蛋白激酶Cα在电针减轻内毒素休克诱发兔急性肾损伤中的作用：与Nrf2/HO-1通路的关系  

Role of PKCα in electroacupuncture-induced reduction of acute kidney injury induced by endotoxic shock in rabbits： the relationship with Nrf2/HO-1 pathway

作　　者：曹新顺[1] 史佳[1] 余剑波[1] 张圆[1] 董树安[1] 吴丽丽[1] 

机构地区：[1]天津医科大学南开临床学院天津市南开医院麻醉科,300100

出　　处：《中华麻醉学杂志》2015年第6期727-731,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81372096）;天津市科技支撑计划重点项目（12ZCZDSY03300）

摘　　要：目的 评价蛋白激酶Cα（PKCα）在电针减轻内毒素休克诱发兔急性肾损伤中的作用及其与核因子E2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）通路的关系.方法 健康清洁级雄性新西兰大白兔80只,2月龄,体重1.5 ～ 2.0 kg,采用随机数字表法分为8组（n=10）：假手术组（S组）、内毒素休克致急性肾损伤组（AKI组）、PKCα抑制剂-白屈菜赤碱＋AKI组（CHA组）、白屈菜赤碱组（Che组）、溶媒-二甲基亚砜组（D组）、电针刺激穴位＋AKI组（EA组）、电针刺激非穴位＋AKI组（SEA组）和电针刺激穴位＋白屈菜赤碱＋AKI组（CEA组）.EA组和CEA组于模型制备前1-4 d及模型制备过程中电针刺激双侧足三里和肾俞穴,疏密波,频率2/15 Hz,刺激电流1～2 mA,波宽0.2～0.6 ms,刺激强度以兔肢体出现轻微肌颤为宜,1次/d,30 min/次.SEA组以同样方法电针刺激足三里和肾俞穴旁开0.5 cm非经非穴处.AKI组、CHA组、EA组、SEA组和CEA组采用耳缘静脉注射脂多糖5 mg/kg的方法制备内毒素休克致急性肾损伤模型,S组、Che组和D组给予等容量生理盐水.于模型制备前30 min,CHA组和CEA组静脉注射白屈菜赤碱5 mg/kg（溶于0.5 ml 0.1％二甲基亚砜）,Che组给予等量白屈菜赤碱,D组给予二甲基亚砜0.5 ml.静脉注射脂多糖或生理盐水后6h时,采集颈内动脉血样,检测血清BUN和Cr浓度,处死大鼠后取肾组织,行病理学观察并进行评分,测定肾组织MDA含量及SOD活性,检测肾组织PKCα、HO-1蛋白、核蛋白及总蛋白中Nrf2的表达水平.结果 与S组比较,AKI组、CHA组、EA组、SEA组和CEA组血清BUN和Cr浓度升高,肾组织MDA含量升高,SOD活性降低,肾组织病理学评分升高,PKCα、HO-1蛋白、核蛋白及总蛋白Nrf2的表达上调（P＜0.05）;与AKI组比较,EA组血清BUN和Cr浓度降低,肾组织MDA含量降低,SOD活性升高,肾组织病理学评分降低,PKCα、HO-1蛋白、核蛋白及总蛋白Nrf2的表达上调,CHA组Objective To evaluate the role of protein kinase Ca （PKCα） in electroacupuncture （EA）-induced reduction of acute kidney injury （AKI） induced by endotoxic shock,and the relationship with nuclear factor E2-related factor 2/heme oxygenase-1 Nrf2/HO-1 pathway in rabbits.Methods Eighty heahhy male New Zealand white rabbits,aged 2 months,weighing 1.5-2.0 kg,were randomly divided into 8 groups （n=10 each） using a random number table：sham operation group （group S）,group AKI,specific PKCα inhibitor chelerythrine ＋ AKI group （group CHA）,chelerythrine group （group Che）,dimethyl sulfoxide （DMSO） group （group D）,EA at acupoints ＋ AKI group （group EA）,EA at non-acupoints ＋ AKI group （group SEA）,and EA at acupoints ＋ chelerythrine ＋ AKI group （group CEA）.Bilateral 30 min EA （disperse-dense wave,wave length 0.2-0.6 ms,frequency 2/15 Hz,intensity 1-2 mA） stimulation of Zusanli and Shenshu acupoints was performed once a day for 4 days before establishment of the model and during the process of establishment of the model in EA and CEA groups.In group SEA,EA was performed at the points 0.5 cm lateral to the acupoints of Zusanli and Shenshu with the same parameters.The animals were anesthetized with iv 20% urethane 5 ml/kg,tracheostomized and kept spontaneous breathing.Lipopolysaccharide 5 mg/kg （in 2 ml of normal saline） was injected via the auricular vein to establish the model of endotoxic shock-induced AKI in AKI,CHA,EA,SEA and CEA groups,while the equal volume of normal saline was given in S,Che and D groups.At 30 min before establishment of the model,chelerythrine 5 mg/kg （in 0.5 ml of 1% DMSO） was injected intravenously in CHA and CEA groups,the equal volume of chelerythrine was given in Che group,while the equal volume of DMSO was given in group D.At 6 h after lipopolysaccharide or normal saline injection,blood samples were taken from the internal carotid artery for determination of serum urea nitrogen （BUN） and creatinine （Cr） concentrations.Th

关 键 词：电刺激疗法 蛋白激酶CΑ NF—E2相关因子2 血红素加氧酶-1 休克 脓毒性 急性肾损伤 

分 类 号：R245[医药卫生—针灸推拿学]