寒喘舒和安喘舒对过敏性支气管哮喘豚鼠细胞因子的影响  

Effects of Hanchuashu and Anchuanshu on Cytokines in A Guinea Pig Model of Allergic Bronchial Asthma

作　　者：张丽玲[1] 张丽萍 朱澄[1] 郭超[1] 顾晓燕[1] 梁卫[1] 周斌[1] 

机构地区：[1]中国人民解放军第四五四医院中医科,南京210002 [2]江西省职业病研究院职业病医院,南昌330006

出　　处：《南昌大学学报（医学版）》2013年第2期45-50,54,共7页Journal of Nanchang University：Medical Sciences

基　　金：南京军区医学科技创新课题(11MBO14)

摘　　要：目的通过对豚鼠整体行为表现、病理组织切片的观察,炎性细胞(嗜酸性粒细胞、中性粒细胞、淋巴细胞)、炎性细胞因子(IL-1β、TNF-α、IL-4、IL-8、IL-13)和抗炎细胞因子(IL-10)在豚鼠支气管肺泡灌洗液(BALF)中含量的变化,初步探讨寒喘舒和安喘舒治疗过敏性支气管哮喘模型豚鼠的作用机制。方法将188只豚鼠按随机数字表法分为6组,应用鸡卵白蛋白雾化吸入法建立稳定的豚鼠过敏性支气管哮喘模型。正常对照组(予生理盐水模拟抗原攻击,A组,n=20);过敏性支气管哮喘模型组(予生理盐水模拟治疗,B组,n=40);寒喘舒组(C组)予低剂量(C1组,n=32)、高剂量(C2组,n=32)寒喘舒治疗;安喘舒组(D组)予低剂量(D1组,n=32)、高剂量(D2组,n=32)安喘舒治疗。肉眼观察各组动物饮食、活动、抓鼻、咳嗽、喷嚏等过敏行为变化;各组于治疗后1、2、8、24h处死动物,观察肺色泽、大小等;用生理盐水行支气管肺泡灌洗,收集BALF,离心后取沉淀细胞作细胞涂片,计数细胞;用Lowrys法测量BALF上清液中总蛋白含量,用双抗夹心ELISA法测定上清液中TNF-α、IL-1β、IL-4、IL-8、IL-10、IL-13的含量,计算各时间点各组细胞因子含量与总蛋白、抗炎因子与致炎因子(IL-10/TNF-α、IL-10/IL-1β)的比值。结果光镜下可见:A组肺泡管和肺泡壁结构完整;B组肺泡管和肺泡壁结构受损,肺泡腔内充满渗出液,可见坏死脱落的肺泡上皮细胞和渗出的白细胞,肺间质水肿,炎性细胞浸润;C1、C2组肺泡管和肺泡壁结构受损程度轻于B组,肺泡腔内见极少量炎症细胞游于其中,支气管管腔及肺泡周围少见炎性细胞浸润。D1、D2组组织结构基本正常,未见明显损害。BALF中细胞学变化:与B组相比,C1、C2、D1、D2组嗜酸性粒细胞、中性粒细胞、淋巴细胞数量显著下降(P<0.05),巨噬细胞显著上升(P<0.05)。BALF中细胞因子含量变化:C1、C2、D1、D2组BALF炎性因子IL-1β、TNFObjective To explore the action mechanisms of Hanchuashu and Anchuanshu in aguinea pig model of allergic bronchial asthma through investigating overall behavior of animals, histopathological changes,and contents of inflammatory cells （eosinophils, neutrophils and lym- phocytes）, inflammatory cytokines （IL-1β, TNF-α, IL-4, IL-8, IL-13） and anti-inflammatory cyto- kines （IL-10） in bronchoalveolar lavage fluid （BALF）. Methods A total of 188 guinea pigs were randomly divided into six groups：normal saline control group （group A,n= 20）, allergic bronchial asthma model group （group B,n=40）, low-dose Hanchuanshu treatment group（group Cl,n= 32）, high-dose Hanehuanshu treatment group（group C2,n=32）,low-dose Anchuanshu treatment group（group D1 ,n=32） and high-dose Anchuanshu treatment group（group Dl,n=32）. Allergic bronchial asthma was induced by chicken ovalbumin inhalation. Animal diet,daily activity, scratc- hing nose, coughing, sneezing and other allergic manifestations were observed. Animals were sac- rificed 1,2,8 and 24 hours after treatment and pulmonary color and size were recorded. In addi- tion,bronchoalveolar lavage was performed with normal saline. BALF was collected for prepara- tion of cell smears after centrifugation and cells were counted. Total protein content in BALF was measured by Lowry＇s method. The contents of TNF-α,IL-1β,IL-4,IL-8,IL-10 and IL-13 were de- termined by double-antibody sandwich ELISA. The ratios of eytokines to total protein, anti-in- flammatory cytokines and inflammatory cytokines were calculated at each time point. Results Optical microscopy showed complete alveolar ducts and alveolar walls in group A,D1 and D2,and destruction of alveolar walls and alveolar ducts in group B, C1 and C2. Furthermore,alveolar space exudate,alveolar epithelial cell necrosis, leukocyte exudation ;pulmonary interstitial edema, in- flammatory cell infiltration, pulmonary interstitial edema and inflammatory cell infiltration were found in group B,and a very smal

关 键 词：支气管哮喘 过敏性 寒喘舒 安喘舒 鸡卵白蛋白 支气管肺泡灌洗液 细胞因子 动物 实验 豚鼠 

分 类 号：R332[医药卫生—人体生理学]