窒息新生儿血清对肾小管上皮细胞内Omi／HtrA2表达的影响  

Effect of Postasphyxial-Serum in Neonate on Expression of Omi/HtrA2 in Renal Tubular Cells

作　　者：张勇[1] 董文斌[1] 邓存良[2] 王明勇[2] 陈枫[2] 许开桂[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]泸州医学院附属医院传染与免疫研究室,四川泸州646000

出　　处：《实用儿科临床杂志》2008年第6期436-437,共2页Journal of Applied Clinical Pediatrics

基　　金：四川省杰出青年学科带头人培养基金资助(04ZQ026-033)

摘　　要：目的探讨窒息新生儿血清对人近曲肾小管上皮细胞(HK-2)内Omi/HtrA2表达的影响。方法以人HK-2为研究对象,将培养在孔板内的细胞随机分为正常对照组、窒息组。从新生儿病房经股静脉收集重度窒息24 h足月新生儿血,经肝素抗凝,3 000 r/min离心20 min,分离血清,56℃恒温水浴30 min灭活补体,微孔滤器过滤除菌,按体积比1∶5配成窒息血清,200 mL/L作为攻击浓度。当细胞生长达到要求后,窒息组加入窒息血清对HK-2细胞进行攻击,正常对照组采用正常培养基进行培养,24 h后终止培养和攻击,固定细胞,采用免疫组织化学法检测二组细胞Omi/HtrA2的表达。采用SPSS 10.0软件进行t检验。结果Omi/HtrA2染色为金黄色或黄褐色分布于细胞质中。与正常对照组阳性细胞率[(9.0±2.5)%]相比,窒息组阳性细胞率[(25.15±3.5)%]明显增加,差异具有显著性意义(t=-15.322 P<0.01)。结论窒息新生儿血清可诱导Omi/HtrA2在HK-2细胞内的表达,Omi/HtrA2可能参与窒息新生儿血清诱导的HK-2凋亡。Objective To explore the effect of postasphyxial - serum in neonate on expression of serine protease Omi/HtrA2 in renal tubular ceils（ HK - 2）. Methods Human renal proximal tubular cell line HK - 2 cell was used as target cell. The cultural cells in orifice were divided into control group and asphyxia - serum attacking group. Blood was cowected from asphyxia newborns by means of femoral venous puncture,then the serum was garthered,anticoagulated by liquemie,3 000 r/min centrifuged 20 min ,abstracted serum, thermostatic waterbathed the serum at 56 ℃ ,so that to inactivate addiment,fihered germ by mieropore fihe,the attacking eoneentrtion of serum was 200 mL/L, the ceils of the asphyxia - serum attacking group were attacked by asphyxia - serum ,and the ceils of control group were cultivated with normal nutritive medium when the ceils was needed. After 24 hours, the ceils were fixed, then the expression of Omi/HtrA2 in cytoplast was detected by the use of immunohistochemical method. Results Omi/HtrA2 was inaurate or yellow brown and localized to the cytoplast. The rate of the cell expressed Omi/HtrA2 was （9.0 ± 2.5 ）% in control group, after stimulated with postasphyxial -serum, in asphyxia group the rate of the cell expressed Omi/HtrA2 was （25.15 ± 3.5 ） %, there was significant difference between 2 groups （ t = - 15. 322 P 〈 0.01 ）. Conclusions The expression of Omi/HtrA2 in HK - 2 ceils might be indued by postasphyxia - serum,Omi/HtrA2 might play an important role in the course of HK - 2 ceils apoptosis induced by postasphyxial - serum of neonate.

关 键 词：婴儿 新生 窒息 血清 凋亡 OMI/HTRA2 

分 类 号：R722.1[医药卫生—儿科]