乙醛脱氢酶1家族成员A2过表达对心肌细胞肥大生长及增殖的影响  

Effect of overexpression of aldehyde dehydrogenase family member A2 on hypertrophic growth and proliferation of cardiomyocytes

作　　者：刘航 刘琪琪 李振华 杨晓 王剑 LIU Hang;LIU Qiqi;LI Zhenhua;YANG Xiao;WANG Jian(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230000,Anhui,China;National Center for Protein Sciences(Beijing),Academy of Military Medical Sciences,Beijing 100850,China;College of Life Sciences,Hebei University,Baoding 071002,Hebei,China)

机构地区：[1]安徽医科大学基础医学院生物化学与分子生物学教研室,安徽合肥230000 [2]国家蛋白质科学中心(北京)军事科学院军事医学研究院,北京100850 [3]河北大学生命科学院,河北保定071002

出　　处：《生物工程学报》2024年第12期4660-4669,共10页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(82270354,82270355);国家重点研发计划(2019YFA0801601)。

摘　　要：维甲酸信号通路在调节脊椎动物发育、细胞分化和维持组织稳态平衡中发挥重要功能。作为催化视黄醛氧化成维甲酸的关键酶,乙醛脱氢酶1家族成员A2(aldehyde dehydrogenase 1 family member A2,Aldh1a2)在调节心脏发育过程中具有重要功能,而其在心脏疾病中发挥的功能尚不明晰。本研究通过构建和包装用于过表达Aldh1a2的腺病毒(Ad-Aldh1a2),将其感染原代心肌细胞后分析Aldh1a2过表达对心肌细胞生物学功能的影响。结果显示心肌细胞感染Ad-Aldh1a2后能够成功实现Aldh1a2的过表达;与感染Ad-GFP的对照组相比,感染Ad-Aldh1a2的心肌细胞尺寸明显增加,胚胎期基因心房利钠肽(atrial natriuretic factor,ANF)、脑利钠肽(brain natriuretic peptide,BNP)和β-肌球蛋白重链(β-myosin heavy chain,β-MHC)表达水平上调;此外,5-乙炔基-2'-脱氧尿苷(EdU)掺入实验结果显示,Aldh1a2过表达使EdU掺入阳性信号的心肌细胞比例升高,检测增殖相关基因的表达发现细胞周期蛋白D2(cycline D2,Ccnd2)和苯并咪唑出芽抑制解除同源物蛋白-1(budding uninhibited by benzimidazole 1,Bub1)的表达水平也明显升高。以上结果表明Aldh1a2过表达可促进心肌细胞肥大生长和增殖。本研究为进一步了解Aldh1a2在心脏疾病中的功能、发展心脏疾病的治疗策略提供了有益的基础。Retinoic acid signaling pathway plays a role in regulating vertebrate development,cell differentiation,and homeostasis.As a key enzyme that catalyzes the oxidation of retinal to retinoic acid,aldehyde dehydrogenase 1 family member A2(Aldh1a2)is involved in cardiac development,while whether it functions in heart diseases remains to be studied.In this study,we infected primary cardiomyocytes with adenovirus overexpressing Aldh1a2(Ad-Aldh1a2)to explore the effects of Aldh1a2 overexpression on the biological function of cardiomyocytes.The results showed that the infection with Ad-Aldh1a2 realized the overexpression of Aldh1a2 in cardiomyocytes.Compared with the control group infected with Ad-GFP,the cardiomyocytes infected with Ad-Aldh1a2 showcased significantly increased size and up-regulated expression levels of the atrial natriuretic factor gene(ANF),brain natriuretic peptide gene(BNP),andβ-myosin heavy chain(β-MHC).In addition,5-ethynyl-2ʹ-deoxyuridine(EdU)incorporation assay demonstrated that Aldh1a2 overexpression increased the proportion of cardiomyocytes with positive EdU signals and upregulated the expression levels of proliferation-related genes cyclin D2(Ccnd2)and budding uninhibited by benzimidazole 1(Bub1).The above data indicated that overexpression of Aldh1a2 induced hypertrophic growth and proliferation of cardiomyocytes.This study provides a basis for further understanding the function of Aldh1a2 in heart diseases and developing therapies for heart diseases.

关 键 词：乙醛脱氢酶1家族成员A2 腺病毒 心肌肥厚 心肌细胞增殖 

分 类 号：R542.2[医药卫生—心血管疾病]