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Sertoli細胞促進大鼠胚胎中腦神經前體細胞分化的作用及其對Sertoli細胞DHH基因的相關研究

發(fā)布時間:2018-03-05 17:40

  本文選題:神經前體細胞 切入點:多巴胺能神經元 出處:《河北醫(yī)科大學》2005年博士論文 論文類型:學位論文


【摘要】:神經前體細胞(Neural Precursor Cells,NPCs)是一種具有自我更新和多向分化能力的原始細胞,在特定因素影響或誘導下能向神經元和膠質細胞分化。深入研究并獲得NPCs對神經系統(tǒng)退行性疾病如帕金森病(Parkinson disease, PD)的細胞移植治療具有應用前景。已有研究發(fā)現(xiàn),移植經誘導分化的多巴胺(Dopamine, DA)能神經元的PD模型大鼠行為學有顯著改善。無論如何,找到穩(wěn)定的體外誘導方法來保證NPCs能夠向DA能神經元分化,即做到人為地操縱細胞分化,是有效地將神經干細胞用于PD臨床細胞治療的關鍵。盡管已經有諸多此類嘗試,但目前尚存在不少問題,包括:①尚未找到成熟的方法來誘導NPCs分化成具有合成特異性遞質能力的神經元; ②尚不清楚決定前體細胞分化的確切機制; ③體內實驗尚不能證明體外實驗已經證實的某些誘導分化因素的作用。 為探索從NPCs誘導出DA能神經元的新途徑,本研究建立了大鼠睪丸Sertoli細胞與大鼠胚胎中腦NPCs的共培養(yǎng)體系,利用Sertoli細胞分泌的各種活性物質來促進NPCs向DA能神經元分化;在此基礎上,本研究又以Sertoli細胞表達的DHH基因為切入點,探討了Sertoli細胞促進前體細胞分化的可能機制。 Sertoli 細胞是睪丸中特有的體細胞,對發(fā)育中的生精細胞提供營養(yǎng)支持和免疫屏障,并能促進精原細胞分化為成熟的精子。Sertoli 細胞具有活躍的分泌功能,能夠分泌100 余種不同的蛋白質,其中許多物質與促進DA 能神經元分化的因子相類似。有實驗證實,將Sertoli 細胞與其他組織來源的細胞共培養(yǎng)可以影響這些細胞的生長、發(fā)育和分化。本研究將Sertoli 細胞和NPCs 共培養(yǎng)后,可以明顯增加NPCs 分化為酪氨酸羥化酶(Tyrosine hydroxylase, TH)陽性DA 能神經元的速度和數(shù)量。 Desert Hedgehog(DHH)是Hedgehog(HH)分泌型信號分子家族的成員之一,該家族除DHH 外,還包括Sonic Hedgehog(SHH)和Indian Hedgehog(IHH)基因,它們分別編碼Shh、Ihh 和Dhh 蛋白。DHH 分布
[Abstract]:Neural Precursor cells (NPCs) are a kind of primordial cells with the ability of self-renewal and multi-differentiation. It can differentiate into neurons and glial cells under the influence or induction of specific factors. Further study and acquisition of NPCs for the treatment of neurodegenerative diseases such as Parkinson disease (PDD) has a promising prospect. The behavior of PD model rats transplanted with differentiated dopaminergic neurons was significantly improved. In any case, a stable in vitro induction method was found to ensure that NPCs could differentiate into DA neurons, that is, to manipulate the differentiation of cells artificially. Is the key to the effective use of neural stem cells in the treatment of PD clinical cells. Although there have been many such attempts, there are still many problems. Including: 1 has not yet found a mature way to induce NPCs to differentiate into neurons with the ability to synthesize specific transmitters; 2 it is not yet clear to determine the exact mechanism of precursor cell differentiation; 3 in vivo experiments have not been able to prove that the in vitro experiment. The role of certain factors in inducing differentiation was confirmed. In order to explore a new way to induce DA neurons from NPCs, a co-culture system between rat testicular Sertoli cells and rat embryonic midbrain NPCs was established, and various active substances secreted by Sertoli cells were used to promote the differentiation of NPCs into DA neurons. On this basis, the possible mechanism of Sertoli cells promoting the differentiation of precursor cells was explored by using the DHH gene expressed in Sertoli cells as the breakthrough point. Sertoli cells are specific somatic cells in the testis, which provide nutritional support and immune barrier for developing spermatogenic cells, and promote the differentiation of spermatogonia into mature spermatozoa. Sertoli cells have active secretory function. It can secrete more than 100 different proteins, many of which are similar to the factors that promote the differentiation of DA neurons. Experiments have shown that co-culture of Sertoli cells with cells from other tissues can affect the growth of these cells. In this study, Sertoli cells co-cultured with NPCs could significantly increase the rate and number of NPCs differentiating into tyrosine hydroxylase (TH3) -positive DA neurons. Desert Hedgehog (Desert) is a member of the secretory signal molecule family of Hedgehog (HH). In addition to DHH, the family also includes Sonic Hedgehog) and Indian Hedgehog (IHH) genes, which encode Shhhh and Dhh protein .DHH distribution, respectively.
【學位授予單位】:河北醫(yī)科大學
【學位級別】:博士
【學位授予年份】:2005
【分類號】:R329

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本文編號:1571249


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