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Effect of Lipopolysaccharide on Pulmonary Vascular Endothelial Cells and Its Mechanism in Neonatal Mice
Deng Sijun, Zhang Han, Guo Chunbao, Liu Xuwei, You Yaoyao, Deng Chun*
Department of Neonatology, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China
Abstract: This study aimed to investigate the effects and mechanisms of lipopolysaccharide (LPS) on pulmonary vascular endothelial cells in neonatal mice. The cells were randomly divided into blank control group and LPS group. The cells were treated with 10 μg/mL LPS， and the cell samples were collected at 0 h， 6 h， 12 h， 24 h， 48 h. The migration of pulmonary vascular endothelial cells was observed by the scratch test; the mRNA expression of Chemokine and inflammatory cytokines in the lung vascular endothelial cells were determined by Real-time PCR (RT-PCR); the protein level of P65， VEGF and VEGFR2 were determined by Western blot. The results showed that the pulmonary vascular endothelial cells were exhibited as polygon and presented typical cobblestonelike morphology after fusion to monolayer with contact inhibition. Immunofluorescence staining revealed that the expressions of CD31 and factor VIII-related antigen were positive. The migration of LPS group cells in the scratch test was higher than that of the control group at 12 h (P<0.001). The mRNA expression of inflammatory factors IL-1β， TNF-α and chemokines MIP-1α and MCP-1 secreted by LPS group was significantly higher than that control group by Real-time PCR (P<0.001); the protein expression of NF-κB-related protein P65 activity increased (P<0.05)， the expression of VEGFR2 protein on the surface of pulmonary vascular endothelium was significantly decreased (P<0.001); the vascular endothelial growth factor (VEGF) in LPS group was decreased. This study demonstrated that the inflammatory response induced by lipopolysaccharide affects the development of pulmonary vascular endothelial cells， and its mechanism might be related to the activation of NF-κB pathway， the induction of inflammatory factors， the increase of chemokine expression and the decrease of VEGF/VEGFR2 expression.
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