血管内皮生长因子及其受体2与被动致敏的人气道平滑肌细胞增殖相关性研究

Correlation between the expression of vascular endothelial growth factor, its receptor 2 and the proliferation of human airway smooth muscle cells passively sensitized with asthmatic serum

目的探讨血管内皮生长因子(VEGF)及其受体2(KDR)在被动致敏的人气道平滑肌细胞(ASMC)中表达变化及其对ASMC增殖的影响。方法培养人ASMC,用支气管哮喘病人血清被动致敏ASMC。用免疫组织化学技术检测ASMC增殖细胞核抗原(PCNA)的表达,MTT法检测细胞代谢活性及流式细胞仪分析细胞周期;用RT-PCR及Western blot方法分别检测VEGF和KDR mRNA及蛋白质在不同组人ASMC的表达程度。结果(1)被动致敏组ASMC较对照组和干预组增殖显著增加(P<0．05)。(2)被动致敏组ASMC的VEGF121、VEGF165和VEGF189的mRNA表达分别较对照组和干预组显著增加(P<0．05或P<0．01)。(3)被动致敏组ASMC VEGF及KDR蛋白质的表达较对照组和干预组显著增加(P<0．05)。直线相关性分析显示,ASMC中PCNA表达与ASMC中VEGF121、165、189及KDR mRNA表达水平呈正相关(r分别为0．73、0．82、0．77、0．70,P<0．05);ASMC中PCNA表达与ASMC中VEGF及KDR蛋白质表达水平也呈正相关(r分别为0．69、0．67,P<0．05)。结论被动致敏的ASMC中VEGF及其受体KDR表达上调,并与ASMC增殖密切相关。该结果提示VEGF及其受体2可能参与了哮喘气道重建中ASMC增殖的过程。

Objective To investigate the correlation between the proliferation of human airway smooth muscle cells passively sensitized with asthmatic serum and the expression of vascular endothelial growth factor （VEGF） and its receptor 2（KDB）. Methods Human airway smooth muscle cells （ASMC） were cultured and passively sensitized with asthmatic serum. The proliferation of ASMC was examed by proliferating cell nuclear antigen （PCNA）, MTT and flow cytometry analysis. Western blot and RT-PCR were used to measure the protein and mRNA exxon of VEGF and its receptor 2 in the ASMC. Results （ 1 ） The proliferation of human ASMC passively sensitized with asthmatic serum were significantly increased compared with that in control group and dexamethasone group （P〈0.05）. （2） The expression of VEGF121,165,189 mRNA and KDR mRNA in ASMC of passively senatized group was higher than that in control group and dexamethasone group（P 〈 0.05 or P 〈0.01）. （3） The expression of VEGF, KDR protein in ASMC of passively sensitized group was higher than that in control group and dexamethasene group （P〈0.05）. （4）The expression of VEGF121, 165, 189 and KDR mRNA were well correlated with the exxon of PCNA in ASMC（r =0.73, 0.82, 0.77, 0.70; P 〈0.05, respectively）. The expression of VEGF and KDR protein were also well correlated with the expression of PCNA in ASMC （ r = 0.69, 0.67; P 〈 0.05）.Conclusion The expression of VEGF and KDB were increased in ASMC passively sensized with asthmatic serum and well correhted with the proliferation of ASMC. VEGF and KDR may take part in the process of airway smooth muscle cells, proliferation in airway remodeling of asthma.