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CRYAB protects cardiomyocytes against heat stress by preventing caspase-mediated apoptosis and reducing F-actin aggregation

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Cell Stress and Chaperones Aims and scope

Abstract

CRYAB is a small heat shock protein (sHSP) that has previously been shown to protect the heart against various cellular stresses; however, its precise function in myocardial cell injury caused by heat stress remains unclear. This study aimed to investigate the molecular mechanism by which CRYAB protects cardiomyocytes against heat stress. We constructed two H9C2 cell lines that stably express CRYAB protein to differing degrees: CRYAB-5 and CRYAB-7. Both CRYAB-5 and CRYAB-7 showed significantly reduced granular degeneration and vacuolar degeneration following heat stress compared to control cells. In addition, CRYAB overexpression in H9C2 cells relieved cell cycle proportion at the G0/G1 phase following heat stress compared to control cells. These protective effects were associated with the level of CRYAB protein expression. Our immunofluorescence analysis showed CRYAB could translocate from the cytoplasm to the nucleus under heat stress conditions, but that CRYAB co-localized with F-actin (which accumulates under stress conditions). Indeed, overexpression of CRYAB significantly reduced the aggregation of F-actin in H9C2 cells caused by heat stress. Furthermore, overexpressing CRYAB protein significantly reduced the apoptosis of cardiomyocytes induced by heat stress, likely by reducing the expression of cleaved-caspase 3. Collectively, our results show overexpression of CRYAB significantly increases the heat resistance of H9C2 cardiomyocytes, likely by reducing F-actin aggregation (thus stabilizing the cytoskeleton), regulating the cell cycle, and preventing caspase-mediated apoptosis.

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Funding

The current study was supported by the grants from the National Natural Science Foundation of China (grant no. 31672520), the Fundamental Research Funds for the Central Universities (grant no. KJQN201709), the National Natural Science Foundation of China (grant no. 31602027), the National Natural Science Foundation of China (grant no. 31372403), Jiangsu Natural Science Foundation of China (grant no. BK20160732), China Postdoctoral Science Foundation (2016M591860), and Shandong Natural Science Foundation of China (grant no. ZR2016CM40).

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  1. Bin Yin and Shu Tang contributed equally to this work.

Authors and Affiliations

  1. College of Veterinary Medicine, Nanjing Agricultural University, Weigang 1, Nanjing, 210095, China

    Bin Yin, Shu Tang, Jiao Xu, Jiarui Sun, Xiaohui Zhang & Endong Bao

  2. College of Agronomy, Liaocheng University, Hunan road 1, Liaocheng, 252000, China

    Yubao Li

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  1. Bin Yin
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  2. Shu Tang
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  3. Jiao Xu
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  4. Jiarui Sun
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  7. Endong Bao
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Correspondence to Endong Bao.

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Yin, B., Tang, S., Xu, J. et al. CRYAB protects cardiomyocytes against heat stress by preventing caspase-mediated apoptosis and reducing F-actin aggregation. Cell Stress and Chaperones 24, 59–68 (2019). https://doi.org/10.1007/s12192-018-0941-y

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  • DOI: https://doi.org/10.1007/s12192-018-0941-y

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