Nrf2信号通路在富氢保存液减轻大鼠供肾低温保存期缺血损伤中的作用  

Role of Nrf2 signaling pathway in attenuation of ischemia injury by hydrogen-rich University of Wisconsin solution during cold storage of rat donor kidneys

作　　者：徐如彬[1] 喻文立[1] 盛明薇[1] 翁亦齐[1] 任恒昌[1] 贾莉莉[1] 杜洪印[1] 

机构地区：[1]天津市第一中心医院麻醉科,300192

出　　处：《中华麻醉学杂志》2017年第4期489-493,共5页Chinese Journal of Anesthesiology

基　　金：基金项目：天津市卫生行业重点攻关项目（13KG105）;天津市卫生计生委科技攻关项目（16KG101）

摘　　要：目的评价转录因子E2相关因子2（Nrf2）信号通路在富氢保存液减轻大鼠供肾低温保存期缺血损伤中的作用。方法健康成年雄性Wistar大鼠40只，体重200～250 g，采用随机数字表法将其分为4组（n＝10）：正常对照组（C组）、保存液（UW液）组（UW组）、富氢UW液组（HRUW组）、Nrf2抑制剂全反式维甲酸组（ATRA组）。ATRA组腹腔注射全反式维甲酸7 mg/kg，1次/d，连续2 d，最后1次给药后8 h时制备肾游离及冷保存期缺血损伤模型，肾脏在4 ℃富氢UW液中保存48 h。UW组和HRUW组腹腔注射等容量生理盐水替代，肾脏分别在4 ℃ UW液和富氢UW液中保存48 h。取肾组织，HE染色观察病理学结果，行肾小管损伤评分；采用ELISA法测定肾组织TNF-α、IL-1β、高迁移率族蛋白1（HMGB1）、IL-10及8-异前列腺素F2α（8-iso-PGF2α）的含量；采用分光光度法测定SOD和过氧化氢酶（CAT）活性；采用Western blot法测定肾组织Nrf2、血红素氧合酶-1（HO-1）、Bcl-2、Bax和caspase-3的表达。结果与C组比较，UW组肾小管损伤评分升高，肾组织TNF-α、IL-1β、HMGB1和8-iso-PGF2α含量升高，IL-10含量降低，Nrf2和HO-1表达上调，SOD和CAT活性降低，Bcl-2表达下调，Bax和caspase-3表达上调（P〈0.05或0.01）；与UW组比较，HRUW组肾小管损伤评分降低，肾组织TNF-α、IL-1β、HMGB1和8-iso-PGF2α含量降低，IL-10含量升高，Nrf2和HO-1表达上调，SOD和CAT活性升高，Bcl-2表达上调，Bax和caspase-3表达下调，ATRA组肾组织Nrf2和Bcl-2表达上调（P〈0.05），其余指标差异无统计学意义（P〉0.05）。与HRUW组比较，ATRA组肾小管损伤评分升高，肾组织TNF-α、IL-1β、HMGB1和8-iso-PGF2α含量升高，IL-10含量降低，HO-1和Bcl-2表达下调，SOD和CAT活性降低，Bax和caspase-3表达上调（P〈0.05）。结论富氢保存液减轻大鼠供肾低温保存期炎性反应、氧化损伤和细胞凋亡，从而减轻缺�Objective To evaluate the role of nuclear factor erythroid 2-related factor 2（Nrf2）signaling pathway in attenuation of ischemia injury by hydrogen-rich University of Wisconsin（HRUW）solution during cold storage of rat donor kidneys.Methods Forty healthy adult male Wistar rats, weighing 200-250 g, were divided into 4 groups（n=10 each）using a random number table： control group（group C）, University of Wisconsin（UW）solution group（group UW）, HRUW solution group（group HRUW）and Nrf2 inhibitor all-trans retinoic acid（ATRA）group（group ATRA）. ATRA 7 mg/kg was intraperitioneally injected once a day for 2 consecutive days, kidneys were isolated and underwent cold storage at 8 h after the last administration, and kidneys were stored in HRUW solution for 48 h at 4 ℃ in group ATRA.In UW and HRUW groups, the equal volume of normal saline was intraperitioneally injected instead, and isolated kidneys were stored in UW solution and HRUW solution for 48 h at 4 °C, respectively.Kidney specimens were obtained for microscopic examination and for determination of tumor necrosis factor-alpha（TNF-α）, interleukin-1beta（IL-1β）, high-mobility group box 1 protein（HMGB1）, IL-10 and 8-iso-prostaglandin F2α（8-iso-PGF2α）contents（by enzyme-linked immunosorbent assay）, superoxide dismutase（SOD）and catalase（CAT）activities（using spectrophotometry）, and expression of Nrf2, heme oxygenase-1（HO-1）, Bcl-2, Bax and caspase-3 in renal tissues（by using Western blot）. The damage to the renal tubules was scored.Results Compared with group C, renal tubular damage scores were significantly increased, TNF-α, IL-1β, HMGB1 and 8-iso-PGF2α contents were increased, IL-10 contents were decreased, the expression of Nrf2 and HO-1 was up-regulated, SOD and CAT activities were decreased, the expression of Bcl-2 was down-regulated, and the expression of Bax and caspase-3 was up-regulated in group UW（P〈0.05 or 0.01）. Compared with group UW, renal tubular damage scores were signific

关 键 词：氢 NF-E2相关因子2 肾移植 低温保存 冷缺血 

分 类 号：R699.2[医药卫生—泌尿科学]