羟考酮对大鼠肠缺血再灌注损伤的影响及自噬在其中的作用  

Effect of oxycodone on intestinal ischemia-reperfusion injury in rats and the role of autophagy

作　　者：杨涛 谢淑华[1] 王蕾[1] 杨学伟[1] 耿立成[1] 王志芬 于泳浩[3] Yang Tao;Xie Shuhua;Wang Lei;Yang Xuewei;Geng Licheng;Wang Zhifen;Yu Yonghao(Department of Anesthesiology, Tianjin Union Medical Center, Tianjin 300191, China;Department of Anesthesiology, Tianjin Children′s Hospital, Tianjin 300134, China;Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China)

机构地区：[1]天津市人民医院麻醉科,300191 [2]天津市儿童医院麻醉科,300134 [3]天津医科大学总医院麻醉科,300052

出　　处：《中华麻醉学杂志》2018年第10期1194-1197,共4页Chinese Journal of Anesthesiology

基　　金：天津市卫计委重点攻关项目(14KG106);天津市人民医院院级基金项目(2017YJ013).

摘　　要：目的评价羟考酮对大鼠肠缺血再灌注损伤的影响及自噬在其中的作用。方法健康成年雄性SD大鼠24只,6~9周龄,体重180~220g,采用随机数字表法分为4组(n=6):假手术组(S组)、肠缺血再灌注组(I/R组)、羟考酮组(O组)和羟考酮+自噬抑制剂3-甲基腺嘌呤组(O+3-MA组)。采用夹闭肠系膜上动脉1h再灌注2h的方法制备肠缺血再灌注损伤模型。O组和O+3-MA组于缺血前15min时尾静脉注射羟考酮0.5mg/kg。O+3-MA组于缺血前10min腹腔注射3-MA30mg/kg。于缺血前即刻胃内灌注异硫氰酸荧光素标记的右旋糖酐(FITC-dextran)。再灌注2h时心尖穿刺采集血样,测定血清FITC-dextran浓度;于再灌注2h时心尖穿刺采集血样,ELISA法测定血清TNF-α、IL-1β浓度;取小肠组织,观察病理学变化并行Chiu评分;Westernblot法测定紧密连接蛋白occludin、自噬相关蛋白Beclin-1和微管相关蛋白1轻链3(LC3)的表达,并计算LC3Ⅱ与LC3Ⅰ表达比值(LC3Ⅱ/Ⅰ比值)。结果与S组比较,I/R组Chiu评分、血清FITC-dextran、TNF-α和IL-1β浓度、LC3Ⅱ/Ⅰ比值升高,Beclin-1表达上调,occludin表达下调(P<0.05);与I/R组比较,O组Chiu评分、血清FITC-dextran、TNF-α和IL-1β浓度降低,LC3Ⅱ/Ⅰ比值升高,occludin和Beclin-1表达上调(P<0.05),O+3-MA组上述指标差异无统计学意义(P>0.05);与O组比较,O+3-MA组Chiu评分、血清FITC-dextran、TNF-α和IL-1β浓度升高,LC3Ⅱ/Ⅰ比值降低,occludin和Beclin-1表达下调(P<0.05)。结论羟考酮可减轻大鼠肠缺血再灌注损伤,其机制可能与增强肠组织自噬水平有关。Objective To evaluate the effect of oxycodone on intestinal ischemia-reperfusion injury in rats and the role of autophagy. Methods Twenty-four healthy adult male Sprague-Dawley rats, aged 6-9 weeks, weighing 180-220 g, were divided into 4 groups(n=6 each)using a random number table method: sham group(group S), intestinal I/R group(group I/R), oxycodone group(group O)and oxycodone plus autophagy inhibitor 3-methyladenine(3-MA)group(group O+ 3-MA). Intestinal I/R was induced via clamping the superior mesenteric artery for 1 h, followed by 2-h reperfusion in all the groups except for group S. Oxycodone 0.5 mg/kg was injected via caudal vein at 15 min before ischemia in group O and group O+ 3-MA.3-MA 30 mg/kg was intraperitoneally injected at 10 min before ischemia in group O+ 3-MA.Rats were gavaged with fluorescein-isothiocyanate-conjugated dextran(FITC-dextran)immediately before ischemia.Blood samples were collected from the cardiac apex at 2 h of reperfusion to detect the level of serum FITC-dextran.Blood samples were collected from the cardiac apex at 2 h of reperfusion to measure the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-1beta(IL-1β)in serum by enzyme-linked immunosorbent assay.Small intestinal tissues were obtained at 2 h of reperfusion for examination of the pathological changes and for determination of the expression of occludin, Beclin-1 and microtubule-associated protein 1 light chain 3(LC3)(by Western blot). Intestinal damage was assessed and scored according to Chiu.The ratio of LC3Ⅱ expression to LC3Ⅰ expression(LC3Ⅱ/Ⅰ)was calculated. Results Compared with group S, the Chiu′s score, levels of serum FITC-dextran, TNF-α and IL-1β and LC3Ⅱ/Ⅰ ratio were significantly increased, the expression of Beclin-1 was up-regulated, and the expression of occludin was down-regulated in group I/R(P<0.05). Compared with group I/R, the Chiu′s score and levels of serum FITC-dextran, TNF-α and IL-1β were significantly decreased, LC3Ⅱ/Ⅰ ratio was increased, and the expre

关 键 词：羟可酮 再灌注损伤 肠 自噬 

分 类 号：R614[医药卫生—麻醉学]