论文部分内容阅读
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 interleukin-4 receptor (IL-4R) on rat asthma model. Methods The gene fragment of srh IL-4R was synthesized based on the Homo sapiens (human) IL-4R gene sequence (XM_005255309) published by Gen Bank. The gene fragment was inserted into p ET-28a (+) to construct the recombinant expression vector p ET-28a ) -srh IL-4R. After restriction analysis, the vector was transformed into E. coli BL21 (DE3) to express the recombinant protein srh IL-4R. The recombinant protein was analyzed by SDS-PAGE and Western-blotting. The recombinant protein was extracted to interfere the asthmatic model rats and the lung tissue was taken for pathological observation. Results The recombinant plasmid p ET-28a (+) - srh IL-4R was digested with restriction endonucleases of 500bp to 750bp. The recombinant plasmid p ET-28a (+) - srh IL-4R was transformed into E. coli BL21 The recombinant protein induced by IPTG showed that the molecular weight of the recombinant protein was about 25 kDa by SDS-PAGE and Western blotting. The number of inflammatory cells in the lung tissue of the asthmatic rats treated with recombinant human soluble IL-4R protein was significantly decreased compared with the model group. Conclusion srh IL-4R has a role in the prevention and treatment of asthma.