虫草素对肿瘤坏死因子-α诱导的大鼠血管平滑肌细胞增殖及syndecan-4蛋白表达的影响

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

目的观察虫草素对肿瘤坏死因子-α(TNF-α)刺激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20 ng/mL TNF-α、20μmol/mL虫草素、40μmol/mL虫草素、20 ng/mL TNF-α+20μmol/mL虫草素、20 ng/mL TNF-α+40μmol/mL虫草素作用24 h,并设对照组进行比较。采用MTS/PMS法确定VSMCs的增殖状态,利用Western blot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),而虫草素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联合虫草素共同作用组与TNF-α组比较,VSMCs增殖均显著减少(P<0.05)。(2)Western blot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),而虫草素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联合虫草素共同作用组与TNF-α组比较,VSMCs的syndecan-4蛋白表达显著降低(P<0.05)。结论虫草素可明显抑制TNF-α诱导的VSMCs增殖及细胞syndecan-4蛋白的表达。

Objective To investigate the effects of cordycepin 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 were cultured in vitro and exposed to 20 ng/mL TNF-α,20 μmol/mL cordycepin,40 μmol/mL cordycepin,20 μmol/mL cordycepin with 20 ng/mL TNF-α,and 40 μmol/mL cordycepin with 20 ng/mL TNF-α,respectively.All groups were treated for 24 h in vitro,in addition to the control group established for comparison.The ratio of proliferation of VSMCs was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was evaluated by Western blotting using anti-syndecan-4 antibody.Results（1） Compared to the control group,TNF-α significantly stimulated the proliferation of rat VSMCs at the concentration of 20 ng/mL in TNF-α group（P〈0.05）.Cordycepin alone had no effect on VSMCs growth（P〉0.05）,but significantly inhibited VSMCs proliferation induced by TNF-α（P〈0.05）.（2） Compared to the control group,the expression of syndecan-4 protein in VSMCs was significantly enhanced by TNF-α at the concentration of 20 ng/ml（P〈0.01）.Cordycepin alone had no effect on the expression of syndecan-4 protein（P〉0.05）,but significantly inhibited the expression induced by TNF-α（P〈0.01）.Conclusion Cordycepin could inhibit cell proliferation and expression of syndean-4 protein in rat VSMCs induced by TNF-α in vitro.