大小鼠气囊炎模型实验方案:1) 随机分组,每组10只;2) 于试验第一天在大鼠背部皮下注射无菌空气20ml(空气经0.22μ过滤器滤过),第3天、第6天分别再次注入10ml无菌空气以维持气囊的膨胀。3) 于试验第一天开始给药到实验结束。4) 第6天在给药1小时后,再次注入空气,并随即向气囊内注入1%角叉菜胶致炎,剂量为2ml/只,正常对照组注入等量的溶媒。5) 在致炎24h后麻醉处死大鼠,用4ml的PBS灌洗气囊,记录灌洗液量,以此计算渗出液量。6) 进行相关指标检测(TNF-a, IL-6等细胞因子,中性粒细胞和巨噬细胞等)。
Brief protocol of air pouch model: After one week acclimation, animals will be randomly divided;1) Day 0: s.c. inflation at the backside of mice with 20ml sterile air;2) Day 3: re-inflation with 2.5 ml sterile air to maintain the pouch;3) Day 6: re-inflation with 2.5 ml sterile air to maintain the pouch;4) 24 hour after stimulation: flush the pouch with 4mL PBS and lavage will be collected for further analysis. Both cytokine profile and cell differentiation will be measured.5) For cytokine profile, TNF-a and IL-6 will be measured by ELISA according to the manufacturers instruction.6) For cell differentiation, total cell count will be recorded by cell counter and cell differentiation will be measured by FACS. Ly-6G and CD11b will be the cell surface marker for Neutrophils, F4/80 and CD11b will be the cell surface marker for Macrophage.