论文部分内容阅读
Background The common pathological characteristics of ceal injury include inflammatory factors activation,vascular endothelial cells or inflammatory cells infiltration into lesions,ceal edema,ceal neovascularization (CNV),and scar formation.PEDF-34 is the functional fragment of pigment epithelium-derived factor (PEDF) that has anti-angiogenic and anti-inflammatory properties and contains an N-terminal 34-amino acid peptide.This study was to investigate the antiinflammatory effects of PEDF-34 on H2O2-induced ceal injury in vitro.Methods After cultured in H2O2 (0.1 mmol/L) for 2 hours,human ceal fibroblasts (HCFs) and human umbilical vein endothelial cells (HUVECs) were treated with PEDF-34-nanoparticles (NPs) at different concentrations (0.1,0.5,1.0,2.0 μg/ml) or 2.0 μg/ml control-NPs for 24 hours.The viable cells were quantified using the MTT assay.West blotting or ELISA analysis was performed for measuring the human vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1) expression of both HCFs and HUVECs.VEGF and nuclear factor κB (NF-κB) mRNA levels of HCFs were semi-quantified by RT-PCR.Results The survival rates of HCFs or HUVECs stimulated by H2O2 did not decrease significantly (P ＞0.05) compared to those in the normal conditions.As compared to control-NP group,PEDF-34-NPs had dose-dependent inhibitive effect on HUVECs with the MTT assay,but not HCFs.West blotting analysis showed that the VEGF and ICAM-1 levels in the HCFs and HUVECs stimulated by H2O2 were significantly higher than those in the normal conditions,which were decreased dramatically in those treated with PEDF-34-NPs.RT-PCR analysis revealed that the VEGF mRNA and NF-κB mRNA levels increased in H2O2-stimulated HCFs,while both of them decreased in PEDF-34-NP groups dose dependently.Conclusions PEDF-34-NPs may play an important role in regulating the NF-κB pathway,inhibiting inflammatory activity.PEDF-34-NPs may be a potential new drug for treating ceal injury in the future.