富氢液通过激活HO-1减轻炎性痛大鼠炎症因子的表达

Hydrogen-rich saline ameliorates the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1

目的 探讨富氢液是否通过激活血红素氧合酶-1（HO-1）减轻炎性痛大鼠的炎症反应.方法 成年雄性SD大鼠80只,体重180 ～ 220 g,采用随机数字表法,将其分为5组（n=16）：对照组（Con组）、炎性痛组（CFA）、炎性痛＋富氢液组（CFA＋ H2）、炎性痛＋HO-1抑制剂Znpp-Ⅸ组（CFA＋ Znpp-Ⅸ）、炎性痛＋富氢液＋HO-1抑制剂Znpp-Ⅸ组（CFA＋H2＋ Znpp-Ⅸ）.分别于模型制作第1天（T1）、第2天（T2）、第3天（T3）、第5天（T4）、第7天（T5）和第14天（T6）检测大鼠疼痛行为学（机械痛敏和热痛敏）;于第7天收集大鼠脊髓组织,应用RT-PCR和Westem blot等方法检测HO-1的mRNA、蛋白表达和活性的变化,应用ELISA法检测炎症因子的表达.结果 与对照组比较,CFA组动物机械痛敏MWT和热痛敏TWL降低,脊髓组织中HO-1 mRNA、蛋白表达和活性增加,炎症因子TNF-α、IL-1β、IL-6和IL-10表达增加（P＜0.05）;与CFA组比较,CFA＋ H2组动物机械痛敏MWT和热痛敏TWL增加,脊髓组织中HO-1 mRNA、蛋白表达和活性进一步增加,炎症因子TNF-α、IL-1β和IL-6表达减少,而IL-10表达进一步增加（P＜0.05）;与CFA＋ H2组比较,CFA＋ H2＋Znpp-Ⅸ组动物机械痛敏MWT和热痛敏TWL降低,脊髓组织中HO-1 mRNA、蛋白表达和活性减少,炎症因子TNF-α、IL-1β和IL-6表达增加,而IL-10表达减少（P＜0.05）.结论 富氢液可减轻炎性痛大鼠的机械痛敏和热痛敏,降低炎症因子释放,与激活HO-1密切相关.

Objective To investigate whether hydrogen-rich saline could ameliorate the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1 （HO-1）.Methods Eighty male Sprague-Dawley （SD） rats,weighing 180 ～ 220 g,were randomly divided into five groups （n =16 in each group）：Control group （Con）,inflammation pain group （CFA）,inflammation pain ＋ hydrogen-rich saline group （CFA ＋ H2）,inflammation pain ＋ HO-1 inhibitor Znpp-Ⅸ group （CFA ＋ Znpp-Ⅸ）,and inflammation pain ＋ hydrogen-rich saline ＋ HO-1inhibitor Znpp-Ⅸ group （CFA ＋ H2 ＋ Znpp-Ⅸ）.The mechanical withdrawal threshold （MWT） and thermal withdrawal latency （TWL） were tested on days 1 （T1）,2 （T2）,3 （T3）,5 （T4）,7 （T5）,and 14 （T6） after inflammation pain.The expressions of spinal HO-1 mRNA and protein were measured with real-time quantitative polymerase chain reaction （RT-PCR） and Western blot,and spinal inflammatory cytokines were measured with enzyme-linked immunosorbent assay （ELISA） on day 7 after inflammatory pain.Results Compared to Con group,MWT and TWL were significantly reduced;the spinal HO-1 mRNA level,protein expression and activity were increased;and the levels of tumor necrosis factor-α （TNF-α）,interleukin （IL）-1β,IL-6 and IL-10 in spinal tissues were also increased in CFA group （P 〈 0.05）.Compared to CFA group,MWT and TWL were significantly increased;the spinal HO-1 mRNA level,protein expression and activity were further increased;and the levels of TNF-α,IL-1β and IL-6 were decreased,while IL-10 was further increased in CFA ＋ H2group （P 〈 0.05）.Compared to CFA ＋ H2 group,MWT and TWL were decreased;the spinal HO-1 mR-NA level,protein expression and activity were decreased;and the levels of TNF-α,IL-1β and IL-6 in spinal tissue were significantly increased,while IL-10 was decreased in CFA ＋ H2 ＋ Znpp-Ⅸ group （P 〈0.05）.Conclusions Hydrogen-rich saline can ameliorate the mechanical and thermal allodynia in a rat model of inflammatory pain,and reduce the release of inflammatory cytokines via activating HO-1.