目的：基于SIRT1/AMPK通路探讨达格列净对心力衰竭的改善作用。方法：在C57BL/6J小鼠中建立动物主动脉弓缩窄（TAC）模型,分为假手术组、假手术+达格列净（DAPA）组、TAC组和TAC+DAPA组。分析器官大小和质量,HE染色、WGA染色、Masson、Sirius Red染色、DHE染色和TUNEL测定分析组织病理学、心肌细胞大小、胶原纤维、ROS的产生和细胞凋亡,qRT-PCR和蛋白印迹检测心房利钠因子（Nppa）、脑利钠肽（Nppb）、肌球蛋白重链7（Myh7）、TGF-β1和SIRT1/AMPK信号通路的表达。结果：与假手术组比较,DAPA的TAC小鼠心脏肥大指标显著降低。通过WGA染色评估,TAC小鼠心肌细胞的平均横截面积大,Nppa、Nppb和Myh7增加。此外,TAC小鼠的ANP和β-MHC水平高,在DAPA给予的TAC小鼠中观察到平均心肌细胞横截面积减少,心脏肥大标志物的mRNA和蛋白水平均显著降低。Masson和天狼星红检查心肌纤维化的程度,与假手术组相比,TAC小鼠心肌纤维化面积显著增加,TAC组心肌中TGF-β1的mRNA和蛋白水平升高,TAC小鼠的心肌ROS水平高,TUNEL染色显示TAC组心肌细胞凋亡率显著增加,SIRT1/AMPK通路表达降低,但给予DAPA的效果相反,差异有统计学意义。结论：DAPA激活了SIRT1/AMPK通路信号传导,从而减轻心脏纤维化、肥大重塑和心肌氧化应激
Abstract
Purpose To investigate the improvement effect of dapagliflozin with heart failure based on SIRT1/AMPK pathway. Methods Animal TAC models were developed in C57BL/6J mice and divided into sham, sham+DAPA, TAC, and TAC+DAPA groups. Gravimetric and histological analysis of organ size and weight, HE staining, WGA staining, Masson, Sirius Red staining, DHE staining and TUNEL assay to analyze histopathology, cardiomyocyte size, collagen fibers, ROS production and apoptosis, qRT- The expressions of atrial natriuretic factor (Nppa), brain natriuretic peptide (Nppb), myosin heavy chain 7(Myh7), TGF-β1 and SIRT1/AMPK signaling pathway were detected by PCR and western blotting. Results CHW/TL, HW/BW and HW/LW were significantly reduced in DAPA TAC mice compared with sham operation group. As assessed by WGA staining, the mean cross-section area of cardiocytes in TAC mice was larger. Increases in Nppa, Nppb, and Myh7, in addition to high levels of ANP and β-MHC in TAC mice, decreased mean cardiomyocyte cross-sectional area was observed in DAPA-administered TAC mice. mRNA and protein levels of markers of cardiac hypertrophy were significantly reduced. Masson and Sirius red were used to examine the degree of myocardial fibrosis. Compared with the sham operation group, the myocardial fibrosis area of TAC mice was significantly increased, the mRNA and protein levels of TGF-β1 in TAC group were increased, and the myocardial ROS levels of TAC mice were higher. TUNEL staining showed that the apoptosis rate of cardiomyocytes in TAC group was significantly increased, and the expression of SIRT1/AMPK pathway was decreased, but DAPA had the opposite effect, the difference was statistically significant. ompared with the sham-operated group, HW/TL, HW/BW and HW/LW were significantly reduced in TAC mice with DAPA, and the mean cross-sectional area of cardiomyocytes in TAC mice was large, Nppa, Nppb and Myh7 as assessed by WGA staining In addition, TAC mice had high levels of ANP and β-MHC, and a reduction in mean cardiomyocyte cross-sectional area and significantly lower mRNA and protein levels of cardiac hypertrophy markers were observed in DAPA-administered TAC mice. Masson and Sirius Red examined the degree of myocardial fibrosis. Compared with the sham-operated group, the myocardial fibrosis area of TAC mice was significantly increased, the mRNA and protein levels of TGF-β1 in the myocardium of the TAC group were increased, and the myocardial ROS level of TAC mice TUNEL staining showed that the apoptosis rate of cardiomyocytes in the TAC group was significantly increased, and the expression of SIRT1/AMPK pathway was decreased, but the effect of DAPA was the opposite, and the difference was statistically significant. Conclusion DAPA activates SIRT1/AMPK pathway signaling, thereby reducing cardiac fibrosis, hypertrophic remodeling, and myocardial oxidative stress.
关键词
小鼠 /
心力衰竭 /
达格列净 /
SIRT1/AMPK通路
Key words
mice /
heart failure /
dapagliflozin /
SIRT1/AMPK
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基金
湖南省卫生健康委科研项目“达格列净通过SIRT1AMPK通路改善心力衰竭的分子机制”（20210113）
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