Probucol抑制氧化低密度脂蛋白诱导的大鼠主动脉平滑肌细胞增殖

Probucol inhibits proliferation of rat aortic smooth muscle cells stimulated with oxidized low-density lipoprotein

目的:研究Probucol抑制ox-LDL诱导RASMCs增殖与信号蛋白分子ERK1/2、MKP-1、HO-1和Trx-1表达之间的关系。方法:采用MTT、流式细胞术和Western blotting观察ox-LDL刺激条件下probucol对细胞周期、细胞增殖和凋亡、ERK1/2、MKP-1、HO-1和Trx-1表达的影响。结果:(1)Probucol抑制ox-LDL刺激RASMCs增殖:100μmol/Lprobucol+35mg/Lox-LDL组与35mg/Lox-LDL组比较,A值下降了34.9%(P<0.01);(2)Probucol通过使RASMCs停滞在G0/G1期和诱导细胞凋亡2种方式抑制ox-LDL刺激细胞增殖。(3)ox-LDL显著抑制MKP-1的蛋白表达,与对照组比较下降了60.0%(P<0.01),同时使p-ERK1/2表达增加了34.7%;Probucol使MKP-1蛋白表达显著增加2倍,p-ERK1/2表达降低了15.7%(P<0.01);(4)35mg/Lox-LDL使细胞内Trx-1蛋白表达下降28.9%(P<0.05),HO-1蛋白表达轻度增加(P<0.05)。与ox-LDL组比较,probucol使Trx-1蛋白表达增加了91.6%(P<0.01),HO-1表达增加31.9%(P<0.01)。结论:Probucol通过增强MKP-1和HO-1蛋白表达、抑制细胞周期运转和诱导细胞凋亡的机制抑制RASMCs增殖。

AIM： To investigate the relationships between antiproliferative mechanisms of probucol and protein expressions of signaling molecules ERK1/2, MKP-1, HO-1 and Trx-1 in rat aortic smooth muscle cells （RASMCs） stimulated with ox-LDL. METHODS： The effects of probucol on cell cycle, cell proliferation and the expressions of ERK1/2, MKP-1, HO-1 and Trx-1 in the presence of ox-LDL were observed by means of MTT test, FCM and Western blotting. RESULTS： （1） Probucol significantly inhibited the proliferation of RASMCs stimulated with ox-LDL. A value in 100 μmol/L probucol＋35 mg/L ox-LDL group was reduced by 34.9% as compared to ox-LDL group （P〈0.01）. （2） Probucol protected against ox-LDL-induced RASMCs proliferation through inducing cell growth arrest at G0/G1 phase and cell apoptosis. （3） ox-LDL increased the expression of p-ERK1/2 by 34.7% （P〈0.01） and decreased MKP-1 by 60.0% （P〈0.01）, respectively, as compared to control. Probucol attenuated the increase in ox-LDL-stimulated p-ERK1/2 level by 15.7%, but increased MKP-1 expression by 2 times （P〈0.01）. （4）ox-LDL at concentration of 35 mg/L decreased the intracellular Trx-1 expression by 28.9% （P〈0.05）, and slightly increased the level of HO-1 expression as compared to control （P〈0.05）. Probucol enhanced the expression of Trx-1 by 91.6% （P〈0.01） and HO-1 by 31.9% （P〈0.01）, respectively as compared to ox-LDL group. CONCLUSION： Probucol inhibits ox-LDL-stimulated the proliferation of RASMCs through increases in MKP-1/HO-1 expression, suppression of cell cycle progression and induction of cell apoptosis.