浙江大学学报（农业与生命科学版）
浙江大學學報（農業與生命科學版）
절강대학학보（농업여생명과학판）
JOURNAL OF ZHEJIANG UNIVERSITY(AGRICULTURE & LIFE SCIENCES)
2014年
5期
482-488
,共7页
陈梦鹿%罗启慧%孙凤娇%陈正礼%李明阳%曾文%龚立%程安春%彭西%方静%唐丽%耿毅%陈晓露
陳夢鹿%囉啟慧%孫鳳嬌%陳正禮%李明暘%曾文%龔立%程安春%彭西%方靜%唐麗%耿毅%陳曉露
진몽록%라계혜%손봉교%진정례%리명양%증문%공립%정안춘%팽서%방정%당려%경의%진효로
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
慶大黴素%腎毒性%腎損傷分子-1%中性粒細胞明膠酶相關脂質運載蛋白
경대매소%신독성%신손상분자-1%중성립세포명효매상관지질운재단백
gentamicin%nephrotoxicity%kidney inj ury molecule-1%neutrophil gelatinase-associated lipocalin
通过研究肾毒性早期生物标志物肾损伤分子-1（kidney inj ury 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7d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.
通過研究腎毒性早期生物標誌物腎損傷分子-1（kidney inj ury molecule-1，KIM-1）和中性粒細胞明膠酶相關脂質運載蛋白（neutrophil gelatinase-associated lipocalin，NGAL）在慶大黴素誘導的大鼠腎毒性模型中的錶達，探討其在氨基糖苷類藥物安全性評價中的應用價值.將45隻SPF級雄性Sprague-Dawley（SD）大鼠隨機分為對照組、慶大黴素低劑量組（50 mg/kg）和慶大黴素高劑量組（100 mg/kg），每組15隻，連續肌內註射7 d，對照組給予等體積0.9%氯化鈉註射液.給藥第1、3和7天分彆處死各組5隻大鼠，腹主動脈採血檢測血清肌酐（serum creatinine， SCr）和血尿素氮（blood urea nitrogen，BUN）水平，腎組織 HE染色觀察病理改變，實時熒光定量 PCR法和免疫組織化學檢測KIM-1和 NGAL的錶達.結果錶明，血清肌酐和尿素氮水平僅在給藥第7天明顯升高；病理組織學觀察髮現，腎損傷嚴重程度隨時間及劑量依賴性增加，高劑量組給藥7d後近耑小管刷狀緣消失，上皮細胞脫落壞死，齣現蛋白管型及間質炎性細胞浸潤.KIM-1和NGAL 的 mRNA 及蛋白錶達水平均在給藥第1天就顯著上調（P<0.05），之後呈時間劑量依賴性升高，與病理組織學改變過程相一緻，且優先于臨床生化檢測指標.支持這2箇生物標誌物作為慶大黴素引起的急性腎損傷的敏感指標.
통과연구신독성조기생물표지물신손상분자-1（kidney inj ury molecule-1，KIM-1）화중성립세포명효매상관지질운재단백（neutrophil gelatinase-associated lipocalin，NGAL）재경대매소유도적대서신독성모형중적표체，탐토기재안기당감류약물안전성평개중적응용개치.장45지SPF급웅성Sprague-Dawley（SD）대서수궤분위대조조、경대매소저제량조（50 mg/kg）화경대매소고제량조（100 mg/kg），매조15지，련속기내주사7 d，대조조급여등체적0.9%록화납주사액.급약제1、3화7천분별처사각조5지대서，복주동맥채혈검측혈청기항（serum creatinine， SCr）화혈뇨소담（blood urea nitrogen，BUN）수평，신조직 HE염색관찰병리개변，실시형광정량 PCR법화면역조직화학검측KIM-1화 NGAL적표체.결과표명，혈청기항화뇨소담수평부재급약제7천명현승고；병리조직학관찰발현，신손상엄중정도수시간급제량의뢰성증가，고제량조급약7d후근단소관쇄상연소실，상피세포탈락배사，출현단백관형급간질염성세포침윤.KIM-1화NGAL 적 mRNA 급단백표체수평균재급약제1천취현저상조（P<0.05），지후정시간제량의뢰성승고，여병리조직학개변과정상일치，차우선우림상생화검측지표.지지저2개생물표지물작위경대매소인기적급성신손상적민감지표.
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,but they were observed in epithelial cells that were affected by gentamicin toxicity as seen by cell degeneration and regeneration. <br> In sum,the changes in gene and protein expressions of KIM-1 and NGAL were found to be correlated with the progressive histopathological alterations and preceded effects on traditional clinical parameters indicative of impaired kidney function.Time and dose-related effects on these two biomarkers expressions were observed at the target site of gentamicin toxicity,supporting the use of these two biomarkers as sensitive indicators of acute kidney inj ury caused by gentamicin.
