转染Kv1.5基因抑制人气道平滑肌细胞的增殖

 目的：转染Kv1.5基因对人气道平滑肌细胞(HASMCs)增殖及凋亡的影响。方法:通过脂质体介导瞬时转染Kv1.5基因于培养的HASMCs中，以转染空载体pRc/CMV的细胞及未转染质粒的细胞为对照；用Western blotting 检测平滑肌细胞Kv1.5蛋白表达；用荧光光度法检测HASMCs胞内钙浓度；用流式细胞术观察细胞周期；用MTT法检测HASMCs 增殖及DNA 末端转移酶介导的原位缺口末端标记技术(TUNEL)检测细胞的凋亡。 结果: (1) 转染质粒组Kv1.5蛋白质的表达明显高于未转染组及空载体转染组(P<0.01); (2) 转染质粒组细胞胞内钙浓度明显低于未转染组及空载体转染组(P<0.05)，且其细胞周期中的G₀/G₁期细胞比例明显高于、细胞增殖率显著低于未转染组及空载体转染组(P<0.01)；同时，转染质粒组细胞的凋亡率明显高于未转染组及空载体转染组 (P<0.01)。 结论: 转染Kv1.5基因能抑制HASMCs的增殖、促进其凋亡，为进一步探讨哮喘气道重塑的机制及其治疗提供实验依据。

Transfection of human Kv1.5 gene inhibits the proliferation of human airway smooth muscle cells

AIM:To investigate the effect of human Kv1.5 gene transfection on the proliferation and apoptosis in human airway smooth muscle cells(HASMCs).METHODS:Kv1.5 gene was transiently transfected into HASMCs with Lipofectamine 2000 and the level of Kv1.5 protein was observed by Western blotting.Ca 2 concentration in HASMCs was investigated by fluorescent quantitation using fluorospectrophotometer.Flow-cytometry,MTT method and TUNEL were used to detect the effects of Kv1.5 gene transfection on proliferation and apoptosis in HASMCs.RESULTS:(1)Western blotting showed that transfection of human Kv1.5 plasmid significantly increased the Kv1.5 protein expression compared to control group(P<0.01).(2)There was a decrease in Ca 2 concentration and the numbers of G_0/G_1 HASMCs in Kv1.5 gene transfection group compared to control group(P<0.05).The proliferation rate in Kv1.5 gene transfection group was significantly decreased,and the apoptosis rate in Kv1.5 gene transfection group was significantly increased compared to control group(P<0.01).CONCLUSION:Transfection of human Kv1.5 plasmid inhibits proliferation and increases apoptosis in HASMCs,which provides the experimental evidence for investigating the mechanism and treatment of airway remodeling in asthma.