Research Article: Hoxb4 upregulation by Xuan Bi Tong Yu Fang confers cardioprotection via repression of the Wnt/?-catenin pathway in myocardial ischemia-reperfusion injury
Abstract:
Myocardial ischemia-reperfusion injury (MIRI) remains a significant challenge in treating cardiovascular diseases, contributing to substantial myocardial damage and a poor prognosis. Traditional Chinese medicine (TCM) offers promising alternatives, with Xuan Bi Tong Yu Fang (XBTYF) being a classical formula known for its protective effects. However, the therapeutic mechanisms of XBTYF in MIRI have not been fully explored.
MIRI was induced in 36 Wistar rats, which were randomly assigned to sham, model, XBTYF (high, medium, low dose), and positive control (Qishen Yiqi Droplet) groups (n = 6). Serum levels of CK, AST, and cTnI were measured, and myocardial injury was examined by TTC, HE, and Masson staining. Bulk RNA-seq was performed to analyze gene expression profiles. The chemical composition of XBTYF was characterized by UPLC-MS/MS, followed by molecular docking to assess interactions between key compounds and Hoxb4. In vitro , H9C2 cardiomyocytes were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) and treated with XBTYF. The role of Hoxb4 in regulating the Wnt/?-catenin pathway was examined through gain- and loss-of-function approaches, together with the Wnt inhibitor IWR-1. Cell viability, proliferation, apoptosis, and the expression of apoptosis-related proteins and Wnt/?-catenin pathway components were assessed.
XBTYF treatment significantly reduced myocardial enzyme levels and myocardial infarction area in the MIRI rats. Histological analysis showed improved myocardial tissue structure and reduced fibrosis. Bulk RNA-seq analysis identified Hoxb4 as a key gene restored by XBTYF and revealed a significant suppressive effect of XBTYF on the Wnt/?-catenin signaling pathway. Chemical analysis identified multiple bioactive compounds in XBTYF, and molecular docking further revealed that ginsenoside Rg3, a primary component, exhibited a strong binding affinity with the Hoxb4 protein. In vitro studies confirmed that XBTYF upregulated Hoxb4, promoting cell proliferation and inhibiting apoptosis under OGD/R condition. Western blot analysis validated that XBTYF inhibited Wnt/?-catenin signaling, which was associated with reduced apoptosis and improved myocardial protection.
XBTYF alleviates MIRI by upregulating Hoxb4 and subsequent inhibition of the Wnt/?-catenin signaling pathway, leading to reduced cardiomyocyte apoptosis and providing cardio protection. Its effects may be linked to direct interactions between bioactive constituents, such as ginsenoside Rg3, and Hoxb4. Collectively, these findings provide mechanistic and experimental evidence supporting XBTYF as a potential adjunctive therapeutic strategy for ischemia-reperfusion-related myocardial injury and establish a molecular basis for its further translational investigation and clinical application.
Introduction:
Myocardial ischemia-reperfusion injury (MIRI) remains a significant challenge in treating cardiovascular diseases, contributing to substantial myocardial damage and a poor prognosis. Traditional Chinese medicine (TCM) offers promising alternatives, with Xuan Bi Tong Yu Fang (XBTYF) being a classical formula known for its protective effects. However, the therapeutic mechanisms of XBTYF in MIRI have not been fully explored.
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