一氧化氮诱导体外人气管平滑肌细胞凋亡  

Effect of nitric oxide on apoptosis of human airway smooth muscle cells in vitro

作　　者：罗雅玲[1] 赖文岩[1] 徐健[1] 袁勇[1] 张荣华[1] 

机构地区：[1]第一军医大学南方医院呼吸科,广东广州510515

出　　处：《中国药学杂志》2002年第3期178-181,共4页Chinese Pharmaceutical Journal

摘　　要：目的　观察一氧化氮 (NO)是否在体外诱导人气管平滑肌细胞 (airwaysmoothmusclecells,ASMCs)凋亡。方法　人ASMCs用SNAP或SNP或 8 溴 环磷酸鸟苷 (8 Br cGMP)处理。通过四甲基偶氮唑盐 (dimethythiazol zyl7 2 ,5diphenylte trazdium ,MTT)法在 6 ,12 ,18,2 4 ,36h检测SNAP或SNP或 8 Br cGMP对细胞生长的作用。电镜、琼脂糖电泳、原位末端标记和免疫组化链霉菌抗生素蛋白 过氧化酶 (streptavidin peroxidase,SP)法技术用于检测凋亡。 结果　①SNAP组和SNP组ASMCs存活的数量降低 ,而对照组和cGMP组无变化 (P <0 .0 1)。②SNAP组和SNP组电镜显示核皱缩 ,染色体浓聚和凋亡小体形成。③用SNAP或SNP处理 ,ASMCs的凋亡指数显著升高 (P <0 .0 1) ,但对照组或 8 Br cGMP组没有不同。④在SNAP或SNP组 ,琼脂糖电泳代表了核苷酸DNA长度整倍数的特征性的DNA梯状条带 (大约 180～ 2 0 0bp)。⑤SNAP或SNP组P5 3或Bax基因表达显著高于对照组或 8 Br cGMP组 ,但Bcl 2基因表达是低于对照组或 8 Br cGMP组。结论　①NO在体外以时间和剂量依赖的方式诱导人ASMCs凋亡。②NO这些作用通过的通路不涉及到鸟苷酸环化酶和cGMP依赖的蛋白激酶。③对人ASMCs的这种负性调节机制也许与治疗哮喘的气道重建有关。OBJECTIVE: To investigate nitric oxide (NO) induced apoptosis of human airway smooth muscle cells (ASMCs) in vitro. METHODS: SNAP or SNP or 8-Br-cGMP was used for the treatment of ASMCs. The effect of SNAP or SNP or 8-Br-cGMP on the growth of the cells was detected by MTT method at 6,12,18,24,30 hours, respectively. Electric microscope, agarase gels electrophoresis, TUNEL and immunohistochemical S-P technique were used to detect the apoptosis. RESULTS: 1 The living quantity of ASMCs decreased in SNAP and SNP groups, which was not observed in the control and cGMP groups (P<0.01). 2 Electron microscopic examination showed nuclear contraction, chromatin condensation and apoptotic bodies formation in SNAP or SNP groups. 3 Apoptotic index of ASMCs was significantly elevated following SNAP or SNP treatment (P<0.01), but no difference was found between control and 8-Br-cGMP groups. 4 Agarose gel electrophoresis showed a characteristic 'ladder' of DNA bands representing integer multiples of the internuclesomal DNA length (about 180-200 bp) in SNAP or SNP groups. 5 The expression of P53 or Bax gene in SNAP or SNP group was significantly higher than in control or 8-Br-cGMP group, but the expression of Bcl-2 gene was lower than in control or 8-Br-cGMP group. CONCLUSION: 1 NO induced apoptosis in human ASMCs in vitro in a time and dose dependent manner. 2 NO exerted these effects through a pathway that does not involve guanylate cyclase and cGMP-dependent protein kinase. 3 The mechanism of negative regulation to human ASMCs may be related to treatment of airway remodeling in asthma.

关 键 词：一氧化氮 细胞凋亡 气道重建 哮喘 气管平滑肌细胞 ASMCs 

分 类 号：R562.25[医药卫生—呼吸系统] R965[医药卫生—内科学]