汉滩病毒感染神经元细胞模型的建立

摘要

目的　利用体外培养的昆明小鼠胚胎脑皮质神经元，感染汉滩病毒(Hantaan virus.HT-NV)，建立病毒感染的神经元细胞模型，供探讨病毒感染神经元后细胞的变化及损伤机制之用。方法　将孕16～19 d的昆明种小鼠胚胎皮质神经元正常培养存活后，感染汉滩病毒A9株造成细胞损伤模型。使用间接免疫荧光方法　检测神经元荧光抗体阳性情况；利用MTT比色试验检测细胞存活情况；通过免疫组化方法　观察神经细胞生长的情况。结果　病毒感染的神经元在继续培养一周时，间接免疫荧光检测反应阳性；M TT比色试验显示病毒感染组神经元活性明显下降；神经元特异性烯醇化酶(Neuron speciafic enblase.HSE)免疫组化显示病毒感染组神经元胞体面积和突起长度均低于阴性对照组。结论　 HTNV对神经元存在直接作用，使细胞在病毒感染后出现损伤性变化，胚胎鼠皮质神经元是敏感的H TNV靶细胞之一。%Objective To establish a neuron cells model for Hantaan virus (HTNV) infection by using thecultures of cortical neurons of mouse embryo. Changes and mechanism of the damage in neurons after HTNV infec-tion were studied. Methods The cortical neurons of 16-19 day old mouse embryo were cultured and infected withHTNV (A9 strain). Indirect immunofluorescente test(IFT), MTT colorimetric analysis and immunohistochemistryassay (IHCA) were used to detect immunofluorescent antibody reaction of neurons, growth and development of theneurons. Results In neurons of HTNV infected mouse embryo, continually cultured for one week, IFT antigen wasdetected positive, the activity of the infected neurons was reduced obviously and the growth of neurons was ob-structed. Conclusion Direct inflection of HTNV on the cultured neurons were identified, indicating that mouseembryonic critical neurons is one of the sensitive target cells for HTNV infecton.