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目的:探讨钙池操纵的Ca2+(SOC)通道在Th2型细胞因子IL-13促进气道平滑肌细胞(ASMCs)增殖中的作用。方法:体外分离培养大鼠ASMCs,IL-13孵育36 h后,以Fluo 3-AM为钙荧光示踪剂,采用激光共聚焦显微镜,测定大鼠ASMCs基础Ca2+水平以及肌浆网钙泵抑制剂Thapsigargin(TPG)诱发的内钙释放和经SOC通道的外钙内流;采用四甲基偶氮唑蓝比色法(MTT法),检测SOC通道阻断剂SKF-96365对IL-13促增殖作用的影响。结果:①IL-13作用36 h后,ASMCs的内钙释放及经SOC通道的钙内流反应明显高于对照组(P<0.05),并且这种钙内流反应能部分被SOC通道阻断剂SKF-96365抑制(P<0.01)。②IL-13能促进气道平滑肌细胞增殖,该作用可被SKF-96365抑制(P<0.01)。结论:IL-13对气道平滑肌细胞的促增殖作用可能部分是通过促进SOC通道钙离子内流反应来实现的。
AIM: To investigate the role of Ca2 + -staining Ca2 + (SOC) channels in the proliferation of airway smooth muscle cells (ASMCs) stimulated by Th2 cytokine IL-13. Methods: ASMCs were isolated and cultured in vitro for 36 h. Fluo 3-AM was used as a calcium fluorescence tracer. Laser confocal microscopy was used to determine the level of basal Ca2 + in ASMCs and the activity of calcium pump inhibitor Thapsigargin (TPG) -induced release of Ca2 + and Ca2 + influx via SOC channels were measured. MTT assay was used to detect the proliferation of IL-13 induced by SOC channel blocker SKF-96365 Effect of the effect. RESULTS: ① After 36 hours of IL-13 treatment, the release of intracellular Ca2 + and the influx of Ca2 + via the SOC channel were significantly higher than those of the control group (P <0.05), and this Ca2 + influx was partially blocked by SOC channel blockers SKF-96365 inhibition (P <0.01). ②IL-13 can promote airway smooth muscle cell proliferation, which can be inhibited by SKF-96365 (P <0.01). CONCLUSION: The promoting effect of IL-13 on airway smooth muscle cells may be partly through the promotion of SOC channel calcium influx reaction.