乌司他丁预处理对肝缺血再灌注大鼠急性肾损伤的预防作用

Preventive effect of ulinastatin preconditioning on acute kidney injury in rats with hepatic ischemia-reperfusion

目的观察乌司他丁(UTI)预处理对肝缺血再灌注(I/R)大鼠急性肾损伤的保护作用,并探讨其可能作用机制。方法 30只Wistar大鼠,随机分为假手术(SO)组、缺血再灌注(I/R)组、UTI组,每组各10只。I/R组、UTI组采用Pringle法制备大鼠肝脏I/R损伤模型(阻断肝脏血流30 min再灌注2 h)。UTI组阻断肝脏血流前30min经隐静脉UTI(3万U/kg)注射。SO组仅解剖分离肝十二指肠韧带。造模后复流2 h处死各组大鼠,采用比色法检测三组血清和肝肾组织肌酐(Cr)、尿素氮(BUN)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)含量。ELISA法测定血清和肾组织白细胞介素18(IL-18)和肿瘤坏死因子α(TNF-α)。行膀胱穿刺并收集测算尿液体积,HE染色法观察三组肝、肾脏组织病理学改变。结果与SO组比较,I/R组大鼠血清Cr、BUN、ALT、AST、IL-18、TNF-α水平升高,GSH、GSH-Px、SOD水平降低,肾组织IL-18、MDA表达升高,GSH、GSH-Px、SOD表达降低(P均<0.05)。与I/R组比较UTI组大鼠血清Cr、BUN、ALT、AST、IL-18、TNF-α水平降低,GSH、GSH-Px、SOD水平升高,肾组织IL-18、MDA表达降低,GSH、GSH-Px、SOD表达升高(P均<0.05)。与SO组比较,I/R组大鼠尿量少,与I/R组比较,UTI组尿量多(P<0.05)。I/R组大鼠肝组织有明显的细胞坏死灶及大量炎性细胞浸润。肾小球及肾间质水肿,肾小管上皮水肿、变形、扩张。UTI组肝脏和肾脏组织结构变化明显轻于I/R组。结论 UTI预处理可有效减轻肝I/R大鼠急性肾损伤,其机制可能与UTI抑制IL-18、TNF-α等表达,促进GSH、GSH-Px、SOD表达有关。

Objective To observe the protective effect of ulinastatin（UTI） preconditioning on acute renal injury in rats with hepatic ischemia-reperfusion（I/R） and to explore its possible mechanism. Methods Thirty Wistar rats were randomly divided into sham operation（SO） group,ischemia reperfusion（I/R） group and UTI group,with 10 rats in each group. I/R was induced by the Pringle occlusion method. Rats in the I/R group and UTI group underwent 30 min of hepatic ischemia and 120 min of reperfusion. The rats in the UTI group were pretreated with UTI（30 000 U/kg） by saphenous vein administration for 30 min before operation. We only dissected the hepatic duodenal ligament of rats in the SO group.Rats in each group were sacrificed 2 hours after model establishment. The levels of creatinine（Cr）,urea nitrogen（BUN）,alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,glutathione（GSH）,glutathione peroxidase（GSHPx）,superoxide dismutase（SOD）,and malondialdehyde（MDA） in the serum and kidney tissues were detected by colorimetric method. IL-18 and TNF-α in the serum and kidney tissues was detected by ELISA. The urine was collected by bladder puncture. HE staining was used to detect the histological changes of liver tissues and kidney tissues. Results Compared with the SO group,the Cr,BUN,ALT,AST,IL-18,TNF-a,and MDA in the rat serum and kidney tissues were significantly higher,and GSH,GSH-Px,and SOD were significantly lower in the I/R group（all P〈0. 05）. Compared with the I/R group,the Cr,BUN,ALT,AST,IL-18,TNF-a,and MDA in the rat serum and kidney tissues were significantly lower,and GSH,GSH-Px,and SOD were significantly higher in the UTI group（all P〈0. 05）. Compared with the SO group,rat urine volume was significantly lower in the I/R group; compared with the I/R group,the rat urine volume was significantly higher in the UTI group（P〈0. 05）. In the I/R group,the liver tissues had obvious liver cell necrotic foci and a large number of inflammatory cell infiltration. The glomerular,renal interstitial and tubular epithelial was swelling,and glomerular and renal interstitial edema,tubular epithelial edema,deformation,and dilatation were found in the I/R group. The liver and kidney tissue changes in the UTI group were significantly lighter than those in I/R group. Conclusion UTI preconditioning alleviates acute renal injury of hepatic I/R rats,and its mechanism may be that UTI inhibits the expression of IL-18,TNF-α,and promotes the expression of GSH,GSH-Px,and SOD.