组织MMP-13蛋白和基因表达在小鼠慢性变应性接触性皮炎中的意义

Tissue expression of protein and mRNA of MMP-13 in murine allergic contact dermatitis

目的检测皮肤组织基质金属蛋白酶-13(MMP-13)蛋白和基因在小鼠慢性变应性接触性皮炎(ACD)中的表达,并判断其表达与皮肤炎性反应表现的关联性及意义。方法 2016年1—9月在哈尔滨医科大学附属第一医院中心实验室进行实验,随机将140只小鼠分为空白对照组、模型组、CL-82198干预1组和干预2组,各35只,并按照实验设计分别予以相应处理,建立小鼠慢性ACD模型,干预1组和干预2组分别应用抑制剂CL-82198浓度0.5 mmol/L和1.25 mmol/L进行干预,观察小鼠皮肤炎性反应表现,并应用免疫组化方法和实时荧光定量PCR检测各组小鼠右耳组织中MMP-13蛋白和mRNA表达水平情况。结果模型组组织中MMP-13蛋白表达及mRNA表达水平显著高于空白对照组(χ~2=18.65,t=3.30,P均<0.01)。干预1组和干预2组应用MMP-13特异性抑制剂CL-82198干预后,小鼠皮肤炎性反应表现及组织病理学变化较模型组均明显减轻,其MMP-13蛋白和mRNA表达水平较模型组均显著减少(χ~2=4.77、16.57,t=2.16、2.95,P<0.01)。干预1组中MMP-13蛋白和mRNA表达水平较干预2组明显增高(χ~2=4.06,t=6.79,P<0.01),提示CL-82198的干预明显降低MMP-13蛋白和mRNA表达。结论(1)组织MMP-13在小鼠慢性ACD中呈高表达趋势,提示MMP-13可能参与了小鼠慢性ACD的发病。(2)MMP-13特异性抑制剂CL-82198能有效降低MMP-13的表达,并且在抑制慢性ACD的发生及发展中起重要作用。

Objective To detect the role of MMP-13 in murine chronic allergic contact dermatitis and determine the relationship and significance between skin inflammation and expression of MMP-13.Methods The experiment was carried out in central laboratory of the first affiliated hospital of Harbin Medical University from January 2016 to September 2016.One hundred and forty of mice were randomly divided into as follows：blank control group,model group,CL-82198 group（group1=CL-82198 0.5 mmol/L,group2=CL-82198 1.25 mmol/L）.Chronic ACD model was established, skin inflammatory re-sponse was observed.The expression of mRNA and protein level of MMP-13 in the skin was detected using RT-PCR and im-munohistochemical staining methods.Results （1）The expression of mRNA level and protein level of MMP-13 was signifi-cantly up regulated in model group as compared with that of the control group（t=3.30,χ2=18.65,all P〈0.001）.（2） The Skin inflammation performance and pathohistological changes in CL-82198 group were significantly decreased than that of model group.The levels of mRNA and protein of MMP-13 in CL-82198 group were significantly decreased than the model group（t=2.16,t=2.95,all P〈0.001,χ2=4.77,P=0.029,χ2=16.57,P〈0.001）.Compared with CL-82198 group 1,the levels of mRNA and protein of MMP-13 were increased significantly in CL-82198 group 2（t=0.79,P〈0.001,χ2=4.06,P=0.044）.Conclusion （1）The MMP-13 was highly expression in murine chronic ACD,suggesting that MMP-13 participate the development of murine chronic ACD.（2）The MMP-13 inhibitor CL-82198 inhibited DNCB induced skin in-flammation and thus may play role in ACD treatment.