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目的:明确乌头汤(Wutoutang,WTT)对神经病理性疼痛(neuropathic pain,NP)小鼠的镇痛药效,并探索其对下行抑制系统的影响机制。方法:ICR小鼠随机分为假手术组,模型组,普瑞巴林(pregabalin,PGB)组,WTT高(WTT-H),中(WTT-M),低(WTT-L)剂量组。除Sham组只剥离但不结扎外,其余各组均对L5脊神经进行结扎,随后分别以生理盐水,12.60,6.30,3.15 g·kg~(-1)WTT及0.025 g·kg~(-1)PGB对各组小鼠进行灌胃,持续21 d。采用Von Frey法检测WTT的镇痛药效;酶联免疫吸附法(ELISA)检测下行抑制系统关键脑区前扣带回皮质(anterior cingulate cortex,ACC)及延髓头端腹内侧核(rostral ventromedial medulla,RVM)中的脑源性神经营养因子(brain derived neurotrophic factor,BDNF)和5-羟色胺(5-hydroxytryptarnjne,5-HT)的蛋白表达水平;蛋白免疫印迹法(western blot)分析磷酸化细胞外信号调节激酶(phosphorylatedextracellular signal-regulated kinase,p-ERK)和突触后致密蛋白95(postsynaptic density protein-95,PSD-95)的蛋白表达量。结果:给药第21天,与假手术组比较,模型组小鼠机械痛敏阈值明显降低(P<0.01),且其RVM和ACC核团内5-HT和BDNF的表达水平也明显下降(P<0.05),而p-ERK和PSD-95蛋白表达却明显升高(P<0.05);与模型组比较,WTT组小鼠机械痛敏阈值显著回升(P<0.01),5-HT和BDNF表达量明显提高(P<0.05),同时p-ERK和PSD-95蛋白表达有所抑制。结论:WTT能缓解小鼠NP,且可能是通过调节下行抑制系统中RVM和ACC内5-HT,BDNF,p-ERK和PSD-95的蛋白表达水平而发挥作用的。
Objective: To clarify the analgesic efficacy of Wutoutang (WTT) on neuropathic pain (NP) mice and to explore its mechanism of action on the downstream inhibitory system. Methods: ICR mice were randomly divided into sham operation group, model group, pregabalin (PGB) group, WTT-H, WTT-M and WTT-L groups. L5 spinal nerves were ligated except for sham group but without ligation, and then saline, 12.60, 6.30 and 3.15 g · kg -1 WTT and 0.025 g · kg -1 PGB mice were intragastrically administered for 21 days. Von Frey was used to detect the analgesic efficacy of WTT. ELISA was used to detect the anterior cingulate cortex (ACC) in the key brain regions of the down-inhibition system and the rostral ventromedial medulla (RVM), brain derived neurotrophic factor (BDNF) and 5-hydroxytryptarnjne (5-HT) were detected by Western blotting. The levels of phosphorylated extracellular The protein expression levels of p-ERK and postsynaptic density protein-95 (PSD-95) were measured. Results: Compared with the sham operation group, the threshold of mechanical hyperalgesia in model group was significantly decreased (P <0.01), and the expression of 5-HT and BDNF in RVM and ACC nucleus also decreased significantly (P <0.05), while the expressions of p-ERK and PSD-95 protein were significantly increased (P <0.05). Compared with the model group, the threshold of mechanical hyperalgesia in WTT group significantly increased (P <0.01) BDNF expression was significantly increased (P <0.05), while p-ERK and PSD-95 protein expression was inhibited. CONCLUSION: WTT can relieve the NP in mice and may play a role in regulating the protein expression of 5-HT, BDNF, p-ERK and PSD-95 in RVM and ACC in the down-regulated system.