右美托咪定对肾缺血再灌注小鼠肾纤维化的影响:Akt在其中的作用  

Effects of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion:the role of Akt

作　　者：李渭敏[1] 周俊[1] 陈奕豪 吕婉娴 王汉兵[1] 杨智慧[1] 仲吉英[1] Li Weimin;Zhou Jun;Chen Yihao;Lyu Wanxian;Wang Hanbing;Yang Zhihui;Zhong Jiying(Department of Anesthesiology,Foshan First People′s Hospital,Foshan 528000,China;Institute,Foshan First People′s Hospital,Foshan 528000,China;Department of Laboratory,Foshan First People′s Hospital,Foshan 528000,China)

机构地区：[1]佛山市第一人民医院麻醉科,528000 [2]佛山市第一人民医院研究所,528000 [3]弗山市第一人民医院检验科,528000

出　　处：《中华麻醉学杂志》2019年第9期1062-1066,共5页Chinese Journal of Anesthesiology

基　　金：广东省医学科学技术研究基金项目(A2017127);广东省自然科学基金项目(2016A030313376)。

摘　　要：目的评价右美托咪定对肾缺血再灌注小鼠肾纤维化的影响及蛋白质丝氨酸-苏氨酸激酶(Akt)在其中的作用。方法雄性C57BL/6小鼠60只,8周龄,体重20~25 g,采用随机数字表法分为5组(n=12):假手术组(S组)、肾缺血再灌注组(I/R组)、肾缺血再灌注+右美托咪定组(I/R+D组)、肾缺血再灌注+右美托咪定+Akt激动剂SC79组(I/R+D+SC组)和肾缺血再灌注+右美托咪定+生理盐水组(I/R+D+NS组)。采用夹闭双侧肾蒂30 min后恢复肾脏灌注的方法制备肾缺血再灌注损伤模型。术前30 min时,I/R+D组、I/R+D+SC组及I/R+D+NS组腹腔注射右美托咪定0.1μg/g。I/R+D+SC组再灌注1 min时腹腔注射SC790.04 mg/kg,后每隔24 h再腹腔注射相同剂量,直至第7天。于再灌注24 h时检测血清BUN和Scr浓度,取肾组织,行肾小管损伤评分。于再灌注14 d时取肾组织检测肾纤维化程度,采用免疫荧光法和Western blot法检测肾组织胶原蛋白1(COL1)、纤维连接蛋白(FN)和α-平滑肌动蛋白(α-SMA)的表达水平。于再灌注24 h和14 d时,采用Western blot法测定肾组织磷酸化Akt(p-Akt)表达水平。结果与S组比较,I/R组血清BUN和Scr浓度、肾小管损伤评分和肾纤维化程度升高,COL1、FN、α-SMA和p-Akt表达上调(P<0.05);与I/R组比较,I/R+D组和I/R+D+NS组血清BUN和Scr浓度、肾小管损伤评分和肾纤维化程度降低,COL1、FN、α-SMA和p-Akt表达下调(P<0.05)。与I/R+D组比较,I/R+D+SC组血清BUN和Scr浓度、肾小管损伤评分和肾纤维化程度升高,COL1、FN、α-SMA和p-AKT表达上调(P<0.05)。结论右美托咪定可减轻肾缺血再灌注小鼠肾纤维化程度,机制与抑制Akt激活有关。Objective To evaluate the effect of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion(I/R)and the role of serine-threonine kinase(Akt).Methods Sixty male C57BL/6 mice,aged 8 weeks,weighing 20-25 g,were divided into 5 groups(n=12 each)using a random number table method:sham operation group(S group),renal I/R group(I/R group),renal I/R plus dexmedetomidine group(I/R+D group),renal I/R plus dexmedetomidine plus Akt agonist SC79 group(I/R+D+SC group),and renal I/R plus dexmedetomidine plus normal saline group(I/R+D+NS group).Renal I/R injury model was established by clamping the bilateral renal pedicle for 30 min followed by reperfusion.Dexmedetomidine was intraperitoneally injected at 30 min before surgery in I/R+D,I/R+D+SC and I/R+D+NS groups.SC79 was intraperitoneally injected as a bolus of 0.04 mg/kg at 1 min of reperfusion,followed by an intraperitoneal injection of the same dose every 24 h until day 7.The serum blood urea nitrogen(BUN)and Scr concentrations were detected at 24 h of reperfusion.Renal tissues were taken,and the damage to the renal tubules was scored.Renal tissues were removed at 14 days of reperfusion to detect the degree of renal fibrosis and expression of collagen 1(COL1),fibronectin(FN),and α-smooth actin(α-SMA)(by immunofluorescence and Western blot).The expression of phosphorylated Akt(p-Akt)in renal tissues was determined by Western blot at 24 h and 14 day of reperfusion.Results Compared with group S,the serum BUN and Scr concentrations,renal tubule damage score and degree of renal fibrosis were significantly increased,and the expression of COL1,FN,α-SMA and p-Akt was up-regulated in group I/R(P<0.05).Compared with I/R group,the serum BUN and Scr concentrations,renal tubular damage score and degree of renal fibrosis were significantly decreased,and the expression of COL1,FN,α-SMA and p-Akt was down-regulated in I/R+D and I/R+D+NS groups(P<0.05).Compared with I/R+D group,the serum BUN and Scr concentrations,renal tubule damage score and degree of renal fibr

关 键 词：右美托咪啶 纤维化 肾 再灌注损伤 蛋白质丝氨酸-苏氨酸激酶 

分 类 号：R614[医药卫生—麻醉学]