本文選題：SIRT1 + CRABPⅡ�。� 參考：《上海交通大學(xué)》2015年博士論文

【摘要】：維甲酸平衡在胚胎發(fā)育中起著至關(guān)重要的作用,維甲酸缺少和維甲酸過多均會導(dǎo)致先天畸形和發(fā)育缺陷。SIRT1,哺乳動物最保守的NAD+依賴去乙酰化修飾酶,是細(xì)胞代謝感受因子。SIRT1參與調(diào)控一系列細(xì)胞功能,延長哺乳動物壽命,并影響代謝和衰老相關(guān)的疾病。SIRT1全身系統(tǒng)性敲除小鼠導(dǎo)致多器官組織的嚴(yán)重發(fā)育缺陷。但是,SIRT1敲除小鼠發(fā)育缺陷的重要表型的分子機(jī)制還十分不清楚。本研究利用細(xì)胞體內(nèi)同位素標(biāo)記技術(shù)(SILAC)結(jié)合細(xì)胞,SIRT1全敲除轉(zhuǎn)基因小鼠及SIRT1組織特異性敲除小鼠等實(shí)驗(yàn)?zāi)Ｐ?發(fā)現(xiàn)SIRT1參與并調(diào)控細(xì)胞內(nèi)維甲酸信號通路,并通過去乙�；揎椉�(xì)胞內(nèi)維甲酸結(jié)合蛋白Ⅱ(CRABPⅡ)調(diào)節(jié)小鼠胚胎干細(xì)胞分化。過度激活的維甲酸信號通路是小鼠胚胎發(fā)育缺陷的重要原因。具體研究結(jié)果如下:1.SILAC結(jié)合質(zhì)譜分析發(fā)現(xiàn),SIRT1 KO MEF細(xì)胞中CRABP蛋白高度乙�；⒋嬖诙鄠�乙酰化位點(diǎn)。2.CRABPⅡ是SIRT1的去乙酰化修飾底物,維甲酸誘導(dǎo)下SIRT1結(jié)合并去乙酰化修飾CRABPⅡ的K102位點(diǎn)。3.通過去乙酰化修飾CRABPⅡ,SIRT1在細(xì)胞水平和動物水平降低CRABPⅡ的細(xì)胞核轉(zhuǎn)移并抑制維甲酸受體(RAR)的轉(zhuǎn)錄活性,從而下調(diào)維甲酸信號通路。4.SIRT1缺失引起維甲酸信號高度激活,從而導(dǎo)致維甲酸誘導(dǎo)的胚胎干細(xì)胞(m ESCs)分化明顯增強(qiáng),伴隨分化基因誘導(dǎo)增加。5.Microarray進(jìn)一步支持SIRT1在維持胚胎干細(xì)胞多功能干細(xì)胞潛能中非常重要。6.在SIRT1缺失m ESCs中進(jìn)一步沉默CRABPⅡ能部分逆轉(zhuǎn)SIRT1缺失m ESCs的高度分化表型。證明SIRT1通過對CRABPⅡ的調(diào)控來調(diào)節(jié)胚胎干細(xì)胞的分化。7.SIRT1缺失小鼠的多種發(fā)育缺陷,包括小鼠軟骨內(nèi)骨化中心礦化延遲,與其維甲酸信號高度增強(qiáng)呈顯著相關(guān)。本課題研究發(fā)現(xiàn)SIRT1一個嶄新的去乙�；揎椀鞍椎孜顲RABPⅡ,揭示了維甲酸信號通路的嶄新分子機(jī)制,并點(diǎn)亮了SIRT1在胚胎干細(xì)胞分化潛能維持和胚胎發(fā)育中的重要作用。SIRT1是一個對細(xì)胞代謝和環(huán)境信號極其靈敏的關(guān)鍵代謝感受因子,因此這條SIRT1/CRABPⅡ/RAR信號通路將為影響動物發(fā)育的基因-環(huán)境相關(guān)研究提供新的理論基礎(chǔ)和方向。
[Abstract]:Retinoic acid balance plays a crucial role in embryonic development. Retinoic acid deficiency and excessive retinoic acid lead to congenital malformations and developmental defects. SIRT1, the most conservative NAD in mammals, relies on deacetylation modifiers. SIRT1 is involved in regulating a series of cell functions, prolonging the life span of mammals, and affecting the metabolism and senescence related diseases. SIRT1 systemic knockout mice lead to serious developmental defects in multiple organs. However, the molecular mechanism of the important phenotypes of developmental defects in SIRT1 knockout mice is still unclear. In this study, it was found that SIRT1 was involved in and regulated the intracellular retinoic acid signaling pathway by using in vivo isotope labeling technique sil acid) and in combination with the experimental models of total knockout transgenic mice and SIRT1 tissue-specific knockout mice. The differentiation of mouse embryonic stem cells was regulated by deacetylated retinoic acid binding protein 鈪，

本文編號：1853505