单纯疱疹病毒1型感染对原代培养鼠皮质神经元凋亡caspase-3途径的影响

目的 了解caspases-3在单纯疱疹病毒1型（herpes simplex virus type1,HSV-1)感染原代培养鼠皮质神经元中对线粒体的调控作用以及与神经元凋亡的关系。方法 用酶标仪检测体外培养小鼠皮质细胞正常组、病毒组、山梨醇加病毒组、对照组在不同条件下caspase-3发光底物吸收值，测定caspase-3底物的含量。用流式细胞术结合TUNEL方法检测各组细胞凋亡情况及线粒体膜电位变化。结果 加入底物后14h测定A值：正常组为0.3942、病毒1组为0.2408、山梨醇组为0.5392、病毒加山梨醇组为0.4650，正常组caspase-3含量较阴性对照组增加，病毒组较正常组明显减低，山梨醇组较正常组明显增加，病毒组较正常组明显减低，山梨醇组较正常组明显增加，病毒加山梨醇组较山梨醇组减低。山梨醇及神经酰胺组细胞线粒体膜电位明显降低，而病毒组及病毒加山梨醇或神经酰胺组均无明显降低。结论 正常体外培养的神经元存在细胞凋亡，病毒能够阻止线粒体膜电位的超极化，山梨醇诱导的神经细胞凋亡是依赖于caspase-3的细胞凋亡，HSV-1阻止依赖于caspase-3的细胞凋亡途径。

Effect of caspase-3 on the primary cultured mice cortical neurons with herpes simplex virus type-1 infection

Objective To study the effect of caspase-3 on the primary cultured mice cortical neuron w ith herpes simplex virus type 1 (HSV-1) infection. Methods A spectrophotometer was used to de tect the absorbance units at 405 nm of the caspase-3 substrate,the chrom o phore p-nitroanilide(pNA ).Comparing with the normal group, virus group, s orbital group,and sorbital group with virus in the absorbance units of chromophore substrate.The changes in inner mitochondrial membrane p otential of the different group cells were analyzed with flow cytometry. Results The released chromphore was measured by determining the absorbance at 405 nm. N group is 0.394 2, V group is 0.240 8, NS group is 0.539 2, VS gr oup is 0.465 0,the control C1 group is 0.290 0, C2 group is 0.235 0 and all groups were analyzed by statistic (P<0.05). The absorbance units in the pNA of n ormal samples were higher than negative control and these units in the sorbital group were higher than those in normal group while these units in the virus g r oup were much lower than those in normal group,and the sorbitals with virus wer e lower than those of the sorbital group. The mock-infected cells exposed to s orbitol or ceramide-2 showed a decreased mitochondrial membrane potential comp aring to untreated or HSV-1-infected cells. Conclusion The apoptosis can be found at the normal fetal mice cor tical neurons primary cultured in vitro. HSV-1 infection prevents the depolariz ation of the inner mitochondrial membrane induced by sorbital. The pathways of programmed cell induced by sorbitol is caspase-3-dependent. HSV-1 may have th e functions to block caspase-3.