PPARβ调控硝化应激参与高糖对内皮细胞的损伤  

PPARβ involved in endothelial cell injury induced by high glucose via nitrative stress

作　　者：阳创 熊宝萍 郑雲丹 李斯曼 周上钧 蒋青松[1] YANG Chuang;XIONG Bao-ping;ZHENG Yun-dan;LI Si-man;ZHOU Shang-jun;JIANG Qing-song(Dept of Pharmacology,Chongqing Key Lab of Biochemistry and Molecular Pharmacology,Chongqing Key Lab of Pharmacometabolism,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学药理学教研室,重庆市生物化学与分子药理学重点实验室,重庆药物代谢学重点实验室,重庆400016

出　　处：《中国药理学通报》2022年第6期848-853,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81100905)。

摘　　要：目的研究PPARβ与硝化应激在高糖损伤人脐静脉内皮细胞(HUVEC)中的作用。方法采用CCK-8检测细胞活力;EdU法检测细胞增殖能力;Western blot检测PPARβ、eNOS、iNOS、3-nitrotyrosine的蛋白表达水平;荧光试剂盒和Griess Reagent法分别检测过氧亚硝酸盐(OONO^(-))水平和一氧化氮(NO)的含量。结果随葡萄糖浓度增加,高糖(30、40、50 mmol·L^(-1))剂量依赖地降低HUVEC活力;30 mmol·L^(-1)葡萄糖使HUVEC细胞增殖能力明显降低,PPARβ、eNOS蛋白表达水平和NO含量下降,iNOS、3-nitrotyrosine蛋白表达以及OONO^(-)水平增加。PPARβ激动剂GW0742(1μmol·L^(-1))干预可增加HUVEC增殖,上调PPARβ、eNOS蛋白表达水平,降低iNOS、3-nitrotyrosine蛋白表达水平,减少OONO^(-),同时增加NO含量;PPARβ拮抗剂GSK0660(1μmol·L^(-1))以及NOS抑制剂L-NAME(10μmol·L^(-1))均可阻断GW0742(1μmol·L^(-1))的上述作用。结论PPARβ下调参与了高糖引起的血管内皮细胞的损伤,该作用可能是通过负性调控硝化应激途径实现的。Aim To investigate the role of PPARβand nitrative stress in human umbilical vein endothelial cells(HUVECs)injury induced by high glucose.Methods The cell viability was detected by CCK-8.The cell proliferation was detected by EdU proliferation detection kit.The protein expression level of PPARβ,eNOS,iNOS,and 3-nitrotyrosine was detected by Western blot.The content of peroxynitrite and nitric oxide(NO)was determined by peroxynitrite kit and Griess Reagent,respectively.Results Glucose(30,40,50 mmol·L^(-1))significantly reduced the cell viability of HUVECs in a dose-dependent manner.Glucose at 30 mmol·L^(-1)(high glucose,HG)significantly reduced the proliferation of HUVECs,down-regulated the expression of PPARβ,eNOS at protein level and NO content,and increased iNOS,3-nitrotyrosine protein expression and peroxynitrite level.The above effects of HG were reversed by PPARβagonist GW0742(1μmol·L^(-1)).Both PPARβantagonist GSK0660(1μmol·L^(-1))and NOS inhibitor L-NAME(10μmol·L^(-1))blocked the protective effects of GW0742.Conclusion The down-regulation of PPARβis involved in the injury of HUVECs induced by high glucose,which may be mediated,at least partly,by the stimulation of nitrative stress.

关 键 词：PPARβ 硝化应激 高糖 内皮细胞 损伤 3-硝基酪氨酸 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]