摘要
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,Westernblot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45μmo L/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(0h),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。
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 （45μmoL/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＆lt;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 〈br〉 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.
出处
《天津医药》
CAS
2016年第9期1073-1077,共5页
Tianjin Medical Journal
基金
天津市卫生局科技基金资助项目(2014KZ016)
