摘要
探讨阿托伐他汀对心房颤动细胞模型中ATBF1/PIAS3/STAT3信号通路的作用。通过高频电刺激（5 ms,25 Hz,7 V/cm）HL-1心房肌细胞系构建房颤细胞模型,并采用CCK-8实验检测梯度浓度阿托伐他汀（0.01,0.1,1,10,50和100μmol/L）干预下心房肌细胞系HL-1细胞活性,以筛选出阿托伐他汀的药物安全浓度,同时应用Western blotting实验测定以上每组p-STAT3和STAT3蛋白表达情况来选择最佳给药浓度,并分别检测在对照组、房颤组和药物实验组3组中ATBF1/PIAS3/STAT3信号通路中各个蛋白表达水平。实验结果证明,CCK-8实验筛选阿托伐他汀的药物安全浓度为0.1-50μmol/L。房颤细胞模型中,ATBF1的蛋白表达量显著下调,PIAS3蛋白水平降低,p-STAT3水平明显升高,差异均具有统计学意义（P〈0.05）,但总STAT3的表达水平并无显著性改变（P〉0.05）。阿托伐他汀药物干预后,p-STAT3蛋白与STAT3蛋白比值随浓度升高,在1μmol/L时达到最大。在此药物浓度下,房颤模型中ATBF1和PIAS3表达上调,p-STAT3表达下调,有统计学差异（P〈0.05）,而总STAT3无显著性差异（P〉0.05）。提示阿托伐他汀抗心房颤动的分子作用机制可能与其抑制ATBF1/PIAS3/STAT3信号通路有关。
This paper is to explore the effect and mechanism of atorvastatin on ATBF1 / PIAS3 / STAT3 signaling of cell model for atrial fibrillation. Atrial（ HL-1 myocytes） cells cultured in the presence of rapid electrical stimulation（ 5 ms,25 Hz,7 V / cm） was constructed the cell model for atrial fibrillation. CCK-8 experiment was carried out to detect the cell activity of HL-1 cell line under the intervention of atorvastatin at different concentrations（ 0. 01,0. 1,1,10,50 and 100 μmol / L）,aiming at screening out the safety concentration of atorvastatin. The p-STAT3 and STAT3 protein expression was probed by Western blot analysis in above groups to determine the optimized drug concentration. The protein expression levels of ATBF1 / PIAS3 / STAT3 signaling were detected by Western blot analysis in the HL-1 cells at the control group,atrial fibrillation group and atrial fibrillation administration with the above optimized concentration of atorvastatin,respectively. The results indicated that the safety concentrations of atorvastatin screened by CCK-8 experiment were 0. 1 - 50 μmol / L. The protein expression of ATBF1 and PIAS3 was significantly decreased and p-STAT3 was markedly increased（ P〈 0. 05）,while the total STAT3 protein level was no significant change（ P〈 0. 05） in the cell model for atrial fibrillation. The ratio of p-STAT3 and STAT3 up-regulated with the increase of atorvastatin concentration at the pharmacologic interventions,and the atorvastatin concentration of maximal-ratio was 1 μmol / L. Compared with the atrial fibrillation model,the protein expression of ATBF1 and PIAS3 was increased and p-STAT3 was significantly decreased（ P〈 0. 05）,while the total STAT3 protein level was no markedly change（ P〈 0. 05） with the administration of atorvastatin in 1 μmol / L. The results suggest that the molecular mechanisms of atorvastatin on anti-atrial fibrillation might be related to inhibit the activation of ATBF1 / PIAS3 / STAT3 signaling.
出处
《药物生物技术》
CAS
2016年第1期17-21,共5页
Pharmaceutical Biotechnology
基金
国家“重大新药创制”科技重大专项课题资助项目(No.2009ZX09103-345)
