基于肠道菌群-宿主代谢交互作用的兔颈动脉粥样硬化模型的“痰、瘀、毒”病理机制
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浙江中医药大学动物实验研究中心/比较医学研究所,杭州 310053,浙江,中国

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Pathological Mechanism of “Phlegm, Blood stasis, Toxin” in a Rabbit Model of Carotid Atherosclerosis Based on Gut Microbiota-host Metabolism Interactions
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Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China

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    摘要:

    背景: 在中医理论中,“痰、瘀、毒”是颈动脉粥样硬化(Carotid Atherosclerosis,CAS)的病机。高胆固醇饮食联合颈动脉球囊损伤诱导的兔颈动脉粥样硬化模型是研究CAS的经典模型。多项研究表明,肠道微生物群和宿主代谢紊乱参与了兔CAS的发病机制。然而,这种经典的兔CAS模型的中医病理特征和证候尚未见报道。目的: 从“痰、瘀、毒”的角度探讨兔CAS模型的发病机制及其中医证候类型。方法: 将12只雄性新西兰大白兔按体重随机分为NC组和CAS组,分别饲喂基础饲料和1%高胆固醇饲料。两周后,CAS组兔行颈总动脉(CCA)球囊损伤,而NC组家兔仅行CCA分离,未行球囊损伤。 两组在术后接受了六周不同饲料的喂养,分析其脂质、血液流变学、炎症、氧化应激和 CAS 表型的变化。 同时,收集了结肠内容物和血清进行16S rRNA测序和1H-NMR代谢组学分析。结果: 观察到CAS兔脂质代谢和血液流变学明显异常,氧化应激水平剧增,炎症因子过度释放,CAS斑块明显形成。此外,在兔CAS中鉴定出10种特定的肠道菌群(如Akkermansia_muciniphila、Barnesiellaceae和Faecalibacterium)和 14 种特征性代谢物(如trimethylamine oxide, acetic acid和L-carnitine),它们与 CAS 表征显著相关。通路功能分析表明,肠道菌群及其代谢产物主要影响胆固醇代谢、能量代谢、炎症和氧化应激。结论: 兔CAS模型符合“痰瘀毒损”理论。CAS兔肠道菌群与宿主代谢紊乱相互作用,促发内外毒邪,加重CAS的进展。该研究为CAS模型在中医药研究中的应用提供了实验依据。

    Abstract:

    Background: In Traditional Chinese Medicine (TCM) theory, "phlegm, blood stasis and toxin" are the pathogenesis of carotid atherosclerosis (CAS). The rabbit carotid atherosclerosis (CAS), which is induced by high-cholesterol diet combined with carotid artery balloon injury, is a classic model for studying CAS. Many studies indicate that gut microbiota and host metabolic disorders are involved in the pathogenesis of rabbit CAS. However, the TCM pathological features and syndromes of this classic rabbit CAS model have not been reported. Objective: To explore the pathogenesis of the rabbit CAS model and its TCM syndrome types from the perspective of "phlegm, blood stasis, and toxin". Methods: Twelve male New Zealand white rabbits were randomly divided into NC group and CAS group according to their body weight, followed by feeding of basic feed and a 1% high cholesterol diet, respectively. After two weeks, the rabbits in the CAS group underwent common carotid artery (CCA) balloon injury, while the rabbits in the NC group underwent only CCA separation without balloon injury. The two groups received differential feed postoperatively for six more weeks, after which, changes in lipids, hemorheology, inflammation, oxidative stress, and CAS phenotypes were analyzed. In addition, the colon contents and serum were collected for 16S rRNA sequencing and 1H-NMR metabonomic analysis. Results: The CAS rabbits were observed to form noticeable abnormalities in lipid metabolism and blood rheology, a sharp increase in oxidative stress levels, excessive release of inflammatory factors and apparent CAS plaque formation. Furthermore, 10 specific gut microbiota (such as Akkermansia muciniphila, Barnesiellaceae and Faecalibacterium) and 14 characteristic metabolites (such as trimethylamine oxide, acetic acid and L-carnitine) were identified in the CAS rabbits, which were significantly related to the CAS phenotypes. The pathway function analysis showed that the gut microbiota and its metabolites mainly affected cholesterol metabolism, energy metabolism, inflammation and oxidative stress. Conclusion: The rabbit CAS model conforms to the “phlegm, blood stasis and toxin damage” theory. The gut microbiota and host metabolic disorders of the CAS rabbits interact and promote internal and external toxins, aggravating the progression of CAS. Our study provided experimental evidence for the application of this model in the TCM-based research of CAS.

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  • 在线发布日期: 2023-11-24
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