灯盏花乙素干预缺血再灌注损伤人脐静脉内皮细胞血管内皮生长因子与蛋白激酶Cε的表达

Effects of scutellarin on vascular endothelial growth factor and protein kinase Cε expression in human umbilical vein endothelial cells after ischemia-reperfusion injury

背景:有关灯盏花乙素的研究多集中在神经细胞、神经元及平滑肌细胞上,但对血管内皮细胞的作用研究尚少。目的:观察灯盏花乙素预处理后对人脐静脉内皮细胞缺血再灌注损伤后血管内皮生长因子与蛋白激酶Cε表达的影响。方法:体外培养的人脐静脉内皮细胞,分别设立对照组,模型组和及灯盏花乙素高、中、低剂量组。将人脐静脉内皮细胞予缺氧缺糖3h/复氧糖5或24h;灯盏花乙素高、中、低剂量组分别加1×10-5,1×10-6,1×10-7mol/L灯盏花乙素孵育30min,然后予缺血再灌注处理。实验结束后取细胞裂解液用Westernblot检测血管内皮生长因子、蛋白激酶Cε的蛋白表达。结果与结论:缺血3h再灌注5及24h,较对照组,模型组血管内皮生长因子的表达降低,细胞颗粒部分蛋白激酶Cε的表达明显增加。灯盏花乙素能促进缺血3h再灌注5h人脐静脉内皮细胞血管内皮生长因子的表达,尤其以灯盏花乙素高剂量和中剂量组明显,但对蛋白激酶Cε的表达没有影响。

BACKGROUND：The effects of scutellatin on ischemia-reperfusion injury to endothelial cells in vitro have not been elucidated.OBJECTIVE：To investigate the effect of scutellatin on human umbilical vein endothelial cells （HUVECs） after ischemia-reperfusion and to study whether this effect is mediated by vascular endothelial growth factor （VEGF） and protein kinase Cε （PKCε）.Methods：Cultured HUVECs,exposed to oxygen and glucose deprivation followed by reperfusion,were used as an in vitro model of ischemia-reperfusion.After incubated with scutellarin in a low （1×10^-7 mol/L）,middle （1×10^-6 mol/L） and high （1×10^-5 mol/L） dose for 30 minutes,cells were treated by ischemia 3 hours/reperfusion 5 to 24 hours.VEGF protein expression was evaluated by Western blotting.The translocation of PKCε was assayed by Western blotting.RESULTS AND CONCLUSION：Ischemia 3 hours/reperfusion 5 hours injury significantly decreased the expression of VEGF,and scutellatin in the high and middle dose significantly increased VEGF expression （P〈 0.05）.Under ischemia 3 hours/reperfusion 24 hours condition,VEGF expression was not changed and scutellatin failed to further increase the expression of VEGF.Compared with the control group,there were significant increases of PKCε expression in particulate fractions in ischemia-reperfusion group and scutellatin pretreatment groups （P 〈0.01）.Compared with ischemia-reperfusion group,there was no further increase of PKCε in scutellatin groups.Scutellarin pretreatment on vascular endothelial cells after ischemia-reperfusion injury may have a protective effect,and the mechanism may be related to the early increase of VEGF.