The Study Of VEGF121 And VEGF165 On Endothelial Cell Permeability

Objective:Vascular endothelial growth factor (VEGF)which was discov- ered in 1989 could regulated angiogenesis on vascular endothelial cell.VEGF family includes seven members, VEGF-A, VEGF-B, PlGF, VEGF-C, VEGF-D, VEGF-E and VEGF-F.their receptors,VEGFR-1,2,3 and Neuropilins.VEGF could induced physiological function when it binded to its receptor.VEGF stimulates vascular endothelial cell divided,leading to angiogenesis.VEGF regulates vascular permeability.of VEGF family,VEGF-A which play a dominant role included eight isoforms, VEGF121, VEGF145, VEGF148, VEGF165, VEGF165b,VEGF183,VEGF189,VEGF206.of eight isoforms, VEGF121 andVEGF165 are mainly secreted proteins.Nitric oxide(NO) was found in plant and animal.NO could exert physiological and pathological functions,it is one of the most strong material which induces blood vessel relaxed.NO could combin to guanylate cyclase that induced cGMP increase.whereas cGMP regulated Ca²⁺ decrease.the result leads to smooth muscle relaxed , angioectasia, vascular permeability increase.The angiogenesis and vascular permeability plays a important role in the emerging and development of rheumatoid arthritis, cardiovascular disease, diabetes complications and other diseases and tumor metastasis.The angiogenesis is closely relative with VEGF. in recent years,reserchers found that,VEGF is lower in normal skin than the lesion of skin disease,such as atopic dermatitis, psoriasis and warts. These results indicated thging and development of skin disease is closely relative with VEGF.It is easy to insight to the pathogenesis of many skin disease. the research of therapeutic targets will provide a new way for treatment. VEGF121 and VEGF165 are mainly detected in epidermal tissue,they both participation in various of Pathology and physiological processes. Various previous research is mianly focuses on the study of VEGF-A, but this reaserches are rarely specializes in subtype.Therefore, this reaserch mainly invested the influence of VEGF isform for Human umbilical vein endothelial cells (HUVEC) perneability and NO relesed content when HUVEC were stimulated ,and indicated that which is main isforms induces endothelial cells permeability .This reaserch provides the Theoretic basis for researching VEGF blockers, targeted therapy atopic dermatitis, psoriasis, erythema inflammatory skin disease and reducing side effect.Method:1 Human umbilical vein endothelial cells were cultured in vitro.Human umbilical vein endothelial cells (HUVEC) were cultured at 37℃in a humidified atmosphere of 5% CO₂ in EGM-2.EGM-2 were replaced by new one after 2-3 days.2 In Dual-chamber permeability assay kit (ECM640) cell monolayer we-re planted to establish permeability detection model.3 Human umbilical vein endothelial cells were stimulated by VEGF 121or VEGF165.4 FITC-dextran were detected by the Microplate fluorescence determinat-ion.These results indicated different vascular permeability5 NO content were determinatied by Griess kit medium .6 The date were analysed by use statistical software of SPSS13.0.Results:1 FITC- dextran were detected in human umbilical vein endothelial cells which were stimulated by VEGF165 or VEGF121.1.1 FITC-dextran were detected by the Microplate fluorescence determination. Statistical evaluation was done with Student's ANOVA of the repeated measure. FITC-dextran were higher in extracell which were stimulated by VEGF121 than VEGF165 or nothing(p<0.05). VEGF121 influenced on vascular permeability more intensely than VEGF165.1.2 FITC-dextran is effected by different time (p<0.05). VEGF165 or VEGF121 induced a marked increase from 5min up to 6h.1.3 FITC-dextran is effected by time and various VEGF(p<0.05).2 NO were detected in human umbilical vein endothelial cells which were stimulated by VEGF165 or VEGF121.2.1 NO were detected by the Griess kit medium. Statistical evaluation was done with Student's ANOVA of the repeated measure. NO which were higher in human umbilical vein endothelial cells were stimulated by VEGF121 than VEGF165 or nothing(p<0.05). VEGF121 influenced on vascular permeability more intensely than VEGF165.2.2 NO is effected by different time (p<0.05). VEGF165orVEGF121 induced NO a marked increase from 0min up to 30min.2.3 NO is effected by time and various VEGF(p<0.05).Conclusion:VEGF165 and VEGF121could regulated vascular permeability. VEGF121 influenced on vascular permeability more intensely than VEGF165. NO which were higher in human umbilical vein endothelial cells were stimulated by VEGF121 than VEGF165.this result indicated .VEGF121 is mainly factor on vascular permeability.