基于PLA2-TRPV1/TRPA1通路探讨苓桂术甘汤对慢性支气管炎的抗炎化痰作用  

Linggui Zhugantang Treats Chronic Bronchitis in Rats via PLA2-TRPV1/TRPA1 Pathway

作　　者：丁薇 汪文来[2] 刘珍洪[3] 陈香云 何湛湛 褚策 袁雨露 许咏琪 张雨欣 赵培彰 杨桢 赵红霞[2] DING Wei;WANG Wenlai;LIU Zhenhong;CHEN Xiangyun;HE Zhanzhan;CHU Ce;YUAN Yulu;XU Yongqi;ZHANG Yuxin;ZHAO Peizhang;YANG Zhen;ZHAO Hongxia(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute for Brain Disorders,Beijing University of Chinese Medicine,Beijing 100700,China;West China School of Medicine,Sichuan University,Chengdu 610041,China)

机构地区：[1]北京中医药大学中医学院,北京102488 [2]中国中医科学院中医基础理论研究所,北京100700 [3]北京中医药大学中医脑病研究院,北京100700 [4]四川大学华西临床医学院,成都610041

出　　处：《中国实验方剂学杂志》2024年第14期1-9,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院科技创新工程黑地黄丸重大攻关项目(CI2021A00606);国家自然科学基金项目(82104822);中国中医科学院自主选题项目(YZX202237,YZX202241)。

摘　　要：目的:研究苓桂术甘汤对香烟烟雾暴露联合脂多糖(LPS)气道滴注诱导的慢性支气管炎(CB)大鼠的疗效作用机制。方法:将60只SPF级SD大鼠按体质量分层法随机分为正常组、模型组、地塞米松组(1 mg·kg^(-1))、苓桂术甘汤高、中、低剂量组(30.06、15.03、7.515 g·kg^(-1))6组,每组10只。各组造模第1天、第14天分别予LPS(1 g·L^(-1))200μL气道滴注,正常组予等体积的生理盐水。除正常组外,其余各组第2~13天、15~30天予香烟烟雾暴露(10支/次/30 min,2次/d)构建大鼠CB模型。给药组分别在造模第2天连续灌胃30 d,观测大鼠精神行为状态与体质量。造模后30 d取大鼠肺组织,测定左肺湿/干质量比(W/D);苏木素-伊红(HE)染色观察肺组织及支气管病理形态学变化;阿利新蓝-过碘酸雪夫(AB-PAS)染色观察支气管黏液分泌情况与杯状细胞增生程度;酶联免疫吸附测定法(ELISA)检测血清中黏糖蛋白5AC(MUC5AC)、白细胞介素-13(IL-13)、白细胞介素-6(IL-6)与肿瘤坏死因子-α(TNF-α)的含量;免疫组化与全自动蛋白免疫印迹法(Western blot)检测肺组织中磷脂酶A2(PLA2)、热敏通道瞬时感受器电位香草素受体1(TRPV1)与瞬时感受器电位锚蛋白1(TRPA1)的表达变化。结果:与正常组比较,模型组大鼠精神憔悴,弓背蜷腹,口鼻有血性分泌物,死亡率为40%,体质量增长趋势显著下降,肺支气管结构损害严重,管壁大量炎性介质与炎症细胞浸润,充血水肿与纤维增生,杯状细胞大量增生,黏液过量分泌堆积,管腔明显狭窄变形,且肺水肿显著加重(P<0.01);血清MUC5AC、IL-13、IL-6、TNF-α及肺PLA2的组织表达与蛋白表达水平显著上调(P<0.01);与模型组比较,苓桂术甘汤高、中、低3组反应灵敏,无异常行为与分泌物,死亡率低,体质量持续增长稳定,肺支气管损伤减轻,杯状细胞增生与黏液分泌减少,肺水肿显著减轻(P<0.01);血清MUC5AC、IL-13、IL-6、TNF-α及肺PLA2、TRPA1、TRObjective:To study the effect and mechanism of Linggui Zhugantang in treating chronic bronchitis(CB)induced by exposure to cigarette smoke combined with tracheal instillation of lipopolysaccharide(LPS).Method:Sixty SPF-grade SD rats were randomly divided into normal,model,dexamethasone(1 mg·kg^(-1)),and high-,medium-,and low-dose(30.06,15.03,7.515 g·kg^(-1),respectively)Linggui Zhugantang groups by the body weight stratification method,with 10 rats in each group.Each group was administrated with 200μL LPS(1 g·L^(-1))by tracheal instillation on days 1 and 14,respectively,while the normal group was administrated with an equal volume of normal saline.Except the normal group,the other groups were exposed to cigarette smoke on days 2-13 and 15-30(10 cigarettes/time/30 min,twice/day)for the modeling of CB.The rats were administrated with corresponding drugs by gavage for 30 consecutive days from day 2 of modeling,and the mental status,behavior,and body weights of the rats were observed and measured.The wet/dry mass ratio(W/D)of the left lung was measured 30 days after modeling.Hematoxylin-eosin staining was employed to observe the pathological changes in the lung and bronchial tissues.The bronchial mucus secretion and goblet cell proliferation were observed by Alcian blue-periodic acid Schiff(AB-PAS)staining.The levels of mucin 5AC(MUC5AC),interleukin(IL)-13,IL-6,and tumor necrosis factor(TNF)-αin the serum were determined by enzyme-linked immunosorbent assay.The expression of phospholipase A2(PLA2),transient receptor potential vanilloid receptor 1(TRPV1),and transient receptor potential ankyrin 1(TRPA1)in the lung tissue was quantitatively analyzed by immunohistochemistry and Western blot.Result:Compared with the normal group,the model group showcased abnormal mental status and behaviors,bloody secretion in the nose and mouth,the mortality rate of 40%,decreased body weight,severe lung bronchial structure damage,a large number of inflammatory mediators and inflammatory cell infiltration in the tube wall,hyperemia,

关 键 词：苓桂术甘汤 慢性支气管炎 气道黏液高分泌 炎症 瞬时感受器电位(TRP)通道 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33