庆大霉素致大鼠肾毒性模型中早期生物标志物的表达  

Expression of early biomarkers in gentamicin-induced nephrotoxicity in rats

作　　者：陈梦鹿 罗启慧[1] 孙凤娇[1] 陈正礼[1] 李明阳[2] 曾文 龚立 程安春[1] 彭西[1] 方静[1] 唐丽[1] 耿毅[1] 陈晓露[4] 

机构地区：[1]四川农业大学动物医学院,动物疾病与人类健康中心四川省重点实验室,四川雅安625014 [2]中国医药集团总公司四川抗菌素工业研究所,成都610052 [3]四川普莱美生物科技有限公司/国家实验用猕猴种源基地,四川雅安625014 [4]四川乐山职业技术学院,四川乐山614000

出　　处：《浙江大学学报（农业与生命科学版）》2014年第5期482-488,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家"重大新药创制"科技重大专项(2011ZX09301-001);四川省应用基础研究计划资助项目(2013JY0160);四川省青年科技创新研究团队资助项目(2013TD0015)

摘　　要：通过研究肾毒性早期生物标志物肾损伤分子-1(kidney injury molecule-1,KIM-1)和中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)在庆大霉素诱导的大鼠肾毒性模型中的表达,探讨其在氨基糖苷类药物安全性评价中的应用价值.将45只SPF级雄性Sprague-Dawley(SD)大鼠随机分为对照组、庆大霉素低剂量组(50 mg/kg)和庆大霉素高剂量组(100 mg/kg),每组15只,连续肌内注射7 d,对照组给予等体积0.9%氯化钠注射液.给药第1、3和7天分别处死各组5只大鼠,腹主动脉采血检测血清肌酐(serum creatinine,SCr)和血尿素氮(blood urea nitrogen,BUN)水平,肾组织HE染色观察病理改变,实时荧光定量PCR法和免疫组织化学检测KIM-1和NGAL的表达.结果表明,血清肌酐和尿素氮水平仅在给药第7天明显升高;病理组织学观察发现,肾损伤严重程度随时间及剂量依赖性增加,高剂量组给药7 d后近端小管刷状缘消失,上皮细胞脱落坏死,出现蛋白管型及间质炎性细胞浸润.KIM-1和NGAL的mRNA及蛋白表达水平均在给药第1天就显著上调(P<0.05),之后呈时间-剂量依赖性升高,与病理组织学改变过程相一致,且优先于临床生化检测指标.支持这2个生物标志物作为庆大霉素引起的急性肾损伤的敏感指标.Summary Gentamicin is a member of aminoglycosides which has represented highly effective antimicrobial agents especially in gram-negative infections despite their toxic effects in a kidney.Rapid diagnosis is vital to preserve renal function and to slow down renal inj ury.Owing to the poor sensitivity and specificity of serum creatinine (SCr) and blood urea nitrogen(BUN),there is a strong need for the identification and validation of more sensitive and reliable biomarkers.The aim of the study is to prove whether kidney inj ury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin(NGAL) could be useful to predict or detect acute kidney injury (AKI) caused by gentamycin. <br> In this study,45 male Sprague-Dawley(SD)rats were injected with a range of doses of gentamicin which was administered at 0,50 or 100 mg/(kg.d)(n=5 rats/dose group/time point) and the animals were necropsied on days 1,3 or 7 for toxicity evaluation.Heparinized blood was analyzed for SCr and BUN using a standard clinical chemistry analyzer.Kidney tissue samples were embedded in paraffin sections processing for hematoxylin-eosin staining and immunohistochemistry.Real-time fluorescent quantitative PCR was used for relative quantitation of KIM-1/NGAL mRNA levels. <br> In the gentamicin dose and time-response study,traditional indicators for nephrotoxicity,SCr and BUN levels in rats were significantly above control values after treatment for 7 days.Tubular cell degenerations,necrosis, tubular dilatation,hyaline cast tubules and inflammation were observed in the proximal tubules in the rats at the highest dose for 7 days.Repeated administration of gentamicin to animals resulted in a dose-and time-dependent increase in the expression of KIM-1 and NGAL which were evident as early as 1 day in both low-dose and high-dose animals(P<0.05).The degrees of mRNA increase were correlated well with the extent of tissue damage in the kidney.Consistent with gene expression analyses,KIM-1 and NGAL were undetectable in proximal tubules of control kidneys,

关 键 词：庆大霉素 肾毒性 肾损伤分子-1 中性粒细胞明胶酶相关脂质运载蛋白 

分 类 号：R965.2[医药卫生—药理学] R361[医药卫生—药学]