摘要
目的研究siRNA对体外培养的大鼠海马神经细胞NgR基因表达抑制作用,为进一步研究中枢神经损伤修复提供实验基础。方法使用阳离子脂质体转染试剂将体外化学合成的针对NgR基因的siRNA转染体外培养的海马神经细胞,GAP43对培养细胞进行鉴定,分别于转染后48小时、1周收集转染细胞,提取总蛋白并定量。结果海马神经细胞成功培养,转染48小时后NgR蛋白的含量为阴性对照组的45.11%,差异显著P<0.05;转染1周后NgR蛋白的含量为阴性对照组的99.18%,差异不明显。结论化学合成的针对NgR基因的siRNA在转染48小时后能显著下调NgR基因表达,但在转染1周后对NgR基因表达影响不明显。
Objective To investigate the inhibition of siRNA on the expression of NgR gene in rat hippocampus cell in vitro. Methods siRNA which were designed against NgR gene transfected rat hippocampus cell with cationic lipid agent, then rat hippocampus cells were identified with GAP43 antibody. Transfection protein was collected and assayed by Western-blot 48h and 1 week after transfection respectively. Results Hippocampus cells were cultured successfully. The content of NgR protein was 45.11% to the negative control group 48h after transfection,the discrepancy was significant （ P 〈 0.05 ）, while 99. 18% 1 week after transfection, and the discrepancy was not significant （ P 〉 0.05 ）. Conclusion siRNA which were designed against NgR gene can significantly inhibit the expression of NgR gene in rat hippocampus cell in vitro 48h after transfection, but the inhibition was not significant 1 week after transfection.
出处
《创伤外科杂志》
2007年第6期544-547,共4页
Journal of Traumatic Surgery
基金
国家自然科学基金资助项目(30572009)
重庆市科委自然科学基金资助项目(2005BB5273)
