电针对内毒素性休克兔急性肾损伤的影响：与Keap1-Nrf2/ARE信号通路的关系  

Effects of electro-acupuncture on endotoxic shock-induced acute kidney injury in rabbits： relationship with Keap1-Nrf2/ARE signaling pathway

作　　者：郭艳辉 宫丽荣[2] 余剑波[2] 史佳[2] 张圆[2] 徐妍[2] 吴丽丽[2] 曹新顺[2] 

机构地区：[1]天津市海河医院麻醉科 [2]天津市南开医院麻醉科,300100

出　　处：《中华麻醉学杂志》2014年第10期1255-1258,共4页Chinese Journal of Anesthesiology

基　　金：天津市科技支撑计划重点项目（12ZCZDSY013300）

摘　　要：目的 评价电针对内毒素性休克兔急性肾损伤的影响及其与Kelch样环氧氯丙烷相关蛋白-1（Keap1）-转录因子NF-E2相关因子2（Nrf2）/抗氧化反应元件（ARE）信号通路的关系.方法 清洁级健康成年雄性新西兰大白兔40只,体重1.5 ～ 2.0 kg,采用随机数字表法,将其分为4组（n=10）：对照组（C组）、内毒素性休克诱发急性肾损伤组（AKI组）、电针＋内毒素性休克诱发急性肾损伤组（EA组）、非穴位电针＋内毒素性休克诱发急性肾损伤组（SEA组）.采用耳缘静脉注射脂多糖（LPS）5 mg/kg（溶于2 ml生理盐水）建立兔内毒素性休克诱发急性肾损伤模型.EA组电针足三里穴和肾俞穴,1次/d,每次持续10 min,连续4d,刺激参数：疏密波,频率2/15 Hz,波宽0.2 ～ 0.6 ms,电流1～2mA,以兔出现轻微肌颤为度,留针1h出针,第5天建立模型,持续电针刺激至处死.SEA组选择穴位旁开0.5 cm部位行电针刺激,其余与EA组相同.于注射LPS后6h时,取尿液2ml,测定尿α1微球蛋白（α1-MG）浓度.HE染色下观察肾脏病理学结果,并行肾脏组织学（HSK）评分.干湿重法测定肾组织含水率,RT-PCR法测定Nrf2 mRNA、血红素氧合酶-1（HO-1） mRNA,Western blot法测定肾组织Nrf2总蛋白、核蛋白及下游HO-1蛋白表达水平.结果 与C组比较,AKI组、EA组、SEA组HSK评分、肾组织含水率、尿α1-MG浓度升高,肾组织Nrf2总蛋白、Nrf2核蛋白、HO-1蛋白、Nrf2 mRNA和HO-1mRNA表达上调（P＜0.05）;与EA组比较,AKI组和SEA组HSK评分、肾组织含水率、尿α1-MG浓度升高,肾组织Nrf2总蛋白、Nrf2核蛋白、HO-1蛋白、Nrf2 mRNA和HO-1 mRNA表达下调（P＜0.05）.AKI组和SEA组各项指标比较差异无统计学意义（P＞0.05）.结论 电针足三里和肾俞穴可减轻内毒素性休克兔急性肾损伤,其机制可能与激活Keap1-Nrf2/ARE信号通路有关.Objective To evaluate the effects of electro-acupuncture （EA） at Zusanli and Shenshu acupoints on endotoxic shock-induced acute kidney injury （AKI） and the relationship with Keap1-Nrf2/ARE signaling pathway in rabbits.Methods Forty pathogen-free male New Zealand white rabbits,weighing 1.5-2.0 kg,were randomly divided into 4 groups randomly （n =10 each） using a random number table：control group （group C） ; endotoxic shock-induced AKI （group AKI） ; EA ＋ endotoxin-induced AKI group （group EA） ; EA at non-acupoints ＋ endotoxin-induced AKI group （group SEA）.Endotoxic shock-induced AKI was induced with LPS 5 mg/kg （in 2 ml of normal saline） injected via the auricular vein.Electro-stimulation （2/15 Hz,0.2-0.6 ms,1-2 mA） of Zusanli and Shenshu acupoints was performed for 10 min once a day for 4 consecutive days,and the model was established on 5th day and then EA was continued until the animals were sacrificed in group EA.In group SEA,EA was performed at the points 5 mm lateral to the acupoints of Zusanli and Shenshu,and the other procedures were similar to those previously described in group EA.At 6 h of LPS,the urine was collected for determination of α1-microglobulin （α1-MG） concentrations.The animals were sacrificed and kidneys were removed for microscopic examination of the pathological changes which were also scored.Dry-wet weight method was used to measure water content in renal tissues.Nrf2 mRNA and HO-1 mRNA expression was detected using RT-PCR.The expression of Nrf2 total protein and nuclear protein and downstream HO-1 in renal tissues was determined by Western blot.Results Compared with group C,pathological scores,water content in renal tissues,and urinary α1-MG were significantly increased,and the expression of Nrf2 total protein and nuclear protein,HO-1,Nrf2 mRNA and HO-1 mRNA was up-regulated in AKI,EA and SEA groups.Compared with group EA,pathological scores,water content in renal tissues,and urinary α1-MG were significantly increased,and the expression of

关 键 词：电刺激疗法 穴 足三里 穴 肾俞 内毒素血症 肾功能衰竭 急性 休克 脓毒性 核因子NF-E2相关因子 

分 类 号：R245[医药卫生—针灸推拿学]