补阳还五汤中激活ALDH2抗心梗后心肌细胞凋亡的物质基础研究

Myocardial apoptosis post myocardial infarction is the initiating factor that promotes the progress of chronic heart failure and cardiac death. In our previous study, we found the resistant effect of Buyang Huaiwu Decoction (BYHWD) on myocardial apoptosis post myocardial infarction, and determine the involvement of Acetaldehyde dehydrogenase 2 (ALDH2) in the anti-apoptosis effects of BYHWD. In our preliminary experiment, we found 14 candidate compounds, such as calycosin, which could bind with ALDH2 using the molecular docking, and confirmed the reliability of these results by cell experiment. Thus, we put forward the hypothesis that ALDH2 is the treatment target of BYHWD on anti-apoptosis, 14 candidate compounds may constitute the chemical basis of BYHWD on the activation of ALDH2. In this study, we will perform following study. ① Firstly, we will further verify the candidate compounds by molecular dynamics, cell experiment and cell free enzyme activity assay. ② Then, we will investigate the anti-apoptosis action of BYHWD in vitro and in vivo, and verify these results by the ALDH2 inhibitor. ③ Finally, we will investigate the role of candidate compounds combination in the protective effects of BYHWD on myocardial apoptosis post myocardial infarction. This study will provide a pharmacological justification for the clinical application of BYHWD in treating myocardial apoptosis post myocardial infarction.

心肌细胞凋亡是心梗患者发生心衰以致心源性死亡的始动因素。我们的既往研究发现补阳还五汤具有防治心梗后心肌细胞凋亡的作用，并证明乙醛脱氢酶2（ALDH2）是补阳还五汤的作用靶点。我们还采用计算机辅助虚拟筛选技术从补阳还五汤有效成分中筛选出毛蕊异黄酮等14个能与ALDH2结合的候选化合物，并采用分子动力学模拟及细胞实验初步证明该结果的可靠性。据此我们提出以下假说：毛蕊异黄酮等14个候选成分可能是补阳还五汤激活ALDH2抑制心梗后心肌细胞凋亡的物质基础。本项目拟①采用分子动力学技术、细胞实验等进一步筛选有效成分；②采用动物和细胞模型探索候选化合物抗心梗后心肌细胞凋亡作用，采用ALDH2抑制剂验证ALDH2在候选成分的抗心梗后凋亡中的作用；③探索候选成分组合在补阳还五汤抑制心梗后心肌细胞凋亡中所占的比重。本项目旨在探索补阳还五汤防治心梗后心肌细胞凋亡的物质基础，为补阳还五汤的临床应用提供依据。

心肌细胞凋亡是心梗患者发生心衰以致心源性死亡的始动因素。补阳还五汤具有防治心梗后心肌细胞凋亡的作用，乙醛脱氢酶2(ALDH2)是补阳还五汤的作用靶点。本项目拟采用分子动力学技术、细胞实验等进一步筛选有效成 分，采用动物和细胞模型探索候选化合物抗心梗后心肌细胞凋亡作用，采用ALDH2抑制剂验 证ALDH2在候选成分的抗心梗后凋亡中的作用。4年来，在国家自然科学基金的资助下，本课题组补阳还五汤激活ALDH2抗心梗后心肌细胞凋亡的物质基础进行研究。通过本项目的研究已发表SCI文章8篇，在投稿SCI文章2篇，申请中国发明专利4项。以下为本研究主要发现：①补阳还五汤能抑制心梗后心肌纤维化。②补阳还五汤能激活ALDH2。③抑制ALDH2能显著抑制补阳还五汤的心肌保护作用。④分子模拟、表面等离子共振（SPRi）、生物膜层干涉（BLI）技术研究发现13个成分可与ALDH2相互作用。⑤细胞酶活性实验发现8个成分可以显著升高ALDH2的活性。⑥Calycosin能显著抑制心肌细胞凋亡。⑦Calycosin能显著抑制心肌梗死小鼠心肌细胞凋亡。⑧抑制ALDH2可以显著抑制Calycosin的抗凋亡作用。⑨Magnolol可以显著抑制心肌成纤维细胞的增殖，抑制ALDH2显著抑制Magnolol的抗纤维化作用。