关键词: Vaspin, 核因子-κB, 磷脂酰肌醇3激酶（PI3K）/Akt, 胰岛素抵抗, 炎症, 血管内皮细胞

Abstract: Objective To investigate the effects of visceral adipose tissue-derived serpin (Vaspin) on nuclear factor-kappa B (NF-κB) and phosphatidylinositol 3-kinase(PI3K)/Akt signal pathways in human umbilical vein endothelial cells (HUVEC) stimulated by tumor necrosis factor-α (TNF-α) to elucidate the role of Vaspin in human endothelial cells of inflammation and insulin resistance. Methods HUVEC were isolated and cultured in vitro. A NF-κB luciferase reporter system was constructed and transiently transfected into HUVEC. Following transfection, HUVEC were pretreated with various concentrations of Vaspin (0~320 μg/L) before 10 μg/L TNF-α stimulation. The transcription activity of NF-κB was determined using luciferase reporter assay. The level of Akt phosphorylation was checked by Western blot. Expression levels of NF-κB downstream inflammatory cytokines IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). mRNA and protein expression levels of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemotactic protein-1 (MCP-1) were determined by quantitative real-time PCR (qRT-PCR) and Western blot respectively. Results Vaspin inhibited TNF-α mediated activation of NF-κB and its downstream molecules (P<0.05). Vaspin significantly increased Akt phosphorylation in TNF-α stimulated endothelial cells (P<0.05). Conculsions Vaspin protected endothelial cells from TNF-α induced inflammation and insulin resistance by combination the inhibition of NF-κB, its downstream molecules and the upregulation of the PI3-kinase/Akt signal pathway.

Key words: Vaspin, NF-κB, phosphatidylinositol 3-kinase(PI3K)/Akt, insulin resistance, inflammation, endothelium