State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Tianjin Key Laboratories of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Tianjin Key Laboratories of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
J Ethnopharmacol. 2021 Apr 6;269:113724. doi: 10.1016/j.jep.2020.113724. Epub 2020 Dec 24.
As a classic prescription of Traditional Chinese Medicine in Correction on the Errors of Medical Works, Buyang Huanwu Decoction (BYHWD) has a good curative effect on prevention of atherosclerosis (AS).
This study aims to elucidate the anti-atherosclerosis mechanism of BYHWD, which may promote the differentiation of regulatory T cells by regulating the TGF-β/Smad2 pathway.
ApoE-/- mice were fed a high-fat diet for 12 weeks, then drugs group were given BYHWD with intragastric administration once a day for 4 weeks. The effect of BYHWD on lipid content in peripheral blood and plaque was evaluated by blood lipid test and oil red O staining. The number of Tregs in peripheral blood was tested by flow cytometry, and that in the spleen was evaluated by immunohistochemistry methods. Gene and protein expression relating with Tregs differentiation pathway in mice were checked by RT-PCR and Western blot experiments. CD4T cells were isolated and interfered by BYHWD drug-loaded serum. The proportion of Tregs was evaluated by flow cytometry. The chemical compositions of BYHWD and rat drug-loaded serum were analyzed by ultra-high performance liquid chromatograph and liquid chromatography-tandem mass spectrometry.
BYHWD significantly reduced plaque area and cholesterol accumulation, increased the number of Tregs in spleen and peripheral blood of ApoE-/- AS mice, raised the proportion of Tregs in CD4T cells, and regulated the levels of inflammatory factors. It also increased the TGF-β and Smad2 mRNA and protein levels relating with Tregs differentiation pathway in vivo. The mRNA levels of Foxp3/TGF-β/Smad2 were enhanced via BYHWD in vitro.
BYHWD regulates TGF-β/Smad2 signaling pathway to promotes the peripheral differentiation of Tregs, increases the number of Tregs, restores the immune balance between CD4T cells, regulates lipid metabolism, inhibits inflammatory reaction and possesses the potential of enhancing plaque stability.
作为《医林改错》的经典方剂，补阳还五汤（BYHWD）对动脉粥样硬化（AS）的防治具有良好的疗效。
本研究旨在阐明 BYHWD 的抗动脉粥样硬化机制，可能通过调节 TGF-β/Smad2 通路促进调节性 T 细胞的分化。
ApoE-/- 小鼠用高脂饲料喂养 12 周，然后药物组给予 BYHWD 灌胃，每天 1 次，共 4 周。通过血脂试验和油红 O 染色评价 BYHWD 对血脂和斑块中脂质含量的影响。通过流式细胞术检测外周血和脾脏中 Tregs 的数量，通过免疫组化方法评价脾脏中 Tregs 的数量。通过 RT-PCR 和 Western blot 实验检测与 Tregs 分化途径相关的基因和蛋白表达。通过 BYHWD 载药血清干预 CD4T 细胞，通过流式细胞术评估 Tregs 的比例。通过超高效液相色谱和液质联用分析 BYHWD 和大鼠载药血清的化学成分。
BYHWD 显著减少斑块面积和胆固醇积累，增加 ApoE-/-AS 小鼠脾脏和外周血中 Tregs 的数量，提高 CD4T 细胞中 Tregs 的比例，并调节炎症因子水平。它还增加了体内与 Tregs 分化途径相关的 TGF-β和 Smad2 mRNA 和蛋白水平。体外 BYHWD 增强了 Foxp3/TGF-β/Smad2 的 mRNA 水平。
BYHWD 通过调节 TGF-β/Smad2 信号通路促进 Tregs 的外周分化，增加 Tregs 的数量，恢复 CD4T 细胞之间的免疫平衡，调节脂质代谢，抑制炎症反应，具有增强斑块稳定性的潜力。
