本文關(guān)鍵詞： 神經(jīng)介素U 樹突細(xì)胞 細(xì)胞因子 細(xì)胞凋亡 細(xì)胞增殖　出處：《南京農(nóng)業(yè)大學(xué)學(xué)報》2017年03期 　論文類型：期刊論文

【摘要】：[目的]本文旨在探討不同濃度神經(jīng)介素U(NMU)對外周血單核源樹突細(xì)胞(DC)刺激T淋巴細(xì)胞增殖和相關(guān)細(xì)胞因子分泌的影響。[方法]采集小梅山豬外周血,用集落刺激因子(GM-CSF)和IL-4聯(lián)合誘導(dǎo)外周血單核細(xì)胞形成未成熟樹突細(xì)胞,再加入脂多糖(LPS)刺激獲得成熟的樹突細(xì)胞,同時觀察DC形態(tài)。加入不同濃度(0.01、0.1、1、10、100、1 000 nmol·L~(-1))NMU并分別培養(yǎng)2、4、8和12 h后,收集細(xì)胞上清液,用ELISA法測定IL-4、IL-5和IL~(-1)3的濃度。加入不同濃度(0.1、1、10、100、1 000 nmol·L~(-1))NMU培養(yǎng)24 h后,收集細(xì)胞,用CCK-8試劑盒和細(xì)胞凋亡檢測試劑盒分別檢測DC刺激混合淋巴細(xì)胞時細(xì)胞的增殖和DC凋亡率。[結(jié)果]NMU(0.1~100 nmol·L~(-1))能夠抑制未成熟樹突細(xì)胞(i DC)的細(xì)胞凋亡(P0.01),且NMU與NMU+LPS作用效果相一致,10 nmol·L~(-1)NMU組與10 nmol·L~(-1)NMU+LPS組都能降低i DC凋亡,凋亡率分別為2.383%和2.360%,與LPS組相比,NMU+LPS組抑制i DC細(xì)胞凋亡效果極顯著(P0.01),說明NMU與LPS協(xié)同發(fā)揮抑制i DC細(xì)胞凋亡作用;與對照組相比,0.1~100 nmol·L~(-1)NMU能極顯著促進(jìn)m DC分泌IL-5(P0.01)和抑制IL-4分泌(P0.01)。NMU對IL~(-1)3的影響呈現(xiàn)多樣性,低劑量(0.01~0.1 nmol·L~(-1))NMU在2、4 h抑制m DC細(xì)胞分泌IL~(-1)3(P0.05),中劑量(1~10 nmol·L~(-1))NMU在2 h先抑制m DC分泌IL~(-1)3(P0.05),4 h后又促進(jìn)其分泌(P0.05),高劑量(100~1 000 nmol·L~(-1))NMU則促進(jìn)m DC分泌IL~(-1)3(P0.05)。經(jīng)NMU誘導(dǎo)后,i DC和m DC均能促進(jìn)T淋巴細(xì)胞增殖(P0.01)。[結(jié)論]在一定濃度范圍內(nèi),NMU能夠抑制豬樹突細(xì)胞的凋亡,并提高其細(xì)胞活性和促進(jìn)樹突細(xì)胞刺激淋巴細(xì)胞增殖的功能,并能影響樹突細(xì)胞細(xì)胞因子分泌,提示神經(jīng)介素U參與了對豬免疫功能的調(diào)節(jié)。
[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.
【作者單位】： 南京農(nóng)業(yè)大學(xué)動物醫(yī)學(xué)院;江蘇省徐州市銅山區(qū)禽病門診部;
【基金】：國家自然科學(xué)基金項(xiàng)目(31372388)
【分類號】：S828

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