依达拉奉对急性百草枯中毒大鼠肾损伤的干预

Intervention of edaravone against renal injury induced by acute paraquat poisoning in rats

目的 研究急性百草枯中毒大鼠肾损伤指标的动态变化及依达拉奉的干预效果和机制.方法 将80只清洁级SD大鼠随机分成空白对照组、百草枯中毒组、依达拉奉治疗组、依达拉奉对照组;空白对照组8只大鼠,给予0.9%氯化钠溶液1 ml,腹腔内注射1次,30 min后再次予以等量相同液体腹腔内注射;百草枯中毒组24只,给予20 mg/kg百草枯溶液1 ml腹腔内注射1次构建百草枯中毒模型,30 min后予以等量0.9%氯化钠溶液腹腔内注射;依达拉奉治疗组24只,构建百草枯中毒模型30 min后予5 mg/kg依达拉奉腹腔内注射;依达拉奉对照组24只,给予0.9%氯化钠溶液1 ml,腹腔内注射1次,30 min后予依达拉奉5 mg/kg腹腔内注射;除空白对照组,其余各组按上述方法处理,每天1次,维持7 d.处理后1、3、7、21 d留取肾组织并抽取动脉血,光学显微镜下观察各组肾脏病理变化,ELISA法检测血肌酐水平、血清与肾脏肾损伤分子1（KIM-1）及中性粒细胞明胶酶相关脂质运载蛋白（NGAL）水平,免疫组织化学染色法观察肾脏热休克蛋白70（HSP70）表达的变化.结果 光学显微镜下可见,百草枯中毒组大鼠肾组织病理损害最严重,在第3天达高峰,而各时间点依达拉奉治疗组病理损害均较百草枯中毒组轻,同时两组在21 d时均未见肾纤维化表现.与空白对照组比较,百草枯中毒组第1天大鼠血及肾脏KIM-1表达明显增加（P〈0.05）,血肌酐及NGAL水平在第7天明显增加（P〈0.05）;百草枯中毒组大鼠肾脏NGAL水平随时间推移逐渐升高,但与对照组差异无统计学意义（P〉0.05）.与百草枯中毒组相比,依达拉奉治疗组大鼠血肌酐、KIM-1、NGAL和肾脏KIM-1表达下降（P〈0.05）,肾脏NGAL差异无统计学意义（P〉0.05）.与百草枯中毒组比较,依达拉奉治疗组第3、21天大鼠肾脏组织中HSP70表达明显增加（P〈0.05）.结论 依达拉奉可促使肾脏组织HSP70表达升高,降低KIM-1和NGAL水平,对急性百草枯中毒所致的肾损伤起到一定保护作用.

Objective To investigate the dynamic change of paraquant-induced kidney injury in rats and the protective effect of edaravone. Methods Eighty SD rats were randomly divided into 4 groups： the normal control group, paraquat poisoning group, edaravone treatment group and edaravone control group. The normal control group of 8 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity , and the same amount of fluid into the abdominal cavity after 30 minutes. The paraquat poisoning group of 24 rats were given 1 ml of paraquat solution（20 mg/kg） through the abdominal cavity to build poisoning models, and the same amount of 0.9%sodium chloride was injected into the abdominal cavity after 30 minutes . The edaravone treatment group of 24 rats were given edaravone （5 mg/kg） through the abdominal cavity after 30 minutes when the poisoning models were set up. The edaravone control group of 24 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity,and edaravone（5 mg/kg） was injected into the abdominal cavity after 30 minutes. In addition to the normal control group, the other groups processed 1 times a day to mantain 7 d. On 1, 3, 7, 21 d several rats in each group were excuted and the kidney tissue and serum samples were collected , then each pathological changes of the kidney were observed with light microscopy. Serum creatinine , KIM-1, NGAL were measured by ELISA, the expression of HSP70 protein in kidney were observed with immunohistochemical staining. Results The pathological examination reveald that the damage of kidney tissue in the paraquat group was the most serious on 3 d, and the damage was consistently alleviated in edaravone treatment group at the same time ,renal fibrosisn was unseen in each group until 21 d. Compared with normal control group,there was no statistically significant in edaravone control group （P〉0.05）. The KIM-1 in blood and kidney in paraquat poisoning group were markedly increased in 1 d（P〈0.05）. The NGAL in blood and creatinine were markedly increased in d7（P〈0.05）. The NGAL in kidney increased over time, but had no statistically difference with the control group （P〉0.05）.Compared with paraquat poisoning group, the serum creatinine, KIM-1 in blood and kidney, the KIM-1 in kidney had decreased significantly in edaravone treatment group （P〈0.05）. The NGAL in kidney has no statistically significant compared with the poisoning group （P〉0.05）. HSP70 expression of kidney tissue in edaravone treatment group had significantly increased in d3 compared with the paraquat poisoning group （P〈0.05）. Conclusion Edaravone can prompt a significant rise of HSP70 in kidney tissue, reduce KIM-1 and NGAL levels, and play a protective role in kidney injury of acute paraquat poisoning.