基于肠道菌群驱动TMA/FMO3/TMAO通路研究针刺抗“痰瘀毒”致动脉粥样硬化的机制

The pathogenesis of AS is not clearly identified yet.Previous studies suggest that intestinal microbiome and its metabolites TMAO are involved in the occurrence of AS, which may be a new breakthrough for the further research. Chinese medicine believes that phlegm,blood stasis and toxin is the main pathogenesis of AS,which has great similarity pathogenetic character with TMA / FMO3 / TMAO pathway driven by intestinal microflora induced AS. Therefore, we will do the research of acupuncture and moxibustion treatment of AS and hope to provide new ideas for its prevention and treatment by combining the theory of phlegm ,blood stasis and toxin with intestinal flora .The previous findings in the study conducted by this research group and suggest that acupuncture and moxibustion can play an effective role in relieving ischemic heart disease and can adjust the composition of intestinal flora. This study proposes the hypothesis that acupuncture can relieve AS by adjusting the intestinal flora and the metabolic product TMAO. Observing the changes of intestinal flora via high-throughput sequencing technology, and the levels of TMA,FMO3 and TMAO in TMA / FMO3 / TMAO pathway by RT-PCR and HPLC-MS/MS methods with intestinal flora and TMAO as targets. The objectives of the study are to investigate the relationship between the onset and progression of AS and intestinal microbiome and its metabolites, and to explore the mechanism of acupuncture in the treatment of AS by removing phlegm ,blood stasis and toxin ,which will provide a new theoretical and experimental evidence for applying acupuncture and moxibustion to the prevention and treatment of AS.

动脉粥样硬化(AS)发病机制尚不明确,以往研究提示,肠道微生物及代谢物TMAO可能参与AS发生,对其进一步研究有可能产生新的突破.中医学认为痰瘀毒是致AS的主要病机.其发病特点与肠道菌群驱动TMA/FMO3/TMAO通路致AS具有极大相似性.故本课题将"痰瘀毒"理论与肠道菌群及代谢物相结合研究AS.并介入针灸治疗.希望为其防治提供新的思路.课题组前期研究提示,针灸能够有效干预缺血性心脏疾患及调整肠道菌群.因此.本课题在以往研究基础上.提出"针刺能够通过调整肠道菌群及代谢物TMAO抗AS”假说.以肠道菌群及TMAO为靶点,通过高通量测序等技术观察肠道菌群的变化,用RT-PCR和HPLC-MS/MS等方法观察TMA/FMO3/TMAO通路中TMA、FMO3、TMAO水平变化.探讨AS发生发展与肠道菌群及代谢物之间的关系,研究化痰祛瘀排毒针刺治法抗AS机制,为针灸防治AS提供新的理论及实验依据.

本项目原计划从针刺可能调节肠道菌群驱动TMA/FMO3/TMAO通路抑制“痰瘀毒”致AS的角度，深入研究针刺防治AS的机制。本项目研究过程较为顺利，如期按时完成全部研究内容。.最终，本研究结果显示，AS病变可以通过改变肠道菌群以驱动TMA/FMO3/TMAO通路，导致宿主血浆中TMAO含量升高；而TMAO能够引起血管壁内巨噬细胞表面CD36和SR-A1活性升高，促使巨噬细胞吞噬大量脂质形成泡沫细胞，加剧AS病变。针刺能够有效调节肠道菌群，减少血浆中TMAO的浓度，降低巨噬细胞表面CD36和SR-A1活性，减少巨噬细胞中的胆固醇含量，抑制巨噬细胞泡沫化；同时在针刺干预下，肠道菌群代谢产物TMAO的合成受到抑制，导致Cyp7a1表达的增加、胆固醇的转运增多、胆汁酸生成与代谢活跃，最终使得巨噬细胞中的胆固醇含量减少，AS病变得到控制。本研究结果提示，针刺防治AS可能是通过调节肠道菌群驱动TMA/FMO3/TMAO通路影响机体胆汁酸代谢实现的，为针刺抗AS机理提供了重要研究线索。本项目执行期间，共获科技进步奖（省部级一等奖）1项，学术成果奖（省部级三等奖）2项；项目总体公开发表了13篇期刊论文，其中1篇SCI，6篇中文核心，2篇科技核心。培养了13名研究生，其中10名硕士（含1名升学为博士仍在本课题组中），3名博士（包括1名已毕业，2名预计2021年毕业答辩）。项目投入经费52.0000万元，支出28.4807万元，各项支出基本与预算相符，剩余经费23.5193万元，剩余经费计划用于本项目后续研究。