本文選題：Hmgn + 子宮��； 參考：《吉林大學(xué)》2016年博士論文

【摘要】：子宮蛻膜化是基質(zhì)細胞大量增殖與分化的過程,對胚胎著床的正常進行以及妊娠的維持是必要的。全基因組微陣列結(jié)果分析顯示,在蛻膜化過程中發(fā)現(xiàn)一些基因表達的上調(diào)或下調(diào)。目前大量的研究結(jié)果顯示,一些結(jié)構(gòu)蛋白如高遷移率族核小體結(jié)合蛋白(Hmgn)家族分子通過調(diào)控染色體結(jié)構(gòu)的改變,從而影響一些基因的表達。Hmgn家族分子主要包括5個成員,分別是Hmgn1、Hmgn2、Hmgn3、Hmgn4和Hmgn5,可特異性地與核小體的核心微粒結(jié)合,通過降低染色質(zhì)的壓縮效應(yīng)改變?nèi)旧|(zhì)結(jié)構(gòu),從而調(diào)節(jié)各種DNA依賴性的生理活動,如基因轉(zhuǎn)錄、復(fù)制和DNA修復(fù)等。盡管大量的研究證明,Hmgn參與基因表達調(diào)控、細胞分化和發(fā)育等過程,但關(guān)于其在小鼠子宮蛻膜化過程中的作用及調(diào)控機制尚未見報道。本研究利用原位雜交、熒光定量PCR、基因過表達、RNA干擾等方法研究Hmgn家族分子在小鼠著床期子宮中的表達、作用及調(diào)控機制。結(jié)果發(fā)現(xiàn),Hmgn1 mRNA在早期妊娠第5天子宮胚泡周圍的子宮基質(zhì)細胞及第6-8天子宮蛻膜區(qū)高表達。在人工誘導(dǎo)蛻膜化和體外誘導(dǎo)蛻膜化模型中,Hmgn1 mRNA也高表達在蛻膜化基質(zhì)細胞中。Hmgn1可誘導(dǎo)子宮基質(zhì)細胞的增殖及Ccna1、Ccnb1、Ccnb2和Cdk1的表達。過表達Hmgn1可增加蛻膜化標(biāo)志分子Prl8a2和Prl3c1的表達,而抑制Hmgn1的表達可顯著減少蛻膜化標(biāo)志分子的表達。進一步研究發(fā)現(xiàn),Hmgn1可介導(dǎo)C/EBPβ對蛻膜化基質(zhì)細胞中Prl8a2和Prl3c1表達的調(diào)節(jié)。在子宮基質(zhì)細胞中,c AMP通過C/EBPβ調(diào)控Hmgn1的表達。干擾Hmgn1可減弱c AMP對子宮基質(zhì)細胞中蛻膜化標(biāo)志分子表達的上調(diào)效應(yīng)。在體外蛻膜化過程中,Hmgn1可能作為C/EBPβ的下游靶基因調(diào)控Cox-2、m PGES-1和Vegf的表達。在卵巢摘除小鼠子宮及體外分離的子宮上皮細胞和基質(zhì)細胞中,孕酮可上調(diào)Hmgn1的表達。敲除C/EBPβ可減弱孕酮對子宮基質(zhì)細胞中Hmgn1表達的上調(diào)效應(yīng)。Hmgn2 mRNA在早期妊娠第2-5天小鼠子宮的腔上皮、腺上皮和基質(zhì)細胞中表達,在妊娠6-8天,Hmgn2 mRNA廣泛表達在子宮蛻膜區(qū),在體內(nèi)人工誘導(dǎo)蛻膜化子宮的蛻膜化基質(zhì)細胞中也可觀察到顯著上調(diào)的Hmgn2 mRNA信號。Hmgn2可能對子宮基質(zhì)細胞分化起重要作用,但對細胞增殖未見影響。在子宮基質(zhì)細胞中,孕酮可誘導(dǎo)Hmgn2的表達明顯升高,Hmgn2可介導(dǎo)孕酮對蛻膜化標(biāo)志分子Prl8a2和Prl3c1表達的影響。Hmgn2可調(diào)控子宮基質(zhì)細胞中蛻膜化基因Hand2的表達,抑制Hmgn2的表達可阻礙孕酮對Hand2表達的調(diào)節(jié)。雌激素可上調(diào)Hmgn2在子宮內(nèi)膜上皮細胞中的表達,而添加雌激素受體抑制劑ICI 182,780可減緩這種誘導(dǎo)效應(yīng)。Hmgn3 mRNA在小鼠子宮蛻膜組織和蛻膜化的基質(zhì)細胞有高水平的表達。在子宮基質(zhì)細胞及蛻膜化基質(zhì)細胞中,過表達Hmgn3的可變體Hmgn3a或Hmgn3b可增強蛻膜化標(biāo)志分子Prl8a2和Prl3c1的表達,而抑制Hmgn3可減少蛻膜化標(biāo)志分子的表達。Hmgn3可介導(dǎo)Hoxa10和c AMP調(diào)控Prl8a2和Prl3c1的表達。進一步研究發(fā)現(xiàn),Hmgn3通過影響Hand2的表達調(diào)控蛻膜化過程。在卵巢摘除小鼠子宮及子宮內(nèi)膜上皮和基質(zhì)細胞中,孕酮可誘導(dǎo)Hmgn3的表達。添加Hoxa10 si RNA可減輕孕酮和c AMP對Hmgn3表達的誘導(dǎo)作用,干擾Hmgn3可減緩孕酮、c AMP和Hoxa10對子宮基質(zhì)細胞中Hand2表達的調(diào)控作用。Hmgn5 mRNA在早期妊娠第5天子宮胚泡周圍的基質(zhì)細胞中高表達,且特異性地表達在第6-8天子宮和人工誘導(dǎo)蛻膜化子宮的系膜側(cè)蛻膜區(qū),同時體外誘導(dǎo)蛻膜化模型中也存在高表達的Hmgn5 mRNA。在體外蛻膜化過程中,Hmgn5可通過調(diào)控Ccnd3和Cdk4的表達影響子宮基質(zhì)細胞增殖,且Hmgn5也可調(diào)節(jié)子宮基質(zhì)細胞蛻膜化標(biāo)志分子的表達。Hmgn5可介導(dǎo)Hoxa10調(diào)控Prl8a2和Prl3c1的表達,c AMP可通過Hoxa10調(diào)控子宮基質(zhì)細胞中Hmgn5的表達,而減少Hmgn5的表達可損壞c AMP對子宮基質(zhì)細胞中蛻膜化標(biāo)志分子的上調(diào)效應(yīng)。Hmgn5可能作為Hoxa10的下游靶基因調(diào)控Cox-2、Vegf和Mmp2在蛻膜化過程中的表達。孕酮可刺激Hmgn5在子宮基質(zhì)細胞和上皮細胞中的表達顯著升高,且Hoxa10可介導(dǎo)孕酮對Hmgn5在子宮基質(zhì)細胞中表達的調(diào)控。綜上所述,Hmgn家族分子可能在小鼠子宮蛻膜化過程中起重要的調(diào)節(jié)作用。
[Abstract]:Decidua of the uterus is a process of proliferation and differentiation of stromal cells. It is necessary to carry out the normal implantation of the embryo and the maintenance of pregnancy. The result of the whole genome microarray analysis shows that some genes are up or down in the process of deciduating. A large number of research results show that some structural proteins such as high mobility groups are found. The nucleosome binding protein (Hmgn) family molecule regulates the changes in the structure of chromosomes, thereby affecting the expression of some genes, which include 5 members of the.Hmgn family, namely, Hmgn1, Hmgn2, Hmgn3, Hmgn4 and Hmgn5, which are specifically associated with the core particles of the nucleosome, and change the chromatin structure by reducing the compression effect of chromatin. In order to regulate various DNA dependent physiological activities, such as gene transcription, replication and DNA repair, although a large number of studies have shown that Hmgn is involved in the process of gene expression regulation, cell differentiation and development, but the role and regulatory mechanism of it in the process of uterine deciduating in mice has not yet been reported. In this study, in situ hybridization, fluorescence quantitative PC R, gene overexpression, RNA interference and other methods were used to study the expression, role and regulation mechanism of Hmgn family in the uterus of mice during the implantation period. The results showed that Hmgn1 mRNA was highly expressed in the uterus stromal cells around the uterine blastocyst and the 6-8 day decidua in the fifth day of early pregnancy. In artificial decidua and in vitro deciduate model, Hmg N1 mRNA also highly expressed that.Hmgn1 could induce the proliferation of uterine stromal cells and the expression of Ccna1, Ccnb1, Ccnb2 and Cdk1 in decidua matrix cells. Over expression of Hmgn1 increased the expression of Prl8a2 and Prl3c1 of the deciduating marker molecules, while inhibition of Hmgn1 expression could significantly reduce the expression of deciduating markers. Further research found that Hmgn1 can mediate C/EBP beta regulates the expression of Prl8a2 and Prl3c1 in deciduated matrix cells. In uterine stromal cells, C AMP regulates the expression of Hmgn1 through C/EBP beta. Interference Hmgn1 can weaken the up-regulated effect of C AMP on the expression of deciduating markers in uterine stromal cells. In the process of deciduating in vitro, Hmgn1 may be regulated as a downstream target gene of C/EBP beta. The expression of X-2, m PGES-1 and Vegf. Progesterone up-regulated the expression of Hmgn1 in uterine epithelial cells and stromal cells isolated from the uterus and in vitro. Knockout C/EBP beta can weaken the up-regulated effect of progesterone on Hmgn1 expression in uterine stromal cells.Hmgn2 mRNA in the uterine cavity epithelium, gland epithelium and matrix of mouse uterus on the 2-5 day of early pregnancy. The expression of Hmgn2 mRNA is widely expressed in the decidua region of the uterus on the 6-8 day of pregnancy. In the deciduated matrix cells of the human deciduated uterus, the significantly up-regulated Hmgn2 mRNA signal.Hmgn2 may play an important role in the differentiation of uterine stromal cells, but it has no effect on the cell proliferation. In uterine stromal cells, progesterone is in the uterus. The expression of inducible Hmgn2, Hmgn2 can mediate the effect of progesterone on the expression of decidua markers Prl8a2 and Prl3c1,.Hmgn2 can regulate the expression of deciduating gene Hand2 in uterine stromal cells and inhibit the expression of Hmgn2, which inhibits the regulation of progesterone on Hand2 expression. The addition of estrogen receptor inhibitor ICI 182780 reduced the induction effect of.Hmgn3 mRNA to a high level of expression in the murine decidua and decidua matrix cells. In uterine stromal cells and deciduate stromal cells, the variant Hmgn3a or Hmgn3b that overexpressed Hmgn3 could increase the table of the deciduate markers Prl8a2 and Prl3c1. Inhibition of Hmgn3 can reduce the expression of deciduating marker molecules.Hmgn3 mediates the regulation of Hoxa10 and C AMP to regulate the expression of Prl8a2 and Prl3c1. Further studies have found that Hmgn3 can regulate the deciduating process by affecting the expression of Hand2. Progesterone can induce the expression of Hmgn3 in the ovary and endometrium epithelium and matrix cells of the ovariectomized mice. 10 Si RNA can reduce the induction of progesterone and C AMP on Hmgn3 expression, interfering Hmgn3 can slow the regulation of progesterone, C AMP and Hoxa10 on the expression of Hand2 in uterine stromal cells..Hmgn5 mRNA is highly expressed in the stromal cells around the uterine blastocyst on the fifth day of early pregnancy, and is specifically expressed in the uterus and induced decidua on day 6-8. There is also a high expression of Hmgn5 mRNA. in the deciduating model of the mesuial lateral decidua of the uterus. Hmgn5 can regulate the proliferation of uterine stromal cells by regulating the expression of Ccnd3 and Cdk4, and Hmgn5 also regulates the expression of the uterine stroma cell deciduating markers..Hmgn5 can mediate Hoxa10 to regulate Prl8a2 and Pr. The expression of l3c1, C AMP can regulate the expression of Hmgn5 in uterine stromal cells by Hoxa10, and the decrease of Hmgn5 expression can damage the up-regulated effect of C AMP on the deciduate markers in uterine stromal cells..Hmgn5 may be used as the downstream target gene of Hoxa10, which may regulate the Cox-2, Vegf, and the expression in the process of decidua. The expression in stromal cells and epithelial cells is significantly elevated, and Hoxa10 can mediate the regulation of progesterone on the expression of Hmgn5 in uterine stromal cells. To sum up, Hmgn family molecules may play an important role in the process of uterine deciduating in mice.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S814
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本文編號：2108789