rAAV-hGDNF对谷氨酸钠诱导原代培养大鼠胚胎脊髓神经元损伤的保护作用

THE EFFECT OF rAAV-hGDNF ON PROTECTING RAT SPINAL CORD NEURONS FROM DEATH INDUCED BY GLUTAMATE

目的 探讨rAAV hGDNF对谷氨酸钠诱导的大鼠脊髓神经元死亡的保护作用。 方法 建立谷氨酸钠诱导体外培养大鼠脊髓神经元损伤模型 ,用自行包装的rAAV hGDNF病毒感染原代培养大鼠脊髓神经元 ,经双醋酸荧光素和碘化丙啶法辨认存活细胞和死亡细胞 ,观察rAAV hGDNF抑制兴奋性氨基酸对脊髓神经元的毒性作用 ;同时运用半定量RT PCR法 ,以 β actin为内对照 ,检测脊髓神经元内NOSmRNA的表达。 结果 rAAV hGDNF感染组的细胞死亡率为 5 0 %± 0 0 2 ,对照组为 5 9 2 5 %± 0 0 2 3,统计分析两组有显著性差异 (P <0 0 5 )。rAAV hGDNF感染后的脊髓神经元NOSmRNA表达为 0 97± 0 0 5 ,未感染rAAV hGDNF的对照组NOSmRNA表达为 1 2 3± 0 10 ,统计分析两组有显著性差异 (P <0 0 1)。 结论 rAAV hGDNF能抑制兴奋性氨基酸诱导的神经元NOS的表达 。

Objective In this experiment,we explored the effects of rAAV\|hGDNF on protecting spinal neurons From death. Methods rAAV\|hGDNF particals were produced by recombinant virus technolgy,and infected the culture spinal neurons which were exposed to glutamate.We counted the mortality rate and detected the expression of NOS mRNA by RT\|PCR. Results In the group transfering rAAV\|hGDNF the death rate was inhibited(50%±0\^02,control 59\^25%±0\^023, P <0\^05) and the expression of NOS mRAN was down regulated in the spinal neuronal cells(0\^97±0\^05),control 1\^23±0\^10, P <0\^01).Conclusion\ The results suggested that the mechanisms of neuronal protection are related with inhibiting the free radicals production by rAAV\|hGDNF.