本文選題：基因芯片 + 腦衰老�。� 參考：《昆明醫(yī)學(xué)院》2010年博士論文

【摘要】： 隨著世界各國(guó)人口老齡化進(jìn)程的到來(lái),研究衰老的機(jī)制,進(jìn)一步防治衰老相關(guān)的疾病已逐漸成為當(dāng)今醫(yī)學(xué)科學(xué)研究新的熱點(diǎn)。為了更深入地進(jìn)行腦衰老及相關(guān)疾病發(fā)生發(fā)展分子機(jī)制的研究,我們利用目前公認(rèn)的腦衰老哺乳動(dòng)物模型——SAMP8(senescence-accelerated mice prone 8,快速老化小鼠-8)——作為研究對(duì)象,利用微點(diǎn)陣技術(shù)(Microarray/Gene Chip/基因芯片),對(duì)12月齡與4月齡SAMP8鼠額葉基因的轉(zhuǎn)錄水平進(jìn)行了對(duì)比研究。 Microarray是將指大量寡核苷酸分子固定于支持物上,然后與標(biāo)記的樣品進(jìn)行雜交,通過(guò)檢測(cè)雜交信號(hào)的強(qiáng)弱進(jìn)而判斷樣品中靶分子數(shù)量的一項(xiàng)新興生物技術(shù)。1995年,Stanford大學(xué)的P Brown實(shí)驗(yàn)室發(fā)明了第一塊以玻璃為載體的基因微點(diǎn)陣芯片,標(biāo)志著基因芯片技術(shù)進(jìn)入了廣泛研究和應(yīng)用的時(shí)期�；蛐酒粡V泛應(yīng)用于高通量基因表達(dá)水平檢測(cè)、基因診斷、藥物篩選及個(gè)性化給藥、新基因發(fā)現(xiàn)等領(lǐng)域,成為21世紀(jì)生物技術(shù)發(fā)展的重要特征。 生物信息學(xué)(Bioinformatics)是20世紀(jì)80年代末隨著人類基因組計(jì)劃(human genome project,HGP)的啟動(dòng)而興起的一門新的邊緣學(xué)科和技術(shù),是分子生物學(xué)與計(jì)算機(jī)科學(xué)的交叉。其任務(wù)之一為發(fā)展有效的信息分析工具,構(gòu)建適合于基因組研究的數(shù)據(jù)庫(kù),搜集、管理和使用人類基因組和模式生物基因組的巨量信息。迄今已發(fā)展完善了GenBank核酸序列、SWISS-PROT蛋白質(zhì)序列和PDB生物大分子結(jié)構(gòu)等著名的數(shù)據(jù)庫(kù),開(kāi)發(fā)了如Blast、NEBcutter、Primer-3等上百種生物信息分析軟件。生物信息學(xué)就如同一個(gè)向?qū)?為科研工作者提供了一個(gè)嶄新的工具,幫助科研人員從紛繁復(fù)雜生物信息中找尋真理。 同源基因通常指不同物種中起源于同一祖先的基因,在遺傳學(xué)中,同源這一概念主要是指序列同源。直向同源序列(Orthologs)被認(rèn)為在不同的物種中具有相近甚至相同的功能、相似的調(diào)控途徑,扮演相似甚至相同的角色,而且,絕大多數(shù)核心生物功能就是由相當(dāng)數(shù)量的直向同源基因所承擔(dān)。因此,研究模式生物中與人同源基因的功能對(duì)揭示人體各種生理過(guò)程及疾病發(fā)生發(fā)展分子機(jī)制具有重要意義。 本研究應(yīng)用基因芯片技術(shù)檢測(cè)12月齡和4月齡SAMP8鼠額葉基因表達(dá)譜,篩選到65個(gè)明顯上調(diào)基因和632個(gè)明顯下調(diào)基因。上調(diào)基因多為促炎癥、應(yīng)激反應(yīng)相關(guān)基因;而下調(diào)基因則多為突觸可塑性、囊泡運(yùn)輸、線粒體功能、蛋白質(zhì)和DNA修復(fù)、神經(jīng)激素等相關(guān)基因。進(jìn)一步經(jīng)生物信息學(xué)分析及RT-PCR、WesternBlot驗(yàn)證,證實(shí)了4個(gè)基因——Gnaq(Guanine Nucleotide Binding Protein-alpha qPolypeptide)、KIF1b(Kinesin Family Member 1b)、Sort1(sortilin-1)、Sst(somatostatin)——是腦衰老相關(guān)人鼠同源基因。這4個(gè)基因隨衰老呈明顯降低的mRNA轉(zhuǎn)錄水平和蛋白表達(dá)水平。這些發(fā)現(xiàn)將有助于更好地在模型動(dòng)物中研究腦衰老及相關(guān)疾病發(fā)生的分子機(jī)制。 基于上述發(fā)現(xiàn),我們進(jìn)一步研究抗腦衰老藥物對(duì)腦衰老相關(guān)人鼠同源基因表達(dá)的影響及這些基因在腦衰老過(guò)程中的功能。首先我們研究了天麻素對(duì)SAMP8鼠Sst表達(dá)水平及相關(guān)衰老表征的影響。結(jié)果顯示天麻素能明顯上調(diào)SAMP8鼠額葉內(nèi)Sst的轉(zhuǎn)錄水平和蛋白表達(dá)水平,并明顯改善SAMP8鼠的學(xué)習(xí)記憶能力和其它衰老相關(guān)表征。這些結(jié)果提示天麻素治療可能通過(guò)增進(jìn)額葉Sst的轉(zhuǎn)錄和表達(dá)水平以延緩SAMP8鼠的腦衰老進(jìn)程,各種原因所致的Sst表達(dá)水平降低可能是導(dǎo)致腦衰老及相關(guān)疾病的分子機(jī)制之一。Gnaq、KIF1b及Sort1的功能研究正在進(jìn)行之中。
[Abstract]:With the coming of the aging process of population in the world, the study of the mechanism of aging and the further prevention and control of aging related diseases have gradually become a new hot spot in the research of medical science. In order to further study the molecular mechanism of brain senescence and related diseases, we use the recognized model of brain senescence mammal. - SAMP8 (senescence-accelerated mice prone 8, fast aging mouse -8) - as an object of study, using microarray technology (Microarray/Gene Chip/ gene chip), the transcriptional level of the frontal lobe genes of 12 month old and 4 month old SAMP8 mice was compared.
Microarray is a new biotechnology that is used to immobilize a large number of oligonucleotide molecules on the support and then hybridize with the labeled samples to determine the number of target molecules in the sample by detecting the strength of the hybridization signals in.1995. The P Brown Laboratory of the University of Stanford invented the first gene microdot matrix with glass as the carrier. Gene chip technology has entered a period of extensive research and application. Gene chip has been widely used in high throughput gene expression level detection, gene diagnosis, drug screening and individualization, new gene discovery and so on. It has become an important feature of the development of biotechnology in twenty-first Century.
Bioinformatics (Bioinformatics) is a new frontier discipline and technology that began with the start of the human genome project (HGP) at the end of the 1980s. It is the intersection of molecular biology and computer science. One of its tasks is to develop effective information analysis tools to build data suitable for genome research. Library, collecting, collecting, managing and using the huge information of the human genome and model biological genome. So far, the GenBank nucleic acid sequence, the SWISS-PROT protein sequence and the PDB biological macromolecular structure have been developed, and a hundred kinds of bioinformatics analysis software, such as Blast, NEBcutter, Primer-3, etc. have been developed. A guide provides a new tool for researchers to help researchers find truth from complex biological information.
The homologous gene usually refers to the gene originating from the same ancestor in different species. In genetics, the concept of homologous is mainly the sequence homology. The direct homologous sequence (Orthologs) is considered to have similar or even identical functions in different species, similar to the regulatory pathway, acting similar or even the same role, and the vast majority of the nuclei. The biological function of the heart is assumed by a considerable number of direct homologous genes. Therefore, it is of great significance to study the function of the homologous genes in model organisms to reveal the physiological processes of the human body and the molecular mechanism of the development of the disease.
In this study, gene chip technology was used to detect the gene expression profiles in the frontal lobe of 12 month old and 4 month old SAMP8 mice, and 65 obviously up-regulated genes and 632 down regulated genes were screened. The up regulation was mostly inflammatory and stress related genes, while the down regulated genes were mostly synaptic plasticity, vesicle transport, mitochondrial function, protein and DNA repair, and nerve repair. 4 genes, Gnaq (Guanine Nucleotide Binding Protein-alpha qPolypeptide), KIF1b (Kinesin Family Member), were confirmed by bioinformatics analysis and RT-PCR, WesternBlot verification. These genes are the homologous genes of brain senescence related human mice. These 4 genes are associated with decline. The mRNA transcriptional level and protein expression levels are obviously reduced, and these findings will help to better study the molecular mechanisms of brain senescence and related diseases in model animals.
Based on the above findings, we further studied the effects of anti brain aging drugs on the expression of homologous genes in brain senescence related human mice and the functions of these genes in the process of brain senescence. First, we studied the effect of gastrodin on the expression level of Sst and related senescence in SAMP8 rats. The results showed that gastrodin could significantly increase the Sst in the frontal lobe of SAMP8 rats. The transcriptional level and protein expression level, and obviously improve the learning and memory ability of SAMP8 mice and other aging related characterization. These results suggest that gastrodin therapy may delay the brain aging process in SAMP8 rats by enhancing the transcription and expression level of the frontal Sst, and the decrease of Sst expression caused by various causes may lead to brain senescence and One of the molecular mechanisms of related diseases is the functional study of.Gnaq, KIF1b and Sort1.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2010
【分類號(hào)】：R339.3

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