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目的:探讨钙库操纵的钙内流(SOCE)在转化生长因子-β1(TGF-β1)促进气道平滑肌细胞(ASMCs)增殖中的作用。方法:体外培养大鼠ASMCs,以Fluo-3/AM为细胞内钙离子浓度示踪剂,观察ASMCs在TGF-β1作用后,细胞内基础钙离子浓度、钙释放和SOCE的变化;四甲基偶氮唑蓝(MTT)微量比色法测定TGF-β1对胎牛血清(FBS)刺激的大鼠ASMCs的增殖的促进作用,以及钙库操纵的钙通道(SOC)抑制剂SKF-96365、NiCl2组和LaCl3对这一作用的影响。结果:①在无钙溶液中,肌浆网钙泵抑制剂thapsigargin诱导ASMCs的钙释放,这一作用在TGF-β1(10μg/L)处理组高于对照组;同时,复钙后的SOCE幅度在TGF-β1处理组也高于对照组。②TGF-β1(10μg/L)可以促进2.5%FBS刺激的ASMCs的增殖(P<0.001),这一效应可以被钙通道抑制剂SKF-96365(10μmol/L)和NiCl2(100μmol/L)抑制,而钙通道抑制剂LaCl3则对TGF-β1的促增殖作用无显著作用(P>0.05)。结论:钙库操纵的钙内流可能参与了TGF-β1促进ASMCs增殖的作用,提示其可能是调控气道重塑的重要靶点。
AIM: To investigate the role of calcium store-operated calcium influx (SOCE) in the proliferation of airway smooth muscle cells (ASMCs) induced by transforming growth factor-β1 (TGF-β1). Methods: Rat ASMCs were cultured in vitro. Fluo-3 / AM was used as intracellular calcium ion tracer to observe the changes of basic calcium concentration, calcium release and SOCE in ASMCs treated with TGF-β1. The effect of TGF-β1 on the proliferation of rat ASMCs stimulated by FBS was measured by MTT assay. The calcium channel inhibitor (SOC) inhibitor SKF-96365 and NiCl2 Effect of LaCl3 on this effect. Results: ① In calcium-free solution, thapsigargin, a calcium-pump inhibitor of sarcoplasmic reticulum, induced calcium release from ASMCs, which was higher in TGF-β1 treated group than in control group. Meanwhile, the SOCE amplitude The TGF-β1-treated group was also higher than the control group. ②TGF-β1 (10μg / L) can promote the proliferation of ASMCs stimulated by 2.5% FBS (P <0.001), and this effect can be inhibited by calcium channel inhibitor SKF-96365 (10μmol / L) and NiCl2 However, calcium channel inhibitor LaCl3 had no significant effect on the proliferation of TGF-β1 (P> 0.05). CONCLUSION: Calcium-infused calcium influx may play a role in the proliferation of ASMCs induced by TGF-β1, suggesting that it may play an important role in the regulation of airway remodeling.