隔姜灸对大鼠脊髓损伤后神经源性膀胱M2、M3乙酰胆碱mRNA及P2X3受体表达的影响

目的探讨隔姜灸改善脊髓损伤大鼠神经源性膀胱症状的作用靶点,为脊髓损伤神经源性膀胱患者临床干预提供理论基础。方法选取SD大鼠为实验动物,随机分组后,按照改良Allen′s法建立T 10脊髓损伤模型,根据评估标准,筛选符合脊髓损伤后神经源性膀胱标准的动物模型纳入模型组和隔姜灸组;假手术组仅切除T 10椎板,不破坏脊髓;各组均纳入10只SD大鼠。三组术后均给予抗感染、创口护理及Crede手法辅助排尿。隔姜灸组同时行隔姜灸治疗,即术后第1天开始,每日1次,每次每个穴位灸10 min,连续12 d。结果隔姜灸组治疗后嘌呤能P2X3受体的相对表达水平及M2、M3乙酰胆碱mRNA的相对表达水平显著高于模型组(均P<0.05)。结论隔姜灸治疗可提高脊髓损伤大鼠脊髓神经节嘌呤能P2X3受体及膀胱组织中M2、M3乙酰胆碱mRNA的表达水平,从而有利于改善神经源性膀胱功能障碍。

高乌甲素对坐骨神经慢性压迫性损伤大鼠背根神经节P2X3受体转运介导神经病理性疼痛的影响

目的探讨高乌甲素(LA)对坐骨神经慢性压迫性损伤(CCI)大鼠背根神经节(DRG)神经元轴突中嘌呤能P2X3受体(P2X3R)转运介导神经病理性疼痛(NPP)的影响。方法选择72只健康SD雄性大鼠,采用手术结扎右侧坐骨神经造成坐骨神经CCI的方法构建NPP模型。按随机数字法表分为CCI组、CCI+LA组和正常对照组,每组24只。正常对照组仅暴露右侧坐骨神经不结扎。CCI+LA组行CCI组相同处理后给予2 g/L的LA(4 mg/kg,腹腔注射,1次/d,共1次),其余两组以同样方式注射等量生理盐水。三组大鼠分别于伤前、伤后2,6,12,24 h测定机械缩足反射阈(MWT)和热缩足反射潜伏期(TWL),评价神经损伤引起的神经痛症状。收集3组大鼠神经结扎点近端和远端的神经片段,通过Western blot检测各组大鼠伤后2,6,12,24 h P2X3R表达量和伤后24 h神经生长因子(NGF)及酪氨酸激酶受体A(TrkA)表达量,以评估LA对P2X3R、NGF和TrkA的影响。结果伤前各组间MWT、TWL差异无统计学意义(P均>0.05);与正常对照组比较,伤后2,6,12,24 h CCI组和CCI+LA组MWT、TWL明显降低(P<0.05或0.01);伤后2,6 h CCI组和CCI+LA组MWT、TWL差异无统计学意义(P均>0.05),伤后12,24 h CCI+LA组MWT、TWL明显高于CCI组(P<0.05或0.01)。Western blot结果显示,伤后2,6,12,24 h在近端坐骨神经片段中,各组P2X3R表达量差异无统计学意义(P均>0.05)。伤后2,6,12,24 h在远端坐骨神经片段中,CCI组P2X3R表达量高于正常对照组(P均<0.01);伤后2,6,12 h CCI+LA组P2X3R表达量高于正常对照组(P均<0.05),伤后24 h与正常对照组差异无统计学意义(P>0.05);伤后2,6 h CCI组和CCI+LA组P2X3R表达量差异无统计学意义(P均>0.05),而伤后12,24 h CCI+LA组P2X3R表达量低于CCI组(P<0.05或0.01)。伤后24 h在近端和远端坐骨神经片段中,正常对照组NGF表达量低于CCI组和CCI+LA组(P<0.05或0.01),CCI组和CCI+LA组NGF表达量差异无统计学意义(P>0.05)。伤后24 h在近端和远端坐骨神经片段中,各组间TrkA表达量差异无统计学意义(P均>0.05)。结论伤后早期给予LA可以缓解NPP大鼠的机械性和热性痛觉过敏,该作用可能与LA减少P2X3R在DRG神经元中的逆向轴突转运有关。

Regulatory effect of mild moxibustion on P2X3 receptors in spinal cord,anterior cingulate cortex and thalamic ventral posterolateral nucleus of rats with IBS visceral hyperalgesia

Objective To observe the therapeutic effect of mild moxibustion on irritable bowel syndrome(IBS)visceral hyperalgesia model rats and its regulatory effect on P2X3 receptors in the spinal cord,anterior cingutate cortex(ACC)and thalamic ventral posterolateral nucleus(VPL).Methods Thirty 8-day-old newborn rats were randomly divided into a normal group(n=6)and a modeling group(n=24)according to the completely random number table method.Rats in the normal group were bred routinely,and those in the modeling group were subjected to preparing IBS chronic visceral hyperalgesia model using colorectal distention(CRD)in stimulation method.Rats successfully modelled were re-divided into a model group,a mild moxibustion group,a P2X3 receptor antagonist group,and a normal saline group according to the completely random number table method with 6 rats in each group.Rats in each group received corresponding interventions from the 37-day old,once a day for 7 consecutive days.Immunohistochemistry and Western blot assays were used to detect P2X3 protein expressions in the spinal cord,ACC and VPL of rats.Results Under different intensities of CRD stimulation,the abdominal withdrawal reflex(AWR)scores of the model group were significantly increased versus the normal group(all P<0.05);the AWR scores of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group(all P<0.01).The P2X3 protein expressions in rat spinal cord,ACC and VPL tissues of the model group were significantly increased versus the normal group(all P<0.01);the P2X3 protein expressions in rat spinal cord,ACC and VPL tissues of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group(all P<0.01).Conclusion Mild moxibustion can inhibit the P2X3 receptor expressions in the spinal cord,ACC,and VPL tissues of IBS visceral hyperalgesia model rats,which may be the mechanism of mild moxibustion in relieving the central sensitization of rats with IBS visceral hyperalgesia.