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目的:研究脱氧鬼臼毒素对大鼠背根神经节神经元膜电位的影响及其与钠通道的关系。方法:分别用终浓度为1、5、25、125μmol/L的脱氧鬼臼毒素作用于荧光染料DiBAC4(3)标记的大鼠背根神经节神经元,在激光共聚焦显微镜上监测神经元膜电位的实时动态变化,并观察钠通道阻断剂TTX(tetrodotoxin)对脱氧鬼臼毒素膜电位效应的影响。结果:加入脱氧鬼臼毒素后大鼠背根神经节神经元膜电位呈去极化改变,5 min后达到最大水平,8 min内趋于稳定。1、5、25、125μmol/L的脱氧鬼臼毒素作用5 min后,所测得的荧光强度值分别为62.3±2.1、63.8±3.6、68.5±3.8、88.1±5.4,和空白对照组相比差异均有显著性(P<0.01)。1μmol/L TTX与大鼠背根神经节神经元共孵育20 min后再加入25μmol/ml脱氧鬼臼毒素所测值为57.5±2.3,与对照组相比无显著差异(P>0.05),说明脱氧鬼臼毒素的膜电位效应可被TTX完全抑制。结论:脱氧鬼臼毒素可引起大鼠背根神经节神经元膜电位去极化,且其效应在1~125μmol/L范围内随浓度增加而增大,钠通道可能参与了这一过程。
Objective: To study the effect of deoxypodophyllotoxin on membrane potential of rat dorsal root ganglion neurons and its relationship with sodium channels. METHODS: The rat dorsal root ganglion neurons labeled with the fluorescent dye DiBAC4(3) were treated with deoxypodophyllotoxin at a final concentration of 1, 5, 25, and 125 μmol/L, respectively, and the neuronal membranes were monitored on a laser confocal microscope. Real-time dynamic changes in potential, and observe the effect of sodium channel blocker TTX (tetrodotoxin) on the membrane potential effect of deoxypodophyllotoxin. Results: After deoxypodophyllotoxin was added, the membrane potential of rat dorsal root ganglion neurons showed a depolarization change, reached the maximum level within 5 minutes, and tended to be stable within 8 minutes. After 5, 25, 125μmol/L deoxypodophyllotoxin action for 5 min, the measured fluorescence intensity values were 62.3±2.1, 63.8±3.6, 68.5±3.8, 88.1±5.4, respectively, compared with the blank control group. The differences were significant (P<0.01). After 1 μmol/L TTX was co-incubated with rat dorsal root ganglion neurons for 20 min and then 25 μmol/ml deoxypodophyllotoxin was added, the measured value was 57.5±2.3, which was not significantly different from the control group (P>0.05). The membrane potential effect of deoxypodophyllotoxin is completely inhibited by TTX. CONCLUSION: Deoxypodophyllotoxin can induce depolarization of membrane potential in rat dorsal root ganglion neurons, and its effect increases with increasing concentration in the range of 1 to 125 μmol/L. The sodium channel may participate in this process.