基于16S rDNA技术探讨痰热清注射液对COPD模型大鼠肺部菌群的影响  

16S rDNA Sequencing Reveals Effect of Tanreqing Injection on Pulmonary Flora in Rat Model of COPD

作　　者：罗倩 符蕤 彭博[1] 陈韡亚 魏晓露[1] 陈腾飞 宋玲 高云航 张广平[1] 侯红平[1] LUO Qian;FU Rui;PENG Bo;CHEN Weiya;WEI Xiaolu;CHEN Tengfei;SONG Ling;GAO Yunhang;ZHANG Guangping;HOU Hongping(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2024年第23期98-103,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82104502);中央级公益性科研院所基本科研业务费专项(ZXKT23006,ZZ13-YQ-060,ZXKT21020);中国中医科学院科技创新工程项目(CI2021A04615)。

摘　　要：目的:本文通过痰热清注射液治疗慢性阻塞性肺疾病(COPD)模型大鼠,初步探讨痰热清注射液对COPD模型大鼠肺部菌群的影响。方法:Wistar大鼠随机分为正常组、模型组和痰热清组(TRQ),除正常组外,其余各组采用烟熏联合气管滴注脂多糖(LPS)制备COPD模型,TRQ组腹腔注射痰热清(2 g·kg^(-1))。实验终点,大鼠腹主动脉取血后采集肺组织进行苏木素-伊红(HE)及苦味酸-天狼猩红(PSR)染色,观察大鼠肺部病理状态;采集肺灌洗液,通过16S rDNA扩增子测序分析探讨肺部菌群在不同组别的多样性和相对丰度。结果:病理染色结果表明,正常组的肺脏正常,模型组肺脏出现中性粒细胞浸润和毛细血管扩张,个别动物还可见肺脏出血,肺气肿等改变,气管的胶原纤维明显增厚,TRQ组的肺脏有明显好转,胶原纤维有所改善。平均线性截距(MLI)分析显示,与正常组比较,模型组肺泡间的间隙增大(P<0.01);与模型组比较,TRQ组的肺泡间隙明显缩小(P<0.05);Wall thickness测量分析显示,与正常组比较,模型组Wall thickness显著升高(P<0.01);与模型组比较,TRQ组的Wall thickness显著降低(P<0.01)。菌群多样性分析结果表明,痰热清注射液升高simpson指数值,改变α多样性,主坐标分析结果显示痰热清注射液改变肺部菌群β多样性,降低β多样性指数。菌群丰富度分析表明,在属水平上,模型组芽孢杆菌属g_Bacillus、短波单胞菌属g_Brevundimonas相对丰度升高,假单胞菌属g_Pseudomonas相对丰度降低;经痰热清注射液治疗后,寡养单胞菌属g_Stenotrophomonas相对丰度大幅度升高,芽孢杆菌属g_Bacillus相对丰度降低。进一步通过LEfSe分析不同组别差异显著的菌群,结果表明,在属水平,模型组毛螺菌属g_Lachnospiraceae_NK4A136_group相对丰度升高,痰热清注射液治疗后红球菌属g_Rhodococcus、寡氧单胞菌属g_Stenotrophomonas相对丰度升高。结论:痰热清注射液调节COPD模型大鼠�Objective:To study the effect of Tanreqing injection(TRQ)on the pulmonary flora in the rat model of chronic obstructive pulmonary disease(COPD).Method:Wistar rats were randomized into control,model,and TRQ groups.The rats in other groups except the control group were treated by smoking combined with intratracheal instillation of lipopolysaccharide for the modeling of COPD.The TRQ group was intraperitoneally injected with TRQ(2 g·kg^(-1)).At the end of the experiment,after blood collection from the abdominal aorta of the rats,the lung tissue was collected for hematoxylin-eosin and picric sirius red staining to reveal the pathological changes.The lung lavage fluid was collected,and the diversity and relative abundance of lung flora in different groups were analyzed by 16S rDNA amplicon sequencing.Result:The lungs of the control group were normal,and those of the model group showed neutrophil infiltration,telangiectasia,lung hemorrhage and emphysema in individual cases,and thickening of collagen fibers in the trachea.Compared with the model group,the TRQ group showed significantly improved lungs and recovered collagen fibers.The MLI analysis showed that compared with the control group,the model group showcased increased alveolar space(P<0.01),which was reduced in the TRQ group(P<0.05).Compared with the control group,the model group showed increased wall thickness(P<0.01),and the increase was attenuated in the TRQ group(P<0.01).TRQ increased the Simpson index and altered theαdiversity of pulmonary flora.The results of principal co-ordinate analysis showed that TRQ changed theβdiversity and reduced theβdiversity index of pulmonary flora.At the genus level,the model group showed increased relative abundance of g_Bacillus and g_Brevundimonas and decreased relative abundance of g_Pseudomonas,compared with the control group.After treatment with TRQ,the relative abundance of g_Stenotrophomonas increased,and that of g_Bacillus decreased.The LEfSe of differential taxa between groups showed that the modeling increased the

关 键 词：痰热清注射液 慢性阻塞性肺疾病(COPD) 肺灌洗液 肺部菌群 微生物多样性 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33