RNA干扰抑制小鼠脾淋巴细胞分泌IL -1α  

RNA interference inhibits the secretion of IL-1α in mice spleen lymphocytes

作　　者：郑宁宁[1] 于艳秋[1] 宋晓宇[1] 丁旭东[2] 张海鹏[1] 

机构地区：[1]中国医科大学基础医学院病理生理教研室,辽宁沈阳110001 [2]中国医科大学附属盛京医院麻醉科,辽宁沈阳110001

出　　处：《中国病理生理杂志》2007年第3期435-437,共3页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No30271444)

摘　　要：目的:确定针对IL-1α的siRNA在细胞内与其具有同源互补序列的靶标位点结合后,特异性降解IL-1α的mRNA,从而抑制其表达的效果。方法:体外PCR扩增合成针对IL-1α靶位点的DNA表达盒,纯化后转染经LPS激活的小鼠脾淋巴细胞,在细胞内源性RNA聚合酶Ⅲ作用下转录形成siRNA,诱发目标mRNA的降解。于转染后48h收集细胞培养上清。用ELISA法测定IL-1α浓度。结果:干扰组IL-1α分泌量明显低于对照组和无关干扰组(P<0.05),抑制率约为15%。结论:RNA干扰技术可以在原代培养的小鼠脾淋巴细胞中发挥特异性干扰作用,产生抑制IL-1α分泌的效果,为IL-1α的基因治疗开拓了新途径。AIM： To decide the effect that selected siRNA degrades mRNA of IL - 1α specifically and suppression of its expression after connected with target site with homology complementary sequence, METHODS： Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase IU and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supematant was collected and the concentration of IL - 1 α was assayed using ELISA. RESULTS ： Compared with blank - control and negative - control, selected sequence decreased the expression of IL - 1α, Rate of the suppression was about 15%, CONCLUSION： RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL - 1 α.

关 键 词：RNA干扰 转录后基因沉默 白细胞介素1 淋巴细胞 

分 类 号：R363[医药卫生—病理学]