小窝蛋白-1反义寡核苷酸在AVP诱导大鼠心肌成纤维细胞增殖erk1/2信号转导中的作用及辛伐他汀的干预效应

Role of caveolin-1 in adult rat CFs proliferation induced by AVP and the intervention of Simvastatin

目的研究小窝蛋白-1反义寡核苷酸(cav1-AS ODN)在血管加压素(AVP)诱导的成年大鼠心肌成纤维细胞(CFs)增殖erk1/2信号转导中的作用及辛伐他汀(Sim)对cav1表达的干预效应。方法采用脂质体导入法将cav1-AS ODN导入离体培养的成年大鼠CFs,用3H-TdR掺入法定量观察CFs增殖,蛋白免疫印迹法分析cav1-AS ODN对AVP诱导大鼠CFs增殖后磷酸化erk1/2(p-erk1/2)、p21和细胞周期蛋白A(cyclin A)表达变化及Sim对cav1表达的影响。结果 cav1-AS ODN与10-7 mol/L的AVP共同干预组,大鼠CFs内3H-TdR掺入率和p-erk1/2蛋白表达量分别相当于空白对照组的(212±6)%和(7.9±0.3)%,10-7mol/L的AVP单独干预组的3H-TdR掺入率和p-erk1/2表达量分别相当于空白对照组的(172±4)%和(5.7±0.2)%,两组比较差异非常显著(P<0.01)。cav1-AS ODN单独干预或与10-7mol/L的AVP共同干预大鼠CFs 24 h后,两组CFs内p21表达丰度均较空白对照组下降,cyclin A表达丰度升高。β-环糊精、黄体酮或Sim可减少CFs胞膜上cav1蛋白表达。用10、15或20μg/ml胆固醇分别与10-7 mol/L Sim共同干预CFs 24 h后,CFs胞膜上cav1蛋白表达量分别相当于对照组的(86±3)%(、91±4)%和(94±5)%,与10-7mol/L Sim单独作用CFs组(66±4)%比较,均有显著的统计学差异(P<0.05,P<0.01)。结论 cav1-AS ODN可增强AVP促CFs增殖的作用,Sim降胆固醇作用影响CFs胞膜上cav1蛋白表达,从而影响CFs增殖。

Objective To study the effect of caveolin-1 antisense oligonucleotides（cav1-AS ODN） on cardiac fibroblasts（CFs） proliferation induced by arginine vasopressin（AVP） and the involvement of simvastatin（Sim） in the modulation of cholesterol.Methods CFs were isolated from adult SD rats and cultured.Phosphorothioate modified oligonucleotides and AVP were added to the CFs.CFs proliferation was assessed by 3H-TdR incorporation.Protein expressions of cav1,erk1/2,p21 and cyclin A were evaluated by western blotting.Results The levels of 3H-TdR incorporation and erk1/2 protein expression were（212 ± 6）% and（7.9 ± 0.3） relative to control,respectively,in cav1-AS ODN ＋ AVP group,being different from those of（172±4）% and（5.7±0.2） in AVP group（both P0.01）.Protein expression of p21 was decreased and cyclin A increased in AS ODN ＋ AVP group.The presence of β-cyclodextrin or progesterone or Sim decreased cav1 protein expression.The levels of cav1 protein expression were（86±3）%,（91±4）% and（94±5）% relative to control,respectively,in the presence of 10,15 or 20 μg/ml cholesterol,being different from that of Sim group（66 ± 4）% relative to control（P0.05,P0.01）.Conclusions cav1-AS ODN potentiates the proliferation-inducing effect of AVP on adult rat CFs,and the intervention of cholesterol-lowing effect of statins may provide new insights into the anti-fibrosis effect of these drugs.