法洛四联症患儿心肌组织HOXA3基因表达及凋亡检测  

Detection of HOXA3 gene expression and apoptosis in myocardial tissues of children with tetralogy of fallot

作　　者：姜红堃[1] 安东[1] 李雷[2] 白英龙[3] 邱广蓉[4] 姜红[1] 孙开来[4] 

机构地区：[1]中国医科大学附属第一医院儿科,沈阳110001 [2]中国医科大学附属盛京医院外科 [3]中国医科大学公共卫生学院儿少卫生教研室 [4]中国医科大学遗传学教研室

出　　处：《中华实用儿科临床杂志》2013年第13期970-972,共3页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81070131,30400485）;辽宁省教育厅基金（2008754,12012271）

摘　　要：目的 通过检测法洛四联症（TOF）患儿心肌组织 HOXA3基因表达及心肌细胞凋亡，探讨其在 TOF发病中的分子遗传学机制。方法 选取22例经产前彩超检查及尸检确诊为 TOF的胎儿右心室流出道心肌组织为 TOF组［胎龄（25.67±7.68）周］，12例同胎龄心脏结构正常胎儿右心室流出道心肌组织为对照组［胎龄（26.55±6.36）周］；提取心肌组织总 RNA，采用实时定量 PCR 法检测 HOXA3 mRNA 表达；提取心肌组织总蛋白并采用W estern blot法检测 HOXA3蛋白表达；原位末端标记法（TUNEL）检测心肌细胞凋亡。采用 SPSS13.0软件比较2组的差异及相关性分析。结果 与对照组比较，TOF组心肌组织 HOXA3 mRNA 表达下调（P 〈0.01），蛋白表达亦呈相同趋势（P 〈0.01）。TOF组凋亡心肌细胞数较对照组增多，差异有统计学意义（P 〈0.01）。TOF组心肌细胞凋亡数与HOXA3 mRNA及蛋白表达水平呈负相关（r= -0.566、-0.759，P均 〈 0.01）。结论 在胚胎心脏发育过程中，HOXA3基因表达降低及心肌细胞过度凋亡可能是 TOF发生的一种潜在致病机制。Objective To explore the potential role of HOXA3 gene in children with tetralogy of fallot （TOF） by detection the expression of HOXA3 mRNA and protein levels, as well as the apoptotie eardiomyoeytes in the develo-ping heart tissues. Methods Twenty-two surgical samples from sporadic cases of TOF determined by prenatal color Doppler ultrasound and autopsy [ gestational age （25.67 ± 7.68 ） weeks, TOF group ] were examined with quantitative real-time PCR and Western blot to evaluate the expression of HOXA3 gene. Twelve age-matched autopsies without heart structural abnormalities [ gestational age（ 26.55 _± 6.36 ） weeks, control group ] were also included. Terminal deoxynu- cleotidyl transferase-mediated dUTP nick end-labeling（TUNEL） assay was performed to clarify the apoptosis of cardio-myocytes. The difference between the 2 groups and the correlation analysis of HOXA3 level and the apoptotic eardio- myocytes were performed with SPSS 13.0 software. Results Compared with control group,HOXA3 mRNA expression of the outflow tract of the right ventricle from the TOF group was significantly reduced（ P 〈 0.01 ）. Western blot also con- firmed that the HOXA3 protein was accordingly reduced（ P 〈0.01 ）. The proportion of apoptotie cardiomyocytes in sam- ples of TOF group was significantly greater than that of control group（ P 〈 0.01 ）. The proportion of apoptotic cells was strongly correlated with the mRNA and protein expression of HOXA3 （ r = - 0.566, - 0. 759, all P 〈 0.01 ）. Conclusions Reduction of HOXA3 gene expression and the increase of apoptotic cardiomyocytes at the crucial stage during heart develooment may olav a ootential role in the onset of TOF.

关 键 词：HOXA3基因 法洛四联症 凋亡 心脏发育 

分 类 号：R725.4[医药卫生—儿科]