摘要
目的观察葛根素对低氧性肺动脉平滑肌细胞（PASMC）增殖与凋亡及电压门控型钾离子通道亚型（Kvl．5）表达的影响，探讨葛根素在改善低氧性肺动脉高压与抑制肺血管重建中可能的作用机制。方法原代培养大鼠PASMCs，随机分为正常对照组（5％C0。常氧），低氧组（5％0：、5％C0：、90％N：三气培养），3个浓度葛根素十预组（在低氧组基础上分别加入终浓度为1×10^5、1×10^4、1×10^3mol／L葛根素，37℃培养24h）。采用CCK一8法和流式细胞仪检测细胞增殖情况，分光光度法检测caspase一3活力，蛋白印迹及实时定量聚合酶链反应（PCR）法分别检测Kvl．5蛋白和mRNA表达。结果与正常对照组（细胞活性：0．940±0．045，S期细胞比例：9.67％±1.28％，Caspase一3活力：0.1073±0．0113，Kv1．5蛋白：0．886±0．038，Kv1.5mRNA0.0377±0．0031）比较，低氧组细胞活性（1．296±0．034）、s期细胞比例（18．19％±1．19％）升高，Caspase-3活力（0．0664_＋0．0049）下降，Kvl．5蛋白（0.602±0.064）及mRNA（0．0108±0．0014）表达下降，差异均有统计学意义（P〈0．05）；与低氧组比较，各剂量葛根素干预组细胞活性和s期细胞比例下降，Caspase-3活力升高，Kvl．5蛋白及rnRNA表达上调，差异均有统计学意义（P〈0．05）。结论葛根素可抑制低氧性PASMCs增殖，促进其凋亡，其作用机制可能通过上调Kvl．5表达抑制PASMCs的增殖。
Objective To study the effects of puerarin on proliferation, apoptosis and Kvl.5 gene ex- pression of rat pulmonary artery smooth muscle cells （PASMCs） induced by hypoxia. Methods The rat PASMCs were divided into 5 groups： control group, hypoxia group, hypoxia plus puerarin （1 xl0^5 tool/L） group, hypoxia plus puerarin （1 x 10^4 mol/L） group and hypoxia plus puerarin （ 1x10-3mol/L） group, and cultured at 37℃ for 24 h. The proliferation of rat PASMCs was detected by CCK-8 assay and flow cytometry, the activity of easpase-3 was measured with spectrophotometrle method, Kvl.5 protein was detected by western blot, Kvl.5 mRNA was detected by real-time PCR. Results The cell viability and proportion of synthesis phase in control group were 0.940±0.045 and 9.67%＋ 1.28%, which were significantly lower than those （ 1.296±0.034 and 18.19%±1.19% ） in hypoxia group （P〈0.05）. The Caspase-3 activity, Kv 1.5 protein and Kv 1.5 mRNA in control group were 0.1073±0.0113, 0.886±0.038 and 0.0377±0.0031, which were significantly higher than those （0.0664±0.0049, 0.602±0.064 and 0.0108±0.0014 ） in hypoxia group （p〈0.05）. As compared with hypoxia group, the cell via- bility and proportion of synthesis phase in 3 hypoxia plus puerarin groups significantly decreased, and the Cas- pase-3 activity, Kv 1.5 protein and Kv 1.5 mRNA in 3 hypoxia plus puerarin groups significantly enhanced （P〈 0.05）. Conclusion Puerarin could decrease the proliferation and increase the apoptosis induced by hypoxia in rat PASMCs, and the up-regulated expression of Kv1.5 gene may be the mechanism of puerarin effects.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2011年第10期731-734,共4页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
国家自然科学基金资助项目（30670933）
