普伐他汀对肿瘤坏死因子-α诱导的大鼠血管平滑肌细胞syndecan-4蛋白表达的影响

Pravastatin inhibits the expression of syndecan-4 protein in tumor necrosis factor-α-induced rat vascular smooth muscle cells in vitro

目的观察普伐他汀对肿瘤坏死因子-α(TNF-α)诱导的离体大鼠血管平滑肌细胞(VSMCs)增殖和syndecan-4蛋白表达的影响。方法体外培养大鼠胸主动脉VSMCs,分别应用终浓度为20ng/mlTNF-α、10μmol/ml普伐他汀、20μmol/ml普伐他汀、20ng/mlTNF-α+10μmol/ml普伐他汀、20ng/mlTNF-α+20μmol/ml普伐他汀分别作用24h,并设立相关对照组进行比较。采用MTS/PMS法确定VSMCs的增殖状态,利用Westernblot蛋白免疫印迹法测定VSMCs中syndecan-4蛋白的表达。结果 (1)与对照组比较,TNF-α组能显著刺激大鼠VSMCs的增殖(P<0.05),而普伐他汀不同剂量组单独应用对大鼠VSMCs的增殖均无明显作用(P>0.05)。TNF-α联合普伐他汀共同作用组与TNF-α组比较,VSMCs增殖均有显著减少(P<0.05)。(2)Westernblot蛋白免疫印迹法测定显示:与对照组比较,TNF-α组能显著刺激VSMCs的syndecan-4蛋白表达(P<0.05),而普伐他汀不同剂量组单独应用对VSMCs的syndecan-4蛋白表达均无明显影响(P>0.05)。TNF-α联合普伐他汀共同作用组与TNF-α组比较,VSMCs的syndecan-4蛋白表达显著降低(P<0.05)。结论普伐他汀可明显抑制TNF-α诱导的VSMCs增殖及细胞syndecan-4蛋白的表达。

Objective To investigate the effect of pravastatin on the proliferation of rat vascular smooth muscle cells （VSMCs） and expression of syndecan-4 protein induced by tumor necrosis factor-α （TNF-α）. Methods VSMCs cultured in vitro were exposed to 20 ng/ml TNF-α, 10 μmol/ml pravastatin, 20 μmol/ml pravastatin, 10 μmol/ml pravastatin with 20 ng/ml TNF-α, or 20 μmol/ml pravastatin with 20 ng/ml TNF-α for 24 h. The proliferation of the VSMCs was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was detected by Western blotting using anti-syndecan-4 antibody. Results Compared to the control group, TNF-α at 20 ng/ml significantly stimulated the proliferation of rat VSMCs （P0.05）. Pravastatin alone produced no obvious effect on VSMCs growth （P0.05）, but significantly inhibited TNF-α-induced VSMC proliferation （P0.05）. The expression of syndecan-4 protein in the VSMCs was significantly enhanced by 20 ng/ml TNF-α （P0.01）. Pravastatin alone did not affect the expression of syndecan-4 protein （P0.05）, but significantly inhibited TNF-α-induced enhancement of syndecan-4 protein expression （P0.01）. Conclusion Pravastatin can inhibit the proliferation and syndean-4 protein expression in rat VSMCs induced by TNF-α in vitro.