HSPA12B inhibits lipopolysaccharide‐induced inflammatory response in human umbilical vein endothelial cells

Journal of Cellular and Molecular Medicine

Jun Wu⁴  Xuehan Li⁴  Lei Huang¹  Surong Jiang⁴  Fei Tu²  Xiaojin Zhang⁴  He Ma¹  Rongrong Li¹  Chuanfu Li⁵  Yuehua Li³  Zhengnian Ding¹ 

[1] Department of Anesthesiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China;Jiangsu Key Laboratory for Molecular and Medical Biotechnology and College of Life Sciences, Nanjing Normal University, Nanjing, China;Department of Pathophysiology, Nanjing Medical University, Nanjing, China;Department of Geriatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China;Department of Surgery, East Tennessee State University, Johnson City, TN, USA

关键词: Heat shock protein A12B;    lipopolysaccharide;    HUVECs;    inflammation;    PI3K/Akt signalling;   

DOI  :  10.1111/jcmm.12464

来源: Wiley

【 摘 要 】

Abstract

Heat shock protein A12B (HSPA12B) is a newly discovered member of the HSP70 protein family. This study investigated the effects of HSPA12B on lipopolysaccharide (LPS)-induced inflammatory responses in human umbilical vein endothelial cells (HUVECs) and the possible mechanisms involved. A HUVECs inflammatory model was induced by LPS. Overexpression of HSPA12B in HUVECs was achieved by infection with recombinant adenoviruses encoding green fluorescence protein-HSPA12B. Knockdown of HSPA12B was achieved by siRNA technique. Twenty four hours after virus infection or siRNA transfection, HUVECs were stimulated with 1 μg/ml LPS for 4 hrs. Endothelial cell permeability ability was determined by transwell permeability assay. The binding rate of human neutrophilic polymorphonuclear leucocytes (PMN) with HUVECs was examined using myeloperoxidase assay. Cell migrating ability was determined by the wound-healing assay. The mRNA and protein expression levels of interested genes were analyzed by RT-qPCR and Western blot, respectively. The release of cytokines interleukin-6 and tumour necrosis factor-α was measured by ELISA. HSPA12B suppressed LPS-induced HUVEC permeability and reduced PMN adhesion to HUVECs. HSPA12B also inhibited LPS-induced up-regulation of adhesion molecules and inflammatory cytokine expression. By contrast, knockdown of HSPA12B enhanced LPS-induced increases in the expression of adhesion molecules and inflammatory cytokines. Moreover, HSPA12B activated PI3K/Akt signalling pathway and pharmacological inhibition of this pathway by Wortmannin completely abrogated the protection of HSPA12B against inflammatory response in HUVECs. Our results suggest that HSPA12B attenuates LPS-induced inflammatory responses in HUVECs via activation of PI3K/Akt signalling pathway.

【 授权许可】

CC BY   
© 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

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