高甘油三酯血清对牛主动脉内皮细胞一氧化氮合酶基因表达的影响及其机制的研究  

Mechanisms and Effects of Hypertriglyceride Serum on the Expression of Endothelial Nitric Oxide Synthase Gene in Cultured Bovine Endothelial Cells

作　　者：王燕[1] 汪道文[2] 王旭[1] 邵一兵[1] 王正忠[1] 张纯全[1] 

机构地区：[1]青岛市市立医院心血管内科,山东青岛266071 [2]华中科技大学附属同济医院心血管内科,湖北武汉430030

出　　处：《临床医学工程》2009年第12期23-27,共5页Clinical Medicine & Engineering

摘　　要：目的研究高甘油三酯血清(HTS)对原代牛主动脉内皮细胞(BVEC)一氧化氮合酶(eNOS)基因表达的影响及其可能机制。方法采用Western印迹法从蛋白质水平检测高甘油三酯血清对牛主动脉内皮细胞eNOS和一氧化氮(NO)活性的影响。在证实其效应的基础上,采用Western印迹法检测经HTS作用后,PKC-MAPK-MAPKK信号转导通路的改变。进一步从Ⅳ型高脂蛋白血症血浆提取极低密度脂蛋白(VLDL)(HTG-VLDL)并采用Western印迹法研究HTG-VLDL对eNOS的蛋白质表达的影响。结果HTS以浓度依赖的方式抑制eNOS的蛋白质表达,减少一氧化氮的产生,并以剂量和时间依赖的方式促进MAPK的磷酸化,而经PKC、MAPKK和MAPK抑制剂预处理后,HTS对eNOS的蛋白质表达的抑制作用更明显;另外发现HTG-VLDL可抑制eNOS的蛋白质表达水平。结论HTS抑制eNOS的蛋白质表达水平,可能部分通过HTG-VLDL对eNOS的影响,但未经PKC-MAPKK-MAPK通路,而且PKC-MAPKK-MAPK可能参与了eNOS正常的蛋白质表达,HTS增加MAPK的磷酸化水平有可能是一种细胞自身保护机制。Objective To study the mechanisms and effects of the hypertriglyceride serum （HTS） on the endothelial nitric oxide synthase （eNOS） expression and nitric oxide （NO） production in cultured primary bovine endothelial cells from HTS. Methods Firstly the effects of HTS on protein expression level of eNOS and NO production were detected by Western blot and quantitative analysis. Secondly based on the above results, signal transduction pathways including PKC-MAPK-MAPKK on which HTS may have effects in cultured BAEC. Finally VLDL were isolated from the type of IV type Hyperlipidemia, then detect the effect of HTG-VLDL on eNOS by Western blot. Result HTS down-regulated significantly eNOS at protein levels and decrease NO production in a concentration-dependment fashion. HTS could induce phosphorlylation of MAPK in time and dose dependment fashion. Then the cells, which were incubated with HTS, were pretreated with inhibitors ofPKC, MAPK, MAPKK （MAPK kinase） （H-7, Apigenin, PD98059）, respectively, and the results showed that the PKC, MAPKK and MAPK inhibitors make the inhibition effect of HTS on eNOS protein expression more significant. In addition, HTG-VLDL could inhibit the eNOS protein expression. Conclusion HTS down-regulating eNOS protein expression may be through path- way dependent of HTG-VLDL, but not pathway mediated by PKC-MAPKK-MAPK which may be participate in the essential protein expression of eNOS. HTS inducing phosphorlylation of MAPK may be the self-protect mechanism.

关 键 词：高甘油三酯血清 内皮细胞 一氧化氮合酶 VLDL 

分 类 号：R543.12[医药卫生—心血管疾病]