氢气对单关节炎大鼠的保护作用

Protective effects of hydrogen on monoarthritis rats

目的观察经呼吸道吸入氢气对单关节炎大鼠的保护作用。方法 40只雄性SD大鼠随机分为5组:单关节炎组、单关节炎+氢气0~14 d组、单关节炎+氢气0~3 d组、单关节炎+氢气4~14 d组、假手术+氢气0~14 d组,每组8只。于大鼠左侧踝关节腔注射完全弗氏佐剂(CFA)建立单关节炎模型。单关节炎组大鼠不吸入氢气,另4组大鼠分别在造模当天及造模后第1~14天每天吸入65%氢气1.5 h、在造模当天及造模后第1~3天每天吸入65%氢气1.5 h、在造模后第4~14天每天吸入65%氢气1.5 h、在假手术当天及造模后第1~14天每天吸入65%氢气1.5 h。采用von Frey纤毛测定单关节炎大鼠双侧后爪在建模后0、1、3、5、7、10、14 d的机械刺激抬腿反应阈值(PWT)。另取15只大鼠分为5组,每组3只,于造模或假手术后第10天取致炎侧脊髓腰膨大组织,用大鼠超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)检测试剂盒分别检测致炎侧脊髓组织中SOD、CAT、MDA的含量。结果单关节炎组大鼠致炎侧后爪PWT在建模后第1天即低于健侧,在第3天达到最低值并维持稳定至第14天(P<0.01)。单关节炎+氢气0~14 d组和单关节炎+氢气0~3 d组大鼠致炎侧后爪PWT均高于单关节炎组致炎侧(P<0.05,P<0.01);单关节炎+氢气4~14 d组大鼠致炎侧后爪PWT与单关节炎组致炎侧相比差异无统计学意义(P>0.05)。单关节炎+氢气0~14 d组致炎侧脊髓组织中SOD、CAT水平均高于单关节炎组(P均<0.05),MDA水平低于单关节炎组(P<0.05),而单关节炎+氢气0~3 d组和单关节炎+氢气4~14 d组致炎侧脊髓组织中SOD、CAT和MDA水平与单关节炎组比较差异均无统计学意义(P均>0.05)。结论经呼吸道吸入65%氢气在大鼠单关节炎模型的起始阶段可减轻机械痛敏及氧化应激,预先给予氢气可抑制单关节炎机械痛敏的形成。

Objective To explore the protective effect of hydrogen(H2) inhalation through respiratory tract on monoarthritis(MA) rats. Methods Forty male SD rats were randomly divided into 5 groups: MA group, MA+H2 0-14 d group, MA+H2 0-3 d group, MA+H2 4-14 d group, and sham+H2 0-14 d group(n=8). Complete Freund’s adjuvant(CFA) was injected into the left ankle joint of rats to construct MA model. The rats in the MA group did not inhale H2, and the rats in other four groups inhaled 65% H2 1.5 h per day on the day of modeling and 1-14 days after modeling, 1.5 h per day on the day of modeling and 1-3 days after modeling, 1.5 h per day on the 4 th-14 th day after modeling, and 1.5 h per day on the day of sham operation and 1-14 days after modeling, respectively. Paw withdrawal threshold(PWT) was detected by von Frey method 0, 1, 3, 5, 7, 10, and 14 d after modeling. Another 15 rats were randomly divided into 5 groups and there were 3 rats in each group. Lumbar enlargement tissues were taken from the spinal cord of the inflammatory side on the 10 th day after modeling or sham operation. The contents of superoxide dismutase(SOD), catalase(CAT) and malondialdehyde(MDA) were detected. Results In the MA group, the posterior paw PWT of the inflammatory side was lower than that of the healthy side on the first day after modeling, reached the lowest on the 3 th day and remained stable until the 14 th day(P<0.01). The PWT of the inflammatory side of the hind claw of rats in the MA+H2 0-14 d group and the MA+H2 0-3 d group was higher than that in the MA group(P<0.05, P<0.01). The PWT of the inflammatory side in the MA+H2 4-14 d group was not significantly different from that in the MA group(P>0.05). The levels of SOD and CAT in the spinal cord of the inflammatory side in the MA+H2 0-14 d group were higher than those in the MA group(all P<0.05), while MDA level was lower than that in the MA group(P<0.05). Compared with MA group, there was no significant difference in the SOD, CAT or MDA level in the MA+H2 0-3 d group and MA+H2 4-14 d group(all P>0.05). Conclusion Inhalation of 65% H2 via the respiratory tract at the initial stage of MA rat modeling can alleviate mechanical hyperalgesia and oxidative stress. Pre-administration of H2 can inhibit the formation of mechanical hyperalgesia in MA.