沙丁胺醇在诱导培养人气管平滑肌细胞凋亡中的作用  

Role of salbutamol in inducing apoptosis of cultured human airway smooth muscle cells

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,广州510515

出　　处：《中华结核和呼吸杂志》2001年第4期219-223,I003,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的　研究沙丁胺醇 (商品名 :舒喘灵 )对于诱导培养人气管平滑肌细胞凋亡的作用。方法　分离人气管平滑肌细胞并且在含有 10 %胎牛血清的达尔伯克改良伊格尔 (DMEM)培养基中培养 ,4～ 6代的细胞用于实验。细胞用沙丁胺醇或色满卡林 (cromakalim)或 8 溴 环磷酸腺苷 (Br Camp)培养 2 4h或 48h ,光镜和电镜观察形态学改变。琼脂糖电泳分析DNA碎片。链亲和素 过氧化物酶(SP)免疫组化染色监测p5 3、Bcl 2和Bax基因表达的改变。通过破碎DNA的原位末端标记技术(TUNEL)检测凋亡细胞的百分数。结果　 (1)沙丁胺醇或 8 Br cAMP降低存活细胞的数目。在 48h、30 0 μmol/L的浓度 ,存活的细胞数量最低 ;(2 )人气管平滑肌细胞用沙丁胺醇 (10 0 μmol/L、30 0 μmol/L)孵化 48h显示出凋亡的形态学特征 (细胞皱缩、染色质浓聚 ) ;(3)琼脂糖电泳显示 ,核酸DNA断裂片呈典型的梯状条带 (大约 180～ 2 0 0bp) ;(4 )沙丁胺醇或 8 Br cAMP组p5 3或Bax基因表达显著高于对照组 ,但BCl 2组基因表达低于对照组 ;(5 )TUNEL显示 ,人气管平滑肌细胞的凋亡阳性率用 10 0、30 0μmol沙丁胺醇或 10 0 μmol 8 Br cAMP处理组与对照组比较差异有显著性 (q分别为 2 4 0 4、5 8 47、2 7 2 8,P均 <0 0 0 0 1)。Objective To study the effect of salbutamol on inducing apoptosis of cultured human airway smooth muscle cells (ASMCs). Methods Human ASMCs were isolated and cultured in DMEM containing 10% fetal bovine serum Cells of Passases 4～6 were used in experiments The cells were cultured with salbutamol, cromakalim, 8 Br cAMP for 24 or 48 hours. Morphological changes were observed by light microscopy and electron microscopy DNA fragmentation was analyzed by agarose gels SP immunohistological staining method was performed to detect the changes of expressions of p53, Bcl 2 and Bax gene. The percentage of apoptosis cell was detected by situ end labeling technique (TUNEL) of fragmental DNA. Results (1) Salbutamol or 8 Br cAMP decreased the number of viable cells At 48 hour, at 300 μmol/L, number of viable cells was the lowest. (2) Human ASMCs incubated with salbutamol (100 μmol/L or 300 μmol/L) for 48 h revealed morphological features of apoptosis (cellular shrinkage, condensation of chromatin). (3) Agarose gel electrophoresis showed a characteristic “ladder” of DNA bands reprenting integer multiples of the internucleosomal DNA length (about 180～200 bp). (4) The expression of p53 or Bax gene in salbutamol or 8 Br cAMP group was significantly higher than that in control group, but the expression of BCl 2 gene was lower than that in control group (5) The TUNEL indicated: apoptotic positive rate of human ASMCs was significantly different following 100 μmol,300 μmol salbutamol or 100 μmol 8 Br cAMP treatment ( q =24 04,58 47,27 78 respectivey, P <0 0001),but cromakalim, propranolol before salbutamol not affected basal level of apoptotic cell ( q =0 12,0 52 respectively, P =0 932,0 717 respectly),as compared with the controls. Conclusions (1) Salbutamol induced apoptosis

关 键 词：沙丁胺醇 气管平滑肌 细胞凋亡 CAMP 蛋白激酶A 平喘药 

分 类 号：R562.25[医药卫生—呼吸系统]