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背景:有研究报道病毒载体运载N-甲基-D天冬氨酸受体1小干扰RNA可有效缓解大鼠炎性疼痛,但病毒载体存在安全隐患。目的:探讨水溶性脂质体运载N-甲基-D天冬氨酸受体1小干扰RNA在体内外沉默N-甲基-D天冬氨酸受体1的效应和治疗神经病理性痛的可行性。方法:将PC12随机分为阴性转染组、对照转染组和水溶性脂质体转染组,分别以N-甲基-D-天冬氨酸受体1小干扰RNA、聚乙烯亚胺与N-甲基-D-天冬氨酸受体1小干扰RNA的复合物及水溶性脂质体与N-甲基-D-天冬氨酸受体1小干扰RNA的复合物转染PC12细胞,检测各组N-甲基-D-天冬氨酸受体1基因mRNA及蛋白水平表达的变化。将48只SD大鼠随机分为假手术组、模型组、聚乙烯亚胺组及水溶性脂质体组,后3组建立大鼠神经病理性疼痛模型,并分别鞘内注射生理盐水、聚乙烯亚胺与N-甲基-D-天冬氨酸受体1小干扰RNA的复合物和水溶性脂质体与N-甲基-D-天冬氨酸受体1小干扰RNA的复合物;假手术组只暴露坐骨神经。结果与结论:水溶性脂质体转染组N-甲基-D-天冬氨酸受体1的mRNA与蛋白表达水平明显低于其他两组(P<0.01)。与假手术组比较,模型组、聚乙烯亚胺组及水溶性脂质体组N-甲基-D-天冬氨酸受体1的mRNA和蛋白表达上调,累积疼痛评分升高(P<0.01);与模型组比较,水溶性脂质体转染组脊髓背角N-甲基-D-天冬氨酸受体1mRNA与蛋白表达及累积疼痛评分下降(P<0.01),聚乙烯亚胺组上述指标无明显变化(P>0.05)。表明在体内条件下水溶性脂质体可有效运载N-甲基-D-天冬氨酸受体1小干扰RNA,抑制N-甲基-D-天冬氨酸受体1的过度表达,还可减轻大鼠神经病理性痛。
Background: It has been reported that viral vector carrying N-methyl-D aspartate receptor 1 small interfering RNA can effectively relieve inflammatory pain in rats, but virus vector has potential safety hazard. Objective: To investigate the effect of water-soluble liposomes carrying N-methyl-D aspartate receptor 1 small interfering RNA silencing N-methyl-D-aspartate receptor 1 in vitro and in vivo and the treatment of neuropathic pain feasibility. Methods: PC12 was randomly divided into negative transfection group, control transfection group and water-soluble liposome transfection group, respectively N-methyl-D-aspartate receptor 1 small interfering RNA, polyethyleneimine A complex with N-methyl-D-aspartate receptor 1 small interfering RNA and a complex of water-soluble liposome with N-methyl-D-aspartate receptor 1 small interfering RNA PC12 cells were used to detect the expression of N-methyl-D-aspartate receptor 1 mRNA and protein in each group. Forty-eight Sprague-Dawley rats were randomly divided into sham operation group, model group, polyethyleneimine group and water-soluble liposome group, the latter three groups were established neuropathic pain model, and were intrathecally injected with saline, polyethylene Complex of imine with N-methyl-D-aspartate receptor 1 small interfering RNA and complex of water-soluble liposome with N-methyl-D-aspartate receptor 1 small interfering RNA The sham operation group only exposed the sciatic nerve. RESULTS AND CONCLUSION: The mRNA and protein expression of N-methyl-D-aspartate receptor 1 in water-soluble liposome transfection group was significantly lower than the other two groups (P <0.01). Compared with the sham operation group, the mRNA and protein expression of N-methyl-D-aspartate receptor 1 in the model group, polyethyleneimine group and water-soluble liposome group were increased, and the cumulative pain score was increased (P < 0.01). Compared with the model group, the expression of N-methyl-D-aspartate receptor 1 mRNA and protein in the spinal cord dorsal horn of water-soluble liposome group decreased and the cumulative pain score decreased (P <0.01) Amine group had no significant change in the above indicators (P> 0.05). Indicating that in vivo conditions water-soluble liposomes can efficiently carry N-methyl-D-aspartate receptor 1 small interfering RNA, inhibit N-methyl-D-aspartate receptor 1 overexpression, Can reduce neuropathic pain in rats.