The Interaction Of Vascular Endothelial Growth Factor And P21-activated Kinase-1Signaling In Psoriasis

Part I Expression and correlation of p-PAKl and VEGF in psoriasisObjectives:To detect the expression of p-PAK1and VEGF in psoriasis lesions and analyze correlation between them. To investigate correlation between p-PAK1and PASI score in order to explore the role of PAK1signaling in psoriasis.Methods:The PASI score of every patient was recorded; the protein of p-PAK1and VEGF were detected by immunohistochemistry and Western blotting, Image-Pro Plus6.0and Quantity One were used to make quantitative expression analysis of protein respectively; Pearson’s correlation analysis was utilized to to analyze the correlations between p-PAK1and PASI score, p-PAK1and VEGF.Results:1. The average density and average positive area of p-PAK1and VEGF are higher in psoriasis lesion than those in normal skin (p<0.05).2. The relative protein levels of p-PAKl and VEGF are higher in psoriasis lesion than those in normal skin (p<0.05).3. The expression of p-PAK1is positively correlative with PASI score in patients (p<0.05).4. The expression of p-PAK1in psoriasis lesions is positively correlative with that of VEGF in psoriasis (p<0.05).Conclusions:Overexpression of p-PAK1may contribute to the development of psoriasis, and the correlation of p-PAK1and VEGF may be involved in the pathogenesis of psoriasis. Part Ⅱ Vascular Endothelial Growth Factor Up-regulation via PAK1Signaling RegulatesObjectives:To investigate whether transfection with plasimds pCMV6M-PAK1-T423E and pCMV6M-PAK1-K299R in Hacat cell can regulate the expression of VEGF and further explore the correlation between PAK1signaling and VEGF.Methods:Hacat cells were transfected with pCMV6M-PAKl-T423E and pCMV6M-PAKl-K299R respectively to activate and inhibit PAK1Signaling. The protein of p-PAKl was detected to observe functions of two plasmids by Western Blotting. After successfully transfection, Western Blotting and Elisa assay were utilized to detect VEGF from protein level and Real-Time PCR was used to detect VEGF mRNA from gene level.Results:1The relative protein levels of p-PAK1were significant increased and decreased after Hacat cells were transfected with pCMV6M-PAKl-T423E and pCMV6M-PAKl-K299R respectively (p<0.05).2The expression of VEGF was up-regulated in Hacat cells transfected with pCMV6M-PAK1-T423E (p<0.05).3The expression of VEGF was down-regulated in Hacat cells transfected with pCMV6M-PAKl-K299R (p<0.05).4The max activative effect of pCMV6M-PAK1-T423E appeared in24h after transfection and the max inhibitory effect of pCMV6M-PAKl-K299R appeared30h after transfection.Conclusions:PAK1signaling regulates the expression of VEGF in hacat cell; this may be the key in pathogenesis of psoriasis. Part Ⅲ Vascular Endothelial Growth Factor activate PAK1Signaling and induce hyperproliferation of Hacat cellsObjectives:To investigate whether VEGF can activate PAK1signaling in Hacat cells and to observe the effects of VEGF on bionomics of Hacat cell, such as proliferation and apotosis. Further explore the correlation between PAK1signaling and VEGF in Hacat cell.Methods:After Hacat cells were cultured with VEGF, the relative protein level of p-PAK1was detected by Western Blotting. Besides Hacat cells were treated with VEGF at different doses, Ong/ml,5ng/ml, lOng/ml and25ng/ml respectively, MTT was used to detect the proliferational activity of cells and Flow cytometry was utilized to detect the rate of apoptosis of cells.Results:1The relative protein levels of p-PAK1were significant increased after Hacat cells were treated with VEGF (p<0.05).2VEGF can promote the growth and proliferation activity of Hacat cells in a dose-dependent manner (p<0.05).3VEGF can decrease the rate of apoptosis of Hacat cells in a dose-dependent manner (p<0.05).Conclusions:VEGF can activate PAK1signaling and PAK1signaling can regulate the expression of VEGF in Hacat cell. Autocrine effect of VEGF via PAKil Signaling may be involved in psoriasis.