登革病毒Ⅱ型对人脐静脉血管内皮细胞通透性的研究

目的 研究登革病毒Ⅱ型(DENV-2)对人脐静脉血管内皮细胞(HUVEC)通透性的影响.方法 DENV-2病毒滴定,DENV-2接种HUVEC单层细胞,用间接免疫荧光法动态观察DENV-2感染HUVEC的情况(30 min,l、3、6、12、24、30、36、42、48、72 h),实时定量荧光PCR方法检测病毒载量,transwell法检测DENV-2对HUVEC通透性的影响,透射电镜观察细胞超微结构的改变.结果 DENV-2对HUVEC通透性的影响30 min时作用最为明显,其次为42 h时.且病毒载量与HUVEC通透性改变呈正相关.透射电镜结果显示有细胞微绒毛脱落,胞质溶解,部分核膜间隙增宽,甚至是细胞核中也存在裂隙,部分线粒体嵴消失甚至空化,线粒体髓样变,包膜不完整.结论 DENV-2对HUVEC通透性有影响,为探究登革病毒的发病机制提供一定的理论依据.

Abstract：

Objective To research Dengue virus type 2 (DENV-2) to human umbilical vein endothelial cells (HUVEC) permeability. Methods The titration of DENV-2 was detected and HUVEC infection DENV-2 layer of cells. At different time points after infection (30 min, 1 h, 3 h, 6 h, 12 h, 30 h,36 h, 24 h, 42 h, 48 h,72 h), the permeability in HUVEC were detected after Dengue virus infection. The virus load in HUVEC was detected by quantitative real-time PCR. And ultrastructure in HUVEC was observed by electron microscope. Results The permeability in HUVEC was changed after Dengue virus infection by time lasting. The most permeability in HUVEC was changed after Dengue virus infection at 30 min and 42 h. And at the same time the virus load were most value in all of time. The results of the cell membrane were changed by electron microscope. The deciduous cell microvillus and dissd endochylema were founded. And some of nuclear membrane blank was wided by Dengue virus. Even the leakage was founded in cell nucleus. The other change included that disappearance mitochondrial and vacuolization crista, elder pith change mitochondria. In addition, the complete of cell membrane were dissolved. Conclusion The permeability of HUVEC was changed by DENV-2. To explore the pathogenesis of Dengue virus provide certain theoretical basis.

Dynamic changes in the monolayer permeability in HUVEC after Dengue virus type 2 infection

Objective To research Dengue virus type 2 (DENV-2) to human umbilical vein endothelial cells (HUVEC) permeability. Methods The titration of DENV-2 was detected and HUVEC infection DENV-2 layer of cells. At different time points after infection (30 min, 1 h, 3 h, 6 h, 12 h, 30 h,36 h, 24 h, 42 h, 48 h,72 h), the permeability in HUVEC were detected after Dengue virus infection. The virus load in HUVEC was detected by quantitative real-time PCR. And ultrastructure in HUVEC was observed by electron microscope. Results The permeability in HUVEC was changed after Dengue virus infection by time lasting. The most permeability in HUVEC was changed after Dengue virus infection at 30 min and 42 h. And at the same time the virus load were most value in all of time. The results of the cell membrane were changed by electron microscope. The deciduous cell microvillus and dissd endochylema were founded. And some of nuclear membrane blank was wided by Dengue virus. Even the leakage was founded in cell nucleus. The other change included that disappearance mitochondrial and vacuolization crista, elder pith change mitochondria. In addition, the complete of cell membrane were dissolved. Conclusion The permeability of HUVEC was changed by DENV-2. To explore the pathogenesis of Dengue virus provide certain theoretical basis.