基底刚度对登革病毒2型感染人原代脐静脉内皮细胞影响的初步分析-免疫学专业论文.docxVIP

万方数据 万方数据 【关键词】 基底刚度；登革病毒；人原代脐静脉内皮细胞；增殖；细胞因子。 III Study on effect of substrate stiffness on the primary human umbilical vein endothelial cells infected with dengue virus serotype 2 Major: immunology Postgraduate:FangFang Yu Supervisor:Professor Li Zuo [Abstract] Objective: To investigate the effects of substrate stiffness on dengue virus infected with human umbilical vein endothelial cells（HUVEC). Methods: Polyacrylamide gels were prepared for cell culture[(0±4)Kpa]. The cell proliferation on different substrate stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazo lium,inner salt(MTS) assay. The cycle and apoptosis of HUVEC were determined by flow cytometry analysis. Dengue virus serotype 2(DENV-2) strains were propagated and identified by conventional methods.Then by comparing the cell proliferation seeded on the traditional plastic basement with hydrogel after dengue virus infection by MTS assay at an MOI of 10. The method of nitric acid reductase was used to measure the concentration of nitric oxide and expression of ET-1 by double antibody sandwich method ELISA. Results:1.The primary HUVEC was isolated from healthy neonate umbilical cords. Adherent cells were spindle type , growth well and clear outline, arranged in the pebbles, The expression of factor Ⅷ was high and the expression of CD31 was up to 98.14% which was detected by Flow Cytometry. They were all the characteristics of HUVEC. 2.Young’s modulus E value of the hydrogels was 3030±144Pa. 3.The cell proliferation rate , the level of NO and ET-1 ,but not cell cycle and cell apoptosis along with substrate stiffness increase.. Conclusion: The stiffness of substrate affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection. The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury. The establishment of hydrogel substrat