度洛西汀对神经病理性疼痛大鼠kindlin/integrin/RhoA通路及脊髓星形胶质细胞活化的影响

Effect of duloxetine on kindlin/integrin/RhoA pathway and activation of spinal astrocytes in rats with neuropathic pain

目的探讨度洛西汀对神经病理性疼痛大鼠脊髓星形胶质细胞活化的影响,以及与kindlin/整合素(integrin)/RhoA信号通路的关系。方法将60只SD大鼠随机分成5组:假手术组、模型组、度洛西汀低剂量组、度洛西汀中剂量组和度洛西汀高剂量组。除假手术组外,其余各组均采用慢性缩窄损伤诱导大鼠神经病理性疼痛。各组分别给予药物处理,检测机械撤退阈值(mechanical withdrawal threshold,MWT)和热撤退潜伏期(thermal withdrawal latency,TWL);采用免疫组织化学检测脊髓GFAP蛋白表达水平;采用ELISA法检测脊髓炎性因子白细胞介素1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;采用Western blot法检测大鼠脊髓kindlin、integrin、RhoA蛋白表达水平。结果与假手术组相比,模型组大鼠MWT、TWL显著降低(P<0.05),炎性因子IL-1β和TNF-α水平、GFAP及kindlin-1、integrin、RhoA蛋白表达水平显著升高(P<0.05);与模型组相比,度洛西汀各剂量组大鼠MWT、TWL显著升高(P<0.05),炎性因子IL-1β和TNF-α水平、GFAP及kindlin-1、integrin、RhoA蛋白表达水平显著降低(P<0.05)。结论度洛西汀可抑制星形胶质细胞活化和炎症反应,减轻神经病理性疼痛,其机制可能与kindlin/integrin/RhoA信号通路有关。

Objective To investigate the effect of duloxetine on the activation of spinal astrocytes in rats with neuropathic pain and its relationship with kindlin/integrin/RhoA signaling pathway.Methods Sixty SD rats were randomly divided into five groups:sham operation group,model group,low-dose duloxetine group,medium-dose duloxetine group and high-dose duloxetine group.Except for the sham operation group,chronic constriction injury was used to induce neuropathic pain in other groups.All of them were treated with drugs,the mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured;the expression level of GFAP protein in spinal cord was detected by immunohistochemistry;the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in spinal cord were measured by ELISA;the protein expression levels of kindlin,integrin and RhoA in spinal cord of rats were measure by Western blot.Results Compared with those in sham operation group,MWT and TWL of model group were significantly lower(P<0.05),while the levels of inflammatory factors IL-1βand TNF-α,protein expression levels of GFAP and kindlin-1,integrin and RhoA were significantly higher(P<0.05);compared with those in the model group,MWT and TWL of duloxetine groups were significantly higher(P<0.05),while the levels of inflammatory factors IL-1βand TNF-α,protein expression levels of GFAP and kindlin-1,integrin and RhoA were significantly lower(P<0.05).Conclusion Duloxetine can inhibit the activation of astrocytes and inflammatory response and relieve neuropathic pain,which may be related to the kindlin/integrin/RhoA signaling pathway.