神经介素U在体外对猪树突细胞活性及功能的影响

发布时间：2018-02-16 00:00

本文关键词： 神经介素U 树突细胞细胞因子细胞凋亡细胞增殖　出处：《南京农业大学学报》2017年03期 　论文类型：期刊论文

【摘要】：[目的]本文旨在探讨不同浓度神经介素U(NMU)对外周血单核源树突细胞(DC)刺激T淋巴细胞增殖和相关细胞因子分泌的影响。[方法]采集小梅山猪外周血,用集落刺激因子(GM-CSF)和IL-4联合诱导外周血单核细胞形成未成熟树突细胞,再加入脂多糖(LPS)刺激获得成熟的树突细胞,同时观察DC形态。加入不同浓度(0.01、0.1、1、10、100、1 000 nmol·L~(-1))NMU并分别培养2、4、8和12 h后,收集细胞上清液,用ELISA法测定IL-4、IL-5和IL~(-1)3的浓度。加入不同浓度(0.1、1、10、100、1 000 nmol·L~(-1))NMU培养24 h后,收集细胞,用CCK-8试剂盒和细胞凋亡检测试剂盒分别检测DC刺激混合淋巴细胞时细胞的增殖和DC凋亡率。[结果]NMU(0.1~100 nmol·L~(-1))能够抑制未成熟树突细胞(i DC)的细胞凋亡(P0.01),且NMU与NMU+LPS作用效果相一致,10 nmol·L~(-1)NMU组与10 nmol·L~(-1)NMU+LPS组都能降低i DC凋亡,凋亡率分别为2.383%和2.360%,与LPS组相比,NMU+LPS组抑制i DC细胞凋亡效果极显著(P0.01),说明NMU与LPS协同发挥抑制i DC细胞凋亡作用;与对照组相比,0.1~100 nmol·L~(-1)NMU能极显著促进m DC分泌IL-5(P0.01)和抑制IL-4分泌(P0.01)。NMU对IL~(-1)3的影响呈现多样性,低剂量(0.01~0.1 nmol·L~(-1))NMU在2、4 h抑制m DC细胞分泌IL~(-1)3(P0.05),中剂量(1~10 nmol·L~(-1))NMU在2 h先抑制m DC分泌IL~(-1)3(P0.05),4 h后又促进其分泌(P0.05),高剂量(100~1 000 nmol·L~(-1))NMU则促进m DC分泌IL~(-1)3(P0.05)。经NMU诱导后,i DC和m DC均能促进T淋巴细胞增殖(P0.01)。[结论]在一定浓度范围内,NMU能够抑制猪树突细胞的凋亡,并提高其细胞活性和促进树突细胞刺激淋巴细胞增殖的功能,并能影响树突细胞细胞因子分泌,提示神经介素U参与了对猪免疫功能的调节。
[Abstract]:[objective] to investigate the effects of different concentrations of interleukin UNMUs on the proliferation of T lymphocytes and the secretion of related cytokines in peripheral blood mononuclear dendritic cells (DCCs). [methods] Peripheral blood samples from Xiaomeishan pigs were collected. Colony stimulating factor (GM-CSF) and IL-4 were used to induce immature dendritic cells from peripheral blood monocytes, and then lipopolysaccharide (LPS) was added to stimulate the formation of mature dendritic cells. At the same time, the morphology of DC was observed. The supernatant of cells was collected after the addition of 1 000 nmol 路L ~ (-1) nmol 路L ~ (-1) nmol 路L ~ (-1) NMU and cultured for 2 h and 12 h, respectively. The concentrations of IL-4, IL-5 and IL~(-1)3 were measured by ELISA assay, and the cells were collected after 24 h culture with different concentrations of 0.110 nmol 路L ~ (-1) ~ (-1) nmol 路L ~ (-1) NMU. CCK-8 kit and apoptosis kit were used to detect the cell proliferation and DC apoptosis rate when DC stimulated mixed lymphocytes, respectively. [results] NMU(0.1~100 nmol 路L ~ (1 +) could inhibit the apoptosis of immature dendritic cells (P0.01), and NMU acted with NMU LPS. The results showed that both 10 nmol 路L ~ (-1) NMU group and 10 nmol 路L ~ (-1) NMU LPS group could decrease the apoptosis of I DC. The apoptotic rates were 2.383% and 2.360, respectively. Compared with LPS group, NMU LPS could inhibit the apoptosis of I DC cells significantly (P 0.01), which indicated that NMU and LPS played a synergistic role in inhibiting the apoptosis of I DC cells. Compared with the control group, 0.1 nmol 路L ~ (-1) nmol 路L ~ (-1) nmol 路L ~ (-1) NMU significantly promoted the secretion of IL-5 ~ (5) P _ (0.01) and inhibited the secretion of IL-4 (P _ (0.01) ~ (-1) 路N ~ (MU)). The effect of NMU on IL~(-1)3 was diverse. Low dose of 0. 01 nmol 路L ~ (1) nmol 路L ~ (1) nmol 路L ~ (-1) NMU inhibited the secretion of IL ~ (1 +) -1 ~ (3) P0. 05 by MDC cells for 2 h, while at a medium dose of 1 ~ 10 nmol 路L ~ (-1) nmol 路L ~ (-1) NMU at 2 h, it inhibited the secretion of IL ~ (1) -1 3P ~ (0. 05) C ~ (-1) by MDC for 2 h. After 4 h of induction by NMU, both MDC and MDC were induced to promote the secretion of IL ~ (-1) ~ (-1) nmol 路L ~ (1) N ~ (-1) NMU. After induction by NMU, both I DC and M DC could promote the secretion of IL ~ (1 +) -13 ~ (3) nmol 路L ~ (-1) NMU. [conclusion] in a certain concentration range, NMU can inhibit the apoptosis of porcine dendritic cells. It also increased the cell activity and stimulated the proliferation of lymphocytes by dendritic cells, and affected the secretion of cytokines of dendritic cells, suggesting that interleukin-U was involved in the regulation of porcine immune function.
【作者单位】：南京农业大学动物医学院;江苏省徐州市铜山区禽病门诊部;
【基金】：国家自然科学基金项目(31372388)
【分类号】：S828

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