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November 3, 2025PLoS ONE3 citationsOpen Access

Heart failure induced by isoproterenol: A comparison of two doses and two delivery methods in C57BL/6J mice

RYRong YangAYAidan YuPYPeng Yang

Key Result

Subcutaneous administration of 5 mg/kg/day isoproterenol for 14 days optimally induced stable heart failure phenotypes with 100% survival in mice, whereas 60 mg/kg/day resulted in 25% mortality.

Structured PICO

P
Population
72 male C57BL/6J mice aged 6-8 weeks were evaluated to determine the optimal dose and administration route of isoproterenol for inducing heart failure over 14 days.
I
Intervention
Isoproterenol (ISO) administered subcutaneously (SC) or intraperitoneally (IP) at 5 mg/kg/day or 60 mg/kg/day for 14 days
C
Comparator
Subcutaneous (SC) saline control and intraperitoneal (IP) saline control
O
Outcome
Cardiac remodeling, systolic dysfunction, left ventricular dilation, mortality, hypertrophy indices, fibrosis, and serum NT-proBNP levelssurrogate

Subcutaneous administration of isoproterenol at 5 mg/kg/day for 14 days is the optimal protocol for inducing a reliable heart failure phenotype in mice with minimal mortality.

Limitations

  • Statistical power was constrained by cohort size and data variability.
  • Observed divergences in select echocardiographic parameters and molecular markers lacked sufficient evidence to conclusively establish route-specific causality.
  • Requires future validation in rat models to enhance translational relevance.

Abstract

Heart failure (HF) modeling requires standardized protocols to ensure translational relevance. Despite the widespread use of isoproterenol (ISO)-a β-adrenergic agonist-in HF modeling, methodological inconsistencies in dosing and administration routes limit reproducibility. This study evaluated the effects of subcutaneous (SC) and intraperitoneal (IP) administration of ISO at two literature-established doses (5 and 60 mg/kg/day for 14 days) on cardiac remodeling in C57BL/6J mice, aiming to identify the optimal protocol for HF modeling. Using a factorial design, male C57BL/6J mice aged 6-8 weeks were divided into six cohorts: (1) SC saline control, (2) IP saline control, (3) SC 5 mg/kg ISO, (4) IP 5 mg/kg ISO, (5) SC 60 mg/kg ISO, and (6) IP 60 mg/kg ISO, with daily administration for 14 days. High-dose ISO (60 mg/kg/day) induced a 25% mortality rate in both SC and IP cohorts, yet IP administration exhibited marked inter-individual variability, undermining model reliability. Echocardiography revealed SC 5 mg/kg group induced stable systolic dysfunction accompanied by left ventricular dilation, while maintaining 100% survival. This cohort also displayed significantly elevated hypertrophy indices. Histopathological quantification suggested that SC 60 mg/kg induced extensive fibrosis. All ISO-treated groups showed upregulated myocardial hypertrophy markers and approximately 2-fold elevation in serum NT-proBNP levels. In summary, SC 5 mg/kg/day regimen not only ensures reliable phenotype induction but also reduces animal attrition, offering a robust platform for investigating CHF mechanisms and accelerating therapeutic development.

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Cite This Study

Yang et al. (2025) studied Heart failure (model induction) (n=72). Isoproterenol vs. Saline was evaluated on Survival and cardiac remodeling (optimal HF model). Subcutaneous administration of 5 mg/kg/day isoproterenol for 14 days optimally induced stable heart failure phenotypes with 100% survival in mice, whereas 60 mg/kg/day resulted in 25% mortality.

synapsesocial.com/papers/6aa24a578c48a028767a34cdhttps://doi.org/10.1371/journal.pone.0334880
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