摘要
目的研究庆大霉素、奈替米星对体外培养的人肾小管上皮细胞(HK-2)的毒性作用,及急性肾损伤标志物表达与分泌的特点和庆大霉素诱导的SD大鼠急性肾损伤模型中血液、尿液生物标志物表达与分泌的特点。方法采用不同浓度的庆大霉素和奈替米星作用于体外培养的HK-2细胞,通过MTT法检测药物对HK-2细胞的抑制率、ELISA法检测细胞上清液中KIM-1和NGAL的分泌与表达。采用连续7d给予庆大霉素,建立SD大鼠的急性肾损伤模型,利用全自动生化分析仪测定血清中肌酐(CRE)和尿素(UREA)的浓度,ELISA法检测血液中Cys C、β2-MG、NGAL和尿液中的KIM-1、NGAL、IL-18、Cys C、β2-MG的分泌与表达。结果庆大霉素和奈替米星对HK-2细胞均具有增殖抑制作用,其IC50分别为2.444和5.366mg/mL;在HK-2细胞发生损伤后的短时间内,细胞上清液中KIM-1和NGAL的浓度会显著升高,不同的是KIM-1的浓度升高后会渐渐下降,而NGAL的浓度会维持在较高水平。肾脏组织病理学检查显示,在给予庆大霉素后的第2天,50mg/kg组、100mg/kg组肾脏无明显病理变化,第4天、50mg/kg组可见少量上皮细胞受损,100mg/kg组比50mg/kg组受损严重,第8天、50mg/kg组上皮细胞受损比较严重,100mg/kg组大量上皮细胞严重受损。在给予庆大霉素后的第8天、100mg/kg组血清中肌酐、尿素、β2-MG的浓度显著高于0.9%NaCl组(P<0.05);但3个剂量组血清中NGAL和Cys C的浓度不具有显著性差异(P>0.05);从给予庆大霉素后的第4天开始,100mg/kg组尿液中KIM-1、NGAL、IL-18、CysC、β2-MG的含量均显著高于0.9%NaCl组(P<0.05)。结论体外试验表明,庆大霉素和奈替米星对体外培养的HK-2细胞具有增殖抑制作用,且能引起KIM-1和NGAL的异常表达和分泌。体内试验表明,与血清肌酐、尿素和组织病理学检查相比,尿液中KIM-1、NGAL、IL-18、Cys C、β2-MG含量的变化能更早预测庆大霉素引起的肾损伤。
Objective To study the cytotoxic effect of gentamicin and netilmicin on human proximal tubular epithelial cells （HK-2）, and the expression and secretion of acute kidney injury markers and the expression and secretion of the bio-markers in serum and urine of the gentamicin induced acute kidney injury rat model. Methods The in vitro cultured HK-2 cells were treated with different concentrations of gentamicin and netilmicin, the IC50 of gentamicin and netilmicin on HK-2 cells was evaluated by MTT assay, and the expression and secretions of KIM-1 and NGAL in the culture supernatants was detected by ELISA. Dose Gentamicin for 7 consecutive days in order to establish the acute kidney injury SD rat model. Detect the serum creatinine and serum urine by the automatic biochemical analyzer. The expression and secretion of Cys C, β2-MG and NGAL in serum, as well as KIM-1, NGAL, IL-18, Cys C, β2-MG in urine are detected by ELISA. Results Gentamicin and Netilmicin significantly inhibited the proliferation of HK-2 cells, the ICs0 values of which were 2.444 and 5.366mg/mL, respectively. In a short period after impairment, the concentration of KIM-I and NGAL in the culture supernatant rise significantly. The only difference is that the concentration of KIM-1 will decreased gradually after the rise, while the concentration of NGAL will maintained in a relatively high level. Shown from the histopathology detection, on the 2nd day after dosage, the pathological change of kidneys of the rats in the 50mg/kg, 100mg/kg groups was inconspicuous. On the 4th day after dosage, the epithelial cells of the kidneys of the rats in the 50mg/kg group were impaired, and the impairment was severer in the 100mg/kg group. On the 8th day after dosage, the epithelial cells of the kidneys of the rats in the 50mg/kg group were impaired relatively severe, and large amount of the epithelial cells of the kidneys of the rats in the 100mg/kg group were impaired badly. On the 8th day after first dosage, the concentration of the serum creatinine, urea and
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2014年第4期316-320,I0001,I0002,共7页
Chinese Journal of Antibiotics
基金
"重大新药创制"科技重大专项--药物安全性评价关键技术研究与平台建设(2011ZX11301)
