1,8-桉叶素对背根神经节内P2X3受体介导神经病理痛的作用研究

发布时间：2018-05-13 22:44

本文选题：神经病理痛 + 1　；参考：《南昌大学》2017年硕士论文

【摘要】：目的:观察1,8-桉叶素对神经病理性疼痛大鼠的治疗作用,探讨背根神经节内P2X3受体在其中的作用及机理。方法:1、建立坐骨神经慢性压迫性损伤(CCI)神经病理痛大鼠模型。2、将所有大鼠无规则划分五组:假手术组(Sham)、慢性坐骨神经结扎损伤组(CCI)、二甲亚砜对照组(CCI+DMSO)、低剂量1,8-桉叶素治疗组(CCI+50mg/kg)、高剂量1,8-桉叶素治疗组(CCI+100mg/kg)。3、在实验大鼠手术后第7天和第14天都分别检测其机械缩足反射阈值(MWT)和热缩足反射潜伏期(TWL)。4、在实验大鼠手术后第7天和第14天,取各组大鼠L4-5背根神经节使用原位杂交技术和逆转录聚合酶链反应(qRT-PCR)技术检测P2X3受体m RNA水平的变化。5、使用免疫组织化学染色法和蛋白印迹法(Western-Blot)于实验大鼠手术后第7天和第14天分别检测L4-5背根神经节P2X3受体蛋白的表达变化。结果:1、术后第7d和14d CCI组大鼠MWT和TWL明显低于Sham组(P0.05);CCI+50mg/kg组和CCI+100mg/kg组较CCI组MWT、TWL均明显升高(P0.05);CCI+DMSO组与CCI组比较无明显差异(P0.05);而CCI+100mg/kg组较CCI+50mg/kg组MWT、TWL升高更明显(P0.05);2、术后第7d和14d L4-5背根神经节P2X3受体mRNA原位杂交及qRT-PCR结果表明:CCI组明显高于Sham组(P0.05);CCI+50mg/kg组和CCI+100mg/kg组较CCI组均明显降低(P0.05);CCI+DMSO组与CCI组比较无明显差异(P0.05);其中CCI+100mg/kg组较CCI+50mg/kg组更低(P0.05);3、术后第7d和14d L4-5背根神经节P2X3受体蛋白的免疫组化和蛋白印迹结果表明:CCI组明显高于Sham组(P0.05);CCI+50mg/kg组和CCI+100mg/kg组较CCI组均明显降低(P0.05);CCI+DMSO组与CCI组比较无明显差异(P0.05);其中CCI+100mg/kg组较CCI+50mg/kg组更低。结论:1,8-桉叶素对背根神经节内P2X3受体介导的神经病理性疼痛有控制效果。
[Abstract]:Aim: to observe the therapeutic effect of 1-eucalypsin on neuropathic pain in rats and to explore the role and mechanism of P2X3 receptor in dorsal root ganglion (DRG). Methods the rat model of neuropathic pain induced by chronic compression injury of sciatic nerve (CCI) was established. All rats were randomly divided into five groups: sham-operated group, chronic sciatic nerve ligation injury group, dimethyl sulfoxide control group, CCI DMSON, low dose 188- CCI 50 mg 路kg ~ (-1) 路kg ~ (-1) and CCI 100 mg 路kg ~ (-1) 路kg ~ (-3) in Eucalypsin treatment group. The mechanical foot reflex threshold (MWTs) and the latent period (TWL.4) of heat contraction foot reflex were measured on the 7th and 14th day after operation in the experimental rats, and TWL.4 on the 7th and 14th day after the operation in the experimental rats. L4-5 dorsal root ganglion (DRG) of rats in each group was used to detect the level of P2X3 receptor m RNA by in situ hybridization and reverse transcriptase polymerase chain reaction (RT PCR). The expression of P2X3 receptor protein in L4-5 dorsal root ganglion was detected on the 7th and 14th day after operation. Results on the 7th and 14th day after operation, MWT and TWL in CCI group were significantly lower than those in Sham group (P 0.05). TWL in 50mg/kg group and CCI 100mg/kg group were significantly higher than that in CCI group. There was no significant difference between DMSO group and CCI group, while that in CCI 100mg/kg group was significantly higher than that in CCI 50mg/kg group. The results of mRNA in situ hybridization and qRT-PCR analysis of P2X3 receptor in dorsal root ganglion on the 7th and 14th days showed that the expression of P2X3 receptor was significantly lower in the qRT-PCR group than that in the Sham group (P 0.05) and the CCI 100mg/kg group was significantly lower than that in the CCI group, and there was no significant difference between the CCI 100mg/kg group and the CCI group, especially in the CCI 100mg/kg group compared with the CCI 50mg/kg group. The immunohistochemical and Western blotting results of P2X3 receptor protein in dorsal root ganglion on the 7th and 14th day after operation showed that the expression of P2X3 receptor protein was significantly higher in the P2X3 group than that in the Sham group and the CCI 100mg/kg group. There was no significant difference between the DMSO group and the CCI group. CCI 100mg/kg group was lower than CCI 50mg/kg group. ConclusionEucalypsin can control neuropathic pain mediated by P2X3 receptor in dorsal root ganglion.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R402

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