摘要
目的建立一种新的人近曲肾小管上皮细胞HK2缺氧/复氧损伤模型。方法本实验使用人近曲HK2。实验分为缺氧4、12和24h及缺氧24h后复氧4、12和24h组,每个实验组均设立空白对照组。采用经高温灭菌的液体石蜡覆盖法造成缺氧环境;苔盼兰摄取法进行细胞计数及检测细胞存活率,生化法检测培养基中的乳酸脱氢酶(LDH)含量。结果人肾小管上皮细胞经过缺氧/复氧处理后,苔盼兰摄取率显著提高,细胞存活率显著下降,LDH含量升高。说明缺血再灌注损伤导致了细胞膜的完整性破坏,甚至细胞发生了不可逆转的损伤。结论采用液体石蜡覆盖法建立人近曲肾小管上皮细胞缺氧/复氧损伤模型,较其他制模方法操作简单、易行、可靠。
Objective To replicate a new model of injury to human renal proximal tubular cells （HK - 2） induced by hypoxia/reoxygenation. Methods Human renal proximal tubular cell line HK - 2 cell was used as the target cell. Tubular cells were divided into six groups： 4 hours of hypoxia, 12 hours of hypoxia, 24 hours of hypoxia, and 24 hours of hypoxia followed by reoxygenation 4, 12 or 24 hours later groups. Each group was accompanied by a control group. Hypoxic culture conditions were produced by covering the culture with liquid paraffin. Trypan blue exclusion was used for cell count and cell viability. The activity of lactate dehydrogenase （LDH） in the culture medium was determined by biochemical method. Results After being challenged by hypoxia followed by reoxygenation, trypan blue exclusion rate was greater, cell count and cell viability were lower, and the activity of LDH was increased. It indicated that the destruction of integrality of cellular membrane was induced by ischemia/reperfusion injury, and the tubular cells may be injuried irrexersibly. Conclusion： A simple model of hypoxic injury of renal tubular cells is replicated by covering the culture cells with liquid paraffin.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2005年第10期619-622,F0005,共5页
Chinese Critical Care Medicine
基金
四川省杰出青年学科带头人培养基金资助项目(04ZQ02633)
四川省中医管理局重点科研基金资助(200239)
