The Hippocampal Proteomics Research Of Cattle Encephalon Glycoside And Ignotin Injection On Transgenic AD Mice

Objective:Alzheimer’s disease(AD) is an insidious onset in progressive development of the nervous system degenerative disease, and correlated with age, the serious influence the quality of life of the elderly, also cause caregiver burden. AD’s pathogenesis and complex affecting factors include gene mutations, epigenetic changes, abnormal protein modification, amyloid deposition, neurotrophic and plasticity change, cell apoptosis, oxidative stress, energy metabolism, cell structure, etc. Pathologic study found that patients or animal models with AD have obvious hippocampal neurons damage, which directly affects the physiological function such as memory and spatial orientation. The hippocampus is the early part of the pathological change in patients with dementia. As the treatment of AD, the current clinical first-line treatment drugs are limited to donnelly and mementine,which can only relieve symptoms and can’t reverse or prevent disease progression. Because of its nerve repair effect, cattle encephalon glycoside and ignotin injection(CEGI) is widely used in all kinds of central and peripheral nervous system traumatic disease. The more commonly used glycosides carnosine treatment for alzheimer’s disease in domestic, and clinical observation has certain curative effect, but its mechanism is not clear.Proteomics was originally defined as proteins expressed by a genome, while the produce proteins of cell organization is dynamic, and easily affected by external environment. So now that protein is a known set of cells in a particular moment including all subtypes and modified protein. Proteomics is the analysis of dynamic change of protein in the cell from the perspective of the overall, including its composition, expression level and modification status, then understanding the interaction between proteins and contact, prompting the protein function and cellular activity.As the pathological changes is relatively single, and the time that produced pathological changes is shorter, transgenic mouse model has become a ideal animal model that research AD’s etiology and pathogenesis and clinical treatment strategies at home and abroad. It can better simulate the AD pathological changes and clinical characteristics.This study compared the normal control group, model group and cerebral glycosides carnosine APPswe/PS1dE9 double transgenic AD after intervention model mice hippocampal tissue and research there differences in proteomics, then determine the expression of different protein and analyze its biological function. It can help us to understand the CEGI on the protective mechanism of brain tissue, then provide theoretical basis and experimental basis for its clinical application.Methods:Select 5 months APPswe/PS1dE9 double transgenic AD model mice, a total of 24 randomized: 12 model group(Tg), 12 cattle encephalon glycoside and ignotin injection intervention group(Tg + CEGI 6.6), and select C57/BL6 J mice 12 months of age with the same genetic background only as control group at the same time. CEGI intervention group was given brain cattle encephalon glycoside and ignotin injection 6.6 mL/kg/d by intraperitoneal injection, model group and the control group given same dose of normal saline. After 30 days, put to death animals respectively, and separate the mice brain hippocampus, then extract protein and dispose them with peptides TMT tags, LC-LTQ-MS/MS analysis and mass spectrum data retrieval. Finally, analyze the biological function of the difference protein.Results:1 We use the combining TMT quantitative LC-MS/MS detection technology, and adopt international popular standard routine analysis of protein expression differences-i.e., the change of more than 1.5 times the amplitude can be trusted. Compared with model group,CEGI intervention group draw a credible differences in protein 122, which include 56 protein up-regulated(Table1)and 66 protein own-regulated(Table2); Compared with normal control group, CEGI intervention group draw a credible differences in protein 143, which include 51 protein up-regulated(Table3) and 92 protein own-regulated(Table4).2 With the GeneCoDis3 analysis of the differences in protein biological processes, we found that the function of CEGI intervention group mice brain hippocampus difference protein mainly relates to biological processes such as the cellular structure to maintain, membrane transport, cell energy metabolism, cell signal transduction, lipid metabolism, protein transport, protein hydrolysis, cell proliferation and apoptosis, inflammation, oxidative stress, immune response and so on.Conclusion:1 The study uses proteomics technology to identify the differentially expressed proteins in hippocampus tissue in APP/PS1 mice and after the intervention of CEGI mice, preliminary builds there proteome database. These proteins are mainly involved in the follow biological processes such as maintaining cell structure, cell membrane transport, cell energy metabolism, cell signal transduction, lipid metabolism, protein transport and hydrolysis, cell proliferation and apoptosis,inflammation, oxidative stress, immune response. These processes closely related to the pathogenesis and the pathological process of neurodegenerative diseases, especially AD, and delve into help us further understand the inherent nature of these diseases.2 Cattle encephalon glycoside and ignotin injection has certain protective effect on APP/PS1 mice with nerve cell damage, its mechanism may be through stable cytoskeletal structure, maintaining synapses stability, slow ganglioside degradation speed, inhibit inflammation and oxidative stress to play a role of the protection of neurons.