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1,8-桉葉素對背根神經(jīng)節(jié)內(nèi)P2X3受體介導神經(jīng)病理痛的作用研究

發(fā)布時間:2018-05-13 22:44

  本文選題:神經(jīng)病理痛 + 1; 參考:《南昌大學》2017年碩士論文


【摘要】:目的:觀察1,8-桉葉素對神經(jīng)病理性疼痛大鼠的治療作用,探討背根神經(jīng)節(jié)內(nèi)P2X3受體在其中的作用及機理。方法:1、建立坐骨神經(jīng)慢性壓迫性損傷(CCI)神經(jīng)病理痛大鼠模型。2、將所有大鼠無規(guī)則劃分五組:假手術組(Sham)、慢性坐骨神經(jīng)結扎損傷組(CCI)、二甲亞砜對照組(CCI+DMSO)、低劑量1,8-桉葉素治療組(CCI+50mg/kg)、高劑量1,8-桉葉素治療組(CCI+100mg/kg)。3、在實驗大鼠手術后第7天和第14天都分別檢測其機械縮足反射閾值(MWT)和熱縮足反射潛伏期(TWL)。4、在實驗大鼠手術后第7天和第14天,取各組大鼠L4-5背根神經(jīng)節(jié)使用原位雜交技術和逆轉(zhuǎn)錄聚合酶鏈反應(qRT-PCR)技術檢測P2X3受體m RNA水平的變化。5、使用免疫組織化學染色法和蛋白印跡法(Western-Blot)于實驗大鼠手術后第7天和第14天分別檢測L4-5背根神經(jīng)節(jié)P2X3受體蛋白的表達變化。結果:1、術后第7d和14d CCI組大鼠MWT和TWL明顯低于Sham組(P0.05);CCI+50mg/kg組和CCI+100mg/kg組較CCI組MWT、TWL均明顯升高(P0.05);CCI+DMSO組與CCI組比較無明顯差異(P0.05);而CCI+100mg/kg組較CCI+50mg/kg組MWT、TWL升高更明顯(P0.05);2、術后第7d和14d L4-5背根神經(jīng)節(jié)P2X3受體mRNA原位雜交及qRT-PCR結果表明:CCI組明顯高于Sham組(P0.05);CCI+50mg/kg組和CCI+100mg/kg組較CCI組均明顯降低(P0.05);CCI+DMSO組與CCI組比較無明顯差異(P0.05);其中CCI+100mg/kg組較CCI+50mg/kg組更低(P0.05);3、術后第7d和14d L4-5背根神經(jīng)節(jié)P2X3受體蛋白的免疫組化和蛋白印跡結果表明:CCI組明顯高于Sham組(P0.05);CCI+50mg/kg組和CCI+100mg/kg組較CCI組均明顯降低(P0.05);CCI+DMSO組與CCI組比較無明顯差異(P0.05);其中CCI+100mg/kg組較CCI+50mg/kg組更低。結論:1,8-桉葉素對背根神經(jīng)節(jié)內(nèi)P2X3受體介導的神經(jīng)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.
【學位授予單位】:南昌大學
【學位級別】:碩士
【學位授予年份】:2017
【分類號】:R402

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2 李冬梅;李小云;;2013年我院多重耐藥菌的分布及耐藥性分析[J];藥物流行病學雜志;2015年06期

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