School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong Province, China.
The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong Province, China.
J Ethnopharmacol. 2021 Feb 10;266:113436. doi: 10.1016/j.jep.2020.113436. Epub 2020 Oct 1.
Dingxin Recipe (DXR) is a traditional Chinese medicine formula that has been reported to be effective and safe treatment for cardiovascular diseases, such as arrhythmias, coronary heart disease. Dingxin Recipe IV (DXR IV) was further improved from the DXR according to the traditional use. However, the mechanism of DXR IV in atherosclerosis is unclear.
This study aimed to illustrate whether DXR IV improve atherosclerosis through modulating the lipid metabolism and gut microbiota in atherosclerosis mice.
40 male ApoE mice were fed on HFD for 12 weeks and were then treated with DXR IV (1.8, 0.9, or 0.45 g/kg/d) for another 12 weeks. The decroation of DXR IV contains four traditional Chinese medicines: the dried rhizome of Coptis chinensis Franch. (15.09%), the root of Salvia miltiorrhiza Bunge (28.30%), the seed of Ziziphus jujuba Mill. (37.74%) and the fruiting body of Ganoderma lucidum (Leyss.ex Fr.) Karst. (18.87%). 8 male c57BL/6 mice fed a normal diet served as control group. The atherosclerotic plaque was quantified by oil-red O staining and masson trichrome staining. Mice feces were collected. The gut micobiota were detected by 16S rRNA gene sequencing and fecal metabolites were analyzed by H NMR spectroscopy. The effect of DXR IV on blood lipids (TG, TC, LDL-C, HDL-C) was investigated. The lipid metabolism related genes were determined by RT-qPCR and western blotting respectively.
DXR IV exerted the anti-atherosclerosis effect by inhibiting the excessive cholesterol deposition in aorta and regulating the level of TG, TC, LDL-C and HDL-C. The composition of gut microbiota was changed. Interestingly, the relative abundance of Muribaculaceae and Ruminococcaceae increased after DXR IV administration, whereas the abundance of Erysipelotrichaceae decreased, which have been beneficial to lipid metabolism. Nine potential metabolic biomarkers, including acetate, butyrate, propionate, alanine, succinate, valerate, xylose, choline, glutamate, were identified, which were related to fatty acid metabolism. Further, the pathway of fatty acid was detected by the RT-qPCR and western blotting. Compared with model group, the level of LXR-α and SREBP1 decreased significantly in DXR IV group while LXR-β, SREBP2 showed no statistical significance. It indicated that DXR IV modulated lipid metabolism by LXR-α/SREBP1 but not LXRβ and SREBP2.
DXR IV exhibits potential anti-atherosclerosis effect, which is closely related to lipid metabolism and the gut microbiota. This study may provide novel insights into the mechanism of DXR IV on atherosclerosis and a basis for promising clinical usage.
定心方（DXR）是一种中药方剂，据报道可有效治疗心律失常、冠心病等心血管疾病。定心方 IV（DXR IV）是根据传统用途从 DXR 进一步改进而来。然而，DXR IV 在动脉粥样硬化中的作用机制尚不清楚。
本研究旨在阐明 DXR IV 是否通过调节动脉粥样硬化小鼠的脂质代谢和肠道微生物群来改善动脉粥样硬化。
40 只雄性 ApoE 小鼠用高脂肪饮食喂养 12 周，然后用 DXR IV（1.8、0.9 或 0.45 g/kg/d）治疗 12 周。DXR IV 的 decoction 含有四种中药：黄连根茎（15.09%）、丹参根（28.30%）、大枣种子（37.74%）和灵芝子实体（18.87%）。8 只正常饮食喂养的雄性 c57BL/6 小鼠作为对照组。用油红 O 染色和 Masson 三色染色定量动脉粥样硬化斑块。收集小鼠粪便。通过 16S rRNA 基因测序检测肠道微生物群，通过 H NMR 光谱分析粪便代谢物。研究 DXR IV 对血脂（TG、TC、LDL-C、HDL-C）的影响。通过 RT-qPCR 和 Western blot 分别测定脂质代谢相关基因。
DXR IV 通过抑制主动脉中胆固醇的过度沉积和调节 TG、TC、LDL-C 和 HDL-C 的水平发挥抗动脉粥样硬化作用。肠道微生物群的组成发生了变化。有趣的是，DXR IV 给药后 Muribaculaceae 和 Ruminococcaceae 的相对丰度增加，而 Erysipelotrichaceae 的丰度减少，这有利于脂质代谢。鉴定出 9 种潜在的代谢生物标志物，包括乙酸盐、丁酸盐、丙酸盐、丙氨酸、琥珀酸盐、戊酸盐、木糖、胆碱、谷氨酸，它们与脂肪酸代谢有关。进一步通过 RT-qPCR 和 Western blot 检测脂肪酸途径。与模型组相比，DXR IV 组 LXR-α 和 SREBP1 水平显著降低，而 LXR-β 和 SREBP2 无统计学意义。这表明 DXR IV 通过 LXR-α/SREBP1 而不是 LXRβ 和 SREBP2 调节脂质代谢。
DXR IV 表现出潜在的抗动脉粥样硬化作用，与脂质代谢和肠道微生物群密切相关。本研究可为 DXR IV 对动脉粥样硬化的作用机制提供新的见解，并为其有希望的临床应用提供依据。
