宣肺解毒方对多重耐药铜绿假单胞菌肺炎的作用机制探讨  

Mechanism of Xuanfei Jiedu Formula in treating multi⁃drug⁃resistant Pseudomonas aeruginosa pneumonia

作　　者：沈婷婷[1,2] 李亚[2] 李素云[3,4] 李高峰[1,2] 韩冰洋 SHEN Tingting;LI Ya;LI Suyun;LI Gaofeng;HAN Binyang(Henan University of Chinese Medicine,Zhengzhou 450046,China;Chinese Medicine Pharmacology(Respiratory)Laboratory,the First Affiliated Hospital of Henan University of Chinese Medicine Henan Key Laboratory of Traditional Chinese Medicine for the Prevention and Treatment of Respiratory Diseases,Zhengzhou 450000,China;Respiratory Department of the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Co-Construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of China,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学,郑州450046 [2]河南中医药大学第一附属医院中心实验室中药药理(呼吸)实验室河南省呼吸病防治中医药重点实验室,郑州450000 [3]河南中医药大学第一附属医院呼吸科,郑州450000 [4]河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,郑州450046

出　　处：《中国实验动物学报》2024年第6期721-732,共12页Acta Laboratorium Animalis Scientia Sinica

基　　金：河南省中医药科学研究专项(2023ZYZD03,2022JDZX124);河南省医学科技攻关计划项目(LHGJ20220573);中医药传承创新专项(2022CCCX011);河南省科技研发计划联合基金(14207908)。

摘　　要：目的探究宣肺解毒方对多重耐药铜绿假单胞菌肺炎的机理研究。方法除空白组外,其余组采用气管插管的方法建立MDR-PA(9×10^(8) CFU/mL,0.5 mL)肺炎大鼠模型,造模成功后随机分为模型组、宣肺解毒方低剂量组、宣肺解毒方中剂量组、宣肺解毒方高剂量组以及亚胺培南西司他丁组,每组12只;以上除空白组与模型组外,剩余给药组统称为干预治疗组。造模1 d后,宣肺解毒方低、中、高剂量组分别给予相应剂量的宣肺解毒方(Xuanfei Jiedu Formula,XFJDF)灌胃,亚胺培南西司他丁组给予亚胺培南西司他丁(imipenem,IPM)腹腔注射,空白组和模型组给予生理盐水灌胃,每天2次,持续7 d。观察大鼠的一般状态、体重变化、肺湿重/肺干重(W/D),采用苏木素-伊红(HE)染色法于光镜下观察大鼠肺组织病理学改变,采用酶免疫吸附测定法检测大鼠血清中IL-1β、TNF-α、TGF-β、IL-6炎症因子水平,采用比色法检测大鼠血清中GSH含量与MPO活性,采用TBA法检测大鼠血清中MDA含量,试剂盒法检测总抗氧化能力T-AOC;采用免疫组化法对肺组织中TLR4、Myd88、NF-κBp65蛋白进行定位与半定量观察,采用qPCR和Western Blot技术检测大鼠肺组织中TLR4、Myd88、NF-κBp65 mRNA和蛋白表达水平。结果与空白组比,模型组大鼠反应迟缓,呼吸频率加快、呼吸杂音增多且出现不同程度寒颤,饮食饮水减少,体重下降;肺W/D(P<0.01)显著升高,肺组织的肺泡腔以及肺支气管周围存有大量炎性细胞浸润,部分肺泡壁出现断裂融合形成气腔并伴有炎性渗出,肺间质增厚,局部可见肺纤维形成等;血清中IL-1β、TNF-α、TGF-β、IL-6水平明显升高(P<0.01),MDA含量增加、MPO活性增强、GSH含量与T-AOC能力降低(P<0.01),肺组织中TLR4、Myd88、NF-κBp65 mRNA和蛋白表达显著升高(P<0.01)。与模型组相比,干预治疗组均不同程度改善上述指标变化(P<0.05,P<0.01),以宣肺解毒方高剂量组以及亚�Objective Study on mechanism of Xuanfei Jiedu Formula in treating multi⁃drug resistant Pseudomonas aeruginosa pneumonia.Methods Except the Control group,the MDR⁃PA(9×10^(8) CFU/mL,0.5 mL)pneumonia rat model was established by tracheal intubation,and an un⁃modeled control group was used.After successful modeling,rats were randomly divided into model group,XFJDF⁃low dose group,XFJDF⁃medium dose group,XFJDF⁃high dose group and IPM group,with 12 animals in each group;In addition to the blank group and the model group,the remaining drug administration groups were collectively referred to as the intervention treatment group.One day after modeling,the XFJDF⁃low dose,XFJDF⁃medium dose,and XFJDF⁃high dose groups were given the corresponding doses by gavage.The imipenem and cilastatin group was given Imipenem intraperitoneal injection,and the control and model groups were given saline gavage twice a day for 7 days.The rats’general status,body weight changes,and wet⁃dry weight ratio(W/D)of the lung tissue were observed.Hematoxylin⁃eosin staining was used to observe the pathological changes to the lung tissue of rats in each group under a light microscope.The serum levels of IL⁃1β,TGF⁃β,TNF⁃α,and IL⁃6 were detected by enzyme immunosorbent assay.Serum GSH content and MPO activity of rats were detected by colorimetry.The serum content of MDA was detected by TBA method.The total antioxidant capacity(T⁃AOC)was detected by kit method.The protein levels of TLR4,Myd88,and NF⁃κBp65 in lung tissues were determined by immunohistochemistry.The mRNA and protein levels of TLR4,Myd88,and NF⁃κBp65 in the lung tissues of each group were detected by qPCR and Western Blot.Results Compared with the control group,the model groups had delayed responses,increased respiratory frequency,increased respiratory noise,and appeared to have different degrees of chills,as well as decreased diet and water intake and decreased body weight.The W/D of lung tissue was significantly increased(P<0.01),and a large numbe

关 键 词：多重耐药菌 铜绿假单胞菌 肺炎 宣肺解毒方 

分 类 号：Q95-33[生物学—动物学]