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Journal of Traditional Chinese Medicine >

2026 , Vol. 46 >Issue 3: 530 - 543

DOI: https://doi.org/10.19852/j.cnki.jtcm.20260214.001

Original Articles

Yiqi Huoxue formula (益气活血方) relieves heart failure with reduced ejection fraction in calcium/calmodulin-dependent protein kinase Ⅱ delta B isoform and calcium/calmodulin-dependent protein kinase Ⅱ delta C isoform transgenic mice by inhibiting myocardial remodeling and enhancing calcium homeostasis

  • XU Jianglin ,
  • SHI Xiaolu ,
  • LI Yan ,
  • ZHOU Kun ,
  • LI Peng ,
  • LI Sinai ,
  • LU Jinjin ,
  • CHEN Yajun ,
  • ZHAN Nannan ,
  • ZHANG Ruotong ,
  • CHENG Bairu ,
  • LIN Qian ,
  • WAN Jie ,
  • LIU Jing
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  • 1 Department of Cardiology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China
    2 Beijing Key Laboratory of TCM Basic Research on Prevention and Treatment of Major Disease, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China
    3 Department of Cardiology, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
    4 Department of Cardiology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100010, China
    5 Beijing Institute of Traditional Chinese Medicine, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, China
First author contact:

XU Jianglin and SHI Xiaolu are co-first authors and contributed equally to this work

LIU Jing, Department of Cardiology, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China. liujingth@sina.com, Telephone: +86-13161161811;
WAN Jie, Department of Cardiology, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China. yuezhijie1226@sina.com, Telephone: +86-19047159498

Received date: 2025-01-11

  Accepted date: 2025-09-29

  Online published: 2026-02-14

Supported by

Natural Science Foundation-funded Project: Underlined Mechanism of Calcium/calmodulin-dependent Protein Kinase II Regulating Heart Failure With Preserved Ejection Fraction and the Effect of Yiqi-Huoxue Formula(81903993);Investigation on the Mechanism of Yiqi-Huoxue Method on Heart Failurebased on Mitochondrial Calcium/calmodulin-dependent Protein Kinase II Overexpression Mediated Energy Shuttle Disturbance-calcium Homeostasis Disorder(82305170);Fundamental Research Funds for the Central Public Welfare Research Institutes National Traditional Chinese: High-throughput Detection Technology for the Study of Excitation-contraction-coupled Calcium Transport(JBGS2021006);National High Level Chinese Medicine Hospital Clinical Research Funding: Mechanism Study on the Reduction of Post-Percutaneous Coronary Intervention Microvascular Obstruction by Yiqi Liangxue Shengji Formula: Exploring the Role of Nicotinamide Adenine Dinucleotide Phosphate Oxidase 2/Peptidylarginine Deiminase 4 Pathway-mediated Neutrophil Extracellular Trap Formation(DFG2RB2024GJR013)

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Abstract

OBJECTIVE: To confirm the efficacy of Yiqi Huoxue formula (益气活血方, YQHX) on heart failure with reduced ejection fraction (HFrEF), and elucidate its potential effect on calcium homeostasis.

METHODS: Cardiac specific calcium/calmodulin-dependent protein kinase II delta C isoform (CaMKⅡδC)-overexpression transgenic mice (C-CaMKⅡδC+/- Tg mice) and calcium/calmodulin-dependent protein kinase II delta B isoform (CaMKⅡδB)-overexpression transgenic mice (C-CaMKⅡδB+/- Tg mice) were established by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 technique, and their genotypes and phenotypes were identified thereafter. C57BL/6 mice and transgenic mice were divided into seven groups in random, each containing eight mice. The groups were Control group, CaMKⅡδB model group, CaMKⅡδB treatment group (B Treated), CaMKⅡδB inhibitor group (B Inhibitor), CaMKⅡδC model group (C Model), CaMKⅡδC treatment group (C Treated), CaMKⅡδC inhibitor group (C Inhibitor). B and C Treated groups were given YQHX once a day for 4 weeks by gavage. The cardiac function and structure were evaluated using echocardiogram. The myocardial cellular status and fibrosis were detected by hematoxylin and eosin staining and Masson staining. Calcium transients and calcium leakage from cardiac ventricular myocytes were detected to reflect calcium homeostasis, and the expressions of CaMKⅡδB, CaMKⅡδC, hypertrophy factors, calcium pathway-related genes, and the level of calcium handling proteins were assessed by western blotting and reverse transcription-quantitative polymerase chain reaction.

RESULTS: We found CaMKⅡδB and CaMKⅡδC transgenic mice showed HFrEF phenotype. YQHX improved the systolic function of the CaMKⅡδB transgenic mice, reduced the cross-sectional area of cardiomyocytes, inhibited the overexpression of CaMKⅡδB and hypertrophy-related genes (B-type natriuretic peptide, myocyte enhancer factor 2, and atrial natriuretic factor), and increased calcium handling protein sarcoendoplasmic reticulum Ca2+ adenosine triphosphatase 2a (SERCA2a) and calcium capacity. YQHX improved the cardiac systolic function of CaMKⅡδC transgenic mice, reduced the ventricular diameter and cardiomyocyte cross-sectional area, alleviated myocardial fibrosis, increased SERCA2a expression and calcium capacity, and reduced calcium leakage.

CONCLUSION: Our results suggest that YQHX improved HFrEF by inhibiting CaMKIIδB overexpression, which may be related to reduced expression of hypertrophy-related factors, and increased SERCA2a expression to increase calcium reserve. In addition, YQHX improved HFrEF by inhibiting CaMKIIδC overexpression, which may be related to reduce calcium leakage, and increase SERCA2a expression to increase calcium reserve.

Key words： heart failure, systolic; calcium homeostasis; Yiqi Huoxue formula; calcium/calmodulin-dependent protein kinase II delta C isoform overexpression; calcium/calmodulin-dependent protein kinase II delta B isoform overexpression

Cite this article

XU Jianglin , SHI Xiaolu , LI Yan , ZHOU Kun , LI Peng , LI Sinai , LU Jinjin , CHEN Yajun , ZHAN Nannan , ZHANG Ruotong , CHENG Bairu , LIN Qian , WAN Jie , LIU Jing . Yiqi Huoxue formula (益气活血方) relieves heart failure with reduced ejection fraction in calcium/calmodulin-dependent protein kinase Ⅱ delta B isoform and calcium/calmodulin-dependent protein kinase Ⅱ delta C isoform transgenic mice by inhibiting myocardial remodeling and enhancing calcium homeostasis[J]. Journal of Traditional Chinese Medicine, 2026 , 46(3) : 530 -543 . DOI: 10.19852/j.cnki.jtcm.20260214.001

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