背景:根据最近的研究,通过抑制G蛋白偶联受体激酶2(GRK2)改善心肌能量代谢来治疗心力衰竭(HF)已被确定为一种潜在的方法。肉桂醛(CIN),苯丙醛化合物,已被证明在心血管疾病中表现出有益的作用。然而,CIN是否抑制GRK2改善HF心肌能量代谢尚不清楚。
目的:本研究探讨了CIN对GRK2和心肌能量代谢的影响,以阐明其治疗HF的潜在机制。
方法:用Sprague-Dawley(SD)大鼠和原代新生大鼠心肌细胞(NRCMs)构建异丙肾上腺素(ISO)诱导的HF体内和体外模型。基于此,研究了CIN对心肌能量代谢和GRK2的影响。此外,在ISO诱导的NRCM中干扰和过表达GRK2后进行验证实验,以验证CIN对GRK2的调节作用。此外,GRK2和CIN之间的结合能力通过细胞热转移分析(CETSA)和微尺度热泳法(MST)进行了研究。
结果:体内和体外,通过逆转心肌损伤标志物的异常变化证明,CIN显着改善了HF,抑制心肌肥厚,减少心肌纤维化。此外,CIN通过激活AMPK/PGC-1α信号通路促进心肌脂肪酸代谢,改善心肌能量代谢紊乱。此外,CIN通过GRK2过表达逆转ISO诱导的NRCM对心肌脂肪酸代谢和AMPK/PGC-1α信号通路的抑制作用。同时,当GRK2被破坏时,CIN对ISO诱导的NRCM中心脏脂肪酸代谢的刺激和AMPK/PGC-1α信号通路没有更好的影响。值得注意的是,CETSA和MST证实CIN与GRK2直接结合。CIN和GRK2的结合促进了鼠双模拟物2介导的GRK2的泛素化降解。
结论:这项研究表明,CIN通过靶向GRK2并促进其泛素化降解以激活AMPK/PGC-1α信号通路,在HF中发挥保护性干预作用,最终改善心肌脂肪酸代谢。
BACKGROUND: According to recent research, treating heart failure (HF) by inhibiting G protein-coupled receptor kinase 2 (
GRK2) to improve myocardial energy metabolism has been identified as a potential approach. Cinnamaldehyde (CIN), a phenylpropyl aldehyde compound, has been demonstrated to exhibit beneficial effects in cardiovascular diseases. However, whether CIN inhibits GRK2 to ameliorate myocardial energy metabolism in HF is still unclear.
OBJECTIVE: This study examines the effects of CIN on GRK2 and myocardial energy metabolism to elucidate its underlying mechanism to treat HF.
METHODS: The isoproterenol (ISO) induced HF model in vivo and in vitro were constructed using Sprague-Dawley (SD) rats and primary neonatal rat cardiomyocytes (NRCMs). Based on this, the effects of CIN on myocardial energy metabolism and
GRK2 were investigated. Additionally, validation experiments were conducted after interfering and over-expressing GRK2 in ISO-induced NRCMs to verify the regulatory effect of CIN on
GRK2. Furthermore, binding capacity between GRK2 and CIN was explored by Cellular Thermal Shift Assay (CETSA) and Microscale Thermophoresis (MST).
RESULTS: In vivo and in vitro, CIN significantly improved HF as demonstrated by reversing abnormal changes in myocardial injury markers, inhibiting myocardial hypertrophy and decreasing myocardial fibrosis. Additionally, CIN promoted myocardial fatty acid metabolism to ameliorate myocardial energy metabolism disorder by activating AMPK/PGC-1α signaling pathway. Moreover, CIN reversed the inhibition of myocardial fatty acid metabolism and AMPK/PGC-1α signaling pathway by GRK2 over-expression in ISO-induced NRCMs. Meanwhile, CIN had no better impact on the stimulation of cardiac fatty acid metabolism and the AMPK/PGC-1α signaling pathway in ISO-induced NRCMs when GRK2 was disrupted. Noticeably, CETSA and MST confirmed that CIN binds to
GRK2 directly. The binding of CIN and
GRK2 promoted the ubiquitination degradation of
GRK2 mediated by murine double mimute 2.
CONCLUSIONS: This study demonstrates that CIN exerts a protective intervention in HF by targeting GRK2 and promoting its ubiquitination degradation to activate AMPK/PGC-1α signaling pathway, ultimately improving myocardial fatty acid metabolism.