Shiwei Liu1, Yanting Dong2, Tong Wang3, Shujun Zhao4, Kun Yang5, Xiaoqin Chen5, Caihong Zheng5. 1. Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China. Electronic address: lswspring6@gmail.com. 2. Department of Biochemistry, Shanxi Medical University, Taiyuan 030001, China. 3. Department of Medical Statistics, School of Public Health of Shanxi Medical University, Taiyuan 030001, China. 4. Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China. 5. Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China.
Abstract
AIMS: In this study, we investigated the effects of visceral adipose tissue-derived serpin (vaspin), a newly discovered adipocytokine, on nuclear factor-kappa B (NF-κB) and its downstream molecules in proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukine-1 (IL-1), stimulated human endothelial EA.hy926 cells to elucidate the role of vaspin in the inflammatory states of endothelium. METHODS: A NF-κB luciferase reporter system was constructed and stably transfected into human endothelial cell line EA.hy926. Following transfection, EA.hy926 cells were pretreated with various concentrations of vaspin (0-320 ng/ml) before TNF-α and IL-1 stimulation. The transcription activity of NF-κB was determined using luciferase reporter assay. Expression levels of NF-κB downstream inflammatory cytokines, TNF-α, IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Expressions of adhesion molecules and chemokines, 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 (RT-PCR) and western blot in mRNA and protein levels, respectively. RESULTS: Results showed that vaspin inhibited TNF-α and IL-1 mediated activation of NF-κB and its downstream molecules in a concentration-dependent manner (P<0.05). CONCLUSIONS: We conclude that vaspin protected endothelial cells from proinflammatory cytokines induced inflammation by inhibition of NF-κB and its downstream molecules.
AIMS: In this study, we investigated the effects of visceral adipose tissue-derived serpin (vaspin), a newly discovered adipocytokine, on nuclear factor-kappa B (NF-κB) and its downstream molecules in proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukine-1 (IL-1), stimulated human endothelial EA.hy926 cells to elucidate the role of vaspin in the inflammatory states of endothelium. METHODS: A NF-κB luciferase reporter system was constructed and stably transfected into human endothelial cell line EA.hy926. Following transfection, EA.hy926 cells were pretreated with various concentrations of vaspin (0-320 ng/ml) before TNF-α and IL-1 stimulation. The transcription activity of NF-κB was determined using luciferase reporter assay. Expression levels of NF-κB downstream inflammatory cytokines, TNF-α, IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Expressions of adhesion molecules and chemokines, 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 (RT-PCR) and western blot in mRNA and protein levels, respectively. RESULTS: Results showed that vaspin inhibited TNF-α and IL-1 mediated activation of NF-κB and its downstream molecules in a concentration-dependent manner (P<0.05). CONCLUSIONS: We conclude that vaspin protected endothelial cells from proinflammatory cytokines induced inflammation by inhibition of NF-κB and its downstream molecules.
Authors: Mohamed I Saad; Taha M Abdelkhalek; Moustafa M Saleh; Maher A Kamel; Mina Youssef; Shady H Tawfik; Helena Dominguez Journal: Endocrine Date: 2015-08-14 Impact factor: 3.633
Authors: Di Qi; Daoxin Wang; Chunrong Zhang; Xumao Tang; Jing He; Yan Zhao; Wang Deng; Xinyu Deng Journal: Int J Mol Med Date: 2017-10-09 Impact factor: 4.101