摘要
目的:研究补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠肠道菌群的影响,探讨中药调控肠道菌群进而恢复肺-肠轴平衡的作用机制。方法:将大鼠随机分为7组,每组12只,即空白组、模型组、粪菌移植(FMT)组、地塞米松组及补肺汤低、中、高剂量组。除空白组外,其余各组采用香烟和锯末烟熏联合气管内滴入脂多糖(LPS)的方法建立COPD肺气虚证大鼠模型。补肺汤低、中、高剂量组分别灌胃补肺汤药液(3.645、7.29、14.58 g·kg^(-1)),FMT组给予粪菌液灌肠(10 mL·kg^(-1)),地塞米松组给予醋酸地塞米松片混悬液灌胃(0.135 mg·kg^(-1)),空白组和模型组大鼠灌胃等量蒸馏水,连续干预28 d后收集新鲜粪便,进行16S rRNA基因测序,取肺脏及结肠组织苏木素-伊红(HE)染色后进行病理形态学观察,酶联免疫吸附测定法(ELISA)检测肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)含量。结果:与空白组比较,模型组肺组织结构重度异常,肺泡萎缩塌陷,伴随炎症细胞重度浸润;与模型组比较,补肺汤高剂量组损伤程度明显改善,肺泡结构基本正常,炎症细胞浸润减少。与空白组比较,模型组结肠组织结构重度异常,黏膜层上皮细胞糜烂脱落,炎症细胞数量增多,黏膜下层水肿,间隙增大;与模型组比较,补肺汤中、高剂量组损伤程度明显改善,黏膜层上皮细胞排列整齐紧密,未见明显变性,黏膜层仅少量炎症细胞浸润。与空白组比较,模型组肺组织TNF-α、IL-8含量显著升高(P<0.01);与模型组比较,补肺汤低、中、高剂量组肺组织TNF-α、IL-8含量显著降低(P<0.01)。补肺汤明显调节模型大鼠的物种数量及Alpha、Beta多样性,纠正肠道菌群回归正常丰度与多样性,正向调节COPD肺气虚证模型大鼠的4种差异菌门(厚壁菌门、变形菌门等)与13种差异菌属(苏黎世杆菌属、乳杆菌属、厌氧螺菌属、肠单胞球菌属等),并下调2种碳水化合物代谢通路功能,包括磷酸戊糖途径(非氧化分支)Ⅰ和还原型戊糖磷酸循环。结论:补肺汤可调整肠道菌群物种丰度和多样性,影响代谢通路功能,修复肺及结肠组织结构,调节炎性因子水平,进而改善COPD肺气虚证,作用机制可能与其调节炎症相关的肠道菌群而恢复COPD肺气虚证肺-肠轴平衡有关。
Objective:To investigate the effect of Bufeitang on intestinal flora of rats with lung Qideficiency syndrome of chronic obstructive pulmonary disease(COPD),and to explore the mechanism of traditional Chinese medicine in regulating intestinal flora and thus restoring the balance of lung-gut axis.Method:A total of 84 rats were randomly divided into 7 groups,including blank group,model group,fecal bacterial transplantation(FMT)group,dexamethasone group and low,medium and high dose groups of Bufeitang,12 rats in each group.Except for the blank group,cigarette and sawdust fumigation combined with intratracheal instillation of lipopolysaccharide(LPS)were used to establish the COPD rat model with lung Qideficiency syndrome in all other groups.The low,medium and high dose groups of Bufeitang were intragastric administrated with Bufeitang(3.645,7.29,14.58 g·kg^(-1)),the FMT group was given fecal bacteria liquid enema(10 mL·kg^(-1)),dexamethasone group was given dexamethasone acetate tablet suspension by gavage(0.135 mg·kg^(-1)),the blank group and model group were given equal amount of distilled water.Fresh feces were collected after 28 d of continuous intervention for 16S rRNA gene sequencing.Lung and colon tissues were stained with hematoxylineosin(HE)for pathomorphological observation,and enzyme-linked immunosorbent assay(ELISA)was performed to detect the contents of tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)in lung tissues.Result:Compared with the blank group,the model group showed severe abnormal lung tissue structure with alveolar atrophy and collapse accompanied by severe inflammatory cell infiltration.Compared with the model group,the extent of injury was significantly improved,and inflammatory cell infiltration was reduced with basically normal alveolar structure in the high dose group of Bufeitang.Compared with the blank group,the model group had severely abnormal colonic tissue structure,the epithelial cells in the mucosal layer were eroded and shed,the number of inflammatory cells increased,the submucosal layer was edematous and the gap was enlarged.Compared with the model group,the extent of damage was significantly improved in the medium and high dose groups of Bufeitang,the epithelial cells in the mucosal layer were neatly and closely arranged,with only a small amount of inflammatory cell infiltration and no significant degeneration.Compared with the blank group,the TNF-αand IL-8 levels of lung tissue in the model group were significantly increased(P<0.01).Compared with the model group,the TNF-αand IL-8 levels of lung tissues in the low,medium and high dose groups of Bufeitang were significantly decreased(P<0.01).Bufeitang significantly modulated the number of bacteria species as well as alpha and beta diversity of model rats,corrected the return of intestinal flora to normal abundance and diversity,and positively regulated 4 differential phyla(such as Firmicutes,Proteobacteria)and 13 differential genera(such as Turicibacter,Lactobacillus,Anaerobiospirillum,Intestinimonas)in COPD model rats with lung Qi-deficiency syndrome,and down-regulated 2 carbohydrate metabolic pathway functions,including the pentose phosphate pathway(non-oxidative branch)Ⅰand the Calvin-Benson-Bassham cycle.Conclusion:Bufeitang can modulate the abundance and diversity of intestinal flora species,affect the function of metabolic pathways,repair the structure of lung and colon tissues,regulate the level of inflammatory factors,and thus improve COPD with lung Qi-deficiency syndrome.The mechanism may be related to its regulation of inflammation-related intestinal flora to restore the balance of lung-gut axis in COPD with lung Qi-deficiency syndrome.
作者
沈俊希
朱星
陈云志
刘怀全
褚璨灿
张钰
苏钢
李文
徐昌君
童平珍
余欣然
杨光勇
邓颖
SHEN Junxi;ZHU Xing;CHEN Yunzhi;LIU Huaiquan;CHU Cancan;ZHANG Yu;SU Gang;LI Wen;XU Changjun;TONG Pingzhen;YU Xinran;YANG Guangyong;DENG Ying(Guizhou University of Traditional Chinese Medicine(TCM),Guiyang 550025,China;Peopleʹs Hospital of Anshun City,Anshun 561000,China;The Second Affiliated Hospital of Guizhou University of TCM,Guiyang 550003,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第7期47-56,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81960830)。
