松針多糖對(duì)小鼠腹腔巨噬細(xì)胞免疫調(diào)節(jié)作用的研究
發(fā)布時(shí)間:2018-03-07 12:04
本文選題:松針多糖 切入點(diǎn):巨噬細(xì)胞 出處:《動(dòng)物營養(yǎng)學(xué)報(bào)》2017年02期 論文類型:期刊論文
【摘要】:本試驗(yàn)旨在研究松針多糖對(duì)正常狀態(tài)及脂多糖(LPS)刺激狀態(tài)下小鼠腹腔巨噬細(xì)胞的免疫調(diào)節(jié)作用。試驗(yàn)采用不同濃度的松針多糖作用于正常的和經(jīng)LPS刺激的小鼠腹腔巨噬細(xì)胞,設(shè)空白對(duì)照組(加入100μL RPMI-1640培養(yǎng)基)、陽性對(duì)照組(加入100μL終濃度為5μg/m L的LPS)、松針多糖組(分別加入100μL 25、50、100、200μg/m L的松針多糖)和LPS+松針多糖組(加入與松針多糖組相同濃度的松針多糖和終濃度為5μg/m L的LPS,液體終體積為200μL)。噻唑藍(lán)(MTT)比色法檢測(cè)細(xì)胞活力,中性紅吞噬試驗(yàn)檢測(cè)巨噬細(xì)胞吞噬能力,Griess法檢測(cè)一氧化氮(NO)的分泌量,酶聯(lián)免疫吸附測(cè)定(ELISA)法檢測(cè)巨噬細(xì)胞培養(yǎng)上清液中白細(xì)胞介素-1β(IL-1β)、腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素-10(IL-10)的分泌量。結(jié)果表明:1)對(duì)空白對(duì)照組相比,各松針多糖組巨噬細(xì)胞相對(duì)增殖率均顯著或極顯著提高(P0.05或P0.01),50、100和200μg/m L松針多糖組巨噬細(xì)胞中性紅吞噬率顯著或極顯著提高(P0.05或P0.01),各LPS+松針多糖組巨噬細(xì)胞中性紅吞噬率均極顯著提高(P0.01),200μg/m L松針多糖組巨噬細(xì)胞NO分泌量極顯著提高(P0.01),50、100和200μg/m L松針多糖組巨噬細(xì)胞TNF-α和IL-1β分泌量顯著或極顯著提高(P0.05或P0.01),50、100和200μg/m L松針多糖組巨噬細(xì)胞IL-10分泌量極顯著降低(P0.01)。2)與陽性對(duì)照組相比,各松針多糖組巨噬細(xì)胞相對(duì)增殖率均極顯著降低(P0.01),100和200μg/m L LPS+松針多糖組巨噬細(xì)胞中性紅吞噬率極顯著升高(P0.01),50、100和200μg/m L LPS+松針多糖組巨噬細(xì)胞NO分泌量極顯著提高(P0.01),各LPS+松針多糖組巨噬細(xì)胞TNF-α分泌量顯著或極顯著提高(P0.05或P0.01),50、100和200μg/m L LPS+松針多糖組巨噬細(xì)胞IL-1β分泌量顯著或極顯著提高(P0.05或P0.01),100和200μg/m L LPS+松針多糖組巨噬細(xì)胞IL-10分泌量極顯著降低(P0.01)。由此可見,松針多糖通過發(fā)揮其促炎作用調(diào)節(jié)巨噬細(xì)胞的免疫功能,進(jìn)而增強(qiáng)機(jī)體抗疾病的能力。
[Abstract]:This experiment was conducted to study the pine polysaccharides on normal and lipopolysaccharide (LPS) stimulation of immune function of peritoneal macrophages in mice under the condition of test. With different concentrations of Polysaccharides from pine needles on normal and stimulated by LPS in mouse peritoneal macrophages, blank control group (adding 100 L RPMI-1640 medium), positive control group (adding 100 L final concentration of 5 g/m L LPS), pine polysaccharide group (100 L 25,50100200 were added to g/m L pine polysaccharide) and LPS+ group (adding polysaccharides from pine needles and pine needle polysaccharides were the same concentration of Polysaccharides from pine needles and a final concentration of 5 g/m L LPS finally, the liquid volume is 200 L). Methyl thiazolyl tetrazolium (MTT) cell vitality assay, neutral red phagocytosis test to detect macrophage phagocytosis, detection of nitric oxide Griess (NO) secretion, enzyme linked immunosorbent assay (ELISA) method for the detection of macrophage cell culture supernatant in white blood cells Cell interleukin -1 beta (IL-1 beta), tumor necrosis factor alpha (TNF- alpha), interleukin -10 (IL-10) secretion. The results show that: 1) compared to the control group, the pine polysaccharide group macrophage relative proliferation rate were significantly increased (P0.05 or P0.01). 50100 and 200 g/m L pine polysaccharide group macrophage neutral red phagocytosis rate significantly higher (P0.05 or P0.01), the LPS+ pine polysaccharide group neutral red phagocytosis rate of macrophages increased significantly (P0.01), 200 g/m L pine polysaccharide group macrophage NO secretion increased significantly (P0.01). 50100 and 200 g/m L pine polysaccharide group macrophage TNF- alpha and IL-1 beta secretion significantly improve (P0.05 or P0.01), 50100 and 200 g/m L pine polysaccharide group significantly decreased the secretion of macrophage IL-10 (P0.01).2) compared with the positive control group, each group of macrophages relative polysaccharides from pine needles the proliferation rate was extremely significant Reduce (P0.01), 100 and 200 g/m L LPS+ pine polysaccharide group neutral red phagocytosis rate of macrophages increased significantly (P0.01), 50100 and 200 g/m L LPS+ pine polysaccharide group macrophage NO secretion increased significantly (P0.01), the LPS+ pine polysaccharide group of macrophage TNF- alpha secretion significantly increased significantly (P0.05 or P0.01), 50100 and 200 g/m L LPS+ pine polysaccharide group IL-1 beta macrophage secretion significantly higher (P0.05 or P0.01), 100 and 200 g/m L LPS+ pine polysaccharide group macrophage IL-10 secretion decreased significantly (P0.01). Thus, polysaccharides from pine needles through its proinflammatory effects regulate macrophage immune function, thereby enhancing the body's ability to resist disease.
【作者單位】: 江西省天然產(chǎn)物與功能食品重點(diǎn)實(shí)驗(yàn)室江西農(nóng)業(yè)大學(xué)食品科學(xué)與工程學(xué)院;江西工業(yè)貿(mào)易職業(yè)技術(shù)學(xué)院糧食工程系;
【基金】:國家科技支撐計(jì)劃(2013BAD10B04-3) 江西省科技廳農(nóng)業(yè)處重點(diǎn)項(xiàng)目(2016BBF60086)
【分類號(hào)】:S816.7
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