本文選題：年齡效應(yīng) + 淋巴細(xì)胞��； 參考：《中國(guó)科學(xué)技術(shù)大學(xué)》2009年碩士論文

【摘要】： 衰老是一種遺傳因素和外界環(huán)境共同影響的,由在細(xì)胞水平上發(fā)生的變化逐步積累并最終表現(xiàn)在組織器官水平上的現(xiàn)象。在衰老的過(guò)程中,有許多因素會(huì)誘發(fā)細(xì)胞內(nèi)遺傳物質(zhì)的損傷(基因突變、端粒縮短、細(xì)胞內(nèi)調(diào)控機(jī)制失調(diào)等)。在這些與衰老有關(guān)的遺傳物質(zhì)的損傷中,最為明顯的表現(xiàn)就是在細(xì)胞分裂過(guò)程中染色體分離異常的年齡效應(yīng)以及非整倍體細(xì)胞的年齡效應(yīng)。 在本次研究中,我們采用cytokinesis-block method分別從染色體分離異常的角度以及非整倍體細(xì)胞命運(yùn)的角度探討在中國(guó)男性外周血淋巴細(xì)胞中的年齡效應(yīng): 1.運(yùn)用cytokinesis-block method,我們首次研究并報(bào)道了在中國(guó)男性的外周血淋巴細(xì)胞中染色體分離異常的年齡效應(yīng)。本文收集了14名老年(60-70歲之間)和10名青年(22-26歲之間)中國(guó)男性的外周血,從中分離得到淋巴細(xì)胞并進(jìn)行培養(yǎng),以松胞素B阻滯細(xì)胞質(zhì)分裂而獲得雙核細(xì)胞,用人X、Y和2號(hào)染色體特異的著絲粒探針進(jìn)行熒光原位雜交,以檢測(cè)X和Y染色體的分離異常。發(fā)現(xiàn):老年人淋巴細(xì)胞中,X染色體(老年組9.2±3.2‰,青年組1.1±0.9‰, P0.001)和Y染色體丟失的頻率(老年組2.5±1.9%,年輕組0.2±0.3‰,P0.001)較青年人顯著升高; X染色體(老年組16.5±3.4‰,年輕組3.5±1.1‰,P0.001)和Y染色體不分離的頻率(老年組7.2±2.6‰,年輕組2.4±1.3‰,P0.001)也隨著年齡增加而顯著升高;在老年人和青年人的淋巴細(xì)胞中,無(wú)論X染色體還是Y染色體,其染色體不分離的頻率均顯著高于丟失的頻率;觀(guān)察到的多種染色體分離異常同時(shí)發(fā)生的頻率也顯著高于相應(yīng)的預(yù)期頻率。這些結(jié)果表明,年齡顯著影響中國(guó)男性淋巴細(xì)胞中X和Y染色體的正常分離;與染色體丟失相比,染色體不分離是中國(guó)男性性染色體分離異常的是主要類(lèi)型;細(xì)胞內(nèi)負(fù)責(zé)染色體精確分離的機(jī)制出現(xiàn)異�？赡軙�(huì)同時(shí)引起X和Y染色體的丟失及不分離。 2.我們分析發(fā)生了染色體分離異常的有絲分裂理論上應(yīng)產(chǎn)生的子細(xì)胞,在理論上應(yīng)產(chǎn)生的子細(xì)的群體中Chr X-細(xì)胞在淋巴細(xì)所占比例值為25.7‰,而我們?cè)隗w內(nèi)的外周血淋巴細(xì)胞群體中實(shí)際并未觀(guān)察到Chr X-細(xì)胞,從這一結(jié)果中,我們發(fā)現(xiàn)X染色體的缺失對(duì)男性淋巴細(xì)胞的存活有極大的影響。此外,通過(guò)比較進(jìn)入分裂形成雙核的非整倍體淋巴細(xì)胞以及未進(jìn)入分裂的單核非整倍體淋巴細(xì)胞,我們探討了不同染色體的缺失或者獲得對(duì)淋巴細(xì)胞分裂能力的影響。結(jié)果表明,在老年人的幾種非整倍體外周血淋巴細(xì)胞中,Y染色體缺失的淋巴細(xì)胞(Chr Y-)的分裂能力最強(qiáng)(P0.001,χ2-test),與之不同的是,在年輕人的非整倍體外周血淋巴細(xì)胞中,Y染色體缺失的淋巴細(xì)胞(Chr Y-)的分裂能力與X染色體獲得的淋巴細(xì)胞(Chr X+)相似(P=0.46,χ~2-test),而Y染色體獲得的淋巴細(xì)胞(Chr Y+)的分裂能力最弱(P0.001,χ~2-test)。這一結(jié)果說(shuō)明了在老年人和青年人中Y染色體對(duì)細(xì)胞分裂能力的不同影響,證實(shí)了Y染色體隨年齡增長(zhǎng)對(duì)細(xì)胞的功能和重要性的變化。綜上所述,不同染色體的缺失或者獲得對(duì)細(xì)胞命運(yùn)的影響是不同的,并且這些染色體在年輕人和老年人細(xì)胞中的作用和影響也不同。
[Abstract]:Aging is a combination of genetic factors and external environment, which gradually accumulates at the level of cell and eventually appears at the level of tissue and organ. In the process of aging, there are many factors that induce the damage of genetic material in cells (gene mutation, telomere shortening, and dysregulation of intracellular regulation mechanism, etc.). The most obvious manifestation of the damage to genetic material related to aging is the age effect of abnormal chromosomal separation during cell division and the age effect of aneuploidy.
In this study, we used cytokinesis-block method to investigate the age effect in Chinese male peripheral blood lymphocytes from the angle of chromosomal segregation and the fate of aneuploidy, respectively.
1. using cytokinesis-block method, we first studied and reported the age effect of abnormal chromosomal segregation in the peripheral blood lymphocytes of Chinese men. This article collected the peripheral blood of 14 elderly (60-70 years old) and 10 young Chinese men (22-26 years old). X, Y, and chromosome 2 specific centromere probes were used for fluorescence in situ hybridization to detect the abnormal separation of X and Y chromosomes by blocking the cytoplasmic division. The frequencies of X chromosomes (aged 9.2 + 3.2%, 1.1 + 0.9 per thousand, P0.001) and Y loss were found in the lymphocytes of the elderly (2.5 + 1.9% in the elderly group). The young group was 0.2 + 0.3 per thousand, P0.001) was significantly higher than that of young people, and the frequency of X chromosome (aged 16.5 + 3.4%, 3.5 + 1.1 per 1000, P0.001) and Y chromosome non separation (aged 7.2 + 2.6 per 1000, young group 2.4 + 1.3 per thousand, P0.001) also increased with age; in the lymphocytes of the elderly and young people, no matter X chromosome or Y The frequencies of chromosomes were significantly higher than those of the lost frequency; the frequencies of abnormal chromosomal abnormalities observed at the same time were also significantly higher than the corresponding expected frequencies. These results suggest that age significantly affects the normal separation of X and Y chromosomes in Chinese male lymphocytes; chromosomes are not compared with chromosome loss. Separation is the main type of male sex chromosome segregation in China, and the abnormal mechanism of chromosomal separation in cells may cause the loss and separation of X and Y chromosomes at the same time.
2. we analyzed the subcells in the theory of mitosis that had occurred in chromosomal separation. In theory, the proportion of Chr X- cells in the subfine group was 25.7 per 1000, and we did not observe Chr X- cells in the peripheral blood lymphocyte population in the body. In this result, we found X The loss of chromosomes has a great effect on the survival of male lymphocytes. In addition, we have explored the loss of chromosomes or the effect on the mitosis of the cells by comparing into the division of the diploid aneuploidy lymphocytes and the mononuclear cells that have not entered the division. The Y chromosome missing lymphocyte (Chr Y-) has the strongest splitting capacity in several aneuploidy peripheral blood lymphocytes (P0.001, X 2-test) in several aneuploidy peripheral blood lymphocytes of the year, which is different from that of the Y chromosome missing lymphocyte (Chr Y-) and the X chromosome obtained lymphocyte (Chr X+) in the young's aneuploidy peripheral blood lymphocytes. Similar (P=0.46, X ~2-test), and the Y chromosome acquired lymphocyte (Chr Y+) is the weakest (P0.001, Chi ~2-test). This result illustrates the different effects of Y chromosome on cell division in the elderly and young people, and confirms the changes in the function and importance of the Y chromosome with age increasing. The effects of chromophore deletion or acquisition on cell fate are different, and these chromosomes play different roles in young and elderly cells.
【學(xué)位授予單位】：中國(guó)科學(xué)技術(shù)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：R392.12

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