title: HSPA12B inhibits lipopolysaccharide-induced inflammatory response in human umbilical vein endothelial cells.
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.
authors.affiliation: Department of Geriatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.forename: Xuehan
authors.initials: X
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
authors.forename: Surong
authors.initials: S
authors.lastname: Jiang
authors.forename: Fei
authors.initials: F
authors.lastname: Tu
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Zhang
authors.forename: He
authors.initials: H
authors.lastname: Ma
authors.forename: Rongrong
authors.initials: R
authors.lastname: Li
authors.forename: Chuanfu
authors.initials: C
authors.lastname: Li
authors.forename: Yuehua
authors.initials: Y
authors.lastname: Li
authors.forename: Zhengnian
authors.initials: Z
authors.lastname: Ding
authors.forename: Li
authors.initials: L
authors.lastname: Liu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000730:Androstadienes / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000235:genetics; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000235:genetics; D002465:Cell Movement; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000276:immunology*; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000276:immunology*; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides; D009504:Neutrophils / Q000276:immunology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins; D015850:Interleukin-6; D008070:Lipopolysaccharides; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D000077191:Wortmannin; C472131:HSPA12B protein, human; C508600:IL6 protein, human
keywords: HUVECs; Heat shock protein A12B; PI3K/Akt signalling; inflammation; lipopolysaccharide