Liu Juan, Li Ya, Bian Xiyun, Xue Na, Yu Jiancai, Dai Shipeng, Liu Xiaozhi
Department of Cardiology, Cangzhou Central Hospital, Cangzhou, Hebei 061000, P.R. China.
Central Laboratory, The Fifth Central Hospital of Tianjin, Tianjin 300450, P.R. China.
Exp Ther Med. 2021 Oct;22(4):1076. doi: 10.3892/etm.2021.10510. Epub 2021 Jul 28.
The present study investigated whether the protective effect and mechanism of astragaloside IV (AS-IV) on heart failure (HF) involves small ubiquitin-like modifier (SUMO)-specific protease 1 (Senp1). Mouse HF was established by aortic constriction, inducing pressure overload. The model was confirmed by echocardiography 6 weeks after surgery. Mice were randomly divided into control, HF, HF+AS-IV, and AS-IV groups. Ventricular function was examined by echocardiography. Morphological changes of myocardial tissues were examined by H&E staining. The protein levels of the apoptosis-related proteins, cleaved caspase-3, caspase-3, Bcl2, Bax, and SUMO-Senp1 were determined by Western blotting. HO in isolated mitochondria and cells was determined by Amplex Red. A reactive oxygen species (ROS) detection kit determined ROS levels in isolated mitochondria and HL-1 cells. JC-1 reagent measured mitochondrial membrane potential (ΔΨm). Apoptosis of HL-1 cells was examined by terminal deoxynucleotidyl transferase dUTP nick end labeling. Compared with the control group, the heart weight and heart mass/body weight ratio increased in the HF group (P<0.05). Furthermore, the ejection fraction and left ventricular shortening fraction decreased (P<0.05), while the left ventricular end-diastolic diameter (LVID;d) and end-systolic diameter (LVID;s) increased (P<0.05). Finally, mitochondrial ROS and HO increased (P<0.05), while the ΔΨm decreased (P<0.05). However, AS-IV improved the cardiac function of HF mice, decreased the level of ROS and HO in the myocardium, suppressed the decrease in ΔΨm, and decreased the apoptosis of myocardial cells (P<0.05). AS-IV also decreased the Senp1-overexpression. Furthermore, in HL-1 cells, Senp1-overexpression significantly inhibited the protective effects of AS-IV. AS-IV decreased oxidative stress in cardiomyocytes, decreased mitochondrial damage, inhibited ventricular remodeling, and ultimately improved cardiac function by inhibiting HF-induced Senp1-overexpression. This mechanism provides a novel theoretical basis and clinical treatment for HF.
本研究探讨黄芪甲苷（AS-IV）对心力衰竭（HF）的保护作用及机制是否涉及小泛素样修饰物（SUMO）特异性蛋白酶1（Senp1）。通过主动脉缩窄建立小鼠HF模型，诱导压力过载。术后6周通过超声心动图确认模型。将小鼠随机分为对照组、HF组、HF+AS-IV组和AS-IV组。通过超声心动图检查心室功能。通过苏木精-伊红（H&E）染色检查心肌组织的形态学变化。通过蛋白质印迹法测定凋亡相关蛋白、裂解的半胱天冬酶-3、半胱天冬酶-3、Bcl2、Bax和SUMO-Senp1的蛋白水平。通过Amplex Red测定分离的线粒体和细胞中的血红素加氧酶（HO）。活性氧（ROS）检测试剂盒测定分离的线粒体和HL-1细胞中的ROS水平。JC-1试剂测量线粒体膜电位（ΔΨm）。通过末端脱氧核苷酸转移酶dUTP缺口末端标记法检测HL-1细胞凋亡。与对照组相比,HF组心脏重量和心脏重量/体重比增加（P<0.05）。此外，射血分数和左心室缩短分数降低（P<0.05），而左心室舒张末期直径（LVID;d）和收缩末期直径（LVID;s）增加（P<0.05）。最后，线粒体ROS和HO增加（P<0.05），而ΔΨm降低（P<0.05）。然而，AS-IV改善了HF小鼠的心脏功能，降低了心肌中ROS和HO的水平，抑制了ΔΨm的降低，并减少了心肌细胞凋亡（P<0.05）。AS-IV还降低了Senp1的过表达。此外，在HL-1细胞中，Senp1过表达显著抑制了AS-IV的保护作用。AS-IV降低了心肌细胞中的氧化应激，减少了线粒体损伤，抑制了心室重塑，并最终通过抑制HF诱导的Senp1过表达改善了心脏功能。该机制为HF提供了新的理论基础和临床治疗方法。
