摘要
目的研究窒息新生儿血清在诱导人近曲肾小管上皮细胞(HK-2)损伤中的作用。方法以HK-2为研究对象,实验分为15组:新生儿窒息后d1、d3、d7不同体积分数(2.5%、5.0%、10.0%、20.0%)血清处理HK-2细胞组,每个体数分数组均有相应的空白对照组。倒置相差显微镜下观察HK-2细胞的形态学变化,四唑盐(MTT)比色法检测HK-2细胞活力,生物化学法检测培养液中LDH漏出率。结果实验组细胞受损明显,形态改变,活细胞数量较空白对照组明显降低(P均<0.05);10.0%、20.0%血清组LDH漏出率较空白对照组明显升高(P均<0.05),且以窒息后d1 20.0%组最为明显,差异具有显著性(P均<0.05)。结论新生儿窒息后不同时间血清可诱导HK-2细胞损伤,以窒息后d1损伤最明显。此细胞模型的建立为进一步研究新生儿窒息后肾损伤的发病机制和防治提供实验基础。
Objective To investigate the role of postasphyxial ,serum of neonate in inducing injury of human renal proximal tubular cells（HK - 2 cells）. Methods HK- 2 cells were used as target cell, The neonatal different concentration postasphyxial - serum of 1,3,7 days after asphyxia were used as attacking means. The experimental groups were divided into 15 groups： the 2.5%, 5.0%, 10, 0%, 20.0 % attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration. The culture medium and concertration of the control group and the experimental groups were the same. The changes of morphology were observed under inverted microscope, the cell viability was measured by 3 - （4,5 - dimethyl- 2 - thiazolyl ） - 2,5 - diphenyl-2H-tetrazolium bromide（MTT） method and the leakage rate of lactate dehydrogenase（LDH） was determined by biochemical methods. Results Compared with control group, the changes in morphology of HK- 2 were most serious and obvious, the cell viability were obviously decreased （all P 〈 0.05 ）, and the leakage rate of LDH were significantly increased in the 10, 0%, 20.0% attacking concertration groups（all P〈 0.05 ）, but those changes reached the peak in 20.0 % attacting concentration group of 24 hours after asphyxia. Conclusions The postasphyxial - serum of neonate is a stimulus that may induce injury of HK-2 cells and those injuries were most serious and obvious at 24 hours after asphyxia. This duplication of the cell model offered the experimental base to further investigate on the mechanism and prevention of postasphyxial renal injury in neonate.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2006年第17期1135-1136,1138,共3页
Journal of Applied Clinical Pediatrics
基金
四川省杰出青年学科带头人培养计划项目资助(04ZQ026-033)
关键词
窒息
血清
肾损伤
婴儿
新生
asphyxia
serum
renal injury
infant, newborn
