肾脏缺血预适应防治脓毒症急性肾损伤的实验研究  

Effect of renal ischemic preconditioning on sepsis-induced acute kidney injury:an experimental study

作　　者：贾平[1] 丁小强[1] 

机构地区：[1]复旦大学附属中山医院肾内科,上海市肾病与透析研究所,上海市肾脏疾病与血液净化重点实验室,上海200032

出　　处：《上海医学》2016年第5期283-286,I0001,共5页Shanghai Medical Journal

基　　金：国家自然科学基金(81471890;81430015);上海市科学技术委员会基金(14DZ2260200)资助项目

摘　　要：目的探讨肾脏缺血预适应(IPC)对脓毒症诱发急性肾损伤(AKI)的防治作用。方法将32只C57BL/6J小鼠随机分为对照组、模型组、假手术组和IPC组,每组8只。模型组小鼠给予一次性腹腔内注射脂多糖(LPS,20mg/kg);对照组小鼠予腹腔内注射等剂量0.9%氯化钠溶液;IPC组小鼠在注射LPS前24h给予双侧肾脏IPC(双侧肾蒂夹闭15min);假手术组操作过程基本同IPC组,但仅分离双侧肾蒂,不夹闭。注射后24h,采用酶法测定血肌酐(sCr)浓度,尿素酶-谷氨酸脱氢酶(GLDH)法测定尿素氮(BUN)浓度;在光学显微镜下观察肾组织形态学变化;采用ELISA法检测小鼠血清和肾组织匀浆中TNF-α和IL-6水平,采用PCR检测TNF-α和IL-6mRNA表达;采用TUNEL法检测肾脏细胞凋亡情况。结果 LPS注射后24h,模型组和假手术组小鼠sCr和BUN浓度、血液和肾组织匀浆中TNF-α和IL-6水平、肾组织中TNF-αmRNA和IL-6mRNA表达、肾脏细胞凋亡数均显著高于对照组(P值均<0.01);IPC组小鼠sCr和BUN浓度、血液和肾组织匀浆中TNF-α和IL-6水平、肾组织中TNF-αmRNA和IL-6mRNA表达、肾脏细胞凋亡数均显著低于模型组和假手术组(P值均<0.01),但仍显著高于对照组(P值均<0.01)。模型组和假手术组小鼠在LPS注射后24h,可见肾脏皮髓交界处散在的肾小管上皮细胞坏死脱落,基底膜断裂,少许管型和明显的炎性细胞浸润等;而IPC组小鼠的肾组织损伤较轻,仅表现为肾脏间质水肿,少许炎性细胞浸润。结论肾脏IPC可以减轻脓毒症诱发的AKI,减轻炎性反应,抑制细胞凋亡。Objective To observe the effect of renal ischemic preconditioning （IPC） on sepsis-induced acute kidney injury （AKI） in mice. Methods Thirty-two 056BL/6J mice were randomly divided into four groups as follows., control group, model group, sham operation group and IPC group （n = 8）. Sepsis was induced by intraperitoneal injection of lipopolysaccharide （LPS, 20 mg/kg） in model group, and equal volume of 0.9% sodium chloride solution was injected intraperitoneally in control group. In IPC group, IPC was induced by bilateral renal pedicle clamping for 15 minutes, followed by LPS （20 mg/kg） injection 24 hours tater. In sham operation group, mice underwent the same surgical procedures, except that the renal pedicles were not clamped. Blood and renal tissues were collected 24 hours after the LPS injection. The concentrations of serum creatinine （sOt） and blood urea nitrogen （BUN） were detected by enzymatic analysis and urea-enzyme-glutamate dehydrogenase （GLDH） method, respectively. Renal histomorphologic changes were observed under optical microscope. Interleukin （IL）-6 and tumor necrosis factor （TNF）-a in blood and tissue homogenate were detected by enzyme-linked immunosorbent assay （ELISA）. Expression of TNF-a mRNA and IL-6 mRNA in kidneys was assayed by polymerase chain reaction （PCR）. Renal cell apoptosis was tested by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） method. Results Levels of sCr and BUN, IL-6 and TNF-a in blood and tissue homogenate, expression of IL-6 mRNA and TNF-a mRNA, as well as renal cell apoptosis were significantly increased 24 hours after LPS injection in mice of model group and sham operation group when compared with those of control group （all P〈0.01）. The parameters reflecting renal injury and inflammation in IPC group were significantly reduced when compared to those of model group and sham operation group （all P 〈0.01 ）, while higher than those of control group （all P^0.01）. Obvious histol

关 键 词：缺血预适应 脓毒症 急性 肾损伤 炎性因子 

分 类 号：R459.7[医药卫生—急诊医学] R692[医药卫生—治疗学]