高原缺氧對(duì)巨噬細(xì)胞CD14、CD64、MHC-Ⅱ類分子表達(dá)及細(xì)胞因子釋放的影響
發(fā)布時(shí)間:2018-01-15 02:25
本文關(guān)鍵詞:高原缺氧對(duì)巨噬細(xì)胞CD14、CD64、MHC-Ⅱ類分子表達(dá)及細(xì)胞因子釋放的影響 出處:《青海大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 低氧 Mφ 吞噬能力 NO 表面分子 IL-6 TNF-α
【摘要】:目的:通過(guò)建立高原低氧環(huán)境的小鼠模型,探索低氧條件如何影響機(jī)體固有免疫細(xì)胞-巨噬細(xì)胞(Mφ)的吞噬殺傷功能、表面分子表達(dá)以及分泌IL-6、TNF-α功能的影響。方法:分別在400 m,2200 m和4200 m的不同海拔條件下暴露小鼠30d后,(1)用流式細(xì)胞儀檢測(cè)小鼠腹腔Mφ對(duì)FITC標(biāo)記金黃色葡萄球菌的吞噬能力;(2)用雙乙;葻晒馑(DCFH-DA)熒光探針?lè)z測(cè)Mφ呼吸爆發(fā)功能;(3)用ELISA試劑盒檢測(cè)小鼠Mφ培養(yǎng)上清中的NO穩(wěn)定氧化代謝產(chǎn)物亞硝酸鹽(NO2-)水平;(4)用流式細(xì)胞術(shù)(Flow cytometry,FCM)分析法觀察其腹腔Mφ表面分子的變化;(5)用ELISA試劑盒檢測(cè)小鼠腹腔Mφ培養(yǎng)上清中的IL-6、TNF-a的濃度。結(jié)果:海拔4200m和海拔2200m低氧暴露30d后,Mφ吞噬能力、呼吸爆發(fā)能力及NO釋放量降低;小鼠腹腔MφCD14、CD64、TLR4、CD11b、CD18、CD80、MHC-Ⅱ分子的表達(dá)量較對(duì)照組(海拔400 m)均顯著下降(P0.05)。此外,海拔4200 m低氧組與海拔2200 m低氧組相比,小鼠腹腔MφCD14、CD64、TLR4、CD11b、CD18、CD80、MHC-Ⅱ分子的表達(dá)量均明顯下降(P0.05);海拔4200 m和2200 m實(shí)驗(yàn)組小鼠與對(duì)照組(海拔400m)相比,培養(yǎng)上清中的IL-6、TNF-a的濃度MφIL-6、TNF-α的濃度明顯升高(P0.05)。結(jié)論:海拔4200 m和2200 m高原低氧暴露30d后,Mφ的吞噬能力和殺傷能力降低;小鼠腹腔Mφ表面分子表達(dá)水平各自發(fā)生顯著改變;小鼠腹腔Mφ分泌細(xì)胞因子的能力發(fā)生改變。說(shuō)明高原低氧在一定程度上抑制Mφ吞噬殺傷功能,促使Mφ的炎性反應(yīng)及改變Mφ的免疫調(diào)節(jié)能力。
[Abstract]:Objective: to investigate the effects of hypoxia on phagocytosis, expression of surface molecules and secretion of IL-6 in innate immune cell-macrophage M 蠁 by establishing a mouse model of high altitude hypoxia. Effects of TNF- 偽 on the function of mice. Methods: mice were exposed to TNF- 偽 for 30 days at different altitudes of 400 m, 2200m and 4200m, respectively. 1) the phagocytosis of M 蠁 on FITC labeled Staphylococcus aureus was detected by flow cytometry. (2) the respiratory burst function of M 蠁 was detected by diacetyldichlorofluorescein (DCFH-DA) fluorescence probe method. (3) ELISA kit was used to detect the no stable oxidation metabolite (nitrite) in the culture supernatant of M 蠁 of mice. (4) flow cytometric (FCM) analysis was used to observe the changes of M 蠁 surface molecules in abdominal cavity. ELISA kit was used to detect the concentration of IL-6 and TNF-a in mouse peritoneal M 蠁 culture supernatant. Results: after exposure to hypoxia at altitude of 4200m and 2200m for 30 days. M 蠁 phagocytosis, respiratory burst and no release decreased. In mice, M 蠁 CD14, CD64, TLR4, CD11b, CD18, CD80. The expression of MHC- 鈪,
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