中文摘要：

目的：以不同浓度以及序列的小干扰RNA（SiRNA）转染,培养大鼠的背根神经节（DRG）,观察对神经元形态、活性变化以及瞬时感受器电位离子4（TRPV4）通道功能的影响,筛选转染这种神经元的SiRNA。方法：取新生大鼠腰背段全部DRG,将处理好的DRG神经元进行培养,将神经元随机分为正常对照组、SiRNA不同浓度组、错配组,进行SiRNA转染,然后培养相应时间后观察各组神经元在荧光显微镜的形态学特点,并用四甲基偶氮唑兰比色法（MTT）分析各组神经元细胞生长活性的改变。另外利用Fluo3荧光探针,观察SiRNA作用下细胞内游离钙对低渗溶液的反应,采用酶联免疫法方法观察KCl刺激下各组细胞释放P物质的改变。结果：荧光显微镜下观察适宜浓度组SiRNA转染DRG神经元形态较正常组无明显差异。MTT法比较10-40nM各组神经元活性的差异无显著性意义。而SiRNA作用下细胞内游离钙对低渗溶液的反应以及细胞释放P物质的改变有显著差异。结论：40-80nMSiRNA可以有效转染DRG细胞,未对DRG神经元活性产生显著影响,这为研究TRPV4通道功能提供了一个有效的方法。

英文摘要：

Objective：To establish a RNA interfere（RNAi） model of cultured rats dorsal root ganglion（DRG） neurons with different small interfering RNA（SiRNA） in vitro and observe the changes of morphology and activity of neurons and the influence on the function of transient receptor potential vanilloid 4（TRPV4）.Method：The cultured DRG cells were assigned to three groups：SiRNA group,mismatch group and normal control group.The changes of morphology and activity were demonstrated by fluorescent microscope and methylthiazdyl tetrazolium（MTT） assay.The effect of free calcium in DRG cells was investigated.The effect of KCL stimulation on substance P（SP） release with different density in DRG was also studied.Result：TRPV4 SiRNA was uptaken into DRG cells.There was no significant difference in neurons morphology among 3 groups under fluorescent microscope.No significant activity change of neurons was detected by MTT method among 3 groups（P〉0.05）.SiRNA dose-dependently decreased SP activities of DRG cells.SiRNA could effectively inhibit the reaction of free calcium to low osmotic solution in DRG cells.Conclusion：SiRNA could transfect into DRG neuron effectively,and didn＇t influence the activity of neurons.The model would provide an effective tool for studying TRPV4 function.