地佐辛靶向TLR4/NF-κB信号通路减轻瑞芬太尼诱导的痛觉过敏机制研究

Dezocine alleviates remifentanil-induced hyperalgesia via targeting the Toll-like receptor 4/nuclear factor kappa-B pathway

目的探讨地佐辛对瑞芬太尼诱导的痛觉过敏的影响及机制。方法将40只健康雄性SD大鼠,随机分为对照组(C组)、切口痛组(I组)、瑞芬太尼输注+切口痛组(R+I组)、瑞芬太尼联合地佐辛输注+切口痛组(R+D+I组),每组10只。其中,C组不做任何处理。R+I组和R+D+I组于造模前静脉输注瑞芬太尼,I组输注等量0.9%氯化钠溶液。随后,基于左后足底切口术对I组、R+I组和R+D+I组大鼠建立切口痛模型。R+D+I组大鼠在瑞芬太尼输注前通过尾静脉注射给予地佐辛预处理。采用痛觉行为学实验评估大鼠在术前和术后不同时间点的机械缩足反应阈(paw withdrawal threshold,PWT)和热缩足反应潜伏期(paw withdrawal latency,PWL);酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)检测4组大鼠脊髓背角相关炎性因子肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)及IL-10的表达水平;实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测4组大鼠脊髓背角TLR4、NF-κB和TRPA1的mRNA表达水平;Western blot检测组大鼠脊髓背角TLR4、NF-κB和TRPA1的蛋白表达水平。结果成功构建瑞芬太尼诱导的大鼠术后痛觉过敏模型,与C组比较,I组在术后PWT值和PWL值均显著降低,脊髓背角促炎因子TNF-α、和IL-1β表达升高,而抗炎因子IL-10表达降低,TLR4、NF-κB、TRPA1蛋白和mRNA水平均明显增加(P<0.05);与I组比较,R+I组在术后PWT值和PWL值均明显下降,脊髓背角促炎因子TNF-α、和IL-1β表达升高,而抗炎因子IL-10表达降低以及TLR4、NF-κB、TRPA1蛋白和mRNA水平均明显增加(P<0.05);与R+I组比,R+D+I组在术后PWT值和PWL值均明显升高,脊髓背角促炎因子TNF-α、和IL-1β表达降低,而抗炎因子IL-10表达升高以及TLR4、NF-κB、TRPA1蛋白和mRNA水平均明显降低(P<0.05)。结论地佐辛通过抑制TLR4/NF-κB信号通路,下调TRPA1蛋白和mRNA表达,减少脊髓背角炎症,达到减轻瑞芬太尼诱导的术后痛觉过敏的效果。

Objective To investigate the effect of dezocine on remifentanil-induced postoperative hyperalgesia and the underlying mechanism.Methods A total of 40 healthy male Sprague-Dawley(SD)rats were randomly divided into Control group(C group),incision pain group(I group),Remifentanil infusion+incision pain group(R+I group),remifentanil+Dezocine infusion+incision pain group(R+D+I group),with 10 rats in each group.Rats in C group were blank controls.Before modeling,rats in R+I group and R+D+I group were intravenously infused with remifentanil,and those in I group were intravenously infused with the same volume of normal saline.Subsequently,the left posterior plantar incision was made to construct the incision pain model in I group,R+I group and R+D+I group.Rats in the R+D+I group were pretreated with dezocine by tail vein injection before remifentanil infusion.Paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)before surgery and at different time points after operation were measured by behavioral tests.The contents of inflammatory factors,including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-10 in spinal cord dorsal horn were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expressions of Toll-like receptor 4(TLR4),nuclear factor kappa-B(NF-κB)and transient receptor potential ankyrin 1(TRPA1)in spinal cord dorsal horn were detected by quantitative real-time PCR(qRT-PCR)and Western blot,respectively.Results The rat model of remifentanil-induced postoperative hyperalgesia was successfully constructed.Compared with those in C group,rats in I group presented significantly lower PWT and PWL,higher contents of TNF-αand IL-1β,lower content of IL-10,and higher mRNA and protein expressions of TLR4,NF-κB and TRPA1 in spinal cord dorsal horn at different time points after operation(P<0.05).Compared with those in I group,rats in R+I group presented significantly lower PWT and PWL,higher contents of TNF-αand IL-1β,lower content of IL-10,and higher mRNA and protein expressions of TLR4,NF-κB and TRPA1 in spinal cord dorsal horn at different time points after operation(P<0.05).Compared with those in R+I group,rats in R+D+I group presented significantly higher PWT and PWL,lower contents of TNF-αand IL-1β,higher content of IL-10,and lower mRNA and protein expressions of TLR4,NF-κB and TRPA1 in spinal cord dorsal horn at different time points after operation(P<0.05).Conclusion Dezocine alleviates remifentanil-induced postoperative hyperalgesia by inhibiting the TLR4/NF-κB pathway,downregulating mRNA and protein expressions of TRPA1 and suppressing the inflammatory response in spinal cord dorsal horn.