Department of Cardiology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang, China.
Zhejiang Key Laboratory of Integrative Chinese and Western Medicine for Diagnosis and Treatment of Circulatory Diseases, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang, China.
Pharm Biol. 2024 Dec;62(1):790-802. doi: 10.1080/13880209.2024.2415666. Epub 2024 Oct 25.
Tanyu Tongzhi Formula (TTF) exhibits potential against atherosclerosis; however, its mechanisms remain unclear.
This study explores the pharmacological mechanisms of TTF in treating atherosclerosis.
Network pharmacology, molecular docking, mendelian randomization (MR), and liquid chromatography-mass spectrometry (LC-MS) analyses were utilized to reveal potential targets and compounds of TTF against atherosclerosis. After exploring the appropriate concentration of TTF to treat HCAECs using Cell Counting Kit-8 (CCK-8), the HCAECs were divided into three groups: control, oxidized low-density lipoprotein (ox-LDL, 50 μg/mL), and ox-LDL (50 μg/mL) + TTF (1 mg/mL). After 24-h incubation, the efficacy of TTF was verified by CCK-8, Oil red O staining, and ELISA. The expression of key targets was detected by real-time polymerase chain reaction (qPCR) and western blotting.
A total of 137 active compounds and 127 potential TTF targets against atherosclerosis were identified. MR identified ALB, TNF, PPARα, and PPARγ as key targets. Molecular docking indicated that baicalin, naringenin, and curcumin exhibited suitable binding activities to these targets, further confirming by LC-MS analysis. The IC of TTF in HCAECs was 18.25 mg/mL. TTF treatment significantly improved atherosclerosis by enhancing cell viability, reducing lipid accumulation, and inhibiting inflammation factors (IL6, IL1B and TNF-α) in ox-LDL-treated HCAECs. Moreover, qPCR or western blotting indicated that TTF could up-regulate PPARα and PPARγ while down-regulate TNF expression.
Our results revealed active compounds, key pathways, and core targets of TTF against atherosclerosis, providing experimental support for its application in treating of atherosclerosis.
痰瘀同治方（TTF）具有抗动脉粥样硬化作用，但作用机制尚不清楚。
本研究探讨 TTF 治疗动脉粥样硬化的药理学机制。
采用网络药理学、分子对接、孟德尔随机化（MR）和液相色谱-质谱（LC-MS）分析方法，揭示 TTF 抗动脉粥样硬化的潜在靶点和化合物。用细胞计数试剂盒-8（CCK-8）法探索 TTF 治疗人脐静脉内皮细胞（HCAEC）的适宜浓度后，将 HCAEC 分为三组：对照组、氧化型低密度脂蛋白（ox-LDL，50μg/mL）组和 ox-LDL（50μg/mL）+TTF（1mg/mL）组。孵育 24 小时后，用 CCK-8、油红 O 染色和 ELISA 验证 TTF 的疗效。用实时聚合酶链反应（qPCR）和 Western blot 检测关键靶点的表达。
共鉴定出 137 种活性化合物和 127 种潜在的 TTF 抗动脉粥样硬化靶点。MR 鉴定出 ALB、TNF、PPARα 和 PPARγ 为关键靶点。分子对接表明黄芩苷、柚皮苷和姜黄素与这些靶点具有合适的结合活性，进一步通过 LC-MS 分析得到证实。TTF 在 HCAEC 中的 IC 为 18.25mg/mL。TTF 治疗可显著改善动脉粥样硬化，增强细胞活力，减少 ox-LDL 处理的 HCAEC 中的脂质积累，并抑制炎症因子（IL6、IL1B 和 TNF-α）。此外，qPCR 或 Western blot 表明，TTF 可上调 PPARα 和 PPARγ，同时下调 TNF 表达。
本研究结果揭示了 TTF 抗动脉粥样硬化的活性化合物、关键途径和核心靶点，为其在动脉粥样硬化治疗中的应用提供了实验依据。
