加味升陷汤对慢性阻塞性肺病大鼠的治疗作用及机制  

Effect of modified Shengxian decoction in treating rats with chronic obstructive pulmonary disease and its mechanism

作　　者：吴艳蕊[1] 王艳琼 李建梅[1] 张孟月 张俊图[1] 刘杰 杨春艳[1] WU Yanrui;WANG Yanqiong;LI Jianmei;ZHANG Mengyue;ZHANG Juntu;LIU Jie;YANG Chunyan(Department of Pulmonary Diseases&Department of Respiratory and Critical Care Medicine,Kunming Hospital of Traditional Chinese Medicine,Kunming 650000,Yunnan,China;Integrated Traditional Chinese and Western Clinical Medicine,Yunnan University of Traditional Chinese Medicine,Kunming 650500,Yunnan,China)

机构地区：[1]昆明市中医医院肺病科/呼吸与危重症医学科,云南昆明650000 [2]云南中医大学,云南昆明650500

出　　处：《贵州医科大学学报》2025年第3期417-424,468,共9页Journal of Guizhou Medical University

基　　金：云南科学技术厅基础研究计划项目(202301AZ070001-093);昆明市科学技术局社会发展与科技惠民项目(2023-1-NS-027);昆明市卫生健康委员会卫生科研课题项目(2022-03-06-011,2022-19-02-004);云南省中医临床优秀人才研修项目(云卫中医综合发[2024]1)。

摘　　要：目的探讨加味升陷汤用于治疗慢性阻塞性肺病(chronic obstructive pulmonary disease,COPD)大鼠的效果及其调节辅助性T细胞17(T helper 17 cell,Th17)/调节性T细胞(regulatory T cell,Treg)平衡和肠道菌群的机制。方法45只雄性SPF级Wister雄性大鼠随机均分为对照组(不造模,吸入室内空气,正常饲养)、COPD组(造模,生理盐水灌胃)及治疗组(造模,加味升陷汤灌胃),造模后各组连续干预14 d,记录同期各组大鼠的一般情况(体质量、毛发的光滑度、脱毛情况、活动状态、呼吸频率、是否有张口呼吸、腹部膨胀及出汗情况);各组大鼠麻醉后取眼眶静脉血,采用流式细胞术检测血清中Th17和Treg的含量,采用ELISA检测血清中促炎因子白细胞介素-17A(interleukin-17A,IL-17A)及抗炎因子IL-10水平;取血后处死各组大鼠,取肺组织和肠道,采用Western blot和免疫组织化学法检测转录因子视黄酸相关孤儿核受体γt(retinoic acid-related orphan receptorγt,RORγt)和叉头框蛋白P3(forkhead box P3,Foxp3)的表达,采用HE染色检测肺组织学特征,采用16s rRNA基因测序分析肠道菌群功能。结果大鼠体质量表现为COPD组<治疗组<对照组(P<0.05),对照组大鼠其余一般情况正常、COPD组表现明显的病症、治疗组大鼠治疗后症状有所改善;大鼠血清Th17/Treg比值和IL-17A水平表现为COPD组>治疗组>对照组(P<0.05),血清IL-10水平表现为COPD组<治疗组<对照组(P<0.05);大鼠肺组织Foxp3蛋白表达表现为COPD组<治疗组<对照组(P<0.05),RORγt蛋白表达为对照组<治疗组<COPD组(P<0.05);肺功能方面,COPD组大鼠支气管周围有炎性细胞浸润、粘膜上皮增厚、肺泡增大,治疗组大鼠肺组织病理特征有所改善;肠道菌群功能分析结果显示,COPD组大鼠结肠微生物群中Sobs指数低于对照组(P<0.05)和治疗组;鼠肠道菌群与肺部炎症反应之间有相关性(P<0.05)。结论加味升陷汤可改善COPD大鼠的症状、降低炎症反应�Objective To investigate the effect of modified Shengxian decoction on rats with chronic obstructive pulmonary disease(COPD)and its mechanisms in regulating the balance between T helper 17(Th17)cells and regulatory T(Treg)cells as well as gut microbiota.Methods Forty-five male SPF-grade Wister rats were randomly divided into three groups:control(no modeling,inhalation of indoor air,normal feeding),COPD(modeling,intragastric administration of saline)and treatment(modeling,intragastric administration of modified Shengxian Decoction).After modeling,each group was continuously intervened for 14 days.The general conditions of the rats in each group at the same period were documented for body weight,hair smoothness,hair loss,activity status,respiratory rate,whether there were mouth breathing,abdominal distension and sweating.After anesthesia,rat retro-orbital sinus blood was collected from each group.Flow cytometry was used to detect the levels of helper T cells 17(Th17)and regulatory T cells(Treg).Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum pro-inflammatory cytokine interleukin-17A(IL-17A)and anti-inflammatory cytokine IL-10.After blood collection,the rats were sacrificed.Lungs and intestines were collected.Western blot and immunohistochemistry were used to detect the expressions of transcription factors retinoic acid receptor-related orphan receptorγt(RORγt)and forkhead box protein P3(Foxp3).Hematoxylin-eosin(HE)staining was applied to detect lung histological characteristics.16S ribosomal RNA(16S rRNA)gene sequencing was used to analyze the function of the intestinal microbiota.Results Rat body weight was as follows:COPD group<treatment group<control group(P<0.05).The rats in control group exhibited normal general conditions,while the rats in COPD group exhibited significant symptoms.The rats in treatment group showed improvement after treatment.Th17/Treg ratio and IL-17A level were as follows:COPD group>treatment group>control group(P<0.05).Serum IL-10 level was as follows:COPD

关 键 词：阻塞性肺病 慢性 大鼠 免疫调节 加味升陷汤 肠道菌群 

分 类 号：R563[医药卫生—呼吸系统]