山西医科大学学报2017，Vol.48Issue(10)：975-981,7.DOI:10.13753/j.issn.1007-6611.2017.10.001
补骨脂酚经蛋白激酶Cε拮抗内皮细胞缺血再灌注损伤
Bakuchiol protects vascular endothelial cells against ischemia reperfusion injury via PKCε signaling pathway
摘要
Abstract
Objective To explore the protective effects of bakuchiol(BAK) against simulated ischemia reperfusion(SIR)-induced injury in human umbilical vein endothelial cells(HUVECs) and its underlying mechanisms.Methods The cultured HUVECs were pretreated with BAK(2,5,10 μmol/L) for 24 h and then subjected to SIR (45 min ischemia,4 h reperfusion).Cell viability and apoptotic index were detected by CCK-8 and TUNEL methods,and the lactate dehydrogenase (LDH) releasing,intracellular malondialdehyde(MDA) content,superoxide dismutase(SOD) were all assessed by specialized kits.The translocation of protein kinase C epsilon (PKCε) and the expression levels of Bcl-2,Bax,cleaved caspase 3 and oxidative stress marker gp91phox were detected by Western blot.Dihydroethidium (DHE),a fluorescence probe,was used to detect the presence of reactive oxygen species (ROS).The εV1-2,a specific inhibitor of PKCε signaling,was used to further study the role of PKCε in this process.Results BAK pretreatment protected HUVECs against SIR injury in a dose-dependent manner,especially at the concentration of 10 μmol/L.BAK pretreatment significantly improved the cell viability,reduced LDH releasing,up-regulated the SOD activity and down-regulated the MDA content of SIR-treated HUVECs (P ＜ 0.05).In addition,BAK supplementation significantly reduced the apoptotic index,increased PKCε translocation and Bcl-2 expression,and decreased the expression levels of cleaved caspase 3,Bax and gp91phox(P ＜ 0.05).However,the protective effects of BAK mentioned above were significantly attenuated by εV1-2 (P ＜ 0.05).Conclusion BAK pretreatment can protect against SIR-induced oxidative stress and apoptosis on HUVECs via PKCε signaling pathway.关键词
补骨脂酚/人脐静脉内皮细胞/缺血再灌注损伤/蛋白激酶C epsilon/氧化应激/凋亡Key words
bakuchiol/human umbilical vein endothelial cells/ischemia reperfusion injury/protein kinase C epsilon/oxidative stress/apoptosis分类
医药卫生引用本文复制引用
王垒,杜朝升,肖军,杨健..补骨脂酚经蛋白激酶Cε拮抗内皮细胞缺血再灌注损伤[J].山西医科大学学报,2017,48(10):975-981,7.基金项目
北京市自然科学基金资助项目(7122178) （7122178）
总参军事医学和老年病科研基金资助项目(ZCWS14C12) （ZCWS14C12）
