摘要
目的观察苦碟子注射液对血管内皮细胞缺氧损伤后的保护作用,并初步研究PKCδ/MARCKS信号通路在血管内皮细胞缺氧损伤后的作用机制。方法建立CoCl_2诱导的人脐静脉血管内皮细胞缺氧损伤模型,分为正常组、模型组、苦碟子注射液组、PKCδ特异性抑制剂Rottlerin组。光镜下观察缺氧损伤24 h后各组细胞形态学改变,MTT比色法检测各组细胞的活力,免疫细胞化学法(IHC)及蛋白印迹法(Western Blot)检测各组细胞p-PKCδ、p-MARCKS蛋白质的分布与表达。结果苦碟子注射液、Rottlerin可显著减轻细胞缺氧损伤形态改变,苦碟子、Rottlerin可明显增强缺氧损伤后的细胞活力(P<0.01)。与正常组相比,模型组p-PKCδ、p-MARCKS蛋白表达显著增加(P<0.05),苦碟子、Rottlerin显著抑制p-PKCδ、p-MARCKS蛋白的表达(P<0.05)。结论苦碟子注射液对缺氧诱导的血管内皮细胞损伤具有保护作用,其作用机制与下调PKCδ/MARCKS信号转导通路的活性有关。
Objective To observe the protective effect of Kudiezi Injection（ KI） on vascular endothelial cells,and study initially the effective mechanism of PKCδ/MARCKS signal pathway after hypoxia injury.Methods The hypoxia injury model was induced by CoCl_2 in human umbilical vein endothelial cells（ HUVEC）,and then divided into normal group,model group,KI group and Rottlerin group. The cytomorphologic changes were observed by using light microscope 24 h after hypoxia injury,vitality of HUVEC was detected by using methyl thiazolyl tetrazolium test（ MTT）,and distributions and expressions of p-PKCδ and p-MARCKS were detected by using immunocytochemical method and Western Blot in all groups. Results KI and Rottlerin alleviated significantly the cytomorphologic changes,and enhanced significantly the vitality of cells after hypoxia injury（ P 〈 0. 01）. The expressions of p-PKCδ and pMARCKS were significantly increased in model group compared with normal group（ P 〈 0. 05）,and were significantly inhibited by KI and Rottlerin（ P 〈 0. 05）. Conclusion KI has a protective effect on vascular endothelial cells after hypoxia injury,and the mechanism is related to down-regulation of vitality of PKCδ/MARCKS signal pathway.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2017年第3期214-218,共5页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家国际科技合作专项项目(No.2015DFA31130)
