摘要
目的探讨Na+/H+交换器-1(NHE1)在新生儿窒息后血清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,以200 mL/L新生儿窒息后血清作为攻击因素处理HK-2细胞。首先将实验分为2组:对照组和窒息组,通过免疫组织化学方法检测其HK-2细胞在损伤前后胞内NHE1的表达;再分为3组:对照组、窒息组和5-N-乙基-N-异丙基-阿米洛利(EIPA)干预组(每组8个复孔)。倒置相差显微镜观察各组细胞形态变化,四甲基偶氮唑蓝(MTT)比色法检测HK-2细胞活力,生物化学法检测细胞培养液中LDH漏出率变化。结果与对照组比较,窒息组细胞内和细胞膜上NHE1表达明显增加;与对照组比较,窒息组细胞形态发生改变;窒息组细胞的活细胞数量较对照组明显降低;LDH漏出率明显升高,差异具有显著性意义(Pa=0);与窒息组细胞比较,EIPA干预组细胞形态明显得到改善,活细胞数量明显增加,LDH漏出率降低,差异具有显著性意义(Pa=0)。结论新生儿窒息后血清诱导HK-2细胞NHE1的表达增强可能是导致HK-2损伤的重要机制之一,通过抑制NHE1活性可使细胞损伤减轻,活性增强。
Objective To explore the role of Na^＋/H^＋ exchanger 1 （ NHE1 ） in injury of human renal tubular cells （ HK - 2 ） induced by postasphyxial - serum of neonates. Methods HK - 2 was used as the target cell. The attacking concentration of postasphyxial - serum of neonates was 200 mL/L. First, the experiment was designed as control group and asphyxia group, the expression of NHE1 in the HK -2 was detected by immunohisto chemical method in the cells. Second, the experiment group was designed as control group, asphyxia group, and pretreatment with 5 - N - Ethyl - N - isopropylamiloride （EIPA） group, then the change of morphology was observed under inverted microscope, and the cell viability was measured by methyl thiazolyl tetrazolium（MTT） method, and the leakage rate of lactate dehydrogenase （LDH） was determined by biochemical methods. Results Compared with blank control group, the expression of NHE1 in the HK - 2 was increased significantly in asphyxia group, the changes of morphology of HK -2 was most serious and obvious, the cell viability decreased, and the leakage rate of LDH increased significantly in asphyxia group. But compared with asphyxia group, the change of morphology of HK - 2 was greatly improved, the cell injury was decreased obviously, the leakage rate. of LDH was increased and viability was decreased in pretreatment group in a dose 2 μmol/L. Conclusions The postasphyxial - serum may induce the expression of NHE1 ,which plays an important role in injury of renal tubular cell induced by postasphyxial - serum in neonates, and inhibiting activity of NHE1 may relieve the injury of renal tubular cells induced by postasphyxialserum in neonates.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2008年第18期1412-1414,共3页
Journal of Applied Clinical Pediatrics
基金
四川省杰出青年学科带头人培养基金项目资助(04ZQ026-033)
四川省科技厅应用基础项目资助(2008JY0015)
<中华儿科杂志>第1届双鹤珂立苏科研基金
