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| CD38基因敲除对小鼠脾脏B细胞炎性因子产生的影响 |
| 焦会园1,2,陈诚1,2,李玲1,2,戴倩倩1,2,宋矿余1,李蓉1,2* |
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(1. 南昌大学基础医学院微生物学教研室, 南昌 330006;
2. 南昌大学转化医学研究院, 南昌 330031
*通信作者) |
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| 摘要: |
| 目的 分析CD38基因敲除小鼠脾脏中B细胞的数量及B细胞中炎性因子和去乙酰化酶1（SIRT1）的表达水平，探讨CD38基因敲除对B细胞中炎性因子的影响及其潜在机制。方法 采用聚合酶链反应（PCR）检测小鼠尾巴CD38和新霉素（Neo）基因的DNA表达水平；磁珠阴选法分选出野生型（WT）C57BL/6和CD38-/-小鼠脾脏中的B细胞，经流式细胞仪鉴定B细胞分选纯度；实时定量PCR检测CD38和炎性因子肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）基因的mRNA表达水平；蛋白质印迹法检测CD38及SIRT1的蛋白表达水平。结果 鉴定CD38-/-小鼠，并成功分选出WT和CD38-/-小鼠脾脏中的B细胞（纯度>95%）。与WT小鼠相比，CD38-/-小鼠脾脏发育障碍，脾细胞总数及其中的B细胞数量均减少（P<0.01），伴炎性因子TNF-α和IL-1β mRNA表达水平下降（P<0.01），SIRT1蛋白表达水平上升（P<0.05）。结论 CD38基因敲除可引起脾脏B细胞数量减少；并可能通过激活SIRT1通路，抑制脾脏B细胞中炎性因子TNF-α和IL-1β的表达。 |
| 关键词: CD38基因 B淋巴细胞 炎性因子 去乙酰化酶 肿瘤坏死因子α 白介素1β |
| DOI：10.16781/j.0258-879x.2016.11.1373 |
| 投稿时间：2016-08-25修订日期：2016-10-18 |
| 基金项目:国家自然科学基金（81302600）. |
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| Effects of CD38 gene knockout on inflammatory cytokine production in murine spleen B-cells |
| JIAO Hui-yuan1,2,CHEN Cheng1,2,LI Ling1,2,DAI Qian-qian1,2,SONG Kuang-yu1,LI Rong1,2* |
(1. Department of Microbiology, Faculty of Basic Medical Sciences, Nanchang University, Nanchang 330006, Jiangxi, China;
2. Institute of Translational Medicine, Nanchang University, Nanchang 330031, Jiangxi, China
*Corresponding author.) |
| Abstract: |
| Objective To analyze the number of spleen B-cells and the expression of inflammatory factors and sirtuin 1 (SIRT1) in spleen B-cells of CD38-/- mice, so as to explore the effects of CD38 gene knockout on inflammatory factors in B-cells and its potential mechanism. Methods The DNA levels of CD38 and Neo gene in mouse tail tissues were detected by polymerase chain reaction (PCR). Spleen B-cells from wide-type (WT) C57BL/6 and CD38-/- mice were sorted by magnetic activated cell sorting (MACS), and the purity of sorting B-cells were identified by flow cytometry. The mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and CD38 gene were detected by real-time PCR, the protein expressions of CD38 and SIRT1 were detected by Western blotting analysis. Results We confirmed the successful establishment of CD38-/- mice and sorted spleen B-cells from WT and CD38-/- mice (purity>95%). Compared with WT mice, the development of spleen was hampered in the CD38-/- mice, the number of spleen cells and spleen B-cells were significantly reduced (P<0.01), the mRNA levels of TNF-α and IL-1β were significantly decreased (P<0.01), and the expression level of SIRT1 was significantly increased in CD38-/- mice (P<0.05). Conclusion CD38 gene knockout can reduce the number of B-cells in the spleen; and it can inhibit the expression of inflammatory factors (TNF-α and IL-1β) in spleen B-cells by activating SIRT1 pathway. |
| Key words: CD38 gene B-lymphocytes inflammation factors sirtuins tumor necrosis factor-α interleukin-1β |
