同型半胱氨酸对大鼠血管平滑肌细胞MMP-2表达、细胞迁移的影响及瑞舒伐他汀的逆转作用

摘要

目的 观察瑞舒伐他汀对同型半胱氨酸(Hcy)诱导大鼠血管平滑肌细胞基质金属蛋白酶-2(MMP-2)的表达以及对血管平滑肌细胞迁移的影响,探讨Hcy致动脉粥样硬化以及他汀药物逆转的可能机制.方法 体外培养大鼠主动脉平滑肌细胞,加入不同浓度的Hcy分别作用 24、48和72h;在1 000μmol/L Hcy浓度下分别加入不同浓度的瑞舒伐他汀干预,采用免疫印迹法和明胶酶谱法检测细胞培养上清液中MMP-2的表达及酶的活性.在Transwell小室外室中加入1 000μmol/L浓度的Hcy,内室加入定量的大鼠平滑肌细胞悬液,培养48h,观察Hcy对平滑肌细胞迁移侵袭力的影响;同时加入不同浓度瑞舒伐他汀溶液,48h,观察瑞舒伐他汀对Hcy诱导平滑肌细胞迁移侵袭力的影响.结果 Hcy浓度在50～5 000μmol/L时促进血管平滑肌细胞MMP-2蛋白的表达(P＜0.05);Hcy浓度在50～1 000μmol/L时增强血管平滑肌细胞MMP-2的酶活性(P＜0.05),浓度在5 000μmol/L时抑制血管平滑肌细胞MMP-2的活性(P＜0.05).瑞舒伐他汀浓度在10-9～10-5mol/L时,能抑制Hcy对血管平滑肌MMP-2表达和酶活性的增强作用(P＜0.05).Hcy在50～1 000μmol/L浓度下能刺激血管平滑肌细胞迁移(P＜0.05);瑞舒伐他汀可以抑制Hcy对血管平滑肌细胞迁移的刺激作用(P＜0.05).结论 Hcy能影响血管平滑肌细胞MMP-2蛋白的表达,瑞舒伐他汀能抑制Hcy对血管平滑肌细胞MMP-2的表达和酶的活性增强作用,并能抑制Hcy对血管平滑肌细胞迁移的刺激作用,这可能是瑞舒伐他汀抗动脉粥样硬化的作用之一.%Objective To investigate the effects of homocysteine (Hcy)- induced matrix metalloproteinase- 2 (MMP- 2) and cell migration in rat vascular smooth muscle cells (VSMCs) and the reverse of rosuvastatin.Methods Cultured rat VSMCs were incubated with different concentration of Hcy, and different concentration of rosuvastatin with Hcy at 1000μmol/L in vitro for 24, 48 and 72h.The protein expression and enzyme activity of MMP- 2 were determined by using the methods of Western blotting and gelatin zymography.Cultured rat VSMCs were also incubated with the same concentrations of Hcy and rosuvastatin in transwell for 48h.The numbers of VSMCs which transited the membrane were counted.Results Hcy (50～1000μmol/L) significantly increased the expression and activity of MMP- 2 in a time- effect manner.Hcy reduced the expression of MMP- 2 at the concentration of 5 000μmol/L.Rosuvastatin inhibited homocysteine- induced expression and activity of MMP- 2.Hcy (50～5 000μmol/L) stimulated the migration of VSMCs, and rosuvastatin inhibited homocysteine- induced cell migration.Conclusion The data suggest that Hcy increases the MMP- 2 expression/activity and the migration of VSMCs, which may be related to its role in pathogenesis of atherosclerosis.Rosuvastatin can inhibit homocysteine- induced MMP- 2 expression/activity and cell migration of VSMCs, which might be associated with its benefits in treatment of chronic heart disease.