重组小鼠IL-36α成熟肽制备及其在IBD中作用的初步研究  

Preparation of recombinant mouse IL-36α mature peptide and its preliminary study on role in IBD

作　　者：富文达 朱俊丰 乔丽[1,3] 孙逊(指导) FU Wen-Da;ZHU Jun-Feng;QIAO Li;SUN Xun(Department of Immunology,China Medical University,Shenyang 110122,China)

机构地区：[1]中国医科大学免疫学教研室,沈阳110122 [2]辽宁大学生命科学院,沈阳110036 [3]北部战区总医院北陵院区,沈阳110031

出　　处：《中国免疫学杂志》2020年第4期457-460,466,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(81771753);辽宁省“兴辽英才计划”项目(XLYC1807108)资助

摘　　要：目的:制备重组小鼠IL-36α成熟肽,并对其在炎症性肠病(IBD)中的作用进行初步研究。 方法:对小鼠IL-36α成熟肽基因进行密码子优化并化学合成,将其插入原核表达载体 pET28a(+),构建pET28a-IL-36α质粒,转化至大肠杆菌BL21(DE3),经IPTG诱导表达,镍亲和层析法纯化制备IL-36α成熟肽;利用Western blot鉴定IL-36α成熟肽,使用制备的重组小鼠IL-36α成熟肽干预DSS诱导的小鼠IBD模型,检测模型小鼠体重、生存率、疾病活动指数、肠管长度及组织病理学等变化情况。 结果:经SDS-PAGE和Western blot分析获得纯度高达95%的IL-36α成熟肽。与对照组相比,IL-36α成熟肽处理组IBD小鼠的体重下降减少、生存率增高、疾病活动指数降低、肠管长度及组织病理学得到明显改善。结论:成功制备重组小鼠IL-36α成熟肽,初步证实了IL-36α对小鼠IBD模型的炎症抑制作用,为进一步研究IL-36在炎症性肠病中的免疫调节功能奠定基础。Objective:To prepare a recombinant mouse IL-36α mature peptide and test its immunological activity in IBD mouse model.Methods:The gene of mouse IL-36α optimized by codon preference analysis was obtained by chemical synthesis.The gene was inserted into the prokaryotic expression vector pET28a (+).The pET28a-IL-36α plasmid was constructed and transformed into E.coli BL21 (DE3).The expression of IL-36α was induced by IPTG.The purification method of IL-36α mature peptide was nickel affinity chromatography.The recombinant IL-36α mature peptide that was identified by Western blot was used to treat the mice with DSS-induced colitis.The body weight,survival rate,disease activity index,colon length and histological score was detected.Results:Recombinant IL-36α mature peptide with purity up to 95% was confirmed by SDS-PAGE and Western blot analysis.Compared with the control group,IL-36α mature peptide treatment group showed a significant alleviation on weight loss,survival,disease activity index,colon length and histological score in DSS-induced colitis.Conclusion:Recombinant mouse IL-36α mature peptide was successfully prepared and the inhibitory effect of IL-36α on DSS-induced colitis is preliminarily confirmed,which lays the foundation for further study of the immune regulation function of IL-36 in inflammatory bowel disease.

关 键 词：白细胞介素36α成熟肽 原核表达 炎症性肠病 抑制作用 

分 类 号：R392.11[医药卫生—免疫学]