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目的 :探讨氨茶碱对大鼠气道平滑肌细胞 (ASMC)增生及原癌基因c fos表达的影响 .方法 :分离大鼠气道平滑肌细胞 ,在含有 1 0 0mL/L胎牛血清的DMEM中培养 ,第 4～ 6代细胞用于实验 ,细胞用不同浓度氨茶碱 (0 ,5 0 ,1 0 0 ,2 0 0 ,30 0 ,4 0 0mg/L)诱导培养 1 2～ 72h ;MTT法观察ASMCs的增殖情况 ;荧光显微镜形态学观察大鼠ASMCs凋亡 ;RT PCR观察ASMCs的c fosmRNA表达状况 .结果 :①氨茶碱以时间和浓度依赖的方式减少存活的细胞数 ;②当氨茶碱浓度大于 5 0mg/L时 ,荧光显微镜下可见深染、浓缩的细胞核 ,随着药物浓度的增加 ,凋亡小体形成 ;③RT PCR显示对照组及 5 0～ 2 0 0mg/L氨茶碱组均有c fosmRNA表达 ,而且随着氨茶碱浓度的提高 ,表达量逐渐减少 ,30 0和 4 0 0mg/L氨茶碱组则未见有c fosmRNA表达 .结论 :氨茶碱可能通过下调原癌基因c fos表达来抑制大鼠ASMC增生
Objective: To investigate the effect of aminophylline on the proliferation of rat airway smooth muscle cells (ASMCs) and the expression of proto-oncogene c fos.Methods: The airway smooth muscle cells of rats were isolated and cultured in DMEM containing 100 mL / L fetal bovine serum After cultured for 4-6 days, the cells were cultured for 1-22 hours with different concentrations of aminophylline (0, 500, 100, 200, 300, 400 mg / L); MTT The proliferation of ASMCs was observed by fluorescence microscope.Apoptosis of ASMCs was observed by fluorescence microscopy and the expression of c fos mRNA was detected by RT PCR.Results ①Aminophylline reduced the number of viable cells in a time- and concentration- When the theophylline concentration was more than 50mg / L, deep-staining and concentrated nuclei were observed under a fluorescence microscope. The apoptotic bodies formed with the increase of the drug concentration. ③ RT PCR showed that the control group and 50 ~ 200 mg / The expression of c fos mRNA was significantly increased in both groups, and the expression decreased gradually with the increase of aminophylline concentration, but no c fos mRNA expression was found in 30 and 400 mg / L aminophylline group.Conclusion: Aminophylline may pass Downregulation of proto-oncogene c fos expression to inhibit rat ASMC proliferation