受体相互作用蛋白激酶3在大鼠神经病理性疼痛模型中的表达及其作用机制

Expression of receptor-interacting protein kinase 3 in an animal model of neuropathic pain

目的观察受体相互作用蛋白激酶3(receptor-interacting protein kinase 3,RIP3)在大鼠脊神经结扎模型中的表达,并探讨其是否参与大鼠神经病理性疼痛的发生。方法 40只大鼠随机分为手术组、生理盐水组、抑制剂组和假手术组,每组10只。手术组、生理盐水组和抑制剂组分别建立腰5脊神经结扎模型,假手术组只做手术,不结扎神经。生理盐水组、抑制剂组分别在建模30 min前鞘内注射生理盐水和GSK'872。记录各组大鼠的行为学改变与机械痛域,采用免疫组化和Western blot检测各组RIP3的表达水平,并通过ELISA法检测各组大鼠肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白介素1β(Interleukin-1β,IL-1β)含量。结果手术组、生理盐水组分别与抑制剂组、假手术组相比,均有明显行为学改变且机械痛域值明显下降(P<0.05),RIP3与TNF-α、IL-1β蛋白表达水平均显著上调(P<0.001);抑制剂组机械痛域值与RIP3含量均小于手术组和生理盐水组(P<0.05)。结论 RIP3在大鼠神经病理性疼痛模型中表达上调,可能参与神经病理性疼痛的发生。

Objective To observe the expression of receptor-mediated protein kinase 3 （RIP3） in a rat model of spinal nerve ligation and explore whether it is involved in the pathogenesis of neuropathic pain. Methods Rats were randomly divided into four groups： operation, saline,inhibitor and sham operation （n=10 rats per group）. In the operation,saline,and inhibitor groups, a lumbar 5 spinal nerve ligation model was established. Animals in the sham operation group underwent surgery without nerve ligation. In the inhibitor group,GSK＇872 was injected intratheeally at 30 min before model establishment;in the saline group,the same volume of saline was intrathecally injected. Behavior and mechanical allodynia were recorded for each group. RIP3 expression was analyzed using immuno- histoehemistry and Western blotting; TNF-α and IL-1β levels were determined using ELISA. Results The operation and saline groups showed significant behavioral differences,significantly lower mechanical allodynia,as well as significantly higher levels of R1P3 protein, TNF-α and IL-1β than the sham operation group （P〈0.05）. The inhibitor group showed lower mechanical pain sensitivity and protein content than the operation and saline groups （P〈0.05）. Expression of RIP3 ,TNF-α and IL-1β negatively correlated with mechanical allodynia. Condusions RIP3 is up-regulated in this animal model of neuropathic pain, so RIP3 may be involved in the development of neuropathic pain.