Table1_Ginsenoside Rc ameliorated atherosclerosis via regulating gut microbiota and fecal metabolites.DOCX

Atherosclerosis (AS) and the accompanied cardiovascular diseases (CVDs) were the leading cause of death worldwide. Recently, the association between CVDs, gut microbiota, and metabolites had aroused increasing attention. In the study, we headed our investigation into the underlying mechanism of ginsenoside Rc (GRc), an active ingredient of ginsenosides used for the treatment of CVDs, in apolipoprotein E-deficient (ApoE−/−) mice with high-fat diet (HFD). Seven-week-old male ApoE−/− mice were randomly divided into four groups: the normal control (NC) group, the HFD group, the GRc group (40 mg/kg/d), and the atorvastatin (Ato) group (10 mg/kg/d). Atherosclerotic injury was evaluated by aortic lesions, serum lipid levels, and inflammatory factors. The composition of gut microbiota and fecal metabolite profile were analyzed using 16S rRNA sequence and untargeted metabolomics, respectively. The results showed that GRc significantly alleviated HFD-induced aortic lesions, reduced serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), tumor necrosis factor-α (TNF-α), and interleukin (IL)-6 and IL-1β, and increased high-density lipoprotein cholesterol (HFD-C) level, as well as the alteration of gut microbiota composition, function, and metabolite profile. GRc also reversed HFD change of Bacteroidetes and Firmicutes at the phylum level, Muribaculaceae, Lactobacillus, Ileibacterium, Bifidobacterium, Faecalibaculum, Oscillibacter, Blautia, and Eubacterium_coprostanoligenes_group at the genus level, and 23 key metabolites involved in taurine and hypotaurine metabolism, arginine biosynthesis, ATP-binding cassette (ABC) transporters, primary bile acid biosynthesis, purine metabolism, tricarboxylic acid (TCA) cycle, and glucagon signaling pathways. Additionally, eight differential intestinal floras at the genus level were associated with 23 key differential metabolites involving atherosclerotic injury. In conclusion, our results demonstrated that GRc ameliorated atherosclerotic injury, regulated microbial and metabolomic changes in HFD-induced ApoE−/− mice, and suggested a potential correlation among gut microbiota, metabolites, and atherosclerotic injury regarding the mechanisms of GRc against AS.

动脉粥样硬化（Atherosclerosis, AS）及其伴随的心血管疾病（cardiovascular diseases, CVDs）是全球范围内的首要致死病因。近年来，心血管疾病、肠道菌群与代谢物之间的关联愈发受到学界关注。本研究聚焦于人参皂苷Rc（ginsenoside Rc, GRc）——一种用于治疗心血管疾病的人参皂苷活性成分——在高脂饮食（high-fat diet, HFD）诱导的载脂蛋白E缺陷型（apolipoprotein E-deficient, ApoE−/−）小鼠中的潜在作用机制。将7周龄雄性ApoE−/−小鼠随机分为四组：正常对照组（normal control, NC）、高脂饮食组、人参皂苷Rc干预组（40 mg/kg/d）以及阿托伐他汀（atorvastatin, Ato）对照组（10 mg/kg/d）。通过主动脉病变程度、血清脂质水平与炎症因子水平评估动脉粥样硬化损伤情况。分别采用16S rRNA测序与非靶向代谢组学技术分析肠道菌群组成与粪便代谢物谱。研究结果显示，GRc可显著缓解高脂饮食诱导的主动脉病变，降低血清总胆固醇（total cholesterol, TC）、甘油三酯（triglyceride, TG）、低密度脂蛋白胆固醇（low-density lipoprotein cholesterol, LDL-C）、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）以及白细胞介素（interleukin, IL）-6、IL-1β水平，升高高密度脂蛋白胆固醇（HDL-C）水平，同时改善肠道菌群组成、功能与代谢物谱的异常改变。GRc还可逆转高脂饮食诱导的菌群丰度异常：在门水平上，可调节拟杆菌门（Bacteroidetes）与厚壁菌门（Firmicutes）的丰度比例；在属水平上，可改善Muribaculaceae、乳杆菌属（Lactobacillus）、Ileibacterium、双歧杆菌属（Bifidobacterium）、Faecalibaculum、Oscillibacter、Blautia以及Eubacterium_coprostanoligenes_group的丰度变化。此外，共有23种关键代谢物参与牛磺酸与次牛磺酸代谢、精氨酸生物合成、ATP结合盒（ATP-binding cassette, ABC）转运体通路、初级胆汁酸生物合成、嘌呤代谢、三羧酸（tricarboxylic acid, TCA）循环以及胰高血糖素信号通路。另有8种属水平差异肠道菌群与23种关键差异代谢物显著相关，且二者共同参与动脉粥样硬化损伤过程。综上，本研究结果表明，GRc可改善高脂饮食诱导的ApoE−/−小鼠的动脉粥样硬化损伤，调节肠道菌群与代谢组的异常变化，并提示在GRc抗动脉粥样硬化的作用机制中，肠道菌群、代谢物与动脉粥样硬化损伤三者间存在潜在关联。