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Chrysin protects cardiac H9c2 cells against H2O2-induced endoplasmic reticulum stress by up-regulating the Nrf2/PERK pathway

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Abstract

Oxidative and endoplasmic reticulum (ER) stress-mediated cardiac apoptosis is an essential pathological process in cardiovascular diseases (CVDs). Chrysin (Chy) is a natural flavonoid that exerts several health benefits, particularly anti-oxidative and anti-apoptotic effects. However, its protective effect against CVDs and its mechanism of action at a molecular level remains unclear. Therefore, the present study aimed to investigate the interaction of ER stress response protein with Chy by computational analysis and molecular action in H2O2-induced oxidative and ER stress in cardiomyoblast cells. H9c2 cells were pre-treated with 50 μM of Chy for 24 h and exposed to H2O2 for 1 h. Explore the Chy-mediated Nrf2 signalling on ER stress reduction, H9c2 cell lines were transfected with Nrf2 siRNA for 48 h and further treated with Chy for 24 h and subjected to H2O2 for 1 h. Chy pre-treatment increased the Nrf2-regulated gene expression, reduced the ER stress signalling genes such as CHOP and GRP78, and increased the PERK and AFT6 expression compared to H2O2-treated cells. Chy preincubation down-regulated the expression of PI3K, NF-κB, and caspase-3. Fluorescence staining revealed that Chy reduced intracellular ROS generation, ER stress, apoptosis, and increased MMP. This beneficial effect of Chy was abolished when silencing Nrf2 in H9c2 cells. Overall, the present study confirmed that Chy showed the cardioprotective effect by attenuating ER stress via the activation of Nrf2 signalling. Therefore, the study concluded that improving Nrf2 signalling by Chy supplementation could provide a promising therapeutic target in oxidative and ER stress-mediated CVDs complications.

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The authors declare that all data supporting the findings of this study are available within the article and its supplementary information files.

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Acknowledgements

S.Yuvaraj expresses his gratitude to the Council of Scientific and Industrial Research Human Resource Development Group, New Delhi, India- for CSIR Senior Research Fellowship and financial support. The authors also thank UGC-CEGS, UGC-CAS, UGC-NRCBS, DST-FIST, and DST-PURSE program for the central instrumentation facility at SBS, MKU.

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Authors and Affiliations

  1. Molecular Cardiology Unit, Department of Biochemistry, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, 625021, India

    Subramani Yuvaraj, Arumugam Kalaiselvi Ajeeth, Shanavas Syed Mohamed Puhari, Albert Abhishek, Varadaraj Vasudevan & Govindan Sadasivam Selvam

  2. Center for Molecular and Translational Medicine, Georgia State University, Atlanta, GA, USA

    Tharmarajan Ramprasath

  3. Department of Medical Biotechnology, Faculty of Allied Health Sciences, Chettinad Academy of Research and Education, Kelambakkam, Chennai, Tamil Nadu, 603103, India

    Narasimman Vignesh

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  1. Subramani Yuvaraj
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Contributions

Conceptualization—SY; Methodology—SY, AKA, SSMP, VV, NV; Formal analysis –SY, TR; Investigation—SY, SSMP, GSS; Writing—Original draft-SY, SSMP; Review and Editing-SY, AA, SSMP, VV, and GSS; Funding acquisition-GSS.

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Correspondence to Govindan Sadasivam Selvam.

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The experimental procedure was approved by the Internal Research and Review Board, Ethical Clearance, Biosafety and Animal Welfare Committee of Madurai Kamaraj University.

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Yuvaraj, S., Ajeeth, A.K., Puhari, S.S.M. et al. Chrysin protects cardiac H9c2 cells against H2O2-induced endoplasmic reticulum stress by up-regulating the Nrf2/PERK pathway. Mol Cell Biochem 478, 539–553 (2023). https://doi.org/10.1007/s11010-022-04531-z

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