Relative Research Between Apoptosis And Alzheimer's Disease-like Patholgical Change Of Cerebral Cortex With HCMV Congential Infection Aged Mouse

Objective The study was on the base of the aged mouse model with HCMV congenital infection. From this model, we observed the pathological changes and apoptosis changes in the cerebral cortex of aged mice with HCMV congenital infection. The study aimed at providing a new idea and appropriate model for studing the etiology and mechanism of Alzheimer's diseases.Methods The study was on the base of the aged mouse model with HCMV congenital infection. After an aged mouse model was built, we breed the mouse in the barrier cage for 18~20 month. 5 mice with HCMV congenital latency infected and 5 mice with HCMV activited were selected randomly.We also selected 5 mice for control group randomly. Congenital HCMV latency infected group mouse were injected 150mg/kg cyclophosphamide to actived viru. We sacrificed the mouse and asepsis fetch the cerebral cortex. 1)We used co-cultivation experiment and polymerase chain reaction (PCR) or RT-PCR to analyze the infection fettle of the mouse.2)We also used electron microscope,FCM and TUNEL to measure the apoptosis in cerebral cortex. 3 )Research the pathology change. Stained the brain slices with hematoxylin-eosin (H.E.) for pathology research;observed the amyloid plaques by Congo red stain ;observed the neurofibrillary tangles and amyloid plaques by silver stain.Results 1) The infection fettle of the mouse. Through RT-PCR, we couldn't detected HCMV IE,UL83 mRNA in HCMV congenital latency infect group.But we could detected HCMV IE, UL83 DNA in HCMV congenital latency infect group through PCR. In HCMV activation mouse ,we could detected HCMV IE, UL83 through PCR and RT-PCR. In the co-cultivation experiment of Cerebral Cortex in mouse with HCMV latency infect group, we observed the CPE in HF, but we couldn't found thous change in control group.These results proved that our model mouse's infection fettle according with device.2)The results of apoptosis. We used electron microscope measure the three group mold mouse found that that the group with congenital latent infected HCMV and active infected HCMV chromosome shrink in nerve cell nucleolus, karyotheca disappear and nucleolus fragment.We also found that the ultrastructures of neuron in Congenital HCMV latency infected group and active infected HCMV group were severely damaged.We used TUNEL measure the three group mold mouse found that the apoptosis cell in control group was 2.0±0.7, the apoptosis cell in congenital latent infected HCMV group was 5.2±1.3, the apoptosis cell in active infected HCMV group was 18.1±3.5 . The results were check up by T test and found that The difference of three group were markedness.We used FCM measure the three group mold mouse found that the apoptosis ratio in control group was 1.75±0.3%.The aged mouse which congenital latent infected HCMV apoptosis ratio was 4.83±0.96%.The aged mouse with active infected HCMV apoptosis ratio was 10.09±1.18%.The results were check up by T test and found that The difference of three group were markedness.3) The result of Pathology change. HE stain found that cerebral cortex in congenital latent infected HCMV group were stained slightly than control group. Neuron stained very slightly by HE stain in the cerebral cortex in active infected mouse group.We also found neuron lost and quantity decreased in active infected mouse group. In control mice, there were no obvious pathological alterations in the cerebral cortex by HE staining. The amyloid plaque exists in the cerebral cortex of infected mice by Congo red stain, The amyloid plaque and abundant neurofibrillary tangles (NFTs) in the cerebral cortexe of Congenital HCMV latency infected group and active infected HCMV group were found by silver stain.Conclusion (1) On the basis of these results, we found HCMV congenital infection may be result in the appearance of AD-like pathologic changes. We may conclusion that HCMV infection may be a risk factor for Alzheimer's disease. It provides a new idea and a technic flat for us to study AD pathogeny and the occurrence mechanism of AD. (2) HCMV congenital latent infection can accelerate apoptosis in cerebral cortex with aged mouse.This could destory nerve center's fuction and induce similarity Alzheimer's patholgical change. This provides us a new evidence to prove congenital HCMV infection could induce Alzheimer's disease.