新生儿窒息后血清对人肾小管细胞抗Bcl-2蛋白、Bcl-2联接X蛋白表达的影响

Effect of Postasphyxial-Serum in Neonate on the Expressions of Bcl-2-Antagonist of Cell Death and Bcl-2-Associated X Protein in Renal Tubular Cells

目的探讨窒息后新生儿血清对人近曲肾小管上皮细胞(HK-2)内促凋亡蛋白抗Bcl-2蛋白(BAD)、Bcl-2联接X蛋白(BAX)表达的影响。方法以人HK-2为研究对象,分为对照、窒息和吡咯烷二硫氨基甲酸(PDTC)干预组。对照组:每组培养液换成不含窒息血清的DMEM;窒息组:每组培养液换成20mL/L窒息血清的DMEM窒息培养液;PDTC干预组:每组培养换成20mL/L窒息血清的DMEM培养液后,加40μmol/LPDTC。采用免疫组织化学(SP)法检测各组细胞BAD、BAX水平的表达。采用单因素方差分析进行统计学处理。结果用HSCORE系数作半定量评分,与对照组BAD、BAX的表达[(1.97±0.26)和(1.77±0.11)]比较,窒息组[(2.73±0.20)和(2.44±0.13)]和PDTC干预组[(2.38±0.13)和(2.17±0.08)]均明显增加,差异具有显著性;但与窒息组比较,PDTC干预组BAD、BAX的表达显著减少,差异有显著性[F(BAD)=28.61,F(BAX)=15.51Pa<0.01)]。结论新生儿窒息后血清可诱导人HK-2细胞BAD、BAX的表达上调,其胞内信号机制涉及到核因子-κB活化。

Objective To explore the effect of postasphyxial- serum in neonate on the expressions of Bcl-2- antagonist of cell death（BAD） and Bcl-2 -associated X protein（BAX） in renal tubular cells（HK-2）. Methods HK-2 cells were used as target cells, The experiment were divided into control group ,asphyxia group and pyrroledine dithiocarbamate （ PDTC ） blocking group, Control group ： DMEM culture fluid was not contained asphyxia blood serum in every greup ;asphxia group：DMEM culture fluid contained 20 mL/L asphyxia blood serum in every group;PDTC blocking group：DMEM culture fluid contained 20 mL/L asphyxia blood serum and 40 μmol/L PDTC in every group. The expressiolls of both BAD and BAX on cytoplast were detected by immunohistochemical method. Results Calculated Points according to HSCORE,compared with controls group [ （ 1.97 ± 0. 26） and（ 1.77 ± 0. 11 ） ],after stimulated with postasphyxial - serum, the expressions of both BAD and BAX of HK - 2 cells of asphyxia group[ （2. 73±0, 20）and（2.44 ±0. 13）] and PDTC blocking group[ （2.38±0. 13）and（2.17 ±0.08）] significantly increased[ F（BAD） =28.61 ,F （BAX） = 15.51 Pa 〈0. 01 ] ,but compared with asphyxia group, the expressions of both BAD and BAX of PDTC blocking group were significantly decreased （Pa 〈 0.01 ）. Conclusions Postasphyxial - serum is a stimulus that can induce the expression of both BAD and BAX in HK - 2 and its in- tracellular signal transduction mechanism is presumably involved in the activation of nuclear factor - κB.