The Study Of Primary Culture Of Human Umbilical Vein Endothelial Cell And The Modulation With Acitretin

Psoriasis is a common, chronic, recurrent, inflammatory skin disease. Since the morbidity of psoriasis is relatively high and it can recur frequently and last for a long period, especially to the youth, this disease has a very significant impact on the physical and mental health of the patient. Accordingly, psoriasis has become an important area in the field of dermatology. Accompanying the achievement of tumor angiogenesis in the study of cancer research and clinical tumor therapeutics in recent years, interest in other angiogenesis-based disorders is consistently increasing. As one of the classic histological pathology features of psoriatic skin is the microvascular expansion and endothelial cell proliferation in the papillary dermis, the study on the relationship between angiogenesis and psoriasis has become a new direction of its pathogenesis research, and many investigators try to cure psoriasis by way of angiogenesis inhibition. In the study, we primarily cultured human umbilical vein endothelial cell (HUVEC), and investigated the modulation effect of acitretin on endothelial cell proliferation or cell cycle in vitro. Thus we further discussed the possibility of psoriasis treatment through angiogenesis inhibition.Firstly, primary cultured HUVEC and directly observed the cell morphology with light microscopic studies. Then cultures were identified by factor â…§ related antigen (â…§RA) immunohistochemical staining. Secondly, with the MTT method, we investigated the modulation of acitretin to the proliferation of endothelial cell, keratinocyte line HaCaT and cell line ECV-304. Lastly, with the flow cytometricmethod, we measured the variation of acitretin to the cell cycle of endothelial cell and HaCaT.The results show: (1) Primary cultured HUVEC were polygonal, and grew in confluent monolayers with a characteristic "cobblestone" morphology. Staining for VIDRA revealed the intracytoplasmic granular staining pattern characteristic for endothelial cells. (2) HUVEC reacted to acitretin treatment with a double-direction of modulation. Along with the gradually increased concentrations, the endothelial cell proliferation rates were enhanced earlier with a peak effect, and then were inhibited. HaCaT and ECV-304 reacted with an obviously inhibitory effect. In a time-course study, the effect of acitretin on cell proliferation was notable gradually.(3) HUVEC and HaCaT cell cycle modulations to acitretin were different completely. The drug induced Gl-phase arrest of endothelial cell, and S-phase arrest of HaCaT.(4) ECV-304 reacted to acitretin treatment with an inhibitory effect, the same as HaCaT, and differed from endothelial cell.Conclusion: The method of HUVEC primary culture is reliably relatively, and achievable easily. It provides a favorable cell culture method to angiogenesis study in vitro. The double-direction modulation of HUVEC with acitretin suggests that, the pharmacologic activity of acitretin is not only inhibitory of keratinocyte proliferation, but also effect directly on vascular endothelial cell proliferation and cell cycle.