Yu L J, Zhang Z C, Wu Y Z, Wang W J, Jian X D, Kan B T
Department of Rheumatology, Qilu Hospital, Shandong University, Jinan 250012, China.
Shandong University Hospital, Jinan 250100, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2025 Jan 20;43(1):62-67. doi: 10.3760/cma.j.cn121094-20231221-00164.
To establish the model of acute kidney injury (AKI), search for more sensitive and reliable biomarkers. In April 2018, 100 male Wister rats aged 6 to 8 weeks were selected and randomly divided into experimental group (n=90) and control group (n=10). The experimental group was given Diachalefin (140 mg/kg body weight) by intragastric administration, while the control group was given saline intragastric administration. Ten rats in the experimental group were killed 0.5 h, 2 h, 6 h, 24 h, 3 d, 7 d, 14 d, 21 d and 28 d after intragastric administration, respectively. Serum creatinine (Cr), urea nitrogen (BUN) and uric acid (UA) were detected by automatic biochemical analyzer with 5 ml of blood from inferior vena cava puncture. Serum neutrophil gelatinase-associated lipid carrier protein (NGAL), kidney damage molecule-1 (KIM-1) and transforming growth factor-β1 (TGF-β1) levels were determined by enzyme-linked immunosorbent assay (ELISA). The data between groups were compared using two independent sample t tests. The renal tissue structure of rats in the control group was not significantly abnormal, while the renal tissue cell damage of rats in the experimental group was obvious, which gradually increased with the extension of time in the early stage, and gradually recovered in the later stage. UA in experimental group reached its peak at 24 h after exposure and was still higher than that in control group at 14 d (<0.05), Cr reached its peak at 7 d, and then gradually decreased, and there was no statistical significance between experimental group and control group at 28 d (>0.05). BUN increased at 6 h after exposure and reached the highest value at 7~14 d (<0.05). Blood NGAL increased at 0.5 h after exposure, reached its peak at 24 h, continued to increase at 3, 7 and 14 days (<0.05), and began to decrease at 21 days. KIM-1 began to increase at 0.5 h, continued to peak at 24 h, 3 and 7 d after exposure, and began to decrease at 14 d, but it was still higher than that in control group (<0.05). There was no significant difference in TGF-β1 at each time point (>0.05). Western blot assay results: Compared with control group, there was no significant difference in the expression level of TGF-β1 in kidney tissue of experimental group (>0.05). NGAL increased gradually from 2 h and was higher at 7 and 14 d, with statistical significance (<0.05). KIM-1 increased at 2 h, decreased at 6 and 24 h, and increased again at 3 and 7 d. NGAL and KIM-1 can be used as early diagnostic biomarkers for diquat-induced acute kidney injury.
为建立急性肾损伤（AKI）模型，寻找更敏感可靠的生物标志物。2018年4月，选取100只6至8周龄的雄性Wistar大鼠，随机分为实验组（n = 90）和对照组（n = 10）。实验组给予敌草快（140 mg/kg体重）灌胃，对照组给予生理盐水灌胃。实验组分别于灌胃后0.5 h、2 h、6 h、24 h、3 d、7 d、14 d、21 d和28 d处死10只大鼠。经下腔静脉穿刺采集5 ml血液，用自动生化分析仪检测血清肌酐（Cr）、尿素氮（BUN）和尿酸（UA）。采用酶联免疫吸附测定（ELISA）法测定血清中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、肾损伤分子-1（KIM-1）和转化生长因子-β1（TGF-β1）水平。组间数据比较采用两独立样本t检验。对照组大鼠肾组织结构无明显异常，而实验组大鼠肾组织细胞损伤明显，早期随时间延长逐渐加重，后期逐渐恢复。实验组UA在染毒后24 h达到峰值，14 d时仍高于对照组（<0.05），Cr在7 d达到峰值，随后逐渐下降，28 d时实验组与对照组无统计学差异（>0.05）。BUN在染毒后6 h升高，7至14 d达到最高值（<0.05）。血液NGAL在染毒后0.5 h升高，24 h达到峰值，3、7和14 d持续升高（<0.05），21 d开始下降。KIM-1在0.5 h开始升高，染毒后24 h、3和7 d持续达到峰值，14 d开始下降，但仍高于对照组（<0.05）。各时间点TGF-β1无明显差异（>0.05）。蛋白质免疫印迹分析结果：与对照组相比，实验组肾组织中TGF-β1表达水平无明显差异（>0.05）。NGAL从2 h开始逐渐升高，7和14 d时更高，具有统计学意义（<0.05）。KIM-1在2 h升高，6和24 h下降，3和7 d再次升高。NGAL和KIM-1可作为敌草快诱导急性肾损伤的早期诊断生物标志物。
