辛伐他汀对血管加压素诱导大鼠心肌成纤维细胞增殖的影响及其与小窝蛋白-1表达的关系

Effect of simvastatin on proliferation of adult rat CFs induced by AVP and its relationship with expression of caveolin-1 protein

目的研究辛伐他汀（simvastatin，Sim）对精氨酸血管加压素（arginine vasopressin，AVP，简称血管加压素）诱导成年大鼠心肌成纤维细胞（CFs）增殖的影响及其与小窝蛋白-1（caveolin-1，cavl）的关系。方法离体培养成年大鼠CFs，以四氮唑盐（MTT）比色法检测CFs的增殖，流式细胞分析仪测定其细胞周期，蛋白免疫印迹法检测cavl蛋白的表达。观察carl在AVP诱导大鼠CFs增殖前后及Sire干预后的变化。结果10^-7mol／LAVP干预24h后，MTT比色法检测CFs的吸光值（A）（0．24±0．03）较对照组（0．15±0．02）显著增高（P〈0．01）；给予10^-8 -10^-5mol／L的Sim和AVP共同干预后，CFs的A值呈递减趋势，分别为0．22±0．03、0．21±0．02、0．19±0．02和0．17±0．02，均较AVP组降低，差异具有统计学意义（P〈0．05，P〈0．01）。以10^-7mol／L的AVP干预24h后，CFsS期的百分率（19．52±1．07）和增殖指数（48．25±1．27）较对照组（分别为7．02±0．27和18．93±3．03）均显著增高（均P〈0．01）；10^-7～10^-5mol／LSim与10^-7 mol／LAVP联合干预组CFs S期的百分率和增殖指数均较AVP单独干预组降低，差异具有统计学意义（P〈0．05，P〈0．01）。10^-7mol／L的AVP分别与10^-8 - 10^-5moFL的Sim共同干预后，cavl蛋白的表达呈浓度依赖性地减少。甲羟戊酸（MVA）可逆转Sim对CFs增殖的抑制效应，并拮抗Sim诱导的cavl蛋白表达的降低。结论Sim可抑制AVP诱导的CFs增殖，Sim的干预效应可能受到cavl蛋白表达的调节。

AIM To study the effect of simvastatin （Sim） on the proliferation of rat cardiac fibroblasts （CFs） induced by arginine vasopressin （AVP） and its relationship with expression of caveolin-1 （carl） protein. METHODS CFs were isolated from adult SD rats and cultured. The cellular proliferation and cell cycle were evaluated by MTT colorimetry （A49o value） and flow cytometry （FCM）, respectively. The expression of carl protein was analyzed by immunobloning. RESULTS ①10 -Tmol/L AVP significantly increased A490 value （0.24 ±0.03 ） of CFs in comparison with that in control group （0. 15 ±0.02, P 〈 0.01 ）. 10^-8 - 10^-5 mol/L Sim plus AVP decreased the A49o value of CFs in a concentration-dependent manner （0.22 ±0.03, 0.21 ±0.02, 0. 19 ±0.02 and 0. 17 ±0.02, respectively, P 〈0.05 -0.01 vs control）. ② The percentage of cells in S phase and proliferation index were markedly increased by 10^-7 mol/L AVP compared with those in control （19.52 _± 1.07 and 48.25 ± 1.27 vs 7.02 ±0.27 and 18.93 ±3.03, both P 〈 0.01 ）. In 10^-7 -10^-5 mol/L Sim plus 10^-7 mol/L AVP group, the percentage of cells in S phase and the value proliferation index were significantly lower than those in AVP single-drug group （P 〈0.05 ,P 〈0.01 ）. ③In the presence of 10-7mol/L AVP, 10^-8 -10^-5 mol/L Sim dose-dependently decreased the expression of carl protein. ④The effect of Sim on CFs proliferation and carl protein expression was reversed by the addition of MVA. CONCLUSION The inhibitory effect of Sim on proliferation of adult rat CFs induced by AVP may be associated with cavl expression.