基于cGMP信号通路探讨地佐辛对神经病理性疼痛的镇痛作用

Study on the analgesic effect of Dizosin on neuropathic pain through cGMP signaling pathway

  • 摘要: 目的:探讨地佐辛对神经病理性疼痛的镇痛作用机制以及对cGMP信号通路活性的影响。方法:选取70只SD大鼠纳入研究,除空白对照组(Sham组,n=10)外,剩余SD大鼠均结扎左侧第5腰神经制备神经病理性疼痛模型(NPP);分别给予NPP组大鼠腹腔注射低、中、高剂量地佐辛溶液,构建Sham组(n=10)、NPP组(n=10)、L-地佐辛组(n=10)、M-地佐辛组(n=10)、H-地佐辛组(n=10)分组。术后两周分别统计各组大鼠机械痛阈值(MWT)、热缩足反射潜伏期(TWL)、脊髓组织内炎性因子〔(白介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)〕、cGMP信号通路关键蛋白cGMP、PKG表达差异性;选取中等浓度地佐辛干预剂量,设置Sham组(n=10)、NPP组(n=10)、NPP+地佐辛组(n=10)、NPP+Avanafil组(NPP模型+cGMP 信号通路激活剂,n=10)、NPP+地佐辛+Avanafil 组(NPP+地佐辛+cGMP 信号通路激活剂,n=10),分别统计各组大鼠MWT、TWL、脊髓组织内炎性因子(IL-6、IL-1β、TNF-α、MCP-1)、cGMP信号通路关键蛋白cGMP、PKG表达差异性。结果:与Sham 组相比,NPP 组TWL、MWT、cGMP、PKG 蛋白表达下调,炎症因子(IL-6、IL-1β、TNF-α、MCP-1)水平上调(P< 0.05);与NPP组相比,不同剂量地佐辛组均可上调TWL、MWT值、cGMP、PKG表达,下调炎症因子(IL-6、IL-1β、TNF-α、MCP-1)水平,呈现剂量依赖性(P< 0.05)。与NPP组相比,地佐辛及cGMP信号通路激活剂Avanafil的干预均可上调cGMP、PKG蛋白表达及TWL、MWT值,下调炎症因子(IL-6、IL-1β、TNF-α、MCP-1)水平(P< 0.05);二者联合可进一步上调cGMP信号通路关键蛋白cGMP、PKG表达及TWL、MWT值,下调炎症因子(IL-6、IL-1β、TNF-α、MCP-1)水平(P< 0.05)。结论:地佐辛可能通过靶向抑制cGMP信号通路活性来减轻神经病理性疼痛大鼠脊髓炎症反应,发挥镇痛作用。

     

    Abstract: Objective:To explore the analgesic mechanism of Dizosin on neuropathic pain and its impact on the activity of cGMP signaling pathway.Methods:A total of 70 SD rats were selected for inclusion in the study; excluding the blank control group(Sham group, n=10), the remaining SD rats were ligated with the left 5th lumbar nerve to prepare a neuropathic pain model (NPP); rats in the NPP group were injected intraperitoneally with low, medium, and high doses of Dizosin solution.Sham group (n=10), NPP group (n=10), L-Dizosin group (n=10), M-Dizosin group(n=10), and H-Dizosin group(n=10)were constructed, and the mechanical pain threshold(MWT), thermal contraction foot reflex latency (TWL), inflammatory factors in spinal cord tissue interleukin(IL)-6, Il-1β, tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1), and differences in expression of key proteins cGMP and PKG in cGMP signaling pathway of rats in each group 2 weeks after surgery were calculated; a moderate concentration of Dizosin intervention dose was selected and the Sham group(n=10), NPP group(n=10), NPP+Dizosin group(n=10), NPP+Avanafil group(NPP model+cGMP signaling pathway activator, n=10), NPP+Dizosin+Avanafil group (NPP+Dizosin+cGMP signaling pathway activator, n=10) were set; MWT, TWL, inflammatory factors (IL-6, IL-1β、TNF-α、MCP-1)in the spinal cord tissue and differences in expression of key proteins cGMP and PKG in cGMP signaling pathways of rats in each group were calculated.Results:Compared with the Sham group, the expression of TWL, MWT, cGMP, and PKG proteins in the NPP group was downregulated, and inflammatory factors (IL-6, IL-1β、TNF-α、MCP-1) were upregulated (P< 0.05); compared with the NPP group, different doses of Dizosin could upregulate TWL and MWT values as well as cGMP and PKG expression, and could downregulate the levels of inflammatory factors (IL-6、IL-1β、TNF-α、MCP-1), reflecting dose-dependency (P< 0.05).Compared with the NPP group, the intervention of Dizosin and the cGMP signaling pathway activator Avanafil could upregulate the expression of cGMP and PKG proteins, as well as TWL and MWT values, and could downregulate the levels of inflammatory factors (IL-6, IL-1β、TNF-α、MCP-1) (P< 0.05); the combination of the two could further upregulate the expression of key proteins in the cGMP signaling pathway, such as cGMP and PKG, as well as TWL and MWT values, and could downregulate the levels of inflammatory factors(IL-6, IL-1β、TNFα、MCP-1)(P< 0.05).Conclusion:Dezosin may alleviate the inflammation of the spinal cord in neuropathic pain rats by targeting the inhibition of cGMP signaling pathway activity, exerting analgesic effect.

     

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