【作者】 樊小农；

【导师】 石学敏；

【作者基本信息】 天津中医学院 ， 针灸推拿学， 2003， 博士

【摘要】 老年性痴呆是多基因协同(拮抗)作用的生命过程，发病率逐年上升，尚未找到确切病因。本研究利用小鼠基因表达谱芯片，应用AD替代研究模型SAM，筛选老年痴呆异常表达基因，探讨其在老年痴呆病生理机制中的作用；用Northern杂交研究周期蛋白依赖性蛋白激酶抑制因子P15和钙调神经磷酸酶CaN mRNA表达水平的增龄性变化，了解不同针刺穴区对其表达的影响。 得到如下主要结论： 1．选择2月龄SAMP8作10月龄SAMP8的实验对照组，筛选到差异表达基因234个，其中下调表达的基因有146个，59个基因能在GenBank中登陆；上调表达的基因有88个，87个基因能在GenBank中登陆。 2．选择10月龄SAMR1作10月龄SAMP8的实验对照组，筛选到差异表达基因有242个，其中下调表达的基因有173个，62个基因能在GenBank中登陆；上调表达的基因有69个，68个基因能在GenBank中登陆。 3．选择2月龄SAMP8和10月龄SAMR1同时作10月龄SAMP8的实验对照组，筛选到差异表达基因110个，其中下调表达的基因有79个，30个基因能在GenBank中登陆；上调表达的基因有31个，并全部能在GenBank中登陆。下调涉及突触囊泡释放、信号传导、细胞骨架、能量代谢等；上调涉及细胞周期，离子通道，蛋白质合成，炎症反应等病生理功能。 4．与主导衰老的P16属同一家族基因，P15 mRNA的表达上调使细胞周期调控异常，推测P15可能是导致老年性痴呆的重要因素；CaN基因表达下调，改变了蛋白磷酸化和去磷酸化的动态平衡，由此可能影响信号传导、基因转录等重要的细胞功能活动和加速NFT的形成，从而促进老年痴呆的发生发展。 5．进一步用Northern杂交研究SAMP10脑P15 mRNA的增龄性变化，发现其在6月龄开始表达上调。两针刺组均有下调8月龄SAMP10脑P15 mRNA表达的趋势，其中滋补肝肾组下调P15 mRNA表达的作用有优于醒脑组的趋势。针刺可能通过抑制P15 mRNA的表达，增强DNA损伤修复能力，减少细胞凋亡，延缓衰老速度。 6．用Northern杂交研究SAMP10脑CaN mRNA的增龄性变化，CaN mRNA在6月龄时表达水平最低。两针刺组均有上调8月龄SAMP10脑CaN mRNA表达的趋势。其中醒脑组上调CaN mRNA表达的作用有优于滋补肝肾组的趋势。针刺可能通过增强CaN mRNA的表达，改善了蛋白磷酸化-去磷酸化的平衡状态和水平，影响离子通道天津中医学院博士研究生毕业(学位)论文中文摘要开关的调控，调节基因转录水平，减少NHF的形成，稳定微管结构。不同针刺穴区对不同基因的调整作用不同。更多还原

【Abstract】 Senile dementia is a life process with multiple genes cooperated (antagonistic) actions. Its incidence is increasing rapidly. The definitive etiological factors are not known up to now. In this study, the mice gene expression profiling is used to screen the genes that are abnormally expressed in the senescence-accelerated-mice (SAM) brain samples, which are regarded as a replacing animal model for studying Alzheimer’s disease. The roles of some genes abnormally expressed in the pathological and physiological mechanisms of senile dementia are also discussed. The mRNA expression changes of two genes, cyclin dependent kinase inhibitor 2b (P15) and calcineurin (CaN), with aging in the SAMP 10 were observed by Northern hybridization. The effects of acupuncture playing in different acupoint regions onto the expression levels of the two genes are investigated. The main results are below:1. The 2-month-old SAMP8 is selected as a control group compared with a 10-month-old SAMPS. We screen out 234 genes that are abnormally expressed, among which 146 genes are down-regulated and 88 genes are up-regulated. Among those down-regulated genes, 87 genes are logging in GenBank while among those up-regulated, 59 genes are logging in GenBank.2. The 10-month-old SAMR1 is selected as a control group compared with the 10-month-old SAMPS. We screen out 242 genes are abnormally expressed, among which 173 genes are down-regulated (among which 62 genes are logging in GenBank), while 69 genes are up-regulated (among which 68 genes are logging in GenBank).3. The 2-month-old SAMP8 and a/the 10-month-old SAMR1 are both selected as control groups compared with the 10-month-old SAMP8. We screen out 110 genes that are abnormally expressed, among which 79 genes are down-regulated (among which 30 genes are logging in GenBank), while 31 genes are up-regulated, and all are logging in GenBank. The down-regulated genes are mostly related to the pathological functions of releasing of synaptic vesicle, signal transduction, cell skeleton, energy metabolism, etc. On the other hand, the up-regulated genes are mainly involved in the pathology functions of cell cycle, ion channel, protein synthesis, inflammatory reaction, and so on.4. As a member of the same family with P16, which has been demonstrated as a major gene ofaging, the up-regulated expression of P15 mRNA disorders the modulation of cell cycle. This suggests that P15 could be a crucial factor leading to senile dementia. Thedown-regulation of CaN mRNA might break the dynamic balance of protein phosphorylation and dephosphorylation, through which CaN accelerates the formation of NHF and affects cells’ normal functions such as signal transduction and gene transcription. So it’s suspected that CaN might activate the occurrence and accelerate the development of senile dementia.5. Northern hybridization is used to study the changes of P1 5 mRNA with aging in the brain of SAMP 10. It is detected that the expression of P15 mRNA is up-regulated from the beginning of 6-month-old. Both acupuncture groups have the capacity of helping to down-regulate the abnormal expression of P15 mRNA in 8-month-old SAMP 10. The down-regulation role of ZBGS on P15 mRNA seems to be stronger than that of XNKQ. By depressing the expression of P15 mRNA, acupuncture might have the capacity of repairing the damaged DNA, lessening the cell apoptosis and delaying the speed of aging..6. By using Northern hybridization, the changes of CaN mRNA with aging is studied in the brain of SAMP 10. The expression level of CaN mRNA is the lowest at 6-month-old sample. Both acupuncture groups have the capacity of helping to up-regulate the abnormal expression of CaN mRNA in 8-month-old SAMP 10. The up-regulation role of XNKQ on CaN seems to be stronger than that of ZBGS. By elevating the expression of CaN mRNA. acupuncture might have improved the state and the level of balance that of phosphorylation and dephosphorylation, influenced the switching modulation of the ion channel, reg更多还原