HO-1/CO 信号通路对切口痛大鼠炎症因子的调控作用

目的 观察脊髓血红素氧合酶-1（HO-1）/CO 信号通路对切口痛大鼠炎症因子的调节作用及可能机制。方法 切口痛模型大鼠 36 只在实验即刻（0 h）， 术后 1、4、8、12 及 24 h 各处死 6 只， 取患侧脊髓腰膨大处， Western blot 法检测 HO-1 蛋白表达， 酶联免疫吸附试验（ELISA）法检测炎症因子肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6 和高迁移率族蛋白 1（HMGB1）。 另外， 对照（C）组 36 只大鼠不做切口痛模型；切口痛模型大鼠 144 只按处理方式不同分为切口痛（IP）组， 切口痛+HO-1 诱导剂（IP+hemin）组， 术前给予腹腔注射大鼠 100 mg/kg 血红素（hemin）； 切口痛+HO-1 抑制剂（IP+Znpp-IX）组， 术前给予腹腔注射大鼠 45 μmoL/kg 锌原卟啉（Znpp）-IX； 切口痛+ CO 释放剂（IP+CORM-2）组， 于术前经腹腔注射给予 10 mg/kg 一氧化碳释放分子（CORM）-2。 各组于实验即刻（0 h）， 术后 1、4、8、12 及 24 h 行机械缩足阈值（PWMT）和热缩足潜伏期（PWTL）的检测， 应用 ELISA 法检测各组 TNF- α、IL-1β、IL-6 和 HMGB1 的表达情况。 结果 与 0 h 比较， 术后 1、4、8、12 及 24 h HO-1 蛋白表达水平和 TNF-α、 IL-1β、IL-6 和 HMGB1 均增高（P＜ 0.05）。 与 C 组比较， 其余组大鼠各时间点 PWMT 值和 PWTL 值减少， TNF-α、 IL-1β、IL-6 和 HMGB1 表达增加（P＜ 0.05）； 与 IP 组比较， IP+hemin 组和 IP+CORM-2 组大鼠 1、4、8、12 及 24 h 时 PWMT 值和 PWTL 值均增高， 而 TNF-α、IL-1β、IL-6 和 HMGB1 表达减少， IP+Znpp-IX 组 PWMT 值和 PWTL 值降低， 但 TNF-α、IL-1β、IL-6 和 HMGB1 表达增加（P＜ 0.05）。 结论 切口痛可诱发大鼠 HO-1 表达增加， 而 HO-1/CO 信号通路在调控切口痛大鼠炎症因子的释放方面发挥重要作用。

The regulatory effect of HO-1 /CO pathway on inflammatory cytokines in a rat model of incisional pain

Objective To investigate the effects of HO/CO pathway on inflammation cytokines in a rat model of incisional pain. Methods Thirty-six rats were executed to collect ipsilateral spinal cord tissues for HO- 1 detection by Western blot assay, and cytokines tumor necrosis factor (TNF)- a, interleukin (IL)-1b, IL-6 and high mobility group box (HMGB)1 were detected by ELISA before and at 1, 4, 8, 12 and 24 h after establishing incisional pain model. Additionally, 36 rats without establishment of incisional pain model were used as control group. A total of 144 model rats of incisional pain were divided into incisional pain (IP) group, IP+hemin group (100 mg/kg hemin was injected by i.p. before operation), IP+ Znpp-IX group (45 μmoL/kg Znpp-IX was injected by i.p. before operation) and IP+CORM-2 group (10 mg/kg CORM-2 was injected by i.p. before operation). Values of paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were detected, and expressions of TNF-a, IL-1 b, IL-6 and HMGB1 were measured by ELISA before and at 1, 4, 8, 12 and 24 h after operation. Results Compared with pre-operation of incisional pain in rats, expression levels of HO-1 protein and cytokines TNF-a, IL-1 b, IL-6 and HMGB1 were increased at 1, 4, 8, 12 and 24 h after operation (P＜ 0.05). Compared with control group, values of PWMT and PWTL were obviously decreased, and expression levels of IL-1β, TNF-α, IL-6 and HMGB1 were increased at 1, 4, 8, 12 and 24 h after operation in IP groups (P＜ 0.05). Compared with IP groups, values of PWMT and PWTL were significantly increased and cytokines TNF- a, IL-1 b, IL-6 and HMGB1 were decreased at 1, 4, 8, 12 and 24 h after operation in IP+hemin group and IP+CORM-2 group (P＜ 0.05). Values of PWMT and PWTL were decreased and cytokines TNF-α, IL-1β, IL-6 and HMGB1 were increased in IP+Znpp-IX group (P＜ 0.05). Conclusion Incisional pain can increase the expression of HO-1, and HO-1/CO pathway exists the regulatory effect on inflammatory cytokines in the rat model of incisional pain.