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HSPA 12B inhibits lipopolysaccharide‐induced inflammatory response in human umbilical vein endothelial cells

2014; Wiley; Volume: 19; Issue: 3 Linguagem: Inglês

10.1111/jcmm.12464

1582-4934

Jun Wu, Xuehan Li, Lei Huang, Surong Jiang, Fei Tu, Xiaojin Zhang, He Ma, Rongrong Li, Chuanfu Li, Yuehua Li, Zhengnian Ding, Li Liu,

Connexins and lens biology

Abstract Heat shock protein A12B ( HSPA 12B) is a newly discovered member of the HSP 70 protein family. This study investigated the effects of HSPA 12B on lipopolysaccharide ( LPS )‐induced inflammatory responses in human umbilical vein endothelial cells ( HUVEC s) and the possible mechanisms involved. A HUVEC s inflammatory model was induced by LPS . Overexpression of HSPA 12B in HUVEC s was achieved by infection with recombinant adenoviruses encoding green fluorescence protein‐ HSPA 12B. Knockdown of HSPA 12B was achieved by si RNA technique. Twenty four hours after virus infection or si RNA transfection, HUVEC s 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 HUVEC s 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 . HSPA 12B suppressed LPS ‐induced HUVEC permeability and reduced PMN adhesion to HUVEC s. HSPA 12B also inhibited LPS ‐induced up‐regulation of adhesion molecules and inflammatory cytokine expression. By contrast, knockdown of HSPA 12B enhanced LPS ‐induced increases in the expression of adhesion molecules and inflammatory cytokines. Moreover, HSPA 12B activated PI 3K/Akt signalling pathway and pharmacological inhibition of this pathway by Wortmannin completely abrogated the protection of HSPA 12B against inflammatory response in HUVEC s. Our results suggest that HSPA 12B attenuates LPS ‐induced inflammatory responses in HUVEC s via activation of PI 3K/Akt signalling pathway.