Chen Yanmin, Yu Jiaying, He Hualing, Xu Wencai, Yi Binghua, Lin Xingyu, Yu Qiulu, Lin Zhonghao, Lin Jiafeng, Lin Cong, Cai Chenchen, Huang Kaiyu
Department of Cardiology, Key Laboratory of Panvascular Diseases of Wenzhou, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, PR China.
Department of Cardiology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, PR China.
Phytomedicine. 2025 Oct;146:157126. doi: 10.1016/j.phymed.2025.157126. Epub 2025 Jul 30.
Acute myocardial infarction (MI) remains a leading cause of morbidity and mortality worldwide. Dihydrocapsaicin (DHC), a natural antioxidant derived from plants, has demonstrated pharmacological effects in various diseases. However, its research in the cardiovascular field remains relatively limited.
This study aimed to explore the anti-oxidative and anti-apoptotic effects of DHC on MI and its underlying mechanisms.
The MI model was established by left coronary artery ligation. In vitro, the H9c2 cardiomyocyte injury model was established via oxygen-glucose deprivation (OGD). Myocardial injury was assessed by echocardiography, infarct size, histopathological changes, and serological testing. TUNEL assay, Annexin V/PI staining assay, DHE, DCFH-DA staining and western blot analysis were used to validate cell apoptosis and oxidative stress level. The target of DHC was investigated using immunofluorescence, biotin pull-down assays, molecular docking, molecular dynamics simulations, cellular thermal shift assays (CETSA), and drug affinity responsive target stability (DARTS). Subsequently, the molecular mechanism of the interaction between Raf-1/ASK1 complex and DHC was investigated. Moreover, Raf-1 silencing or ASK1 overexpression H9c2 cardiomyocytes were used for further validation.
DHC was found to exert anti-oxidative and anti-apoptotic effects, attenuating cardiac dysfunction and injury in the MI model and protecting against OGD-induced injury in H9c2 cardiomyocytes. Mechanistically, Raf-1 was identified as a direct target of DHC. The protection of DHC against superoxide production and apoptotic cell death was compromised with Raf-1 silencing. Additionally, we found that DHC enhance the interaction between Raf-1 and ASK1, inhibiting the activation of the p-ASK1/p-JNK/p-P38 signaling pathway during OGD insult. The overexpression of ASK1 abolished the above protective effects of DHC.
Our results demonstrated that DHC may serve as a promising therapeutic agent for attenuating cardiac dysfunction after MI by alleviating oxidative stress and apoptosis.
急性心肌梗死（MI）仍是全球发病和死亡的主要原因。二氢辣椒素（DHC）是一种从植物中提取的天然抗氧化剂，已在多种疾病中显示出药理作用。然而，其在心血管领域的研究仍然相对有限。
本研究旨在探讨DHC对心肌梗死的抗氧化和抗凋亡作用及其潜在机制。
通过左冠状动脉结扎建立心肌梗死模型。在体外，通过氧糖剥夺（OGD）建立H9c2心肌细胞损伤模型。通过超声心动图、梗死面积、组织病理学变化和血清学检测评估心肌损伤。采用TUNEL检测、Annexin V/PI染色检测、DHE、DCFH-DA染色和蛋白质印迹分析来验证细胞凋亡和氧化应激水平。使用免疫荧光、生物素下拉试验、分子对接、分子动力学模拟、细胞热位移试验（CETSA）和药物亲和力响应靶点稳定性（DARTS）研究DHC的靶点。随后，研究Raf-1/ASK1复合物与DHC相互作用的分子机制。此外，使用Raf-1沉默或ASK1过表达的H9c2心肌细胞进行进一步验证。
发现DHC具有抗氧化和抗凋亡作用，可减轻心肌梗死模型中的心脏功能障碍和损伤，并保护H9c2心肌细胞免受OGD诱导的损伤。机制上，Raf-1被确定为DHC的直接靶点。Raf-1沉默会损害DHC对超氧化物产生和凋亡细胞死亡的保护作用。此外，我们发现DHC增强了Raf-1与ASK1之间的相互作用，在OGD损伤期间抑制了p-ASK1/p-JNK/p-P38信号通路的激活。ASK1的过表达消除了DHC的上述保护作用。
我们的结果表明，DHC可能是一种有前景的治疗药物，可通过减轻氧化应激和凋亡来减轻心肌梗死后的心脏功能障碍。
