红霉素抑制烟草烟雾提取物诱导人巨噬细胞炎症及其机制的研究  

The inhibitory effect of erythromycin on inflammation induced by cigarette smoke extract in human macrophages and its mechanism

作　　者：霍健君 李梅华[1] 马南[1] 邱居烽 冷子杨 王绍霜 钟小宁[1] 何志义[1] Huo Jianjun;Li Meihua;Ma Nan;Qiu Jufeng;Leng Ziyang;Wang Shaoshuang;Zhong Xiaoning;He Zhiyi(Department of Respiratory And Critical Care Medicine,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院呼吸与危重症医学科,南宁530021

出　　处：《广西医科大学学报》2021年第4期716-720,共5页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81360012);广西自然科学基金资助项目(No.2016GXNSFAA380269);广西卫生健康委员会自筹经费科研课题资助项目(No.Z20201205)。

摘　　要：目的:初步探讨红霉素(EM)对烟草烟雾提取物(CSE)诱导的人巨噬细胞炎症的抑制作用及其可能机制。方法:将体外培养的人单核细胞U937用佛波脂(PMA)诱导分化为人巨噬细胞。CCK-8法检测不同浓度、不同时间CSE及EM对细胞活性的影响。细胞分为对照组、CSE组(1%CSE刺激24 h或48 h)、CSE+EM组(1μg/mL EM预孵育24 h或48 h后加1%CSE刺激24 h或48 h)、CSE+核因子(NF)-кB抑制剂吡咯烷二硫氨基甲酸(PDTC)组(20 nmol/L PDTC预孵育24 h或48 h后加1%CSE刺激24 h或48 h)、I型组蛋白去乙酰化酶(HDAC3)抑制剂曲古霉素(TSA)组(100 ng/mLTSA孵育24 h)。采用酶联免疫吸附试验(ELISA)法检测细胞培养上清中炎症介质白介素(IL)-8浓度,应用凝胶迁移率阻滞实验(EMSA)检测NF-кB的活性,Western blotting法检测HDAC3和NF-кB蛋白表达。结果:与对照组比较,CSE组IL-8浓度、NF-кB转录活性明显增高,HDAC3蛋白表达下降(P<0.05),TSA组HDAC3蛋白表达下降,NF-кB蛋白表达升高;与CSE组比较,CSE+EM组和CSE+PDTC组IL-8浓度、NF-кB转录活性及NF-кB蛋白表达明显下降,HDAC3蛋白表达升高(P<0.05)。结论:EM能够抑制CSE诱导的人巨噬细胞炎症介质IL-8释放,其抗炎活性可能与抑制NF-кB的转录活性和表达以及上调HDAC3表达有关。Objectives:To investigate the inhibitory effect of erythromycin on inflammation induced by cigarette smoke extract in human macrophages and its mechanism.Methods:Human monocytic cellsU937 cultured in vitro were differentiated into macrophages by using phorbol 12-myristate 13-acetate(PMA).The concentration and time of cigarette smoke extract(CSE)and erythromycin(EM)were screened by CCK-8 method.The cells were divided intocontrol group,CSE group(1%CSE stimulation for 24 or 48 h),CSE+EMgroup(pre-incubation 1μg/mL EM for 24 or 48 h before adding CSE),CSE+nuclear factor-κB(NF-κB)inhibitor pyrrolidine dithiocarbamate(PDTC)group(pre-incubation 20 nmol/L PDTC for 24 or 48 h before adding CSE),HDAC inhibitor trichostatin A(TSA)group(pre-incubation100 ng/mL TSA for 24 h).The IL-8 level in culture supernatants was measured by enzyme linked immunosorbent assay(ELISA),and activity of NF-κB in macrophages was evaluated by Electrophoretic mobility shift assay(EMSA).HDAC3 and NF-κB proteinslevels were determined by western blotting.Results:Compared with the control group,the IL-8 concentration andNF-κB activity were increased,while the HDAC3 protein expression was significantly decreased in CSE group(P<0.05).The HDAC3 protein expression was decreased and NF-κB protein expression was increased in TSA group.Compared with CSE group,IL-8 concentration,NF-κBtranscription activity and NF-κBprotein expression were significantly decreased in CSE+EM group and CSE+PDTC group,while HDAC3 protein expression was increased(P<0.05).Conclusion:Erythromycin can inhibit the release of IL-8 induced by CSE.The mechanism is related tothe inhibition of the transcriptional activity and expression of NF-κB,and the up-regulation of HDAC3 expression.

关 键 词：烟草烟雾提取物 HDAC3 NF-кB IL-8 红霉素 

分 类 号：R563.9[医药卫生—呼吸系统]