中文摘要：

目的观察siRNA—pool对大鼠脊髓背角神经细胞（ratneurons—dorsalspinalcordcells，RNdsc）上T细胞死亡偶联基因8（T—celldeath—associatedgene8，TDAG8）表达的影响。方法设计并合成针对TDAG8的siRNA3对（siR—NA。siRNA341、siRNA897）及一条带绿色荧光标记的阴性对照siRNA。在阳离子聚合物纳米粒jetPEITM介导下转染RNdsc。RNdsc细胞随机分为正常（normal）组，载体（vehicle）组，阴性对照（mismatch）组，siRNA50、100、200pmol组。转染24h后，荧光显微镜下计算转染复合物的转染效率，用MTT法检测转染复合物的细胞毒性，采用real—timePCR及Westernblot测定siRNA—pool的干扰效率。结果采用jet—PEI…作为转染介质转染效率高达98．9％。转染复合物的细胞毒性低，各组细胞成活率差异无统计学意义。siRNA50、100、200pmol可剂量依赖性地抑制TDAG8的表达，siRNA200pmol组对TDAG8mRNA表达的抑制率达88％，对TD—AG8蛋白的表达抑制率达73％。结论siRNA—pool能高效地抑制RNdsc上TDAG8的表达。

英文摘要：

Aim To investigate the ability of siRNA- pool to knockdown TDAG8 in rat neurons-dorsal spinal cord （RNdsc） ceils. Methods Four siRNAs were chemically synthesized ： three of them were used to in- hibit TDAG8 expression in RNdsc cells, and the rest was fluorescence-labeled mismatch siRNA as a negative control. They were all transfected into RNdsc cells with jetPEITM, respectively. RNdsc cells were randomly di- vided into 5 groups： normal, vehicle, mismatch, and siRNA 50, 100, 200 pmol group. After transfection with siRNA for 24 h, the rate of transfection was calcu- lated under fluorescence microscope,and the cytotoxic- ity of complex was detected using MTT. The expressionof TDAG8 was detected blot analysis. Results by real-time PCR and Western The transfection rate was high enough to reach 98.9%, and the cytotoxicity of com- plex was very low. Meanwhile, TDAG8-siRNA 50, 100, 200 pmol could dose-dependently inhibit the ex- pression of TDAG8 mRNA and protein levels. The TD- AG8 siRNA 200 pmol reduced the expression of TD- AG8 mRNA and protein up to 88% and 73%, individ- ually. Conclusion SiRNA-pool can effectively inhibit the expression of TDAG8 in RNdsc cells.