和厚朴酚抑制CFP-10、ESAT-6诱导的人肺泡Ⅱ型上皮细胞炎症因子表达  

Honokiol inhibits expression of inflammatory cytokines induced by CFP-10 and ESAT-6 in human AEC II alveolar epithelial cells

作　　者：沈颖[1] 王叶萍 程林芳[3] 姚航平[3] 

机构地区：[1]浙江医学高等专科学校临床医学系内科教研室,杭州310053 [2]长兴县人民医院检验科 [3]浙江大学医学院附属第一医院传染病诊治国家重点实验室

出　　处：《浙江医学》2012年第8期583-586,共4页Zhejiang Medical Journal

基　　金：国家＂十二五＂重大科技专项（2012ZX10001003-002）;浙江省医药卫生科学研究基金资助项目（2009B003,2011KYB119）

摘　　要：目的 研究和厚朴酚（HNK）对早期分泌抗原靶蛋白6（ESAT-6）和培养滤液蛋白10（CFP-10）作为特异性抗原刺激人肺泡Ⅱ型上皮细胞（A549）炎症因子表达的抑制作用,探讨抗炎在结核病治疗中的潜在价值.方法 用MTS法检测HNK（0～80μmol/L）对A549细胞的毒性反应,确定合适的药物实验浓度.用ELISA法检测以CFP-10、ESAT-6（0～40μg/ml）为抗原刺激的A549表达IL-8水平,选择最适刺激浓度.将A549细胞与CFP-10,ESAT-6及不同浓度的HNK（0～20μmol/L）共同培养24h后收集培养上清液和细胞,用ELISA法检测各组培养上清液中IL-1β、IL-6、IL-8、TNF-α及Rantes的表达水平,分析HNK对细胞因子的剂量依赖抑制.结果 20μmol/L及以下的HNK浓度对A549细胞无明显毒性反应.抗原最适刺激浓度为5μmol/L.CFP-10,ESAT-6刺激可显著诱导A549细胞炎症因子IL-1β、IL-6、IL-8、TNF-α及Rantes的产生.HNK（0～20μmol/L）剂量依赖性地抑制CFP-10,ESAT-6诱导的炎症因子的表达.结论 HNK能有效抑制CFP-10,ESAT-6诱导的人肺泡Ⅱ型上皮细胞炎性细胞因子的表达.Objective TO investigate the inhibitory effects of honokiol （HNK） on inflammatory cytokines production induced by ESAT-6 or CFP-10 in human AEC II alveolar epithelial A549 cells. Methods The safe dosage of HNK for A549 cells was ini- tially identified by MTS assay. The expression level of IL-8 stimulated by CFP-10 or ESAT （0-40μg/ml） was determined by ELISA for selection of the optimal inducing concentration. A549 cells treated with honokiol （0-20μmol/L） were incubated in the presence or absence of CFP-10 or ESAT-6 for 24 h. The expression levels of IL-1 β, IL-6, IL-8, TNF-α and Rantes in supernatant were determined by ELISA. Results HNK did not significantly change HRMC viability when used at a concentration of 〈20μmol/L. CFP-10 or ESAT-6 treatment led to a marked up-regulation of the levels of IL-1β, IL-6, IL-8,TNF- α and Rantes in A549 cells, especially at the dose of 5 μ mol/L. The up-regulation of these molecules was significantly abolished by HNK in a dose-depen- dent manner. Conclusion HNK can inhibit CFP-10 or ESAT-6-induced expression of inflammatory cytokines in human AEC II alveolar epithelial cells.

关 键 词：和厚朴酚 炎症因子 人肺泡Ⅱ型上皮细胞 早期分泌抗原靶蛋白6 培养滤液蛋白10 结核 

分 类 号：R563[医药卫生—呼吸系统]