庆大霉素致大鼠肾毒性模型中早期生物标志物的表达

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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. To investigate the effects of gemcitabine on the expression of neutrophil gelatinase-associated lipocalin (neutrophil gelatinase-associated lipocalin), an early biomarker of renal toxicity (KIM-1) Induced rat model of nephrotoxicity and explore its value in the safety evaluation of aminoglycosides.Methods Forty-five SPF male Sprague-Dawley rats were randomly divided into control group, gentamicin Low dose group (50 mg / kg) and gentamicin high dose group (100 mg / kg), 15 rats in each group were injected intramuscularly for 7 days, while the control group was given 0.9% sodium chloride injection On the 1st, 3rd, and 7th day after treatment, 5 rats in each group were sacrificed, serum creatinine (SCr) and blood urea nitrogen (BUN) levels were measured by abdominal aorta. The pathological changes were observed by HE staining , Real-time fluorescence quantitative PCR and immunohistochemistry were used to detect the expression of KIM-1 and NGAL.The results showed that serum creatinine and urea nitrogen levels were significantly increased only on the 7th day after administration.Pathological examination showed that the severity of renal injury increased with Time and dose-dependent increase in high-dose group after 7 d after proximal tubule brushing Disappearance of epithelial cells, epithelial cells shedding and necrosis, protein tube and interstitial inflammatory cell infiltration.KIM-1 and NGAL mRNA and protein expression levels were significantly increased on the first day of administration (P <0.05), followed by time - A dose-dependent increase consistent with histopathological changes and precedence over clinical biochemical markers Supporting these 2 biomarkers as a sensitive indicator of gentamicin-induced acute kidney injury.
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