阿片受体激动剂对肠缺血再灌注损伤大鼠NADPH氧化酶活性及eNOS-NO通路的作用机制  

Effects of opioid receptor agonist DADLE on NADPH oxidase activity and the eNOS-NO pathway in rats with intestinal ischemia-reperfusion injury

作　　者：汪婷婷 梁翀[1] 孔繁丽 王燕[1] Wang Tingting;Liang Chong;Kong Fanli;Wang Yan(Wuhan Children’s Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei 430030,China)

机构地区：[1]华中科技大学同济医学院附属武汉儿童医院,湖北武汉430030

出　　处：《遵义医科大学学报》2022年第6期748-753,共6页Journal of Zunyi Medical University

基　　金：湖北省卫生健康委员会联合基金项目(NO:WJ2019H368);武汉市医学科研项目(NO:WX19Q21)。

摘　　要：目的 探究阿片受体激动剂对肠缺血再灌注损伤大鼠NADPH氧化酶活性及eNOS-NO通路的作用机制。方法 选取40只SPF级SD雄性大鼠,随机分为正常(N)组,模型(M)组,白介素-1受体拮抗剂(I)组,DADLE(D)组,每组10只,对M、I、D组采用夹闭肠系膜上动脉法建立肠缺血再灌注损伤模型,N组不建立该模型,建模成功后,对I组尾静脉注射30 mg/kg的白介素-1受体拮抗剂,对D组尾静脉注射3 mg/kg的δ阿片受体激动剂DADLE (D-Ala2-D-Leu5-enkephalin),N组、M组同期静脉注射同体积生理盐水,HE染色法检测肠组织病理形态,Chiu’s评分评价肠组织损伤程度,ELISA法测定D-乳酸、内毒素含量,细胞色素C还原法检测NADPH氧化酶活性,比色法检测eNOS含量,硝酸还原酶法检测NO含量。结果 N组大鼠肠组织结构正常,腺体完整清晰,未见绒毛尖端上皮脱落及炎性细胞浸润,M组肠组织结构遭到破坏,黏膜上皮损伤脱落现象严重,可见明显腺体受损及大量裸露的绒毛,出现大量出血灶和溃疡,且有大量炎性细胞浸润,与M组相比,I组、D组病理状态明显,黏膜损伤明显减轻,炎性细胞明显减少;与N组相比,M组Chiu’s评分显著升高(P<0.05),与M组比较,I组、D组Chiu’s评分显著降低(P<0.05),且D组比I组降低显著(P<0.05);与N组相比,M组血清D-乳酸、内毒素含量显著升高(P<0.05),与M组比较,I组、D组血清D-乳酸、内毒素含量显著降低(P<0.05),且D组比I组降低显著(P<0.05);与N组比较,M组肠组织中NADPH氧化酶活性显著升高(P<0.05),与M组比较,I组、D组肠组织中NADPH氧化酶活性显著降低(P<0.05),且D组比I组降低显著(P<0.05);与N组比较,M组血清eNOS、NO含量显著降低(P<0.05),与M组比较,I组、D组血清eNOS、NO含量显著升高(P<0.05),且D组比I组升高显著(P<0.05)。结论 阿片受体激动剂具有显著疗效,可显著降低肠缺血再灌注损伤大鼠NADPH氧化酶活性,并激活eNOS-NO通路。Objective To investigate the effect of opioid receptor agonist D-Ala2-D-Leu5-enkephalin(DADLE) on the activity of NADPH oxidase and the eNOS-NO pathway in rats with intestinal ischemia-reperfusion injury.Methods Male SD rats were randomly divided into normal,model,interleukin 1 receptor antagonist(IL-1 RA),and DADLE groups,10 rats each.The intestinal ischemia-reperfusion injury was produced by clipping mesenteric artery except for normal group.IL-1 RA(30 mg/kg) and DADLE(3 mg/kg) were injected through the caudal vein,while normal and model groups received the same volume of saline.Intestinal pathology was examined by HE staining,the degree of intestinal tissue injury was evaluated by Chiu’s score;D-lactic acid and endotoxin were determined by ELISA;the activity of NADPH oxidase,the level of eNOS,and the content of NO were detected with commercial kits.Results Compared to normal,model mice exhibited pathology lesions,including destroyed intestinal structure,mucosal epithelial injury and sloughing,damaged glands and a large number of damaged villi,inflammatory cell infiltration,bleeding and ulcers.These lesions were ameliorated in IL-1 RA and DADLE groups.Compared with model group,IL-1 RA and DADLE significantly decreased Chiu’s score(P<0.05).IL-1 RA and DADLE also decreased contents of D-lactic acid and endotoxin in serum.The activity of NADPH oxidase in intestinal tissue of model group was significantly increased,which was alleviated by IL-1 RA and DADLE(P<0.05).The decreased eNOS and NO in Model mice were rescued by IL-1 RA and DADLE(P<0.05).Conclusion Opioid receptor agonist DADLE can significantly reduce the activity of NADPH oxidase and activate the eNOS-NO pathway in rats with intestinal ischemia-reperfusion injury.

关 键 词：阿片受体激动剂 肠缺血再灌注损伤 NADPH氧化酶活性 eNOS-NO通路 

分 类 号：R916.4[医药卫生—药学]