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Journal of Traditional Chinese Medicine >

2025 , Vol. 45 >Issue 2: 291 - 302

DOI: https://doi.org/10.19852/j.cnki.jtcm.2025.02.016

Original articles

Mechanism of Huayu Qutan recipe (化瘀祛痰方) anti-atherosclerosis mediates lipophagy via mammalian target of rapamycin complex 1/ transcription factor EB signaling pathway in ApoE-/-mice

  • Yue LI ,
  • Jiaxiang PAN ,
  • Guanlin YANG ,
  • Jiajia YU ,
  • Xize WU ,
  • Dongyu MIN ,
  • Meijia CHENG ,
  • Dongdong YU ,
  • Minghua NAN ,
  • Xiaoyu GAO ,
  • Linlin PANG ,
  • Lihong GONG ,
  • Lianqun JIA
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  • 1 Department of Cardiology, the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China
    2 Liaoning Provincial Key Laboratory of TCM Geriatric Cardio-Cerebrovascular Diseases, Shenyang 110032, China
    3 Innovation Engineering Technology Center of Traditional Chinese Medicine, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    4 Postdoctoral Program of Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    5 Graduate School of Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    6 Experimental Center of Traditional Chinese Medicine, the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China
    7 Department of Osteology, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China
    8 Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications of Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China
    9 Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110000, China
    10 Department of Oncology department, Shengjing Hospital affiliated to China Medical University, Shenyang 110000, China

Received date: 2023-12-12

  Accepted date: 2024-05-15

  Online published: 2025-03-10

Supported by

Science and Technology Research Project of Education Department of Liaoning Province: the Mechanism of Regulating Lipophagy and Affecting Cholesterol Efflux in Human Promyelocytic Leukemia Cell Line Macrophages by Huayu Qutan Recipe based on the Theory of "Laoxia Shengtan"(L202048);Youth Project of Basic Scientific Research Project of Education Department of Liaoning Province: Mechanism of Mitochondrial Autophagy and Apoptosis in L02 Hepatocytes Regulated by miR-7043-3p based on the Theory of "Spleen Qi Dispersing Essence" of Huayu Qutan Recipe(LJKQZ2021064)

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Abstract

OBJECTIVE: To investigate the effects of Huayu Qutan recipe (化瘀祛痰方, HYQT) on the atherosclerosis (AS) model of ApoE-/- mice with a high-fat diet and to illustrate the underlying mechanisms from modern patho-physiological conceptualizations.

METHODS: High performance liquid chromatography of quadrupole time of flight-tandem mass spectrometry (HPLC-Q-TOF-MS/MS) analysis was used to identify the active compounds in the recipe. The mice were randomly allocated into 7 groups: control (CTRL) group, normal diet (ND) group, high-fat diet (HFD) group, HYQT groups (low dose, medium dose, and high dose), and simvastatin (SIM) group. Deferent doses of HYQT were gavaged twice a day, and then the protective effect of HYQT on plaque formation in ApoE-/- mice with a high-fat diet was verified viahematoxylin-eosin (HE) staining and oil red o (ORO) staining. We observed the co-localization in aortic macrophages and lipid droplets (LDs) by CD68 and the Bodipy fluorescence probe. Light chain 3 phosphoprotein class Ⅱ/light chain 3 phosphoprotein class Ⅰ (LC3Ⅱ/LC3Ⅰ) was examined by western blotting, and sequestosome 1 (SQSTM1/p62), Beclin1, Lamp1, mammalian target of rapamycin (mTOR), phosphorylated mammalian target of rapamycin (p-mTOR), and ATP-binding cassette transporter A1 (ABCA1) were examined by real-time polymerase chain reaction (RT-PCR) and Western blotting. Transcription factor EB (TFEB) nuclear translocation was determined by immunofluorescence analysis.

RESULTS: Five active compounds were identified using HPLC-Q-TOF-MS/MS analysis: ferulic acid, chlorogenic acid, calycosin, formononetin, and 8,2'-dihydroxy-7,4'-dimethoxy-isoflavane. The effect of HYQT on atherosclerotic plaque formation in ApoE-/- mice was investigated. These findings showed that HYQT decreased the co-localization of CD68 and Bodipy and increased the co-localization of CD68 and LC3B. Medium and high doses of HYQT increased autophagosome formation and promoted the maturation of LC3Ⅱ/LC3Ⅰ. Additionally, HYQT decreased the expression of SQSTM1/p62. Medium and high doses of HYQT also increased the expression of Beclin1 and Lamp1. RT-PCR and Western blot results suggested that HYQT enhanced the expression of ABCA1 mRNA and protein and regulated the mTORC1/TFEB signaling pathway.

CONCLUSION: The results indicate that HYQT is an effective traditional Chinese herbal remedy for the treatment of AS. HYQT mitigates macrophage-derived foam cell formation by activating autophagy in atherosclerosis. The mTOR/TFEB signaling pathway and ABCA1 are therapeutic targets of HYQT for the treatment of AS.

Key words： atherosclerosis; foam cells; lipid metabolism; diet, high-fat; Huayu Qutan recipeSupporting information

Cite this article

Yue LI , Jiaxiang PAN , Guanlin YANG , Jiajia YU , Xize WU , Dongyu MIN , Meijia CHENG , Dongdong YU , Minghua NAN , Xiaoyu GAO , Linlin PANG , Lihong GONG , Lianqun JIA . Mechanism of Huayu Qutan recipe (化瘀祛痰方) anti-atherosclerosis mediates lipophagy via mammalian target of rapamycin complex 1/ transcription factor EB signaling pathway in ApoE-/-mice[J]. Journal of Traditional Chinese Medicine, 2025 , 45(2) : 291 -302 . DOI: 10.19852/j.cnki.jtcm.2025.02.016

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