Xuefu Zhuyu Decoction Reduces Excessive Collagen Deposition and Crosslinking in Myocardial Fibrosis via CTR1-Copper-LOX Axis

WANG Ya-fen; YUE Shi-jun; LI Jia-yao; MENG Si-tong; ZHENG Min; ZHANG Yu-fang; GU Ying; HOU Bin; BIAO Ya-ning

doi:10.1007/s11655-026-4042-y

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Xuefu Zhuyu Decoction Reduces Excessive Collagen Deposition and Crosslinking in Myocardial Fibrosis via CTR1-Copper-LOX Axis

Original Article | Updated：2026-11-13

- Xuefu Zhuyu Decoction Reduces Excessive Collagen Deposition and Crosslinking in Myocardial Fibrosis via CTR1-Copper-LOX Axis
- Chinese Journal of Integrative Medicine Vol. 32, Issue 11, Pages: 1005-1015(2026)
- Affiliations：
  
  School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang (050091), China
  Resource Utilization and Quality Evaluation of Traditional Chinese Medicine of Hebei Province, Shijiazhuang (050091), China
  Shijiazhuang Yiling Pharmaceutical Co., Ltd., Shijiazhuang (050035), China
- Author bio：
  
  Correspondence to: Dr. BIAO Ya-ning, E-mail: biaoyaning@hebcm.edu.cn; Prof.YUEShi-jun, E-mail:leshijun@hebcm.edu.cn
- Funds：
- DOI：10.1007/s11655-026-4042-y
  CLC：
- Accepted：15 April 2026，
  
  Published：2026-11
- Accepted：
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WANG Ya-fen, YUE Shi-jun, LI Jia-yao, et al. Xuefu Zhuyu Decoction Reduces Excessive Collagen Deposition and Crosslinking in Myocardial Fibrosis via CTR1-Copper-LOX Axis[J]. Chinese Journal of Integrative Medicine, 2026, 32(11): 1005-1015.
DOI：

WANG Ya-fen, YUE Shi-jun, LI Jia-yao, et al. Xuefu Zhuyu Decoction Reduces Excessive Collagen Deposition and Crosslinking in Myocardial Fibrosis via CTR1-Copper-LOX Axis[J]. Chinese Journal of Integrative Medicine, 2026, 32(11): 1005-1015. DOI： 10.1007/s11655-026-4042-y.

摘要

Abstract

Objective: To explore the molecular mechanism of Xuefu Zhuyu Decoction (XFZYD) against myocardial fibrosis. Methods: The active components of XFZYD were identified by high performance liquid chromatography (HPLC). Network pharmacology

molecular docking

and affinity dot-mass spectrometry (AD-MS) were used to predict and validate the binding interaction between XFZYD components and copper transporter 1 (CTR1). Sixty male Kunming mice were randomly divided into 6 groups using a random number table. Control

isoproterenol hydrochloride (ISO)

captopril

low-

medium-

and high-dose XFZYD (5.69

11.38

and 22.76 g/kg) groups

with 10 mice in each group. Except the control group

mice received daily subcutaneous injection of ISO (10 mg/kg) for 28 d to induce myocardial fibrosis

concurrently treated with respective drugs by gavage. Cardiac function was assessed by echocardiography (ECG) and electrocardiography (UCG). Serum myocardial injury markers and transforming growth factor beta 1 (TGF-β1) were measured by a biochemical analyzer and enzyme-linked Immunosorbent assay (ELISA). Pathological changes and collagen deposition was observed using HE

Masson and sirius red staining methods. Copper content in cardiac tissue was determined by inductively coupled plasma mass spectrometry (ICP-MS)

and protein expressions of collagen Ⅰ

Ⅲ

alpha-smooth muscle actin (α-SMA)

lysyl oxidase (LOX)

CTR1

copper chaperone for superoxide dismutase (CCS)

and cytochrome c oxidase 17 (COX17) were detected by Western blot

immunohistochemistry

and immunofluorescence. In vitro

TGF-β1 (5 ng/mL)-stimulated NIH-3T3 fibroblasts were treated with XFZYD (60 mg/L)

with or without CuCl2 (20 μmol/L) or SLC31A1 overexpression for 24 h

and the same indicators were evaluated. Results: HPLC identified 7 major components in XFZYD. Network pharmacology revealed that XFZYD targets copper metabolism related genes. Molecular docking showed strong binding affinities between the 7 compounds and CTR1

and AD-MS confirmed direct binding of paeoniflorin and amygdalin to CTR1. In vivo

XFZYD could ameliorate the collagen deposition

cardiac dysfunction

and elevated levels of collagen Ⅰ/Ⅲ

α-SMA

LOX

and copper content exhibited in ISO-induced myocardial fibrosis mice (P＜0.05 or P＜0.01). In vitro

XFZYD had the effects of suppressing copper overload and the upregulation of LOX

collagen Ⅰ/Ⅲ

and α-SMA in TGF β1 stimulated NIH-3T3 fibroblasts

whereas co-treatment with CuCl2 exacerbates these fibrotic responses and attenuates the protective effect of XFZYD (P＜0.05 or P＜0.01). XFZYD had the effect of partially reversing the enhanced copper accumulation

LOX activation

and collagen deposition induced by SLC31A1 (CTR1) overexpression in TGF β1 stimulated fibroblasts

and SLC31A1 overexpression reduces its efficacy (P＜0.05 or P＜0.01). Conclusions: XFZYD exerted a protective effect against ISO-induced myocardial fibrosis. The mechanism may be related to regulating the CTR1- copper-LOX axis

thereby reducing excessive collagen deposition and cross-linking in cardiac fibroblasts.

关键词

Keywords

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