ITK蛋白调节小鼠脾淋巴细胞增殖分化的研究

Research of ITK regulation on mouse spleen lymphocytes proliferation and differentiation

目的应用RNA干扰技术特异性抑制Itk基因的表达，观察Itk蛋白在淋巴细胞增殖和相应免疫因子合成分泌中的作用，进行Itk作为药物靶标的确认。方法设计合成针对Itk基因的siRNA，将siRNA电转染至小鼠脾淋巴细胞，WestemBlot检测Itk蛋白的表达、MTS方法检测细胞增殖率、ELISA方法检测细胞因子的含量。结果实验组Itk—siRNA在细胞水平上有效抑制了Itk蛋白的表达；Itk受抑制后细胞增殖率与对照组相比明显降低，差异有统计学意义，P〈0．05；细胞因子IL-2，IL-4，IL-5，IFN-γ的分泌水平下降，P〈0．05。结论Itk表达受抑制后有效降低脾淋巴细胞的增殖率及细胞因子分泌的减少，研究验证了Itk在细胞增殖和炎性细胞因子分泌方面的重要免疫调节作用，为将其作为药物的靶基因提供实验依据。

Objective By using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target. Methods Designed siRNA aims at Irk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory eytokines by ELISA. Results Itk protein can be suppressed by hk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-7. They all have statistical differenc （ P 〈 0.05 ）. Conclusion Irk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokioes. This can supply an experimental basis to regard Itk as drug target for inflammation therapy.