摘要
目的 探讨核因子E2相关因子2(Nrf2)激动剂对肺缺血再灌注损伤大鼠的保护作用.方法 30只SD大鼠按照随机数字表法分为对照组、再灌注组和Nrf2激动剂组,每组10只.再灌注组和Nrf2激动剂组大鼠采用在体左肺门夹闭法复制缺血再灌注损伤模型,对照组仅作切口,不夹闭.Nrf2激动剂组大鼠建模成功后每天口服富马酸二甲酯100 mg/kg,对照组和再灌注组大鼠每天给予等体积生理盐水.治疗7 d后,取左肺测定组织干湿比和总肺含水量;苏木精-伊红(HE)染色分析大鼠肺泡损伤状况;测定3组大鼠肺组织铁离子、丙二醛(MDA)和谷胱甘肽氧化酶的水平;采用酶联免疫吸附实验分析3组大鼠肺组织炎性因子[肿瘤坏死因子(TNF)、白细胞介素(IL)-6和IL-1β]的水平;蛋白质免疫印迹分析大鼠肺组织铁死亡相关蛋白[转铁蛋白受体1、谷胱甘肽过氧化物酶4(GPX4)和溶质载体家族7成员11]蛋白表达水平.组间计量数据比较采用单因素方差分析.结果 Nrf2激动剂组大鼠肺泡损伤百分比[(31.20±5.37)％]明显低于再灌注组[(65.34± 5.60)％],差异有统计学意义(t=13.920,P＜0.05).Nrf2激动剂组大鼠肺干湿比(2.44±0.13)明显低于再灌注组(3.85±0.39),差异有统计学意义(t=10.950,P＜0.05).Nrf2激动剂组大鼠血清TNF、IL-6 和 IL-1β[(33.44±4.72)、(27.61±1.73)、(23.87±2.64)pg/ml]明显低于再灌注组[(78.85±9.63)、(43.12±10.46)、(39.79±8.78)pg/ml],差异有统计学意义(t=13.390、4.625、5.488,P＜0.05).Nrf2激动剂组大鼠肺组织铁离子浓度[(0.62±0.12)μmol/mg]明显低于再灌注组[(1.01±0.10)μmol/mg],差异有统计学意义(t=7.823,P＜0.05).Nrf2激动剂组大鼠肺组织MDA水平[(1.06±0.07)μmol/mg]明显低于再灌注组[(1.68±0.14)μmol/mg],差异有统计学意义(t=12.370,P＜0.05).Nrf2激动剂组大鼠谷胱甘肽过氧化物酶(GSH-Px)活性[(137.70± 11.27)U/mg]明显低于再灌注组[(102.90±10.48)U/mg],差异有统计学意义(t=7.148,P＜0.05).Nrf2激动剂组大鼠肺组织转铁蛋白受体1表达水平(1.24±0.10)明显低于再灌注组(1.71±0.11),差异有统计学意义(t=10.090,P＜0.05).Nrf2激动剂组大鼠肺组织GPX4和溶质载体家族7成员11表达水平(0.95±0.12、0.79±0.06)明显高于再灌注组(0.76±0.06、0.53± 0.06),差异有统计学意义(t=5.877、9.394,P＜0.05).结论 Nrf2激动剂可抑制缺血再灌注损伤大鼠肺组织铁死亡相关蛋白表达水平,抑制肺组织炎性因子的释放,减轻肺组织缺血再灌注损伤.
Abstract
Objective To investigate the protective effect of nuclear factor erythroid 2-related fac-tor 2(Nrf2)agonist on lung ischemia-reperfusion injury in rats.Methods Totally,30 SD rats were divid-ed into control group,reperfusion group and Nrf2 agonist group with 10 rats in each group according to the random number table method.The rats in the reperfusion group and the Nrf2 agonist group were subjected to the left hilar clamp method to replicate the ischemia-reperfusion injury model,while the control group was subjected to incision without clamp.After successful modeling,rats in the Nrf2 agonist group were giv-en dimethyl fumarate 100 mg/kg orally every day,and those in the control group and reperfusion group were given equal volume of normal saline every day.After 7 days of treatment,the dry-wet ratio and total lung water content were measured in the left lung.Hematoxylin and eosin(HE)staining was used to ana-lyze the alveolar damage in rats.The levels of iron ion,malondialdehyde(MDA)and glutathione oxidase in lung tissues were determined.The levels of lung inflammatory factors[tumor necrosis factor(TNF),in-terleukin(IL)-6 and IL-1 β]were analyzed by enzyme-linked immunosorbent assay(ELISA).The expres-sion levels of iron death-related proteins(transferrin receptor 1,glutathione peroxidase 4 and solsolic carri-er family 7 members 11)in rat lung tissues were analyzed by Western blotting.One-way analysis of vari-ance was used to compare the measurement data between groups.Results The percentage of alveolar damage in the Nrf2 agonist group[(31.20±5.37)％]was significantly lower than that in the reperfusion group[(65.34±5.60)％,t=13.920,P＜0.05].The dry/wet lung ratio in the Nrf2 agonist group(2.44±0.13)was significantly lower than that in the reperfusion group(3.85±0.39,t=10.950,P＜0.05).Serum TNF,IL-6 and IL-1[3 levels in the Nrf2 agonist group[(33.44±4.72),(27.61±1.73),(23.87±2.64)pg/ml]were lower than those in the reperfusion group[(78.85±9.63),(43.12± 10.46),(39.79±8.78)pg/ml,t=13.390,4.625,5.488,P＜0.05].The concentration of iron ion in lung tissue of the Nrf2 agonist group[(0.62±0.12)μmol/mg]was significantly lower than that of the reperfusion group[(1.01±0.10)μmol/mg,t=7.823,P＜0.05].The level of MDA in lung tissue of the Nrf2 agonist group[(1.06±0.07)μmol/mg]was significantly lower than that of the reperfusion group[(1.68±0.14)μmol/mg,t=12.370,P＜0.05].The glutathione peroxidase(GSH-px)activity in the Nrf2 agonist group[(137.70±11.27)U/mg]was significantly lower than that in the reperfusion group[(102.90±10.48)U/mg,t=7.148,P＜0.05].The expression level of transferrin receptor 1 in lung tissue of the Nrf2 agonist group(1.24±0.10)was significantly lower than that of the reperfusion group(1.71±0.11,t=10.090,P＜0.05).The expression levels of glutathione peroxidase 4 and solute carrier family 7 members 11 in lung tissue of the Nrf2 agonist group(0.95±0.12,0.79±0.06)were significant-ly higher than those in the reperfusion group(0.76±0.06,0.53±0.06,t=5.877,9.394,P＜0.05).Conclusion Nrf2 agonist can inhibit the expression level of iron death-related protein,suppress the re-lease of inflammatory factors in lung tissue,and alleviate lung ischemia reperfusion injury in rats.
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