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-2ʹ-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.
Effect of overexpression of aldehyde dehydrogenase family member A2 on hypertrophic growth and proliferation of cardiomyocytes
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),and β-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.
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