乌司他丁对糖尿病脓毒症大鼠急性肾损伤的保护作用

Protections of ulinastatin to acute kidney injury in diabetic and septic rats

目的探讨乌司他丁(UTI)对糖尿病脓毒症大鼠肾损害的作用及其机制。方法采用高脂饲料喂养+腹腔小剂量注射链脲佐菌素方法制备大鼠2型糖尿病模型,取造模成功的大鼠40只随机分为4组,每组10只:糖尿病组(D组)、糖尿病假手术组(S组)、糖尿病脓毒症组(DS组)、糖尿病脓毒症UTI预处理组(U组)。另随机抽取同批次大鼠10只,作C组空白对照。采用盲肠结扎穿孔术(CLP)构建脓毒症模型。U组于CLP前1 h予尾静脉注射UTI 100 kU·kg^(-1)。S组拉出盲肠至腹外放置1 min。于CLP术后12 h,检测各组大鼠肾组织病理情况,定量尿微量白蛋白(UMA),内生肌酐清除率(Ccr),血清肿瘤坏死因子-α(TNF-α)、白细胞介素-18(IL-18)含量,血清丙二醛(MDA)、超氧化物歧化酶(SOD)水平,以及肾组织低氧诱导因子-1α(HIF-1α)的蛋白表达水平。结果与S组相比,DS组大鼠肾脏病理损害明显,UMA、IL-18、TNF-α、MDA、HIF-1α水平增高,Ccr和SOD活性降低(均P<0.05)。DS组相比,U组大鼠肾脏病理损害减轻,UMA、IL-18、TNF-α、MDA含量、HIF-1α水平降低,Ccr和SOD活性增高(均P<0.05)。结论 UTI可有效改善糖尿病脓毒症所致急性肾损伤大鼠的肾脏功能,其机制可能与抑制炎性反应、降低氧化应激、改善肾脏缺氧情况有关。

Objective To investigate the protective effects and explore the mechanisms of ulinastatin to the acute kidney injury(AKI)in diabetic and septic rats. Methods A model for type 2 diabetes mellitus( T2 DM) was established by the feeding of high-fat diet-induced insulin resistance and intraperitoneal streptozocin 30 mg · kg^(-1). Fourty successfully diabetic rats were randomly divided into 4 groups(n=10 each) :diabetes mellitus group(group D),DM sham operation group(group S),DM septic group(group DS) and DM septic group with advance-using ulinastatin(group U). Another 10 healthy nondiabetic rats were chosen as the normal control group(group C) with the normal diet. The septic model of the cecal ligation and puncture(CLP) was induced by puncturing the cecum and extruding the intestinal thing of 0. 1 mL for two times. Ulinastatin 100 kU · kg^(-1) was injected via the caudal vein one hour before CLP in U group. The cecums were pulling out of abdomens for one minute in S group. Collecting twelve-hour total urine was to detect urinary micro albumin(UMA) with ELISA. We detected and calculated the creatinine clearance rate(Ccr) by basic picric acid method. Then the serum levels of MDA and SOD were also measured. The serum levels of IL-18 and TNF-a were determined by ELISA. The hematoxylin-eosin staining was used to observe renal pathological changes. Western blot was used to determine the expression of HIF-la. Results Compared with group S,the renal pathological changes in group DS were more obvious,and the levels of UMA,IL-18,TNF-a,MDA and the expression of HIF-1α in kidney were obviously increased,Ccr and SOD were decreased(all P<0.05). Compared with group DS,the renal pathological changes in group U were obviously alleviated and the levels of UMA,IL-18,TNF-a,MDA and the expression of HIF-la in kidney were obviously decresed,Ccr and SOD were increased(all P <0.05). Conclusion Ulinastatin could effectively improve the renal function and reduce the acute kidney injury in diabetic and septic rats, probably based on those mechanisms :the suppression of inflammatory response, the reduction of oxidative stress and the improvement of hypoxemia in kidney.