高浓度棕酸条件下奶牛子宫内膜细胞基因表达谱榈差异分析

发布时间：2018-03-27 19:18

本文选题：棕榈酸　切入点：子宫内膜细胞　出处：《沈阳农业大学》2017年硕士论文

【摘要】：目的:本研究通过Illumina HiSeq高通量测序技术全面分析高浓度棕榈酸条件下对奶牛子宫内膜细胞功能基因表达差异,从而探究高浓度棕榈酸对奶牛子宫内膜细胞基因表达水平的影响,为进一步研究棕榈酸影响奶牛子宫内膜细胞功能以及探讨奶牛酮病等代谢性疾病与繁殖障碍性疾病之间的关系提供理论基础。方法:本试验首先通过对奶牛子宫内膜细胞体外培养,利用MTT法分别测得棕榈酸在不同浓度下对该细胞相对存活率,筛选棕榈酸添加浓度。之后利用高通量测序的方法,研究棕榈酸影响奶牛子宫内膜细胞基因表达谱差异,筛选出差异表达基因。最后利用荧光定量PCR技术对筛选出来的差异基因进行验证,进一步证实棕榈酸对奶牛子宫内膜细胞基因表达差异的影响。结果:1)成功筛选并鉴定366个显著差异表达基因,涉及差异显著通路202条,得到上调基因188个,下调基因178个。这些基因主要参与炎性细胞因子信号通路、B细胞受体信号通路、HIF-1信号通路、胰岛素分泌等多个生理过程。2)荧光定量PCR验证结果表明,高浓度棕榈酸可显著上调奶牛子宫内膜细胞TGFB1、IL6 基因表达,下调 VAV1、CCL28、BLNK、TGFB3、IGF1R、GPX1、SOD2等基因表达,与高通量测序结果基本一致。表明高浓度棕榈酸可通过改变上述相关功能基因表达水平从而引起奶牛子宫内膜细胞炎症信号与氧化应激等相关功能发生改变。结论:高浓度棕榈酸可引起奶牛子宫内膜细胞炎症反应、氧化应激等途径相关基因表达差异,提示棕榈酸可能是诱发产后奶牛子宫炎性疾病导致奶牛繁殖性能降低的重要因素。
[Abstract]:Objective: to investigate the effect of high concentration palmitic acid on the expression of genes in dairy cow endometrial cells by Illumina HiSeq high-throughput sequencing. In order to further study the effect of palmitic acid on the function of endometrium cells and the relationship between metabolic diseases such as ketosis and reproductive disorders in dairy cows. Endometrial cells were cultured in vitro. MTT method was used to determine the relative survival rate of palmitic acid on the cells at different concentrations and to screen palmitic acid addition concentration. Then the difference of gene expression profiles of dairy cow endometrial cells affected by palmitic acid was studied by high-throughput sequencing. The differentially expressed genes were screened. Finally, the differentially expressed genes were verified by fluorescence quantitative PCR. Results: a total of 366 differentially expressed genes were successfully screened and identified, involving 202 differentially expressed pathways, and 188 up-regulated genes were obtained. These genes are mainly involved in inflammatory cytokine signaling pathway, B cell receptor signaling pathway, HIF-1 signaling pathway, insulin secretion and other physiological processes. High concentration of palmitic acid could significantly up-regulate the expression of TGFB1nil 6 gene in bovine endometrial cells, and down-regulate the expression of VAV1 CCL28 / BLNKT TGFB3IGF1RGGFF1GPX1 / SOD2 gene. The results of high throughput sequencing indicated that high concentration of palmitic acid could change the inflammatory signal of endometrium cells and oxidative stress by changing the expression level of the above related functional genes. Conclusion: high concentration of palmitic acid can induce inflammatory reaction of dairy cow endometrial cells. The difference of gene expression related to oxidative stress suggests that palmitic acid may be an important factor in inducing postpartum inflammation of uterus in dairy cows.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23

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