瑞舒伐他汀对血管平滑肌细胞基质金属蛋白酶2表达和细胞迁移的影响

Effects of rosuvastatin on matrix metalloproteinase 2 expression and cell migration of rat vascular smooth muscle cells

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（MMP2）表达以及对血管平滑肌迁移的影响，探讨Hey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24h、48h和72h，在1000μmol／LHey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MMP2的表达及酶的活性。在Transwell小室外室中加入1000μmol／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，分别培养24h、48h和72h，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50～5000μmol／L时促进血管平滑肌细胞MMP2蛋白的表达，Hey浓度在50～1000μmol／L时增强血管平滑肌细胞MMP2的酶活性，浓度在5000μmol／L时抑制MMP2的活性（F值分别为9．31、6．44、5．97，均P〈O．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MMP2表达和酶活性的增强作用（F值分别为3．92、4．78、2．14，均P％0．05）。Hcy在50～5000μmol／L浓度下能刺激血管平滑肌细胞迁移，Hcy浓度为0、50、100、500、1000、5000μmol／L时平滑肌细胞迁移分别为（18．32±2．17）个、（32．68±4．34）个、（44．75±4．08）个、（61．39±5．21）个、（79．74±5．54）个和（90．78±5．83）个（F=5．31，P〈0．05）；而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，在瑞舒伐他汀浓度为10^-9、10、10^-8、10^-7、10^-5mol／L时平滑肌细胞迁移分别为（79．74±5．54）个、（62．53±6．41）个、（48．37±5．66）个、（31。41±4．79）个、（19．27±3．62）个和（11．17±2．33）个（F=4．99，P〈0．05）。结论Hcy能影响血管平滑肌细胞MMP2蛋白的表达，瑞舒伐他汀能抑制Hcy对血管平滑肌细胞MMP2的表达和酶活性的增强作用，并能抑制Hey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机制之一。

Objective To observe the effects of rosuvastatin on the homocysteine （Hcy）- induced expression of matrix metalloproteinase 2 （MMP 2） and cell migration in rat vascular smooth muscle cells （VSMCs）, and to explore the possible mechanism of Hcy-induced atherosclerosis and the role of statins in reversing atherosclerosis. Methods In one cell culture plate, the cultured rat VSMCs were incubated with different concentrations of Hcy （0, 50, 100, 500, 1000μmol/L and 5000 μmol/L） in vitro for 24 h, 48 h and 72 h. And in another cell culture plate, the different concentrations of rosuvastatin （10^-9 , 10^-8, 10^-7, 10^-6 , 10 ^-5mol/L and 0 mol/L） were added to the cultured rat VSMCs （while the concentration of Hcy was 1000 μmol/L）. The MMP 2 expression and enzyme activity were determined by gelatin zymography and Western blotting. The effects of Hcy and rosuvastatin on cell migration and invasiveness of VSMCs were observed. Results Hey （50-5000μmol/L） increased the protein expression, and Hey （50 1000 μmol/I.） increased enzyme activity of MMP 2 significantly. But Hcy （5000 μmol/L） inhibited activity of MMP 2 （F= 9.31, 6.44 and 5.97, all P〈0.05）. Rosuvastatin （10^-9-10^-5 mol/L） inhibited Hcy-induced expression and enzyme activity increasing o{ MMP 2. The counts of cell migration of VSMCs were 18.32±2.17, 32.68± 4.34, 44.75±4.08, 61.39±5.21, 79.74±5.54 and 90.78±5.83, while the concentration of Hcy was 0, 50, 100, 500, 1000 μmol/L and 5000 μmol/I. respectively （F=5.31, P〈0.05）. The counts of cell migration of VSMCs were 79.74±5.54, 62.53±6.41, 48.37±5.66, 31.41±4.79, 19.27±3.62 and 11. 17±2.33, while the concentration of rosuvastatin was 10 ^-9 , 10^-8, 10^-7, 10^-6 and 10^-5 mol/L respectively （F 4.99, P〈0.05）. Rosuvastatin could decrease the stimulation of Hcy-induced migration of VSMCs. Conclusions Hcy can influence the MMP 2 protein expression/activity in VSMCs, and rosuvastatin can inhibit augmentation of Hcy-induced MMP 2 expression/activity and migration of VSMCs. It may be one of the multiple-effects of rosuvastatin reducing atherosclerosis.