维生素D3通过VDR-p38途径抑制内皮细胞VCAM-1表达的研究  

Study on vitamin D3 inhibiting expression of VCAM-1 in endothelial cells through VDR-p38 pathway

作　　者：索荣 刘行 赵辉[1] SUO Rong;LIU Xing;ZHAO Hui(Department of Cardiology,Tianjin Hospital,Tianjin 300211,China;Department of Cardiology,Tianjin Hospital,Tianjin Medical University/Tianjin Institute of Cardiology,Tianjin 300211,China)

机构地区：[1]天津医院心内科,300211 [2]天津医科大学第二医院心脏科/天津心脏病学研究所,300211

出　　处：《重庆医学》2020年第24期4045-4049,4054,共6页Chongqing medicine

基　　金：天津市教委科研计划项目(2017KJ205)。

摘　　要：目的细胞水平探究维生素D(VD)对血管内皮细胞的保护作用。、方法采用肿瘤坏死因子α(TNF-α)诱导内皮损伤和黏附分子表达;维生素D采用最具活性形式的骨化三醇;利用流式细胞术检测血管细胞黏附分子-1(VCAM-1)表达水平;荧光显微镜观察绿色荧光加载的THP-1单核细胞与内皮细胞的黏附作用;Western blot检测VD受体(VDR)、p38、磷酸化p38蛋白水平。结果与对照组比较,TNF-α组内皮细胞VCAM-1表达增高(P<0.05)、内皮细胞与单核细胞黏附作用增加(P<0.05)。当内皮细胞预孵育10-9、10-8、10-7、10-6mol/L骨化三醇后,TNF-α诱导的内皮细胞VCAM-1的表达降低,与单核细胞之间的黏附作用减弱(P<0.05)。特异性siRNA敲低VDR蛋白水平后,骨化三醇对内皮细胞VCAM-1表达的抑制作用减弱,内皮细胞与单核细胞的黏附作用减弱(P<0.05)。TNF-α促进内皮细胞p38蛋白磷酸化、而骨化三醇抑制TNF-α诱导的内皮细胞p38蛋白磷酸化(P<0.05)。特异性siRNA敲低VDR蛋白水平后,骨化三醇对TNF-α诱导的内皮细胞p38蛋白磷酸化及VCAM-1的表达和单核细胞黏附抑制作用均降低(P<0.05)。结论维生素D3通过VDR-p38信号途径抑制TNF-α诱导的血管内皮细胞VCAM-1蛋白表达,以及抑制内皮细胞与单核细胞黏附作用。Objective To study the protective effect of vitamin D(VD)on vascular endothelial cells in cellular level.Methods The tumor necrosis factor-α(TNF-α)was used to induce the endothelial injury and the expression of adhesion molecules.Vitamin D adopted the most active form of calcitriol.The flow cytometry was used to detect the expression level of VCAM-1;the fluorescence microscopy was used to observe the adhesion effect of THP-1 monocytes loaded with green fluorescence and endothelial cells;the Western blot was used to detect the levels of vitamin D receptor(VDR),p38,phosphorylated p38 protein.Results Compared with the control group,the endothelial cellular VCAM-1 expression level in the TNF-αgroup was increased(P<0.05)and the adhesive effect of endothenial cells to monocytes was increased(P<0.05).When endothelial cells preincubating 10-9,10-8,10-7,10-6 mol/L calcitriol,the expression level of endothelial cellular VCAM-1 induced by TNF-αwas decreased,and its adhesive effect to monocytes was weakened(P<0.05).After specific siRNA knocking down the VDR protein level,the inhibiting effect of calcitriol on endothelial cellular VCAM-1 expression was weakened and the adhesive effect of endothelial cells to monocytes was weakened(P<0.05).TNF-αpromoted the phosphorylation of p38 protein in endothelial cells,while calcitriol inhibited the phosphorylation of endothelial cellular p38 protein induced by TNF-α(P<0.05).After specific siRNA knocking down VDR protein level,the effect of calcitriol on p38 protein phosphorylation induced by TNF-αand VCAM-1 expression as well as monocytes adhesive inhibiting effect were decreased(P<0.05).Conclusion Calcitriol inhibits TNF-αinduced endothelial cellular VCAM-1 protein expression and inhibits the adhesive effect of endothelial cells to mononcytes through VDR-p38 signaling pathway.

关 键 词：维生素D 骨化三醇 动脉粥样硬化 内皮细胞 血管细胞黏附分子-1 

分 类 号：R541.4[医药卫生—心血管疾病]