp91^（phox）and apoptosis related proteins such as Bcl-2and Bax were determined by Western blot. Results MIN6 islet cells exposed to palmitic acid at 0. 5 mmol·L^（-1）for 48 h showed a decrease in their viability and an increase in the expression of NADPH oxidase subunits（ p22^（phox）、p47^（phox）、p67^（phox）and gp91^（phox）） and Bax（ P 0. 05）,while Bcl-2 expression was significantly reduced. the pretreat with NADPH oxidase inhibitor diphenyliodonium（ DPI,10 μmol · L^（-1）） significantly inhibited high fat-induced Bcl-2 and Bax expression（ P 0. 05）. Conclusion Activated NADPH oxidase might play an important role in the treatment of high fat-induced injury in MIN6 islet cells.
作者
李晶
陈丹
周宇
鲍翠玉
LI Jing CHEN Dan ZHOU Yu BAO Cui-yu(Hubei Province Key Laboratory on Cardiovascular, Cerebrovascular, and Metabolic Disorders Nursing College,Hubei University of Science and Technology,Xianning Hubei 437100,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第9期1317-1321,共5页
Chinese Pharmacological Bulletin
基金
湖北省教育厅科学研究计划项目(No B2016197)
湖北省自然科学基金计划项目(No 2015CFC773)
