Effect of DACT2 on markers of heart failure in H9C2 cardiomyocytes

Abstract

Objective \nTo investigate the regulation of Dishevelled-binding antagonist of β-catenin 2 (DACT2) overexpression on H9C2 cardiomyocyte heart failure markers and its molecular mechanism by infecting H9C2 cardiomyocytes. \n \n \nMethods \nAd-DACT2-green fluorescent protein (GFP) and its control empty vector Ad-CON177-GFP adenovirus were infected with H9C2 cardiomyocytes. The infection efficiency of H9C2 cardiomyocytes was observed by fluorescence microscope. The overexpression efficiency of DACT2 was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting; RT-qPCR for detecting DACT2 overexpression of heart failure markers α-myosin heavy chain (α-MHC), β-myosin heavy chain (β-MHC), atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and natriuretic peptide type A (ANF) expression changes; Western blotting was used to detect key protein expression changes of Wnt/β-Catenin and transforming growth factor-β (TGF-β)/Smad pathway after DACT2 overexpression. Independent sample t test was used for comparison between the two groups. \n \n \nResults \nThe green fluorescence showed successful cell infection. Compared with the empty group, the mRNA and protein levels of the DACT2 overexpression group were significantly increased (t=11.291, t=31.469, P 0.05). The expression of Smad4 was significantly up-regulated after DACT2 overexpression (t=9.510, P<0.05), and the expression of other factors was not significantly changed. \n \n \nConclusion \nDACT2 inhibits the expression of heart failure markers by up-regulating Smad4, suggesting that DACT2 may be a potential therapeutic target for heart failure diseases. \n \n \nKey words: \nDishevelled-binding antagonist of β-catenin 2;\xa0H9C2 cardiomyocytes;\xa0Overexpression;\xa0Heart failure;\xa0Smad4