脊髓Pellino1在瑞芬太尼诱导的大鼠痛觉过敏中作用及机制

The role of spinal Pellino1 in Remifentanil-induced Hyperalgesia in Rats and its mechanism

摘要:

摘 要 目的：观察脊髓Pellino1(Peli1)在瑞芬太尼诱导的痛觉过敏中作用及可能机制。方法：36只成年雄性SD大鼠随机平均分为6组，每组6只，生理盐水组（S 组），瑞芬太尼组(R 组)，生理盐水+ scrambled shRNA组（S+shscr 组），瑞芬太尼+ scrambled shRNA组（R+shscr 组），生理盐水+ Peli1 shRNA组（S+shPeli1），瑞芬太尼+ Peli1shRNA组（R+shPeli1组）。R、 R+shscr、R+ shPeli1组大鼠通过尾静脉连续输注瑞芬太尼4 μg/（kg?min）2 h，建立瑞芬太尼诱导的痛觉过敏模型；S+shscr、S+shPeli1、R+shscr、R+shPeli1组于瑞芬太尼给药前连续3天鞘内注射shscr或Peli1shRNA；分别采用机械缩足反射阈值（MWT）和热缩足反射潜伏期评价大鼠机械痛敏和热痛敏；Western blotting法和RT-PCR法检测Peli1、Iba1、GFAP蛋白及mRNA表达；ELISA法检测TNF-α、IL-6、IL-1?。结果：与S组相比，R组瑞芬太尼输注后6h、1天、3天MWT和TWL明显降低（P < 0.001）；与R+shscr组相比，R+ shPeli1组大鼠瑞芬太尼输注后6h、1天、3天MWT和TWL明显升高（P < 0.05, P < 0.001）；与瑞芬太尼输注前相比，R组大鼠瑞芬太尼输注后6h、1天、3天脊髓Peli1蛋白及mRNA表达显著升高（P < 0.001）；与S+shscr组相比，R+ shscr组大鼠瑞芬太尼输注后1天Iba1蛋白及mRNA以及TNF-α、IL-6、IL-1?表达明显升高（P < 0.001）；与R+shscr组相比，R+ shPeli1组大鼠瑞芬太尼输注后后1天Iba1蛋白及mRNA以及TNF-α、IL-6、IL-1?表达明显降低（P < 0.001）。结论：Peli1参与瑞芬太尼诱发的痛觉过敏，其机制可能与脊髓小胶质细胞激活及其炎症反应有关。

Abstract:

Abstract Objective: To observe the role of and of spinal Pellino1(Peli1) in hyperalgesia induced by remifentanil and its possible mechanism. Methods: 36 adult male SD rats were randomly divided into 6 groups with 6 rats in each group: saline group(S), remifentanil group (R), saline+scrambled shRNA group (S+shscr), remifentanil+scrambled shRNA group (R+shscr), saline+Peli1shRNA group (S+shPeli1), remifentanil+ Peli1shRNA group (R+shPeli1). The model of remifentanil-induced hyperalgesia was established by continuous infusion of remifentanil 4?g/(kg ?min) for 2 hours in R, R+shscr and R+shPeli1 group, and intrathecal injection of shScr or Peli1shRNA were performed in S+shscr, R+shscr, S +shPeli1 and R+shPeli1 for 3 consecutive days before remifentanil administration. Mechanical allodynia and heat hyperalgesia were evaluated by mechanical withdrawal threshold( MWT) and thermal withdrawal latency( TWL) respectively, Western blotting and PT-PCR were used to observe the protein and mRNA expression of Peli1, Iba1and GFAP, the expression TNF- α, IL-6 and IL-1were determined by ELISA. Results: compared with S group, MWT and TWL were significantly decreased in R group on the 1st day and 3rd day after infusion of remifentanil(P < 0.001), compared with R+ shscr group, MWT and TWL were markedly increased in R+shPeli1 group on the 1st day and 3rd day after infusion of remifentanil(P < 0.001),compared with the baseline, the protein and mRNA expression level of Peli1 in spinal cord were significantly increased in group R at 6h, 1 day and 3day after infusion of remifentanil (P < 0.001). compared with S+shscr group, the protein and mRNA expression level of Iba1 and the expression of TNF- α, IL-6 and IL-1? in spinal cord were obviously increased in group R at 1 day after infusion of remifentanil(P < 0.001), while, compared with R+shscr group, the protein and mRNA expression level of Iba1 and the expression of TNF- α, IL-6 and IL-1? in spinal cord were significantly decreased in group R+ shPeli1 at 1st day after infusion of remifentanil(P < 0.001). Conclusion: Peli1 is involved in the process of hyperalgesia induced by remifentanil, and its mechanism may be related to the activation of microglia and inflammatory reaction in rat spinal cord.