Hmgn在小鼠子宫蜕膜化过程中的作用及调节机制

发布时间：2018-07-08 21:28

  本文选题：Hmgn + 子宫　； 参考：《吉林大学》2016年博士论文

【摘要】：子宫蜕膜化是基质细胞大量增殖与分化的过程,对胚胎着床的正常进行以及妊娠的维持是必要的。全基因组微阵列结果分析显示,在蜕膜化过程中发现一些基因表达的上调或下调。目前大量的研究结果显示,一些结构蛋白如高迁移率族核小体结合蛋白(Hmgn)家族分子通过调控染色体结构的改变,从而影响一些基因的表达。Hmgn家族分子主要包括5个成员,分别是Hmgn1、Hmgn2、Hmgn3、Hmgn4和Hmgn5,可特异性地与核小体的核心微粒结合,通过降低染色质的压缩效应改变染色质结构,从而调节各种DNA依赖性的生理活动,如基因转录、复制和DNA修复等。尽管大量的研究证明,Hmgn参与基因表达调控、细胞分化和发育等过程,但关于其在小鼠子宫蜕膜化过程中的作用及调控机制尚未见报道。本研究利用原位杂交、荧光定量PCR、基因过表达、RNA干扰等方法研究Hmgn家族分子在小鼠着床期子宫中的表达、作用及调控机制。结果发现,Hmgn1 mRNA在早期妊娠第5天子宫胚泡周围的子宫基质细胞及第6-8天子宫蜕膜区高表达。在人工诱导蜕膜化和体外诱导蜕膜化模型中,Hmgn1 mRNA也高表达在蜕膜化基质细胞中。Hmgn1可诱导子宫基质细胞的增殖及Ccna1、Ccnb1、Ccnb2和Cdk1的表达。过表达Hmgn1可增加蜕膜化标志分子Prl8a2和Prl3c1的表达,而抑制Hmgn1的表达可显著减少蜕膜化标志分子的表达。进一步研究发现,Hmgn1可介导C/EBPβ对蜕膜化基质细胞中Prl8a2和Prl3c1表达的调节。在子宫基质细胞中,c AMP通过C/EBPβ调控Hmgn1的表达。干扰Hmgn1可减弱c AMP对子宫基质细胞中蜕膜化标志分子表达的上调效应。在体外蜕膜化过程中,Hmgn1可能作为C/EBPβ的下游靶基因调控Cox-2、m PGES-1和Vegf的表达。在卵巢摘除小鼠子宫及体外分离的子宫上皮细胞和基质细胞中,孕酮可上调Hmgn1的表达。敲除C/EBPβ可减弱孕酮对子宫基质细胞中Hmgn1表达的上调效应。Hmgn2 mRNA在早期妊娠第2-5天小鼠子宫的腔上皮、腺上皮和基质细胞中表达,在妊娠6-8天,Hmgn2 mRNA广泛表达在子宫蜕膜区,在体内人工诱导蜕膜化子宫的蜕膜化基质细胞中也可观察到显著上调的Hmgn2 mRNA信号。Hmgn2可能对子宫基质细胞分化起重要作用,但对细胞增殖未见影响。在子宫基质细胞中,孕酮可诱导Hmgn2的表达明显升高,Hmgn2可介导孕酮对蜕膜化标志分子Prl8a2和Prl3c1表达的影响。Hmgn2可调控子宫基质细胞中蜕膜化基因Hand2的表达,抑制Hmgn2的表达可阻碍孕酮对Hand2表达的调节。雌激素可上调Hmgn2在子宫内膜上皮细胞中的表达,而添加雌激素受体抑制剂ICI 182,780可减缓这种诱导效应。Hmgn3 mRNA在小鼠子宫蜕膜组织和蜕膜化的基质细胞有高水平的表达。在子宫基质细胞及蜕膜化基质细胞中,过表达Hmgn3的可变体Hmgn3a或Hmgn3b可增强蜕膜化标志分子Prl8a2和Prl3c1的表达,而抑制Hmgn3可减少蜕膜化标志分子的表达。Hmgn3可介导Hoxa10和c AMP调控Prl8a2和Prl3c1的表达。进一步研究发现,Hmgn3通过影响Hand2的表达调控蜕膜化过程。在卵巢摘除小鼠子宫及子宫内膜上皮和基质细胞中,孕酮可诱导Hmgn3的表达。添加Hoxa10 si RNA可减轻孕酮和c AMP对Hmgn3表达的诱导作用,干扰Hmgn3可减缓孕酮、c AMP和Hoxa10对子宫基质细胞中Hand2表达的调控作用。Hmgn5 mRNA在早期妊娠第5天子宫胚泡周围的基质细胞中高表达,且特异性地表达在第6-8天子宫和人工诱导蜕膜化子宫的系膜侧蜕膜区,同时体外诱导蜕膜化模型中也存在高表达的Hmgn5 mRNA。在体外蜕膜化过程中,Hmgn5可通过调控Ccnd3和Cdk4的表达影响子宫基质细胞增殖,且Hmgn5也可调节子宫基质细胞蜕膜化标志分子的表达。Hmgn5可介导Hoxa10调控Prl8a2和Prl3c1的表达,c AMP可通过Hoxa10调控子宫基质细胞中Hmgn5的表达,而减少Hmgn5的表达可损坏c AMP对子宫基质细胞中蜕膜化标志分子的上调效应。Hmgn5可能作为Hoxa10的下游靶基因调控Cox-2、Vegf和Mmp2在蜕膜化过程中的表达。孕酮可刺激Hmgn5在子宫基质细胞和上皮细胞中的表达显著升高,且Hoxa10可介导孕酮对Hmgn5在子宫基质细胞中表达的调控。综上所述,Hmgn家族分子可能在小鼠子宫蜕膜化过程中起重要的调节作用。
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S814
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本文编号：2108789