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 Nrf2 total protein and nuclear protein,HO-1,Nrf2 mRNA and HO-1 mRNA was down-regulated in AKI and SEA groups.There was no significant difference in each parameter between AKI and SEA groups.Conclusion EA at Zusanli and Shenshu can attenuate AKI induced by endotoxic shock in rabbits,and activation of keap1-Nrf2/ARE signaling pathway may be involved in the mechanism.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
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）
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
Electric stimulation therapy
POINT ST36 （ZUSANLI）
POINT SL23 （SHENSHU）
Endotoxemia
Shock, septic
Kidney failure, acute
Nuclear factor erythroid 2-related factor2
