普伐他汀抑制肿瘤坏死因子α介导的人脐动脉血管平滑肌细胞成骨样钙化  

Pravastatin inhibits ossific calcification of human umbilical artery vascular smooth muscle cells induced by tumor necrosis factor α

作　　者：李振勇[1] 倪兆慧[1] 钱家麒[1] 戴慧莉[1] 顾乐怡[1] 郭永平[1] 孙明姝[1] 

机构地区：[1]上海交通大学医学院附属仁济医院肾脏科,200127

出　　处：《中华肾脏病杂志》2008年第12期915-919,共5页Chinese Journal of Nephrology

基　　金：基金项目：国家自然科学基金（30370653）;上海市教育委员会科研项目（04BC35）

摘　　要：目的研究普伐他汀干预对肿瘤坏死因子α（TNF-α）介导的人脐动脉血管平滑肌细胞（hUASMC）成骨样分化标志蛋白骨特异性碱性磷酸酶（BAP）、骨桥蛋白（OPN）和骨形成蛋白2（BMP-2）表达以及细胞外钙盐沉积的影响，并探讨普伐他汀在血管钙化防治中的潜在作用。方法植块贴壁法原代培养人hUASMC。予以TNF-α刺激和普伐他汀干预，甲O-酚酞络合酮方法测定细胞外基质钙盐沉积；实时定量PCR法观察血管平滑肌细胞BAP和OPNmRNA表达水平；免疫印迹法观察血管平滑肌细胞BAP、OPN和BMP-2蛋白表达水平。结果-定浓度范围的普伐他汀呈浓度依赖方式抑制TNF—α对hUASMC的促增殖作用（r=-0．946，P〈0．01）；一定浓度范围的普伐他汀呈浓度依赖方式下调TNF-仅介导上调的BAP、OPNmRNA表达（r=-0．972，P〈0．01）与蛋白的表达水平（BAP蛋白，r=-0．820，P〈0．01；OPN蛋白，r=-0．972，P〈0．01；BMP-2蛋白，r=-0．928，P〈0．01），抑制血管平滑肌细胞成骨样分化，减少细胞外基质钙盐的沉积（r=-0．973，P〈0．01）。结论普伐他汀可抑制TNF—α对hUASMC的促增殖作用，抑制细胞成骨样分化，减少细胞外基质钙盐的沉积。Objective To investigate the effects of pravastation intervention on tumor necrosis factor （TNF）-α-induced ossific calcification in human umbilical artery smooth muscle cells （hUASMCs）. Methods hUASMCs were cultured by tissue explant in vitro, hUASMC were treated with TNF-α 50μg/L and pravastatin of three different concentrations. The calcium deposition was determined by O-cresolphthalein complexone method. The mRNA expression of BAP and OPN was determined by real time-PCR. The protein expression of BAP, OPN and BMP-2 was determined by Western blotting. Results Pravastatin inhibited the proliferation of hUASMC （r=-0.946, P〈0.01） and decreased the cell calcium deposition （r=-0.973, P〈O.01） in a dose- dependent manner. Pravastatin down-regulated the expression of BAP, OPN and BMP-2 induced by TNF-αin a dose-dependent manner （mRNA, r=-0.972, P〈0.01; BAP protein, r=-0.820, P〈0.01; OPN protein, r=-0.972, P〈0.01; BMP-2 protein, r=-0.928, P〈0.01）. Conclusion Pravastatin can inhibit the proliferation of hUASMC, decrease the cell calcium deposition and inhibit the ossific calcification of hUASMC induced by TNF-α.

关 键 词：肿瘤坏死因子α 普伐他汀 钙质沉着症 碱性磷酸酶 人脐 动脉血管平滑肌细胞 成骨样钙化 

分 类 号：R96[医药卫生—药理学]