TPA致小鼠耳肿胀模型的中性粒细胞聚集及IL-1β、IL-6、TNF-α、IL-17A的表达水平研究  

Neutrophil aggregation and the expression levels of IL-1β,IL-6,TNF-α,and IL-17A in TPA-induced mouse ear edema model

作　　者：周娟[1] 张梦军[1] 郭嘉伟[1] 杨超[2] 田阳[2] 张惠静[1] 李滨 

机构地区：[1]第三军医大学药学院药物分析与分析化学教研室,重庆400038 [2]第三军医大学药学院学员旅13队 [3]重庆市科学技术研究院,401123

出　　处：《免疫学杂志》2012年第10期872-876,共5页Immunological Journal

基　　金：重庆市科技攻关计划项目(CSTC;2010AB5099);国家自然科学基金项目(31100653);国家科技重大专项(2012ZX09103-101-080)

摘　　要：目的从蛋白水平和mRNA水平探讨佛波酯(TPA)诱导的小鼠炎症耳组织中IL-1β、IL-6、TNF-α和IL-17A的表达情况。方法通过HE染色观察TPA致小鼠耳组织的炎症水肿和淋巴细胞浸润,测定髓过氧化物酶(myeloperoxidase,MPO)活性判断中性粒细胞的聚集情况,ELISA法检测IL-1β、IL-6、TNF-α、IL-17A的蛋白含量以及实时荧光定量PCR法检测它们的mRNA水平。结果与正常组相比,模型组小鼠耳组织有明显的炎症水肿和淋巴细胞浸润,中性粒细胞聚集严重,并且该组小鼠耳组织中IL-1β、IL-6、TNF-α的含量和mRNA水平显著地增高,而IL-17A没有明显地变化;阳性药物组与模型组相比,能有效地抑制小鼠耳组织的炎症水肿,淋巴细胞浸润,中性粒细胞聚集以及下调各炎症细胞因子的蛋白含量和mRNA水平。结论 TPA诱导的小鼠耳肿胀严重程度除了与淋巴细胞浸润和中性粒细胞聚集有关外,还与靶组织中显著上调的IL-1β、TNF-α和IL-6蛋白含量和mRNA水平有关。In this study,we aim to explore the expression of IL-1β,IL-6,TNF-α,and IL-17A in 12-O-tetradecanoylphorbol-13 acetate（TPA）-induced mouse ear edema tissue at protein level and mRNA level.Transverse sections of mouse ears were stained with hematoxylin-eosin for examining the edema and lymphocyte infiltrate under light microscopy.Aggregation of these activated neutrophils was determined by assaying the activity of myeloperoxidase.Then we measured the levels of protein and mRNA of IL-1β,IL-6,TNF-α,and IL-17A by ELISA and real-time RT-PCR.Compared with normal group,model group had obviously edema and lymphocyte infiltration,and the infiltration of activated neutrophils was serious.At the same time,we found that the levels of protein and mRNA of IL-1β,IL-6,TNF-α,and IL-17A had increased obviously,but IL-17A did not change.Furthermore,compared with model group,positive control（treated with dexamethasone） effectively inhibited the TPA-induced ear edema,lymphocyte infiltrate,neutrophils gather,as well as IL-1β,IL-6,and TNF-α cytokine levels.These results indicated that mouse ear edema induced by TPA may be not only due to the lymphocyte infiltrate and neutrophils gather,but also associate with the protein and mRNA levels of IL-1β,IL-6,and TNF-α.

关 键 词：佛波酯诱导的小鼠耳肿胀模型 白细胞介素-1Β 白细胞介素-6 肿瘤坏死因子-α 白细胞介素-17A 

分 类 号：R-332[医药卫生]