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Luhong Formula and Hydroxysafflor yellow A protect cardiomyocytes by inhibiting autophagy
Jiling Feng1, Jiaying Guo2, Jirong Yan2
1Institute of Cardiovascular Disease of Integrated Traditional Chinese and Western Medicine, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, No. 528, Zhangheng Road, Shanghai 201203, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, No. 1200, Cailun Road, Shanghai 201203, China; Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, No. 1200, Cailun Road, Shanghai 201203, China; Precision Research Center for Refractory Diseases, Institute for Clinical Research, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201620, China.
Insights
Luhong formula (LHF) and hydroxysafflor yellow A (HYSA) protect heart cells from damage by inhibiting autophagy through reduced reactive oxygen species (ROS) production, offering a new cardiovascular therapy.
Area of Science:
- Cardiovascular Research
- Cellular Biology
- Pharmacology
Background:
- Heart failure (HF) involves irreversible cardiomyocyte injury, where autophagy balance is crucial for cell survival.
- Luhong formula (LHF) is a traditional treatment known to improve heart function and alleviate angina.
- Understanding LHF's mechanism in protecting against cardiac injury is essential.
Purpose of the Study:
- To investigate how LHF and its key component, hydroxysafflor yellow A (HYSA), protect ischemic cardiomyocytes.
- To elucidate the role of autophagy inhibition in LHF's cardioprotective effects.
- To identify the molecular pathways involved in LHF's action.
Main Methods:
- In vitro studies used CCK-8 assays and immunofluorescence to assess cell viability and autophagic flux in response to LHF/HYSA under hypoxic conditions.
- Western blotting was employed to measure Beclin 1 and HIF1α protein expression.
- In vivo studies involved administering LHF to Wistar rats post-myocardial infarction, followed by echocardiography, immunohistochemistry, and histological staining.
Main Results:
- LHF and HYSA enhanced cell viability and inhibited autophagy by reducing Beclin 1 expression and suppressing cellular reactive oxygen species (ROS).
- The protective effects were linked to the inhibition of HIF1α, which reduced ROS production.
- In a rat model, LHF improved cardiac function, reduced fibrosis, and decreased Beclin 1 and HIF1α expression in infarcted heart tissue.
Conclusions:
- Hydroxysafflor yellow A is identified as the primary bioactive component of LHF responsible for regulating autophagy.
- LHF and HYSA protect cardiomyocytes from hypoxia by inhibiting HIF1α-mediated ROS production, thereby suppressing cardiac autophagy.
- This research provides a mechanistic basis for LHF's development as a novel cardiovascular therapeutic agent.
Background:
Heart failure (HF) is the terminal stage of all heart diseases that is characterized by irreversible cardiomyocyte injury. Equilibrium of autophagy is essential for cardiac cell survival. The Luhong formula (LHF) has been clinically applied for decades, and has exhibited significant efficacy in improving heart function and alleviating the symptoms of angina pectoris.
Purpose:
To clarify the mechanism of action of LHF and one of its main constituents, hydroxysafflor yellow A (HYSA), in protecting ischemic cardiomyocytes by inhibiting autophagy.
Methods:
Cell viability was detected by CCK-8 assay with LHF or HYSA pretreatment followed by hypoxic damage. Immunofluorescence of GFP-LC3-H9C2 and GFP-LC3-HeLa cells was used to observe autophagic flux. Beclin 1 and HIF1α protein expression were assessed using western blotting. LHF was orally administered to Wistar rats following myocardial infarcion. Echocardiography was performed before the rats were sacrificed; immunohistochemistry and western blotting were used to evaluate Beclin 1 and HIF1α expression in the myocardial tissue. Hematoxylin and eosin staining as well as Masson's trichrome staining were used to measure cardiac structure and myocardial fibrosis.
Results:
LHF and HYSA reversed the hypoxia-induced decrease in cell viability in vitro. LHF and HYSA induced the aggregation of GFP-LC3 puncta and reduced the expression of Beclin 1 protein in H9C2, suggesting that LHF and HYSA may inhibit autophagy activity. Pretreatment with reactive oxygen species (ROS) inducers and inhibitors revealed that LHF and HYSA inhibited autophagy by suppressing cellular ROS. Further studies demonstrated that LHF and HYSA reduced the ROS levels by inhibiting HIF1α. LHF delayed fibrosis and protected heart function in vivo in a rat model of HF following myocardial infarction. Western blotting and immunohistochemistry revealed that LHF effectively reduced the expression of Beclin 1 and HIF1α in the infarcted area of the rat heart.
Conclusion:
These results demonstrate that hydroxysafflor yellow A is the representative bioactive compounent of Luhong Formula on regulating autophagy to protectect cardiomyocytes from hypoxia injury. LHF and HYSA inhibit cardiac autophagy by suppressing HIF1α-mediated ROS production. This study helps to further clarify the underlying mechanism of LHF and provide a scientific basis for its development as a novel cardiovascular therapeutic agent.
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