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Heat stress prevents lipopolysaccharide-induced apoptosis in pulmonary microvascular endothelial cells by blocking calpain/p38 MAPK signalling

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Abstract

Pulmonary microvascular endothelial cells (PMECs) injury including apoptosis plays an important role in the pathogenesis of acute lung injury during sepsis. Our recent study has demonstrated that calpain activation contributes to apoptosis in PMECs under septic conditions. This study investigated how calpain activation mediated apoptosis and whether heat stress regulated calpain activation in lipopolysaccharides (LPS)-stimulated PMECs. In cultured mouse primary PMECs, incubation with LPS (1 μg/ml, 24 h) increased active caspase-3 fragments and DNA fragmentation, indicative of apoptosis. These effects of LPS were abrogated by pre-treatment with heat stress (43 °C for 2 h). LPS also induced calpain activation and increased phosphorylation of p38 MAPK. Inhibition of calpain and p38 MAPK prevented apoptosis induced by LPS. Furthermore, inhibition of calpain blocked p38 MAPK phosphorylation in LPS-stimulated PMECs. Notably, heat stress decreased the protein levels of calpain-1/2 and calpain activities, and blocked p38 MAPK phosphorylation in response to LPS. Additionally, forced up-regulation of calpain-1 or calpain-2 sufficiently induced p38 MAPK phosphorylation and apoptosis in PMECs, both of which were inhibited by heat stress. In conclusion, heat stress prevents LPS-induced apoptosis in PMECs. This effect of heat stress is associated with down-regulation of calpain expression and activation, and subsequent blockage of p38 MAPK activation in response to LPS. Thus, blocking calpain/p38 MAPK pathway may be a novel mechanism underlying heat stress-mediated inhibition of apoptosis in LPS-stimulated endothelial cells.

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Acknowledgments

This work was supported by Grants from the National Natural Science Foundation of China [No. 81571940], the Project Team of Natural Science Foundation of Guangdong Province [No. s2013030013217], and the Project of Medical Research of PLA [No. BWS12J108], the Guangzhou Science and Technology Planning Project of China [No. 201504281714528], and the Heart & Stroke Foundation of Canada (Awarded to T.P.). T.P. is a recipient of a New Investigator Award from the Canadian Institutes of Health Research.

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Authors and Affiliations

  1. Department of Intensive Care Unit, General Hospital of Guangzhou Military Command, Guangzhou, 510010, China

    Zhi-feng Liu & Lei Su

  2. Departments of Medicine and Pathology, Critical Illness Research, Lawson Health Research Institute, University of Western Ontario, London, ON, N6A 4G5, Canada

    Zhi-feng Liu, Dong Zheng & Tianqing Peng

  3. Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH, 45267-0575, USA

    Guo-chang Fan

  4. Key Laboratory of Hot Zone Trauma Care and Tissue Repair of PLA, General Hospital of Guangzhou Military Command, Guangzhou, 510010, China

    Lei Su

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  1. Zhi-feng Liu
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Liu, Zf., Zheng, D., Fan, Gc. et al. Heat stress prevents lipopolysaccharide-induced apoptosis in pulmonary microvascular endothelial cells by blocking calpain/p38 MAPK signalling. Apoptosis 21, 896–904 (2016). https://doi.org/10.1007/s10495-016-1263-0

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