发动相关蛋白在窒息新生儿血清诱导人近曲肾小管上皮细胞损伤中的作用及重组人促红细胞生成素干预的影响  

Roles of Dynamin-Related Protein in Human Renal Tubular Cells Injury Induced by Postasphyxial-Serum in Neonates and Effects of Recombinant Human Erythropoietin on Them

作　　者：雷小平[1] 邹礼乐[2] 董文斌[1] 李清平[1] 翟雪松[1] 康兰[1] 

机构地区：[1]泸州医学院附属医院新生儿科,四川泸州646000 [2]泸州医学院组胚教研室,四川泸州646000

出　　处：《实用儿科临床杂志》2012年第2期95-97,共3页Journal of Applied Clinical Pediatrics

基　　金：四川省科技厅应用基础项目(2008JY0015);中华儿科杂志第1届双鹤珂立苏科研基金

摘　　要：目的探讨发动相关蛋白1(Drp-1)在新生儿窒息后血清诱导人近曲肾小管上皮细胞(HK-2细胞)损伤中的作用及重组人促红细胞生成素(rhEPO)干预的影响。方法将培养的HK-2细胞分为对照组、窒息组和rhEPO干预组,以新生儿重度窒息(Apgar评分<4分)后24 h血清(20%体积比)作为攻击因素。光镜下观察其线粒体形态变化,活细胞计数试剂盒(CCK-8)法检测细胞活力,免疫组织化学检测其Drp-1表达,投射电子显微镜下观察其线粒体的形态变化。结果 1.HK-2细胞形态学变化:对照组及rhEPO干预组细胞生长良好,窒息组细胞发生凋亡相关改变。2.细胞活力变化:与对照组细胞活力(0.74±0.08)相比,窒息组细胞活力(0.41±0.06)明显降低,rhEPO干预组细胞活力(0.60±0.08)较窒息组好转,但未恢复至对照组水平(P<0.05)。3.免疫组织化学法测定Drp-1表达:与对照组(0.24±0.03)相比,窒息组细胞Drp-1表达(0.51±0.03)显著升高,rhEPO干预组(0.38±0.03)较窒息组降低,但未恢复至对照组水平(P<0.05)。4.线粒体形态变化:对照组和rhEPO干预组线粒体结构完整,可见线粒体嵴;窒息组线粒体嵴消失,膜肿胀,空泡变性。结论 rhEPO可减轻新生儿窒息后血清诱导的HK-2细胞凋亡,其机制可能是通过减少Drp-1的表达抑制线粒体的分裂而实现。Objective To investigate the role of dynamin-related protein 1（Drp-1）in human renal tubular cells（HK-2 cells） injury induced by postasphyxial-serum in neonates and the effects of recombinant human erythropoietin（rhEPO） on it. Methods The experiment was designed with control group,asphyxia group and the group treated with rhEPO.The serum of neonates at 24 hours after asphyxia（Apgar scoring less than 4）,whose concentration was 20%（volume fraction）,was applied as attacking factor.The following indicators were detected：the changes of morphology were observed under inverted microscope,the cell viability was measured by cell counting kit-8（CCK-8） methods,the expression of Drp-1 in cytoplast was detected by using immunohistochemical method,and the morphological changes of mitochondria were observed under transmission electron microscope（TEM）. Results 1.Under inverted microscopy,HK-2 cells in control group and the group treated with rhEPO were growing well,while the changes in morphology of HK-2 cells were most serious and obvious in asphyxia group.2.The cell viability was measured by CCK-8 methods：compared with control group（0.74±0.08）,the cell viability（optical density,A） was obviously decreased in the asphyxia group（0.41±0.06,P0.05）.Compared with asphyxia group,the cell viability（A） was obviously increased in the group treated with rhEPO（0.60±0.08,P0.05）,but still was not as much as that in the control group（P0.05）.3.Compared with control group（0.24±0.03）,the expression of Drp-1（A）was significantly increased in asphyxia group（0.51±0.03,P0.05）.Compared with asphyxia group,the expression of Drp-1 was obviously decreased in the group treated with rhEPO（0.38±0.03,P0.05）,but was not as much as that in the control group（P0.05）.4.Under TEM,mitochondria in asphyxia group appeared extensive swelling and vacuolar degeneration with less matrix and obscure or vanished mitochondria cristae;but in control group and rhEPO group,mitochondrial structure

关 键 词：发动相关蛋白 促红细胞生成素 新生儿窒息 血清 肾小管上皮细胞 凋亡 线粒体 形态变化 

分 类 号：R722.1[医药卫生—儿科]