CMKLR1基因敲除对去睾丸小鼠骨质疏松的相关性研究

发布时间：2018-06-23 02:19

本文选题：CMKLR1-ko小鼠 + BMSCs　；参考：《山东中医药大学》2015年硕士论文

【摘要】：目的:1.为了研究CMKLR1基因敲除对雄性小鼠骨髓间充质干细胞(BMSC)的分化能力的影响。2.阐明CMKLR1基因敲除对雄性小鼠体内骨组织代谢的影响。方法:实验主要分为体内实验和体外实验。对于体外实验,我们主要分离培养CMKLR1-ko来源和WT来源的雄性小鼠BMSC,在成骨、成脂诱导剂的作用下观察BMSC的分化能力变化。同时检测相关成骨、成脂基因的表达情况。对于体内实验,我们对CMKLR1-KO和WT来源的雄性小鼠进行去除睾丸去势手术,人为制造骨质疏松模型,探究,在小鼠体内骨质疏松的病理条件下,CMKLR1基因缺失与否对骨代谢调控情况。结果:1.CMKLR1基因敲除型小鼠在生理条件下扫CT示骨质丢失,HE染色发现骨小梁明显减少,骨小梁厚度变细。而且检测血清睾酮含量明显减少;2.CMKLR1基因敲除型小鼠来源的BMSC在成脂分化、成骨分化过程中明显受到抑制。并且下调相关marker基因的表达。结论:1.CMKLR1基因的敲除可以抑制骨髓间充质干细胞(BMSC)的成骨分化,并且可能是通过下调Runx2、ALP等与成骨分化相关的maker基因来实现的。而且CMKLR1基因的缺失,chemerin/CMKLR1信号通路受到抑制,还会妨碍的adiponectin、Fabp4等基因的表达,而对BMSC的成脂分化起到了一定的抑制作用。2.CMKLR1基因的缺失在生理的条件下,雄性小鼠就明显出现骨质丢失,然而当我们对小鼠进行去势手术处理造成骨质疏松的病理条件后,就发现WT型小鼠骨质明显丢失,说明造模是成功的,而且KO型小鼠在已发生骨质疏松的基础上没有明显差异性变化。3.在生理条件下,雄性小鼠CMKLR1基因缺失会导致血清睾酮及其受体的含量明显下降,这说明CMKLR1基因敲除会影响生殖系统分泌雄性激素从而直接影响到骨质代谢。
[Abstract]:Purpose 1. To study the effect of CMKLR1 knockout on the differentiation of bone marrow mesenchymal stem cells (BMSC) in male mice. To elucidate the effect of CMKLR1 knockout on bone metabolism in male mice. Methods: the experiment was mainly divided into in vivo and in vitro experiments. For the in vitro experiment, we mainly isolated and cultured male mice BMSCs from CMKLR1-KO and WT origin, and observed the differentiation ability of BMSC under the action of osteogenesis and lipogenic inducer. At the same time, the expression of adipogenic gene and osteoblasts were detected. For in vivo experiments, we performed orchiectomized male mice from CMKLR1-KO and WT sources, created osteoporosis model artificially, and explored whether CMKLR1 gene deletion or absence of CMKLR1 gene regulated bone metabolism under the pathological condition of osteoporosis in vivo. Results 1. CMKLR1 knockout mice showed bone loss and bone trabecular thickness decreased under physiological conditions. Furthermore, the serum testosterone level was significantly decreased in BMSC derived from CMKLR1 knockout mice during adipogenic differentiation and osteogenic differentiation. And down-regulated the expression of related marker gene. Conclusion: 1. The knockout of CMKLR1 gene can inhibit the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and it may be achieved by down-regulating the maker genes related to osteogenic differentiation, such as Runx2ALP. Moreover, the loss of CMKLR1 gene chemerin / CMKLR1 signaling pathway was inhibited, and the expression of adiponectinine Fabp4 and other genes was inhibited. However, the deletion of CMKLR1 gene inhibited the adiponectinine Fabp4 gene expression in BMSC. 2. The loss of CMKLR1 gene in physiological condition resulted in bone loss in male mice. However, when we treated the mice with ovariectomized conditions resulting in osteoporosis, we found that the bone loss of WT mice was obvious, which indicated that the model was successful. And KO mice had no significant difference on the basis of osteoporosis. 3. 3. Under physiological conditions, the loss of CMKLR1 gene in male mice resulted in a significant decrease in serum testosterone and its receptor levels, which indicated that CMKLR1 gene knockout could affect the secretion of androgen in the reproductive system and thus directly affect bone metabolism.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580

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