Modern Research Center for Traditional Chinese Medicine, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, No. 92, Wucheng Road, Taiyuan, 030006, Shanxi, China; Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources (Guangxi Normal University), Guilin, 541004, China.
Modern Research Center for Traditional Chinese Medicine, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, No. 92, Wucheng Road, Taiyuan, 030006, Shanxi, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 2):118860. doi: 10.1016/j.jep.2024.118860. Epub 2024 Sep 26.
According to traditional Chinese medicine, Anxiety-induced cardiac blood insufficiency leads to palpitations and restlessness. Suanzaoren Decoction (SD) is effective in replenishing blood and promoting blood circulation. Clinical practice has shown that it has a better therapeutic effect on cardiac insufficiency. However, its mechanism of action is still unclear.
The study aims to determine the mechanism by which SD treats chronic restraint stress (CRS)-induced anxiety-induced cardiac insufficiency (ACI).
SD was orally administered to mice with CRS-induced ACI. Firstly, we constructed an anxiety model in mice by CRS. Subsequently, SD was investigated to assess cardiac function and pathological changes through echocardiography, H&E staining, and Masson staining. Thirdly, the function of sympathetic and parasympathetic nerves was evaluated using enzyme-linked immunosorbent assay (ELISA) and enzyme activity assays. Network pharmacology and molecular docking were employed to predict potential targets for SD treatment of cardiac insufficiency. CaMKII expression was scrutinized utilizing publicly accessible databases. CaMKII was identified as a target through immunohistochemistry and Western Blot analysis in mouse hearts. Finally, the therapeutic mechanism of SD was confirmed in injured cardiomyocytes via Western Blot and quantitative PCR.
SD exerted anxiolytic effects by increasing the frequency of entries into and the duration spent in open arms while reducing the time spent in the light chamber and increasing the number of transitions between light and dark chambers. Additionally, it mitigated cardiac insufficiency, as evidenced by the enhancement of left ventricular ejection fraction (LVEF) and attenuation of cardiomyocyte damage and inflammatory infiltration. However, SD did not alleviate the elevated norepinephrine (NE) and decreased Acetylcholine (Ach) in anxiety states. To investigate the mechanism of action of SD, we constructed a Drug-Component-Target-Disease network, identifying 13 potential active compounds. Additionally, leveraging bioinformatics analysis and molecular docking targeting heart diseases characterized by clinical left ventricular ejection fraction (LVEF), we focused on the CaMKII target. The ability of SD to modulate CaMKII expression and phosphorylation in the mouse heart was investigated using immunohistochemistry and Western blotting. SD was found to alleviate NE-injured cardiomyocytes by modulating the Ca/CaMKII/MEF2 and GATA4 pathways.
SD is a potential formula for the treatment of chronic restraint stress (CRS)-induced ACI that ameliorates cardiomyocyte injury and improves cardiac function. Its efficacy is associated with the inhibition of the Ca/CaMKII/MEF2 and GATA4 signaling pathways.
民族药理学相关性：根据中医理论，焦虑导致心血不足会引起心悸和烦躁不安。酸枣仁汤（SD）具有补血活血的功效，临床实践表明，它对心功能不全有较好的治疗作用。但其作用机制尚不清楚。
研究目的：本研究旨在确定 SD 治疗慢性束缚应激（CRS）诱导的焦虑性心功能不全（ACI）的机制。
材料与方法：用 CRS 诱导的 ACI 小鼠给予 SD 口服治疗。首先，我们通过 CRS 构建了一种焦虑模型。随后，通过超声心动图、H&E 染色和 Masson 染色评估 SD 对心脏功能和病理变化的影响。然后，通过酶联免疫吸附测定（ELISA）和酶活性测定评估交感神经和副交感神经的功能。采用网络药理学和分子对接技术预测 SD 治疗心功能不全的潜在靶点。利用公共可用数据库研究 CaMKII 的表达情况。通过免疫组织化学和 Western Blot 分析鉴定出小鼠心脏中的 CaMKII 为靶点。最后，通过 Western Blot 和 qPCR 确认 SD 在受损心肌细胞中的治疗机制。
结果：SD 通过增加进入开放臂的频率和在开放臂中花费的时间，同时减少在亮室中的时间并增加亮暗室之间的转换次数，发挥抗焦虑作用。此外，它减轻了心功能不全，表现为左心室射血分数（LVEF）增加，心肌细胞损伤和炎症浸润减轻。然而，SD 并没有减轻焦虑状态下去甲肾上腺素（NE）的升高和乙酰胆碱（Ach）的降低。为了研究 SD 的作用机制，我们构建了一个药物-成分-靶点-疾病网络，确定了 13 个潜在的活性化合物。此外，利用生物信息学分析和针对以临床左心室射血分数（LVEF）为特征的心脏病的分子对接靶点，我们聚焦于 CaMKII 靶点。利用免疫组织化学和 Western blot 研究 SD 对小鼠心脏中 CaMKII 表达和磷酸化的调节作用。结果发现，SD 通过调节 Ca/CaMKII/MEF2 和 GATA4 通路来缓解 NE 损伤的心肌细胞。
结论：SD 是一种治疗慢性束缚应激（CRS）诱导的 ACI 的潜在方剂，可减轻心肌细胞损伤，改善心功能。其疗效与抑制 Ca/CaMKII/MEF2 和 GATA4 信号通路有关。
