摘要
目的探讨重组可溶性白细胞介素-4受体(IL-4R)对大鼠哮喘模型的影响。方法根据Gen Bank公布的Homo sapiens(human)IL-4R基因序列(XM_005255309),合成srh IL-4R基因片段,将合成基因片段插入p ET-28a(+)构建重组表达载体p ET-28a(+)-srh IL-4R,酶切鉴定后,将载体转入表达菌株BL21(DE3)进行诱导表达重组蛋白srh IL-4R,并用SDS-PAGE和Western-blotting方法分析重组蛋白。提取重组蛋白干预哮喘模型大鼠,取肺组织进行病理切片观察。结果重组载体p ET-28a(+)-srh IL-4R酶切获得长度为500bp^750bp的目的片段,表达载体p ET-28a(+)-srh IL-4R转入表达菌BL21(DE3)后经IPTG诱导表达出的重组蛋白经SDS-PAGE和Westernblotting方法可见重组蛋白分子量约为25k Da;重组人可溶性IL-4R蛋白干预哮喘大鼠可见肺组织炎细胞数量较模型组明显减少。结论 srh IL-4R防治哮喘具有一定作用。
Objective To investigate the effect of recombinant soluble human interleukin-4 receptor（srh IL-4R）in the asthma model. Methods The Homo sapiens（human） interleukin-4 receptor gene was synthesized according to the sequence of the Gen Bank（XM_005255309）, the synthesis of gene fragment was cloned into p ET-28a（＋）vector to construct the recombinant expression vector p ET-28a（＋）-srh IL-4R. The expression plasmid was induced into E. coli BL21（DE3） and expression strain was selected, the recombinant was analyzed by SDS-PAGE and Western-blotting. Rat using an ovalbumin（OVA）-induced model of allergic asthma was conducted with the recombinant soluble human interleukin-4 receptor proteins, and lung tissue from model of allergic asthma was taken for pathology observation. Results 500bp-750 bp target fragment was obtained by restricted enzyme, SDS-PAGE and western-blotting analysis results of the recombinant protein was about 25 k Da. Compared with model group, allergic asthma intervention decreased significantly in inflammatory cells. Conclusion Srh IL-4R has a certain effect in the prevention and treatment of asthma.
出处
《浙江中西医结合杂志》
2016年第4期314-316,F0002,共4页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
2011年贵州省社会发展科技攻关项目[No.黔科合SY字(2011)3020]
2014年度贵阳中医学院"2011协同创新中心"和贵州省委组织部高层次人才科研条件特助经费项目(No.TZJF-2011年28号)
