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Gpr84基因敲除小鼠的构建及对巨噬细胞分泌TNF-α功能的影响
发表时间：2017-02-16 浏览量：2603 下载量：538
| 全部作者： | 王绍莹，陈莉莉，殷诚聪，杜冰，钱旻，任华 |
| 作者单位： | 华东师范大学生命科学学院，上海市调控生物学重点实验室 |
| 摘 要： | GPR84在免疫应答中发挥了重要作用。利用CRISPR/Cas9技术构建Gpr84敲除小鼠，分别提取WT小鼠和Gpr84敲除小鼠的骨髓细胞并定向诱导为巨噬细胞。检测在脂多糖（lipopolysaccharides，LPS）刺激下Gpr84-/-巨噬细胞中TNF-α细胞因子的表达变化，从而探讨Gpr84敲除对巨噬细胞功能的影响。结果显示，Gpr84敲除后对骨髓来源并分化的巨噬细胞比例无显著影响。在LPS刺激时，Gpr84敲除小鼠来源的巨噬细胞中TNF-α在mRNA上的表达水平与野生型相比明显降低。由此提示，GPR84能增强巨噬细胞中TNF-α炎症因子的表达从而加重炎症反应。 |
| 关 键 词： | 细胞免疫学；Gpr84；巨噬细胞；CRISPR/Cas9；TNF-α |
| Title： | Establishment of Gpr84 knockout mice and its effect on secretion of TNF-α of macrophage |
| Author： | WANG Shaoying, CHEN Lili, YIN Chengcong, DU Bing, QIAN Min, REN Hua |
| Organization： | Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University |
| Abstract： | GPR84 plays an important role in the immune response. Here we used CRISPR/Cas9 system to build Gpr84 knockout mice. The bone marrow cells of WT mice and Gpr84 knockout mice were extracted respectively to be induced to macrophages. To investigate the effect of Gpr84 knockout on macrophage, we tested the expression of TNF-α in Gpr84-/- macrophages stimulated by lipopolysaccharides (LPS). We found that Gpr84 had no significant effects on the proportion of macrophages differentiated from bone marrow cells. After LPS treatment, mRNA expression levels of TNF-α in macrophages derived from Gpr84 knockout mice were significantly lower than that in the wild type. The results suggest that GPR84 may enhance the mRNA expression of TNF-α in macrophages and regulate the inflammatory response. |
| Key words： | cellular immunology; Gpr84; macrophage; CRISPR/Cas9; TNF-α |
| 发表期数： | 2017年2月第3期 |
| 引用格式： | 王绍莹，陈莉莉，殷诚聪，等. Gpr84基因敲除小鼠的构建及对巨噬细胞分泌TNF-α功能的影响[J]. 中国科技论文在线精品论文，2017，10（3）：242-247. |
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