R59949对完全弗氏佐剂诱导的大鼠炎性痛行为的影响

The effects of R59949 on inflammatory pain induced by complete Freund’s adjuvant in rats

目的:探究甘油二酯激酶(DGK)参与大鼠持续性炎性痛调节的作用和机制。方法:左侧足底注射完全弗氏佐剂(CFA)建立大鼠炎性痛模型,并给予DGK抑制剂R59949进行干预。通过热缩足潜伏期(PWTL)和机械缩足阈值(PWMT)行为学实验观察各组大鼠的痛行为变化,RT-qPCR检测大鼠脊髓背角DGKα、DGKβ和DGKγmRNA的表达,Western Blot法检测TNF-α和IL-6的表达,免疫荧光染色观察DGKα的表达与细胞定位。结果:与对照组相比,炎性痛大鼠PWTL和PWMT显著降低(均P<0.001),并且脊髓中DGKαmRNA(P<0.001)和蛋白表达显著增高(P<0.05),TNF-α和IL-6的蛋白表达显著增高(P<0.001)。炎性痛大鼠给予R59949处理后,PWTL和PWMT明显降低(均P<0.01),TNF-α和IL-6在脊髓的表达显著增多(P<0.05,P<0.001)。免疫荧光染色结果显示,DGKα在炎性痛大鼠脊髓神经元出现表达。结论:DGK抑制剂R59949增强大鼠的炎症性痛并促进TNF-α和IL-6的表达,DGKα参与炎症性痛的调控。

Objective:To preliminarily explore the role and mechanism of diacylglycerol kinase(DGK)involved in the regulation of persistent inflammatory pain in rats.Methods:Complete Freund’s adjuvant(CFA)was injected into the left soles of rats to establish the pain model,and the DGK inhibitor R59949 was given for intervention.The behavioral changes about pain were detected by the paw withdrawal thermal latency(PWTL)and paw withdrawal mechanical threshold(PWMT),the expressions of DGKα,DGKβ,DGKγmRNA in the dorsal horn of spinal cord of rats were analyzed by RT-qPCR,the expression changes of inflammatory factors TNF-αand IL-6 in the spinal cord of rats were examed by Western Blot,and the expression and cell localization of DGKαwas observed by immunofluorescence staining.Results:Compared with control group,the PWTL and PWMT of CFA-induced rats were decreased significantly(P<0.001),Notably,DGKαmRNA(P<0.001)and protein(P<0.05)expression levels were significantly elevated in the spinal cord,as were the protein expressions of TNF-αand IL-6(all P<0.001).After the model rats were treated with DGK inhibitor R59949,PWTL and PWMT decreased significantly(both P<0.01).while the expressions of TNF-αand IL-6 in the spinal cord increased significantly(P<0.05,P<0.001).Immunofluorescence results showed that DGKαwas expressed in spinal cord neurons of model rats.Conclusion:DGK inhibitor R59949 enhances the inflammatory pain and promotes the expression of TNF-αand IL-6 in the model rats,and DGKαis involved in the regulation of inflammatory pain.