Related Experiment Video
Updated: Mar 13, 2026
Author Spotlight: Exploring the Relationship Between Lipotoxicity and HFpEF
Published on: March 29, 2024
Optimized murine HFpEF models for translational preclinical studies
Bailey McIntosh1,2,3,4, Ali Ali Mohamed Elbassioni5, Anmar Raheem5
1School of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Camperdown, New South Wales 2050, Australia.
Introduction:
The most clinically representative murine models of heart failure with preserved ejection fraction (HFpEF) include a '2-hit' model combining nitrosative stress with metabolic perturbation and a '3-hit' model that also includes ageing. Both models have important limitations with regard to substrain and sex.
Methods:
The 2-hit model protocol was modified to reproduce HFpEF in both C57BL/6N and 6J mice by increasing L-NAME doses (0.5 g/L to 1.75 g/L) and protocol lengths (7 weeks to 13 weeks). For the 3-hit model, in addition to deoxycorticosterone pivalate (DOCP), we added 1% NaCl drinking water to enhance and prolong the effect of DOCP ('4-hit'). To maintain the phenotype, a second bolus of DOCP was administered after 8 weeks.
Results:
HFpEF was successfully induced in C57BL/6J mice when exposed to a 13-week 2-hit L-NAME protocol with gradually increasing dosage from 1.0 to 1.75 g/L. For the 4-hit mice, a clear HFpEF phenotype was observed in C57BL/6N and 6J mice in both male and females, and maintained for up to 12 weeks.
Conclusion:
These modifications ensure the 2-hit model is induced in J substrain of C57BL/6 mice. The 4-hit model prevents aldosterone escape and enhances reproducibility across sexes and substrains.
Insights
Modified murine models now better represent heart failure with preserved ejection fraction (HFpEF) in both sexes and mouse strains. These enhanced models improve reproducibility for HFpEF research.
Area of Science:
- Cardiovascular Research
- Animal Models
- Translational Medicine
Background:
- Clinically relevant murine models of heart failure with preserved ejection fraction (HFpEF) are crucial for research.
- Existing "2-hit" (nitrosative stress + metabolic perturbation) and "3-hit" (ageing included) models have limitations regarding mouse substrain and sex.
- Improvements are needed to enhance the translatability of these HFpEF models.
Purpose of the Study:
- To modify and validate existing murine models for HFpEF.
- To improve the "2-hit" model's applicability to C57BL/6J mice.
- To develop an enhanced "4-hit" model for HFpEF that is reproducible across sexes and substrains.
Main Methods:
- Modified the "2-hit" model by increasing L-NAME dosage and protocol duration for C57BL/6N and C57BL/6J mice.
- Developed a "4-hit" model by adding 1% NaCl to deoxycorticosterone pivalate (DOCP) treatment and administering a second DOCP bolus.
- Evaluated HFpEF phenotype induction and maintenance over time in different mouse strains and sexes.
Main Results:
- Successfully induced HFpEF in C57BL/6J mice using an extended "2-hit" L-NAME protocol.
- The enhanced "4-hit" model demonstrated a clear HFpEF phenotype in both male and female C57BL/6N and C57BL/6J mice.
- The HFpEF phenotype was successfully maintained for up to 12 weeks in the "4-hit" model.
Conclusions:
- The modified "2-hit" model is now effective in the C57BL/6J substrain.
- The novel "4-hit" model overcomes aldosterone escape, ensuring sustained HFpEF phenotype.
- These refined models offer enhanced reproducibility across sexes and mouse substrains, advancing HFpEF research.
