基于TGF-β/RORγt/FoxP3通路探讨扶正透邪方对铜绿假单胞菌肺炎大鼠肺损伤的影响  

Effects of Fuzheng Touxie Prescription on Lung Injury of Pseudomonas Aeruginosa Pneumonia Rats Based on TGF-β/RORγt/FoxP3 Pathway

作　　者：李奕璇 刘清泉[2] 孔令博[1] 张迪 晏军[1] LI Yixuan;LIU Qingquan;KONG Lingbo;ZHANG Di;YAN Jun(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]首都医科大学附属北京中医医院,北京100010

出　　处：《中国中医药信息杂志》2025年第3期98-104,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：首都卫生发展科研专项(首发2020-2-4192);北京中医药大学东直门医院科技创新专项(DZMKJCX-2020-027)。

摘　　要：目的观察扶正透邪方对铜绿假单胞菌肺炎(PAP)大鼠肺损伤的影响并探讨其可能的作用机制。方法将36只SD大鼠按随机数字表法分为空白组、模型组、哌拉西林组和扶正透邪方低、中、高剂量组,每组6只,除空白组外,各组气管滴注3×10~9 CFU/mL铜绿假单胞菌悬液0.1 mL,空白组滴注等量无菌生理盐水。造模后24 h,扶正透邪方低、中、高剂量组分别予0.43、0.86、1.73 g/mL扶正透邪方药液1.6 mL灌胃,空白组、模型组予等体积蒸馏水灌胃,哌拉西林组予0.11 g/mL哌拉西林钠他唑巴坦钠溶液0.53 mL腹腔注射,均每日1次,连续3 d。HE染色观察肺组织形态,进行肺损伤评分并测量支气管壁厚度,流式细胞术检测外周血Th17/Treg比例,ELISA检测血清维甲酸相关核孤儿受体γt(RORγt)、叉头框蛋白P3(FoxP3)、转化生长因子-β(TGF-β)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-10含量,RT-qPCR检测肺组织RORγt、FoxP3及炎症因子mRNA表达,Western blot检测肺组织RORγt、FoxP3及炎症因子蛋白表达。结果与空白组比较,模型组大鼠肺组织可见大量炎性细胞浸润,伴支气管管壁增生肥厚、肺泡扩张、肺间质水肿,肺损伤评分和支气管壁厚度明显增加(P<0.01),外周血Th17比例升高(P<0.01),Treg比例降低(P<0.01),Th17/Treg比值升高(P<0.05),血清RORγt、TNF-α、TGF-β、IL-6含量显著升高(P<0.01),FoxP3、IL-10含量显著下降(P<0.01),肺组织RORγt、TNF-α、TGF-β、IL-6 mRNA和蛋白表达显著升高(P<0.01),FoxP3、IL-10 mRNA和蛋白表达显著降低(P<0.01);与模型组比较,各给药组大鼠肺泡相对完整、结构较清晰,炎性细胞浸润、肺间质水肿充血减轻,肺损伤评分和支气管壁厚度减少(P<0.05,P<0.01),扶正透邪方高剂量组和哌拉西林组外周血Th17比例降低(P<0.05),Treg细胞比例升高(P<0.05),Th17/Treg比值降低(P<0.05),扶正透邪方中、高剂量组和哌拉西林组大鼠血清RORγt、TNF-α、TGF-β、IL-Objective To investigate the effects of Fuzheng Touxie Prescription on pulmonary injury of Pseudomonas aeruginosa pneumonia(PAP)in rats;To discuss its possible mechanism.Methods A total of 36 SD rats were divided into blank group,model group,piperacillin group and Fuzheng Touxie Prescription low-,medium-and high-dosage groups according to random number table method,with 6 rats in each group.Except for the blank group,0.1 mL of Pseudomonas aeruginosa suspension with a concentration of 3×109 CFU/mL was injected through oral catheterization.In the blank group,0.1 mL sterile saline was injected by oral catheterization.Fuzheng Touxie Prescription low-,medium-and high-dosage groups were gavaged 1.6 mL of Fuzheng Touxie Prescription solution with the concentration of 0.43,0.86 and 1.73 g/mL respectively,the blank group and model group were gavaged equal volume of distilled water,piperacillin group was intraperitoneal injected 0.53 mL piperacillin tazobactam sodium solution with a concentration of 0.11 g/mL,once a day,for consecutive 3 days.HE staining was used to observe the pathological changes of lung tissue,lung injury score was performed and bronchial wall thickness was measured,flow cytometry was used to detect Th17/Treg ratio in peripheral blood,the serum contents of RORγt,FoxP3,TGF-βand inflammatory factors TNF-α,IL-6 and IL-10 were detected by ELISA,the mRNA expression of RORγt,FoxP3 and inflammatory factors in lung tissue were detected by RT-qPCR,the protein expressions of RORγt,FoxP3 and inflammatory factors in lung tissue were detected by Western blot.Results Compared with the blank group,a large amount of inflammatory cell infiltration was observed in lung tissue of the model group rats,accompanied by bronchial wall hyperplasia,alveolar dilation,interstitial edema,and a significant increase in lung injury score and bronchial wall thickness(P<0.01),the proportion of Th17 in peripheral blood increased(P<0.01),the proportion of Treg decreased(P<0.01),and the Th17/Treg ratio increased(P<0.05),the contents

关 键 词：扶正透邪方 铜绿假单胞菌肺炎 TGF-β/RORγt/FoxP3 大鼠 

分 类 号：R285.5[医药卫生—中药学]