ALK3基因在心肌细胞凋亡中的作用研究  

Effect of ALK3 gene on cardiac myocyte apoptosis

作　　者：来丹丹[1] 章佳颖[1] 高瞻[1] 褚茂平[1] 王赛斌[1] 施翔翔[1] 黄晓燕[1] 张怀勤[1] 杨德业[1] 

机构地区：[1]温州医学院附属第一医院心血管内科,温州医学院心血管生物和基因研究所,浙江温州325000

出　　处：《解放军医学杂志》2009年第2期179-182,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助项目(30571050)

摘　　要：目的研究心脏特异性骨形态形成蛋白受体IA(BMPR1 A,又名ALK3)基因敲除小鼠心肌组织中的细胞凋亡情况,探讨ALK3基因在心肌细胞凋亡过程中的作用及细胞凋亡与室间隔缺损的关系。方法实验分为纯合子小鼠(α-MHC Cre+/-,ALK3 F/-),杂合子小鼠(α-MHC Cre+/-,ALK3 F/+)和野生型小鼠(C57)3组,每组取5只。20只雌性α-MHC Cre+/-,ALK3+/-小鼠和20只雄性ALK3 F/F小鼠交配获得心脏特异性ALK3基因敲除的纯合子小鼠(α-MHC Cre+/-,ALK3 F/-)和杂合子小鼠(α-MHC Cre+/-,ALK3 F/+);雌性和雄性C57小鼠各10只交配获得子代野生型小鼠;均取13.5d胚胎。提取胎膜DNA,PCR鉴定基因型。光镜下HE染色显示心脏组织形态,透射电镜观察心肌细胞的超微结构和凋亡情况,并以脱氧核糖核苷酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测心肌细胞凋亡率。结果光镜显示α-MHC Cre+/-,ALK3 F/-小鼠存在室间隔缺损现象,α-MHC Cre+/-,ALK3 F/+小鼠和C57小鼠心脏形态正常;透射电镜和TUNEL提示α-MHC Cre+/-,ALK3 F/-小鼠心肌细胞凋亡现象较α-MHC Cre+/-,ALK3 F/+小鼠和C57小鼠严重。结论ALK3基因在调控心脏发育和心肌细胞凋亡过程中起重要作用。心脏特异性ALK3基因敲除小鼠中,室间隔缺损可部分归因于心肌细胞的过度凋亡。Objective To investigate cardiac myocyte apoptosis in cardiac-specific deletion of bone morphogenetic protein receptor type IA （BMPR-IA, also named ALK3） in mice, and explore the effect of ALK3 gene on the cardiac myocyte apoptosis and the relationship between apoptosis and ventricular septum defect. Methods Mice were divided into 3 groups （5 each） ： a-MHC Cre ＋/-, ALK3 F/- （homozygote） group, α-MHC Cre ＋/-, ALK3 F/＋ （heterozygote） group and C57 group （wild type）. 13. 5-day embryos （E13.5d） of the mice in homozygote group and heterozygote group were generated by mating 20♀♀ of α-MHC Cre ＋/-, ALK3 ＋/- mice and 20 ♀♀ ALK3 F/F mice, as well as C57 mice （10♀♀ 10 ♀♀ ）. The genomic DNA of embryolemma was extracted, and then gene identi- fication was performed by PCR analysis. The pathologic abnormality of myocardium of cardiac-specific deletion of ALK3 in mice was observed by light microscope and transmission electron microscope. Cardiac myocyte apoptosis in mice was determined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. Results It was shown under light microscope that defects in the interuentricular septurn were present in the α-MHC Cre ＋/-, ALK3 F/- mice, while the α-MHC Cre ＋/-, ALK3 F/＋ mice and C57 mice were normal. The cardiac myocyte apoptosis of the mice in homozygote group was more remarkable than that of the mice in heterozygote group and C57 group as demonstrated by transmission electron microscope and TUNEL assay. Conclusions ALK3 gene may participate in regulating cardiac development and cardiac myocyte apoptosis. Furthermore, ventricular septum defect may partially be attributable to cardiac myocyte apoptosis excessively.

关 键 词：骨形态形成蛋白受体IA 室间隔缺损 细胞凋亡 

分 类 号：R541.12[医药卫生—心血管疾病]