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Burn-induced apoptosis of cardiomyocytes is survivin dependent and regulated by PI3K/Akt, p38 MAPK and ERK pathways

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Abstract

Survivin belongs to the family of genes known as inhibitors of apoptosis, and although it has been implicated in the prevention of cancer, its potential role in burn-induced cardiac injury is unknown. In this study, we investigated the effects of survivin blockade on burn-induced cardiac apoptosis. Using a standardized Sprague-Dawley rat model of third-degree burn injury over 40% of total body surface area, apoptosis was measured in vivo followed by in vitro assessment of burn serum-stimulated cardiomyocytes. Based on the Western blot analyses, real-time PCR, ELISA, and TUNEL, apoptosis and caspase activation both in vivo and in vitro were significantly increased after severe burn injury, while survivin expression was increased (up to 2.90-fold) during the early stage of burn injury and was almost completely abolished 8 h after the burn. Survivin-deficient cardiomyocytes, as well as hearts from rats treated with the survivin inhibitor YM155, exhibited increased caspase-3 protein and mRNA expression and apoptosis ratio at different times after the burn. Furthermore, inhibition of ERK, phosphoinositol 3-kinase contributed the burn serum-induced increase in apoptosis and caspase-3 protein expression, and decreased survivin expression, whereas burn serum-induced increase in apoptosis was attenuated by P38 mitogen-activated protein kinase inhibition. These data identify survivin as a critical anti-apoptotic regulator of cardiomyocytes after burn injury. ERK, P38 MAPK and PI3K were found to be upstream regulators of survivin.

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Acknowledgments

This work was supported by the Grant from the National Natural Science Foundation of China (30800433 to X.Q. Li) and Science and Technology Innovation Project of Shaanxi Province in China (2010K01-177 to S.W. Wang).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, 169 West Changle Road, Xi’an, 710032, China

    Wei Cao, Yan-Hua Xie, Xiao-Kai Zhang, Yue-Tao Chen, Rong Kang, Xi Chen, Shan Miao & Si-Wang Wang

  2. Department of Pharmacology, School of Pharmacy, Fourth Military Medical University, 169 West Changle Road, Xi’an, 710032, Shaanxi, China

    Xiao-Qiang Li

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  1. Wei Cao
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395_2011_199_MOESM1_ESM.tif

Supplementary Fig. 1 Induced survivin expression in the cardiac sections after burn injury. Survivin protein expression in the heart tissues at 0 - 8 h after burn injury was visualized by immunohistochemistry. Figures are representative of two separate and independent experiments. (TIFF 10856 kb)

395_2011_199_MOESM2_ESM.tif

Supplementary Fig. 2 The effect of sham serum on cardiomyocyte apoptosis and survivin expression. Neonatal rat cardiomyocytes were treated with sham serum for various periods of time, as indicated. a The extent of DNA fragmentation was quantified using ELISA (N=6). b Survivin and cleaved caspase-3 levels were measured by Western blot analysis. A typical display is depicted (upper panel) along with the statistical analysis of the changes of the protein (lower panel). Data are expressed as mean±S.E. (N = 3). (TIFF 12916 kb)

395_2011_199_MOESM3_ESM.tif

Supplementary Fig. 3 The effect of burn injury on P38 MAPK, JNK, ERK and Akt phosphorylation in the cardiac sections. The rats were exposed to a 40% TBSA burn (burn injury) or 25°C water (sham). The p-ERK1/2, p-P38, p-JNK and p-Akt levels at 0, 0.5, 1, 2, 4, 6, and 8 h after burn injury were measured by Western blot analysis. a The blots are representative of five rats. b The statistical analysis of the changes of the protein. Data are expressed as mean±SE *P < 0.05 vs. 0 h (TIFF 6949 kb).

395_2011_199_MOESM4_ESM.tif

Supplementary Fig. 4 The effect of sham serum on the expression of six cytokines commonly used to evaluate the inflammatory response. The levels of IL-1β, IL-6, IL-10, IL-12, TNF- α and CINC-1 were determined by ELISA according to the protocol of the manufacturer. Data are expressed as mean±S.E. (N=5, *P < 0.05 vs. time-matched control). (TIFF 10772 kb)

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Cao, W., Xie, YH., Li, XQ. et al. Burn-induced apoptosis of cardiomyocytes is survivin dependent and regulated by PI3K/Akt, p38 MAPK and ERK pathways. Basic Res Cardiol 106, 1207–1220 (2011). https://doi.org/10.1007/s00395-011-0199-3

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