细胞间黏附分子-1在红霉素抗炎机制中的作用  

Expression of intercellular adhesion molecule 1 mRNA in the anti-inflammatory molecular mechanism of erythromycin

作　　者：李梅华[1] 钟小宁[1] 柳广南[1] 何志义[1] 朱敏[2] 周刚[2] 

机构地区：[1]广西医科大学第一附属医院呼吸科,广西南宁530027 [2]中南大学湘雅医学院分子生物学研究中心

出　　处：《中国老年学杂志》2009年第18期2287-2289,共3页Chinese Journal of Gerontology

基　　金：广西自然科学基金项目(桂科自0229024)

摘　　要：目的从分子水平上探讨红霉素(EM)的抗炎作用机制。方法体外培养人支气管上皮细胞(16HBE),将细胞随机分为8组,先加入EM干预,后加入肿瘤坏死因子α(TNF-α)刺激。分组如下:①空白对照组;②TNF-α(20ng/ml,16h)组;③EM(0.3μg/ml,24h)+TNF-α(20ng/ml,16h)组;④EM(3μg/ml,24h)+TNF-α(20ng/ml,16h)组;⑤EM(30μg/ml,24h)+TNF-α(20ng/ml,16h)组;⑥EM(0.3μg/ml,48h)+TNF-α(20ng/ml,16h)组;⑦EM(3μg/ml,48h)+TNF-α(20ng/ml,16h)组;⑧EM(30μg/ml,48h)+TNF-α(20ng/ml,16h)。然后收集各组细胞分别提取RNA,应用逆转录聚合酶链反应(RT-PCR)方法检测细胞间黏附分子-1(ICAM-1)mRNA的表达。因ICAM-1RT-PCR产物分子大小约为700bp,与原设计产物分子大小490bp不相符,进一步作基因克隆测序了解基因之间是否具有同源性。结果ICAM-1的RT-PCR产物即为目的基因:ICAM-1基因。TNF-α刺激人支气管上皮细胞(16HBE)后,细胞ICAM-1mRNA表达增高。先加入不同浓度及作用时间的EM,后加入TNF-α刺激人支气管上皮细胞(16HBE),各组细胞ICAM-1mRNA表达均降低,且提示有浓度与时间依赖性。结论TNF-α能激活人支气管上皮细胞使其ICAM-1基因表达增高,促进炎症的发生发展。EM能抑制人支气管上皮细胞ICAM-1基因表达,可能是EM的抗炎作用机制之一。Objective To investigate the anti-inflammatory molecular mechanism of erythromycin （EM） in order to find a new way to prevent and treat chronic obstructive pulmonary disease （COPD）. Methods Human bronchial epithelial cell line 16HBE was cultured with 10% serum DMEM （high glucose）. The cells were randomly divided into eight groups： blank control group, TNF-α （20 ng/ml, 16 h）, EM（0.3 μg/ml,24 h） ＋ TNF-α （20 ng/ml,16 h）, EM（3 μg/ml,24 h） ＋ TNF-α （20 ng/ml,16 h）, EM（30 μg/ml,24 h） ＋ TNF-α （20 ng/ml, 16 h ）, EM （ 0. 3 μg/ml, 48 h ） ＋ TNF-α （ 20 ng/ml, 16 h ）, EM （ 3 μg/ml, 48 h） ＋ TNF-α （ 20 ng/ml, 16 h ）, and EM （30 μg,/ml,48 h） ＋TNF-α （20 ng/ml,16 h）. Then total cellular RNA was extracted to detect the expression of ICAM-1 mRNA by RTPCR. The molecular size of ICAM-1 RT-PCR production was not correspondent for what it was designed, therefore, clone sequencing was applied to confirm it. Results ICAM-1 RT-PCR production was ICAM-1 mRNA. ICAM-1 mRNA was increased by the addition of TNF-ot, which was inhibited by the preincubation with EM in dose-dependent and time-dependent fasion. Condusions ICAM-lmRNA can be increased by the stimulation of TNF-α, which indicate that EM modify inflammation presumably by suppressing ICAM-1 mRNA in human bronchial epithelial ceils.

关 键 词：人支气管上皮细胞 细胞间黏附分子-1 肿瘤坏死因子Α 红霉素 

分 类 号：R284[医药卫生—中药学]