The Anti-angiogenic Effect Of Photochemotherapy, All-trans Retinoid And Angelica In Vitro

Part 1 The anti-angiogenic effect of photochemotherapy in vitro Objective Photochemotherapy has been widely used in the treatment of psoriasis,howeverthe mechanism of which has not been completely elucidated. Psoriasis is nowregarded as an angiogenesis-related inflammatory disease. Recent studies indicatedthat the inhibition of angiogenesis by photochemotherapy could be an underlyingmechanism. It was found that photochemotherapy can downregulate the expression ofpro-angiogenic factors in keratinocytes. However, the direct effects of PUVA onendothelial cells have not been studied. The objective of this study is to investigate theeffect of photochemotherapy on endothelial cells and elucidate the anti-angiogenicmechanism of it. Methods 1. MTT assay and cell cycle analysi were applied to determine the effect of photochemotherapy on the proliferation of endothelial cells through s. 2. The migration assay and in vitro tube formation assay were used to investigate the migration properties and tube formation ability of human microvascular endothelial cells after been treated with UVA and psoralen. 3. Apoptosis of endothelial cells elicited by photochemotherapy was analyzed with Fluorescence-activated cell sorting analysis (FACS). 4. To investigate the effect of photochemotherapy on the expression of angiogenic factors, the expression level of integrin αⅤβ3,gelatinases and angiopoietin-2 mRNA induced by basic fibroblast growth factor(bFGF) and phorbol 12-myristate-13-acetate (PMA) were detected through Semi-quantitative reverse chain reaction. 5. The expression level of integrin αⅤβ3 and angiopoietin-2 protein were determined through FACS and Enzyme-Linked Immunosorbent Assay respectively. While the proteolytic activities of gelatinases were assessed by zymography. Results 1. UVA (0.8-5.0 J/cm2) irradiation with the presence of 8-MOP (300 ng/ml) resulted in a dose dependant reduction on cell viability of endothelial cells. 2. FACS data showed an accumulation of cells in G0/G1 phase of cell cycle and 5博士研究生学位论文 英文摘要 apoptotic features of cell death after UVA irradiation(0.8-5.0 J/cm2) with psoralen. 3. The migration properties and tube formation ability of endothelial cells were dramatically inhibited by photochemotherapy. 4. UVA (2.0-5.0 J/cm2) irradiation with the presence of 8-MOP (300 ng/ml) can downregulated the expression level of integrin αⅤβ3 mRNA and protein. At 5.0 J/cm2, the inhibition ratio of integrin αⅤ mRNA were (23.43±8.64) % (bFGF induced) and (26.73±7.99) % (PMA induced) respectively; the inhibition ratio of integrin β3 mRNA were (31.15±11.25) % (bFGF induced) and (10.75±8.87) %( PMA induced) respectively. The inhibition ratio of integrin αⅤβ3 protein were (38.75±10.22)% (bFGF induced) and (72.21±5.47 )%(PMA induced). 5. Photochemotherapy can downregulated the expression level and activities of gelatinases. At 5.0 J/cm2, the inhibition ratio of gelatinases A mRNA were (25.48±8.90) % (bFGF induced) and (38.28±12.66) % (PMA induced) respectively; the inhibition ratio of gelatinases B mRNA were (32.68±12.31) % (bFGF induced) and (27.36±19.24) %( PMA induced) respectively. The inhibition ratio of gelatinases A activity were (77.50±5.96)% (PMA induced) and (61.57±4.64) % (PMA induced) respectively. 6. Photochemotherapy can downregulated the expression level of Ang-2 mRNA and protein. At 5.0 J/cm2, the inhibition ratio of angiopoietin-2 mRNA were (30.28±10.36) % (PMA induced); the inhibition ratio of angiopoietin-2 protein were (60.14±6.45)% (bFGF induced) and (61.17±4.79) % (PMA ind...