氢水对脓毒血症大鼠急性肾损伤的保护作用

摘要

Objective To investigate the effects of hydrogen-rich saline (HRS) on acute kidney injury (AKI) in a rat model of sepsis.Methods Twenty-four male SD rats were randomly (random number) assigned to the sham operation group,the cecal ligation and puncture (CLP) group,and the CLP + HRS group (n =8).The rats in the sham operation group were treated with lapa(r ly) without CLP,while the rats in the CLP group and CLP + HRS group were made to be CLP-induced sepsis model.At 0,6 and 18 h after sham operation or CLP,rats in the CLP + HRS group received an intraperitoneal injection of HRS of 5 ml/kg,and 5 ml/kg of normal saline was given to the rats in the sham group and CLP group.Plasma levels of TNF-α,IL-6,IL-10,and HMGB1 were determined by enzyme linked immunosorbent assay (ELISA),plasma BUN and Cr were assayed by enzymatic assays,renal malondialdehyde (MDA) levels and superoxide dismutase (SOD) activities were determined by xanthine oxide method,renal tissue injury was dertermined by hematoxylin-eosin staining (liE),and renal cell apoptosis was assessed by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) at 24 h after sham operation or CLP.Statistical analysis carried out by using one way analysis of variance followed by Student-Newman-Keuls (SNK) test.Results Compared with the sham group,rats in the CLP and CLP + HRS groups had increases in plasma levels of TNF-α,IL-6,IL-10,HMGB1,BUN,Cr,renal MDA,severe renal injury and apoptosis,and a decrease in renal SOD activities (P ＜ 0.05).Compared with the CLP group,the CLP + HRS group had decreases in plasma levels of HMGB1,BUN,renal MDA,mild renal injury and apoptosis,and an increase in renal SOD activities (P ＜ 0.05).Conclusions HRS reduces inflammatory reaction and oxidative stress,improving renal function during AKI in the rat model of sepsis.%目的 研究氢水(HRS)对脓毒血症大鼠急性肾损伤(AKI)的影响.方法 24只雄性SD大鼠随机(随机数字法)均分为3组(n=8),Sham组、盲肠结扎穿孔(CLP)组及CLP+ HRS组.Sham组采用假手术,CLP组及CLP+HRS组采用CLP脓毒症模型.假手术或CLP后0、6、18 h,CLP+HRS组于腹腔注射HRS 5 ml/kg,Sham组和CLP组则分别注射生理盐水5 ml/kg.假手术或CLP后24h,利用酶联免疫吸附试验(ELISA)检测血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)及高迁移率族蛋白1(HMGBl)水平；利用酶法检测血浆尿素氮(BUN)、肌酐(Cr)水平；利用黄嘌呤氧化酶法检测肾组织丙二醛(MDA)及超氧化物歧化酶(SOD)活性；利用苏木精-伊红染色法(HE)评估肾组织损伤程度,原位末端转移酶标记法(TUNEL)检测肾组织细胞凋亡程度.组间比较采用单因素方差分析,两两比较采用Student-Newman-Keulsa (SNK)法.结果 与Sham组相比,CLP组及CLP+ HRS组血浆TNF-α、IL-6、IL-10、HMGB1、BUN、Cr及肾组织MDA升高,肾组织损伤程度加重,细胞凋亡增多,而肾组织SOD降低,差异具有统计学意义(P＜0.05).与CLP组相比,CLP+ HRS组血浆HMGB1、BUN、Cr、肾组织MDA降低,肾组织损伤程度减轻,细胞凋亡减少,而SOD升高,差异具有统计学意义(P＜0.05).结论 HRS抑制脓毒血症大鼠的炎性反应和氧化应激,从而改善AKI时的肾功能.