通络类中药提取物对肺炎支原体感染A549细胞干预作用的研究  

Study on the intervention effect of extract of Chinese medicine for unblocking the collaterals on A549 cells infected by mycoplasma pneumoniae

作　　者：姜永红[1] 吕佳佳 赵晓阳 JIANG Yonghong;LYU Jiajia;ZHAO Xiaoyang(Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《现代中西医结合杂志》2021年第2期115-119,132,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金面上项目(81674024);上海市“十三五”临床重点专科中医儿科项目(shslczdzk04102);上海市市属博士学科点建设基金(B201503)。

摘　　要：目的研究通络类中药虎杖、半枝莲、蝉蜕及其药对和小复方的提取物对肺炎支原体(MP)的体外抑菌作用以及对MP感染A549细胞分泌炎症因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、核因子kappa B(NF-κB)的影响。方法将虎杖、半枝莲、蝉蜕分别用水提法和醇提法提取,液体培养法培养MP菌液,微量稀释法测定最小抑菌浓度;MTT法检测虎杖、半枝莲、蝉蜕、虎杖+半枝莲、虎杖+蝉蜕、半枝莲+蝉蜕、小复方提取物对A549细胞的毒性作用,并计算半数抑制浓度(IC50);挑选抑菌效果较好的组,实验分为正常组,模型组,虎杖醇提低、中、高浓度组,半枝莲醇提低、中、高浓度组,虎杖+半枝莲低、中、高浓度组,小复方低、中、高浓度组,MTT法检测各组培养12 h、24 h、36 h、48 h的MP感染A549细胞活力,ELISA法检测各组MP感染A549细胞分泌IL-6、TNF-α、NF-κB的水平。结果虎杖、半枝莲、蝉蜕的水提物和醇提物均有一定的体外抑制MP生长的作用,小复方醇提物最佳,药对醇提物优于药对水提物;单味药醇提物优于单味药水提物,蝉蜕水提物和醇提物最差;单味药、药对和小复方醇提物对细胞毒性的IC50值均大于水提物。模型组在MP感染A549细胞12 h、24 h、36 h、48 h后细胞活力均明显低于正常组(P均<0.05),各药物组(除虎杖醇提低剂量组外)干预48 h细胞活力均明显高于模型组(P均<0.05);模型组TNF-α、IL-6、NF-κB水平均明显高于正常组(P均<0.05),各药物组TNF-α、IL-6、NF-κB水平均明显低于模型组(P均<0.05),且呈量效关系。结论虎杖、半枝莲、蝉蜕及其药对和小复方均具有一定体外抑制MP的作用,且醇提物的效果优于水提物,小复方的效果优于单味药和药对,推测中药的抑菌作用与中药提取方法和药物的相互配伍产生的协同作用有关;中药提取物对于MP感染后的A549细胞具有保护作用,且对MP引起的炎症反应有调节作�Objective It is to study the antibacterial effect of extracts from Giant Knotweed,Barbed Slullcap,Cicada Slough and its herb pairs and small compound decoction on mycoplasma pneumonia(MP)in vitro and secretion of inflammatory factors interleukins-6(IL-6),tumor necrosis factor-α(TNF-α),nuclear factor kappa B(NF-κB)in MP infected A549 cells.Methods Giant Knotweed,Barbed Slullcap,Cicada Slough were extracted with water and alcohol extraction respectively,and MP bacteria liquid was cultured by liquid culture method,and the minimum inhibitory concentration was determined by micro-dilution method.The toxic effect of extracts of Giant Knotweed,Barbed Slullcap,Cicada Slough,Giant Knotweed+Barbed Slullcap,Giant Knotweed+Cicada Slough,Barbed Slullcap+Cicada Slough and small compound decoction on A549 cells was detected by MTT,and the half inhibitory concentration(IC50)was calculated;the group with better antibacterial effect was selected,and the experiment was divided into normal group,model group,Giant Knotweed low,medium,and high ethanol extract groups,Barbed Slullcap low,medium,and high ethanol extract groups,Giant Knotweed+Barbed Slullcap low,medium,and high ethanol extract groups,small compound low,medium,and high concentration group,the viability of MP-infected A549 cells cultured for 12 h,24 h,36 h,48 h were detected by MTT in each group,and the levels of IL-6,TNF-α,NF-κB secreted by MP-infected A549 cells in each group were detected by ELISA.Results The water extracts and alcohol extracts of Giant Knotweed,Barbed Slullcap,Cicada Slough had a certain inhibiting effect on the growth of MP in vitro,the alcohol extract of small compound formula was the best,and the alcohol extract of drug pair was better than the water extract of drug pair,the alcoholic extract of single herbal medicine was better than the water extract,and the water extract and alcohol extract of Cicada Slough were the worst;the IC50 value of single medicine,medicine pair and small compound alcoholic extract on cytotoxicity was greater than that

关 键 词：肺炎支原体 中药提取物 体外抑菌 炎症因子 

分 类 号：R375.2[医药卫生—病原生物学]