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The Research of FAK and Akt Signal Pathways in HUVEC after Vegf121 or VEGF165 Isoforms Treatment in Vitro

Author: ZhangJunXiu
Tutor: GaoShunQiang
School: Hebei Medical University
Course: Dermatology and Venereology
Keywords: VEGF165 VEGF121 FAK Akt FAK phosphorylation Phosphorylated Akt
CLC: R758.2
Type: Master's thesis
Year: 2011
Downloads: 107
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Abstract

Objective: Vascular endothelial growth factor (Vascular endothelial growth factor, VEGF), also known as vascular permeability factor (Vascular permeability factor, VPF), mainly composed of keratinocytes (keratinocyte, KC) produced the strongest effect is found in the specificity highest angiogenic factor, with angiogenesis, vascular permeability increasing activity of vascular endothelial cell specific. VEGF mainly VEGF121, VEGF145, VEGF148, VEGF165, VEGF165b, VEGF183, VEGF189, VEGF206, 8 subtypes. Normal adult skin dermis of VEGF has maintained the original microvessel density and the maintenance of vascular permeability and other transport nutrients needed stability in their physiological role. Many inflammatory skin diseases have a common pathological features: papillary dermis microvessels, dilation and wall thickening. There are Japanese researchers found that, atopic dermatitis lesions, vascular endothelial growth factor levels than non-lesional skin of healthy people, and significantly higher subtype analysis showed mainly VEGF 121 and VEGF 165, VEGF therefore consider 121 and VEGF 165, and the pathogenesis of atopic dermatitis, but the mechanism is unknown. This study was designed to study different subtypes of vascular endothelial growth factor (VEGF) is a kind of biological by activating signal transduction pathways and cause biological effects, and further clarify the vascular endothelial growth factor and the development of atopic dermatitis in relation to different bit dermatitis clinical prevention and treatment of a new target to provide a theoretical basis for the final states of atopic dermatitis and other diseases and the development of molecular mechanisms. Methods: a cultured human umbilical vein endothelial cells. 2 VEGF165 or VEGF121 on endothelial cells were stimulated cells after different time for recycling. 3 Western blot Western Blot method on vascular endothelial cells FAK signaling pathway in the phosphorylation of FAK FAK and relative quantification, analysis of its phosphorylation of FAK expression changed. 4 The same Western blot Western Blot method, Akt signal transduction pathways Akt and phosphorylated Akt were detected, analysis of phosphorylated Akt expression and its changes. 5 using statistical software SPSS13.0 was used for statistical analysis. Results: 1 VEGF121 and VEGF165 promote human umbilical vein endothelial cells FAK phosphorylation Analysis 1.1VEGF121 promote human umbilical vein endothelial cells FAK phosphorylation analysis capabilities VEGF121 stimulated by vascular endothelial cells, phosphorylation of FAK FAK the percentage of the total stimulus within 30 minutes of the time-dependent changes, increased rapidly over time, five minutes was 0.893 ± 0.048,30 minutes when peaked at 1.192 ± 0.151,60 minute level has decreased to 0.966 ± 0.069, between different stimuli phase phosphorylation of FAK significant differences exist between the percentage, P lt; 0.05. 1.2VEGF165 promoting human umbilical vein endothelial cells FAK phosphorylation analysis capabilities VEGF165 stimulated by vascular endothelial cells, phosphorylation of FAK FAK percentage of the total 10 minutes in stimulating within a time-dependent changes, increased gradually with time, five minutes was 0.469 ± 0.042,10 minute peak 1.015 ± 0.120, between the phases of different stimulation of FAK phosphorylation significant differences exist between the percentage, P lt ; 0.05. 1.3VEGF121 and VEGF165 promote human umbilical vein endothelial cells FAK phosphorylation capacity VEGF121 and VEGF165 compared to the comparative analysis, the percentage of phosphorylated FAK trend was statistically significant difference (P lt; 0.05), stimulated by VEGF121 endothelial cells rapidly phosphorylated FAK, and higher levels of FAK phosphorylation. 2 VEGF121 and VEGF165 promote human umbilical vein endothelial cells Akt phosphorylation Analysis 2.1 VEGF121 promote human umbilical vein endothelial cells capable of phosphorylation of Akt VEGF121 stimulated by vascular endothelial cells, phosphorylated Akt percentage of total Akt changed significantly over time changes in the statistical analysis phase between different stimuli percentage of phosphorylated Akt were significantly different, P lt; 0.05. 2.2 VEGF165 promote human umbilical vein endothelial cells capable of phosphorylation of Akt VEGF165 stimulated by vascular endothelial cells, phosphorylation the percentage of total Akt Akt significant changes over time, the statistical analysis phase between different stimuli percentage of phosphorylated Akt were significantly different, P lt; 0.05. 2.3VEGF121 and VEGF165 promote human umbilical vein endothelial cells Akt phosphorylation capacity Comparative analysis of VEGF121 and VEGF165 compared to the percentage of phosphorylated Akt trend was no statistically significant difference (P gt; 0.05). (Above conclusions satisfy covariance matrix symmetry conditions, P gt; 0.05) Conclusion: 1 VEGF 121 and VEGF 165 can activate FAK signaling pathway, but the role of VEGF 121 compared with VEGF165 significantly and rapidly; 2 VEGF 121 and VEGF 165 can activation of Akt signaling pathway, but both these were no significant differences in terms of capacity; 3 combined with previous research results: VEGF 121 reason than VEGF165 easily cause increased vascular permeability, is likely to promote it through FAK and Src signal strengths Protein phosphorylation pathway achieved.

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CLC: > Medicine, health > Dermatology and Venereology > Dermatology > Allergic and toxic skin diseases
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