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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 expression changes of vascular endothelial growth factor (VEGF) and its receptor 2 (KDR) in passively sensitized human airway smooth muscle cells (ASMCs) and their effects on the proliferation of ASMCs. Methods Human ASMC was cultured and serum was sensitized to ASMC by bronchial asthma. The expression of proliferating cell nuclear antigen (PCNA) in ASMC was detected by immunohistochemistry, the cell metabolic activity was assayed by MTT assay and the cell cycle was analyzed by flow cytometry. The mRNA and protein of VEGF and KDR were detected by RT-PCR and Western blot respectively in different groups Human ASMC expression level. Results (1) Compared with the control group and the intervention group, the ASMC in the passive sensitized group increased significantly (P <0.05). (2) The mRNA expressions of VEGF121, VEGF165 and VEGF189 in passive sensitized group were significantly increased compared with the control group and the intervention group (P <0.05 or P <0.01). (3) The expressions of VEGF and KDR protein in passive sensitized group were significantly increased compared with the control group and the intervention group (P <0.05). The linear correlation analysis showed that the expression of PCNA in ASMC was positively correlated with the expression of VEGF121, 165, 189 and KDR mRNA in ASMC (r = 0.73,0.82,0.77,0.70, P <0.05 ). The expression of PCNA in ASMC was also positively correlated with the protein expression of VEGF and KDR in ASMC (r = 0.69, 0.67, P <0.05, respectively). Conclusions The expression of VEGF and KDR in passively sensitized ASMCs is up-regulated and closely related to the proliferation of ASMCs. The results suggest that VEGF and its receptor 2 may be involved in the process of ASMC proliferation during airway remodeling in asthma.