柯萨奇病毒B组3型感染胰岛细胞的体外研究

目的 观察柯萨奇病毒B组3型(Coxsackie virus B3,CVB3)对体外培养胰岛细胞的感染情况,初步探讨CVB3损伤胰岛细胞的机制.方法 分离并培养骨髓间充质干细胞,用尼克酰胺和β-巯基乙醇进行胰岛细胞诱导并鉴定.用CVB3感染胰岛细胞,RT-PCR检测胰岛细胞内的CVB3特异性片段.结果 骨髓间充质干细胞经药物诱导后,呈半悬浮状并聚集成用.双硫腙特异性染色后细胞变成红棕色,RT-PCR也证明诱导细胞内具有表达胰岛素的mRNA.CVB3感染胰岛细胞,细胞出现皱缩,折光性下降等典型病变,RT-PCR法成功扩增出胰岛细胞内的CVB3特异性片段.结论 CVB3可以在体外直接损伤胰岛细胞,最终引起胰岛细胞的裂解.

In vitro study on Coxsackie virus B3 infection on islet cells

Objective To study the effect of Coxsackie virus B3 (CVB3) infection on islet cells in vitro, and to explore the mechanism of islet cells caused by CVB3. Methods Bone marrow mesenchymal stem cells( BMSCs) were separated from the bone marrow and cultured. Then they were induced to differentiate into islet-like cells using nicotinamide and mercaptoethanol. Differentiated cells were detected by morphology , special staining and RT-PCR. Observe CVB3 infection on islet cells under inverse microscope and detect the specific gene fragment by RT-PCR. Results BMSCs showed half suspended shape and gathered to form a cluster after induction. Cells became red brown by dithizone specific staining. RT-PCR also proved the existence of mRNA expressing insulin. Infected islet cells appeared typical pathological changes like shrinks, refraction decreases. RT-PCR detected the desired specific gene fragment of 299 bp in infected islet cells. Conclusion CVB3 can directly injury islet cells, and damage the function of islet cells of secreting insulin.