Skip to main content
SpringerLink
Log in

Effects of herbal cake-partitioned moxibustion on plasma trimethylamine, trimethylamine-N-oxide, and flavin-containing monooxygenase 3 in atherosclerotic rabbits

隔药饼灸对动脉粥样硬化兔血浆三甲胺、氧化三甲胺及肝黄素单加氧酶3的影响

  • Basic Study
  • Published:
Journal of Acupuncture and Tuina Science Aims and scope Submit manuscript

Abstract

Objective

To investigate the effects of herbal cake-partitioned moxibustion on the plasma levels of trimethylamine (TMA), trimethylamine-N-oxide (TMAO), and flavin-containing monooxygenase 3 (FMO3) in rabbits with atherosclerosis (AS), as well as to explore the possible mechanism of herbal cake-partitioned moxibustion in treating AS.

Methods

After 1-week adaptive feeding, 28 male New Zealand rabbits were divided into a blank group, a model group, an antibiotic group, and a herbal cake-partitioned moxibustion group according to the random number table method, with 7 rabbits in each group. Rabbits were fed with a basic diet in the blank group, while with a basic diet plus 1% choline in the remaining groups to prepare the AS model. Rabbits were given drinking water with broad-spectrum antibiotics in the antibiotic group, and herbal cake-partitioned moxibustion in the herbal cake-partitioned moxibustion group for 12 weeks. The atherosclerotic plaques by hematoxylin-eosin (HE) staining, the blood lipid levels, the plasma TMA and TMAO levels by liquid chromatography-mass spectrometry were detected for rabbits in each group at the end of interventions. Liver FMO3 protein expression was detected by Western blotting. Liver FMO3 mRNA expression was detected by real-time fluorescence quantitative polymerase chain reaction.

Results

HE staining showed that the arterial wall was rough, the intima was significantly thickened, and more foam cells and lipid deposits were seen in rabbits of the model group. Arterial wall thickening was not obvious with a few foam cells and lipid deposits in the herbal cake-partitioned moxibustion group. Compared with the blank group, the serum levels of low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) were increased (P<0.01), the plasma levels of TMA and TMAO were increased (P<0.05, P<0.01), and the expression levels of liver FMO3 protein and mRNA were all increased (P<0.05, P<0.01); while the serum high-density lipoprotein cholesterol (HDL-C) level was decreased in the model group (P<0.05). Compared with the model group, the LDL-C and TC levels were decreased (P<0.01 or P<0.05), the HDL-C levels were increased (P<0.01), the TMA and TMAO levels were decreased (P<0.05), while the protein and mRNA expression levels of FMO3 were decreased without statistical significance in the herbal cake-partitioned moxibustion group and the antibiotic group.

Conclusion

Herbal cake-partitioned moxibustion can slow atherosclerotic plaque formation and regulate lipid levels in AS rabbits, and the mechanism may be related to the down-regulation of TMA and TMAO expression in the plasma.

摘要

目的

观察隔药饼灸对动脉粥样硬化(AS)兔血浆三甲胺(TMA)、氧化三甲胺(TMAO)水平以及肝黄素单氧化酶 3(FMO3)含量的影响, 探讨隔药饼灸治疗AS的可能机制。

方法

将28只雄性新西兰兔适应性喂养1周后根据随机数字 表法分为空白组、模型组、抗生素组和隔药饼灸组, 每组7只。空白组予基础饲料喂养, 其余各组予基础饲料加1% 胆碱喂养, 制备AS模型。抗生素组予添加广谱抗生素的饮水, 隔药饼灸组予隔药饼灸, 持续12周。干预结束后, 采用 苏木素-伊红(HE)染色观察动脉粥样硬化斑块, 检测各组兔血脂水平, 液相色谱-质谱联用检测血浆TMA及TMAO含量。 免疫印迹法检测肝脏FMO3蛋白表达。实时荧光定量聚合酶链反应法检测肝脏FMO3 mRNA表达。

结果

HE染色结果 显示模型组动脉管壁粗糙, 内膜明显增厚, 可见较多泡沫细胞、脂质沉积; 隔药饼灸组动脉壁增厚不明显, 有少量泡 沫细胞和脂质沉积。与空白组比较, 模型组血清中低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)水平升高(P<0.01), 血浆 中 TMA及TMAO 水平升高(P<0.05, P<0.01), 肝脏FMO3蛋白及mRNA表达升高(P<0.05, P<0.01); 血清高密度脂蛋白胆固 醇(HDL-C)水平下降(P<0.05)。与模型组比较, 隔药饼灸组及抗生素组LDL-C及TC水平下降(P<0.01或P<0.05), HDL-C水平 上升(P<0.01), TMA及TMAO水平下降(P<0.05), FMO3蛋白及mRNA表达下降, 但差异无统计学意义。

结论

隔药饼灸能 减缓动脉粥样硬化斑块形成, 调节AS兔的血脂水平, 其机制可能与下调血浆中TMA及TMAO表达相关。

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. XU H L, JIANG J X, CHEN W Z, LI W L, CHEN Z G. Vascular macrophages in atherosclerosis. J Immunol Res, 2019, 2019: 4354786.

    Article  PubMed  PubMed Central  Google Scholar 

  2. WANG Z, TANG W H, BUFFA J A, FU X, BRITT E B, KOETH R A, LEVISON B S, FAN Y, WU Y, HAZEN S L. Prognostic value of choline and betaine depends on intestinal microbiota-generated metabolite trimethylamine-N-oxide. Eur Heart J, 2014, 35(14): 904–910.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. WU P, CHEN J N, CHEN J J, TAO J, WU S Y, XU G S, WANG Z, WEI D H, YIN W D. Trimethylamine N-oxide promotes apoE−/− mice atherosclerosis by inducing vascular endothelial cell pyroptosis via the SDHB/ROS pathway. J Cell Physiol, 2020, 235(10): 6582–6591.

    Article  CAS  PubMed  Google Scholar 

  4. WANG Z N, KLIPFELL E, BENNETT B J, KOETH R, LEVISON B S, DUGAR B, FELDSTEIN A E, BRITT E B, FU X M, CHUNG Y M, WU Y P, SCHAUER P, SMITH J D, ALLAYEE H, TANG W H W, DIDONATO J A, LUSIS A J, HAZEN S L. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature, 2011, 472(7341): 57–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. RATH S, HEIDRICH B, PIEPER D H, VITAL M. Uncovering the trimethylamine-producing bacteria of the human gut microbiota. Microbiome, 2017, 5(1): 54.

    Article  PubMed  PubMed Central  Google Scholar 

  6. TANG W H W, BÄCKHED F, LANDMESSER U, HAZEN S L. Intestinal microbiota in cardiovascular health and disease: JACC state-of-the-art review. Am Coll Cardiol, 2019, 73(16): 2089–2105.

    Article  Google Scholar 

  7. HUANG Y P, ZHAO B X. Research progress of different moxibustion methods in treatment of hyperlipidemia. Yixue Zongshu, 2019, 25(23): 4720–4725.

    Google Scholar 

  8. YANG J J, ZHAO Z, SHI J, GE J Y, OUYANG L Z, CHANG X R, LIU M L, HUANG J. Effect of herbal cake-separated moxibustion on the expression of serum TNF-α and LNF-γ in rabbits with hyperlipidemia and atherosclerosis. Hunan Zhongyiyao Daxue Xuebao, 2018, 38(11): 1258–1261.

    Google Scholar 

  9. YANG J. Effect of Drug-separated Cake Moxibustion on TLR2/4-NF-κB Signaling Pathway in Vulnerable Plaques of AS Rabbits. Changsha: Master Thesis of Hunan University of Chinese Medicine, 2015.

    Google Scholar 

  10. ZOU Y F, MA M Z, ZHAO Z, TAN J, YANG J J, SHI J, LIU M, LIU M L, CHANG X R. Effect of herbal-cake-separated moxibustion on blood lipid levels and expression of hepatic PPARγ and SR-B 1 proteins and genes in hyperlipidemia atherosclerosis rabbits. Zhen Ci Yan Jiu, 2018, 43(2): 86–91.

    PubMed  Google Scholar 

  11. CHEN M L. Resveratrol Attenuates Atherosclerosis via Gut Microbiota Remodeling. Chongqing: Doctor Thesis of the Third Military Medical University, 2016.

    Google Scholar 

  12. SHEN J, LIU T, LIU X, GE J Y, WANG H L, GUO A L, LIU M L, CHANG X R. Effects of herbal-cake-separated moxibustion on the repair of vascular endothelial and SDF-1 in rabbits with atherosclerosis. Zhongguo Zhen Jiu, 2019, 39(2): 173–179, 184.

    PubMed  Google Scholar 

  13. YU S G, XU B. Experimental Acupuncture Science. Beijing: People’s Medical Publishing House, 2016.

    Google Scholar 

  14. ZHENG Z H, ZHOU S P, PENG S Q. Medical Laboratory Animal Sciences. Beijing: Peking University Medical Press, 2008: 217.

    Google Scholar 

  15. BI D Y, NING J J, XU Y, LUO Y, LI M Q, WANG Y H, YI Z. Effects of different doses of ginger-partitioned moxibustion on trefoil factor 1, mucin 5AC and epidermal growth factor receptor in rats with spleen deficiency syndrome. J Acupunct Tuina Sci, 2018, 16(1): 1–7.

    Article  CAS  Google Scholar 

  16. WANG Z, ROBERTS A B, BUFFA J A, LEVISON B S, ZHU W, ORG E, GU X, HUANG Y, ZAMANIAN-DARYOUSH M, CULLEY M K, DIDONATO A J, FU X, HAZEN J E, KRAJCIK D, DIDONATO J A, LUSIS A J, HAZEN S L. Non-lethal inhibition of gut microbial trimethylamine production for the treatment of atherosclerosis. Cell, 2015, 163(7): 1585–1595.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. SELDIN M M, MENG Y, QI H, ZHU W, WANG Z, HAZEN S L, LUSIS A J, SHIH D M. Trimethylamine N-oxide promotes vascular inflammation through signaling of mitogen-activated protein kinase and nuclear factor-KB. J Am Heart Assoc, 2016, 5(2): e002767.

    Article  PubMed  PubMed Central  Google Scholar 

  18. ZHU W W, GREGORY J C, ORG E, BUFFA J A, GUPTA N, WANG Z N, LI L, FU X M, WU Y P, MEHRABIAN M, SARTOR R B, Mcintyre T M, SILVERSTEIN R L, TANG W H W, DIDONATO J A, BROWN J M, LUSIS A J, HAZEN S L. Gut microbial metabolite TMAO enhances platelet hyperreactivity and thrombosis risk. Cell, 2016, 165(1): 111–124.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. KOETH R A, WANG Z, LEVISON B S, BUFFA J A, ORG E, SHEEHY B T, BRITT E B, FU X, WU Y, LI L, SMITH J D, DIDONATO J A, CHEN J, LI H, WU G D, LEWIS J D, WARRIER M, BROWN J M, KRAUSS R M, TANG W H, BUSHMAN F D, LUSIS A J, HAZEN S L. Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis. Nat Med, 2013, 19(5): 576–585.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. SHIH D M, WANG Z N, LEE R, MENG Y H, CHE N, CHARUGUNDLA S, QI H, WU J, PAN C, BROWN J M, VALLIM T, BENNETT B J, GRAHAM M, HAZEN S L, LUSIS A J. Flavin containing monooxygenase 3 exerts broad effects on glucose and lipid metabolism and atherosclerosis. Lipid Res, 2015, 56(1): 22–37.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Project of National Natural Science Foundation of China (国家自然科学基金 项目, No. 81874509); Opening Fund Project of Domestic First-class Discipline Construction of Traditional Chinese Medicine of Hunan University of Chinese Medicine (湖南 中医药大学中医学一流学科开放基金项目, No. 2018ZYX31); Scientific and Research Innovative Projects for Postgraduate Students of Hunan Province (湖南省研究 生科研创新项目, No. CX20190556, No. CX20190557).

Author information

Authors and Affiliations

  1. Hunan University of Chinese Medicine, Changsha, 410208, China

    Lizhi Ouyang  (欧阳里知), Binbin Zhang  (张彬彬), He Huang  (黄河), Jing Wang  (王晶), Jian Luo  (罗坚), Huijuan Liu  (刘惠娟) & Xiaorong Chang  (常小荣)

Authors
  1. Lizhi Ouyang  (欧阳里知)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Binbin Zhang  (张彬彬)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. He Huang  (黄河)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing Wang  (王晶)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jian Luo  (罗坚)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Huijuan Liu  (刘惠娟)
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiaorong Chang  (常小荣)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaorong Chang  (常小荣).

Additional information

Conflict of Interest

The authors declare that there is no potential conflict of interest in this article.

Statement of Human and Animal Rights

The treatment of animals conformed to the ethical criteria in this experiment.

]Co-first Authors: OUYANG Lizhi, M.D.; ZHANG Binbin, M.M.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ouyang, L., Zhang, B., Huang, H. et al. Effects of herbal cake-partitioned moxibustion on plasma trimethylamine, trimethylamine-N-oxide, and flavin-containing monooxygenase 3 in atherosclerotic rabbits. J. Acupunct. Tuina. Sci. 21, 10–17 (2023). https://doi.org/10.1007/s11726-023-1353-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11726-023-1353-9

Keywords

关键词

中图分类号

文献标志码

Search

Navigation