巴氯芬对神经病理性痛大鼠DRG神经元上GABA_A受体的调节作用

发布时间：2018-12-31 15:40

【摘要】：目的：研究GABAA受体γ2亚基和α2亚基在神经病理性痛(Chronic Constriction Injury, CCI)大鼠DRG神经元上的表达，以及巴氯芬干预后在CCI模型大鼠背根神经节(Dorsal RootGganglion, DRG)神经元上GABAA受体的功能和GABAA受体γ2亚基表达的变化。方法： (1)制备SD大鼠CCI模型，其中部分模型鼠以灌胃方式给予巴氯芬干预；(2)热板实验检测给予巴氯芬干预前后CCI模型鼠热刺激缩足反射潜伏期(Thermal WithdrawalLatency, TWL)；(3)膜片钳技术观察CCI模型组及巴氯芬干预组，DRG神经元上GABAA受体介导的内向电流的变化；(4)应用Western Blot技术检测CCI组及巴氯芬组GABAA受体γ2亚基表达变化；(5)应用免疫荧光化学技术检测CCI术后GABAA受体α2亚基的在DRG神经元上的表达分布情况。结果： (1) CCI模型大鼠的TWL比正常组明显缩短，，给予巴氯芬干预后大鼠TWL逐渐延长，至一周趋近于正常(p 0.05, n=10/group)；(2)膜片钳实验发现CCI手术后，手术侧组GABAA受体介导的内向电流较正常组明显减小，对侧组GABAA受体介导的内向电流较正常组明显增强；巴氯芬抑制DRG神经元GABAA受体介导的内向电流(p 0.05, n=6)；(3) WB检测发现CCI手术侧和手术对侧DRG神经元上GABAA受体γ2亚基的表达量较对照组明显下调，但是磷酸化GABAA受体γ2亚基表达量明显上调(p 0.05, n=6)；(4)WB检测发现巴氯芬干预组的GABAA受体γ2亚基表达较CCI组进一步下调，磷酸化的GABAA受体γ2亚基表达较CCI组进一步上调；(5)免疫荧光显示CCI术侧组GABAA受体α2亚基表达量较正常组下调，对侧组表达量上调(p 0.05, n=6)。结论：神经病理性痛状态下，GABAA受体γ2亚基参与痛觉的形成过程，巴氯芬作为GABAB受体的激动剂，通过增加GABAA受体γ2亚基的磷酸化水平抑制GABAA受体的功能。
[Abstract]:Objective: to study the expression of GABAA receptor 纬 2 subunit and 伪 2 subunit in DRG neurons of (Chronic Constriction Injury, CCI) rats with neuropathic pain and (Dorsal RootGganglion, in dorsal root ganglion of CCI model rats after the intervention of baclofen. The function of GABAA receptor and the expression of GABAA receptor 纬 2 subunit in DRG neurons. Methods: (1) the CCI model of SD rats was established, and some of them were treated with baclofen by gavage; (2) the latent period of thermal stimulation of paw reflex was detected by hot plate test in CCI model rats before and after the treatment with baclofen. (3) the changes of inward currents mediated by GABAA receptor in DRG neurons were observed by patch clamp technique in CCI model group and baclofen intervention group, (4) the expression of GABAA receptor 纬 2 subunit in CCI group and baclofen group were detected by Western Blot technique. (5) Immunofluorescence technique was used to detect the expression of GABAA receptor 伪 2 subunit in DRG neurons after CCI. Results: (1) the TWL of the CCI model rats was shorter than that of the normal group. The TWL of the rats treated with baclofen was prolonged gradually and reached to the normal level in one week (p 0.05, n=10/group). (2) after CCI operation, the inward current mediated by GABAA receptor in the operation side group was significantly lower than that in the normal group, and the inward current mediated by the GABAA receptor in the contralateral group was significantly higher than that in the normal group. Baclofen inhibited inward currents mediated by GABAA receptor in DRG neurons (p0.05, nong6). (3) WB showed that the expression of GABAA receptor 纬 2 subunit was significantly down-regulated in the DRG neurons of the operative and contralateral sides of CCI compared with the control group, but the expression of phosphorylated GABAA receptor 纬 2 subunit was significantly up-regulated (p 0.05, nong 6). (4) the expression of GABAA receptor 纬 2 subunit was further down-regulated and phosphorylated GABAA receptor 纬 2 subunit expression was further up-regulated in baclofen intervention group than in CCI group. (5) Immunofluorescence showed that the expression of GABAA receptor 伪 2 subunit was down-regulated in CCI side group and up-regulated in contralateral group (p 0.05, nong 6). Conclusion: in neuropathic pain state, GABAA receptor 纬 2 subunit is involved in the formation of pain. As an agonist of GABAB receptor, baclofen inhibits the function of GABAA receptor by increasing the phosphorylation level of GABAA receptor 纬 2 subunit.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R741

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