CXCR4抑制剂AMD3100对2型登革热病毒诱导血管内皮细胞株Eahy926凋亡的影响

目的: 探讨趋化因子受体CXCR4抑制剂AMD3100对2型登革热病毒(DV2)诱导人脐静脉血管内皮细胞株 Eahy926凋亡的影响。方法: 免疫组织化学法检测Eahy926细胞的Ⅷ因子。Eahy926细胞分成未感染组和DV2感染组,流式细胞术检测两组细胞不同时点(24 h、36 h、48 h和60 h)CXCR4的表达水平。流式细胞术分析未感染组、DV2感染组及DV2+AMD3100组不同时点的细胞凋亡率。免疫荧光法检测细胞膜表面磷脂酰丝氨酸(PS)。结果: Eahy926细胞有Ⅷ因子表达。在DV2感染Eahy926后的4个时点中,CXCR4的表达均有上调,其中以48 h感染组最明显(66.13%±10.30%,P<0.05)。DV2感染能诱导Eahy926细胞凋亡,其中36 h感染组凋亡率出现高峰(29.85%±15.78%,P<0.05)。应用AMD3100后在各时点均能上调DV2感染组的凋亡率,免疫荧光观察到DV2感染组及DV2+AMD3100组绿色荧光标记的细胞增多。结论: DV2感染能诱导血管内皮细胞Eahy926凋亡并上调CXCR4的表达,CXCR4抑制剂AMD3100促进DV2诱导Eahy926细胞凋亡的发生。

Effect of AMD3100 on dengue virus type 2-induced apoptosis in Eahy926 cells

AIM: To investigate the role of AMD3100 (an inhibitor of CXCR4) in dengue virus type 2 (DV2)-induced apoptosis in human umbilical vein endothelial cell line Eahy926. METHODS: The expression of factor Ⅷ in Eahy926 cells was examined by immunohistochemistry staining. The cells were divided into untreated group, DV2 infection group and DV2+AMD3100 group. Flow cytometric analysis was used to detect the expression of CXCR4 in Eahy926 cells 24 h, 36 h, 48 h and 60 h after DV2 infection. In addition, the percentage of apoptotic cells was also analyzed by flow cytometry. Immunofluorescence was performed to detect the phosphatidylserine (PS) on the surface of Eahy926 cells.RESULTS: Eahy926 cells were factor Ⅷ-positive. Compared with untreated group, the expression of CXCR4 increased in DV2 infection group, most markedly 48 h after infection (66.13%±10.30%, P<0.05). The percentage of apoptotic Eahy926 cells after DV2 infection was the highest at 36 h (29.85%±15.78%, P<0.05). The percentage of DV2-induced apoptotic cells in DV2+AMD3100 group was higher than that in DV2 infection group. The green fluorescence-labeled cells in DV2 infection group and DV2+AMD3100 group were more than those in untreated group. CONCLUSION: DV2 infection induces apoptosis and increases the expression of CXCR4 in Eahy926 cells. AMD3100, the inhibitor of CXCR4, may be a promoter of apoptosis in Eahy926 cells after DV2 infection.