小鼠主动脉平滑肌细胞及肌源性泡沫细胞与格列本脲共孵育的反应

Glibenclamide co-cultured with smooth muscle cells and vascular smooth muscle cells-derived foam cells

目的:观察磺酰脲类药物格列本脲对平滑肌细胞及肌源性泡沫细胞内酰基辅酶A:胆固醇酰基转移酶1(ACAT1)酶活性的影响。方法:实验于2003-06/2005-07在华中科技大学同济医学院实验中心完成。①用组织贴块法培养小鼠主动脉平滑肌细胞,采用两步超速离心法制备低密度脂蛋白后,运用CuCl2氧化法制备氧化低密度脂蛋白,采用100mg/L的氧化低密度脂蛋白作用于平滑肌细胞72h,建立泡沫细胞模型。②在平滑肌细胞和泡沫细胞分别加入格列本脲,使终浓度为200μmol/L,孵育24h,用放射性同位素标记法检测ACAT1酶活性。结果:①平滑肌细胞转变为泡沫细胞后,ACAT1酶活性上升(P<0.05)。②平滑肌细胞被格列本脲干预后,ACAT1酶活性无明显变化(P>0.05);肌源性泡沫细胞加入格列本脲后,ACAT1酶活性下降(P<0.05)结论:格列本脲能抑制肌源性泡沫细胞内ACAT1的酶活性,抑制平滑肌细胞向泡沫细胞的转化,有可能成为防止动脉粥样硬化的有效药物。

AIM： To investigate the effect of Glibenclamide （sulfonylurea medicine） on activity of acyl-CoA： cholesterol acyltransferase enzyme-1 （ACAT1） in foam cells （FC） derived from vascular smooth muscle cells （VSMC）. METHODS： The experiment was performed at the Laboratory Center of Tongji Medical College, Huazhong University of Science and Technology from June 2003 to July 2005. （1）Mice aortic vascular smooth muscle cells （VSMCs） was cultured by tissue culture method, Low density lipoprotein was prepared using two-step ultracentrifugation method, and oxidized by CuCl2. VSMCs were stimulated with 100 mg/L oxidized low-density lipoprotein for 72 hours to form FC model. （2）Both VSMCs and FCs were incubated with Glibenclamide （200 μmol/L） for 24 hours, and ACAT1 activity was detected using radioisotope marking. RESULTS： While VSMCs transformed into FCs, the activity of ACAT1 increased （P 〈 0.05）. Glibenclamide did not significantly alter the activity of ACAT1 in VSMCs （P 〉 0.05）, but remarkably inhibited the activity of ACAT1 in FCs （P 〈 0.05）. CONCLUSION： Glibenclamide can inhibit the activity of ACAT1 in VSMC-derived FCs, prevent the formation of FCs derived from VSMCs. Glibenclamide can be used as an inhibitor of atherosclerosis.