高原低氧环境对小鼠巨噬细胞杀伤活性及分泌IL-6、TNF-α的影响  

Effect of hypobaric hypoxia on killing functions and secreting IL-6,TNF-α of peritoneal macrophages in mice

作　　者：蒲小燕[1] 胡方杰[1] 李积东[1] 刘燕[1] 乔丽娟 许玉珍 刘珺 永胜[1] PU Xiao-Yan;HU Fang-Jie;LI Ji-Dong;LIU Yan;QIAO Li-Juan;XU Yu-Zhen;LIU Jun;YONG Sheng(Qinghai University Medical College,Xining 810001,China)

机构地区：[1]青海大学医学院,西宁810001

出　　处：《中国免疫学杂志》2018年第3期344-348,共5页Chinese Journal of Immunology

基　　金：青海省应用基础研究计划项目(2014-ZJ-715;2015-ZJ-741);国家自然科学基金项目(81060249);教育部"春晖计划"科研基金项目(z2016072)

摘　　要：目的:通过建立高原低氧环境的小鼠模型,研究低氧条件对机体固有免疫细胞巨噬细胞(M_φ)吞噬杀伤功能的影响。方法:分别将小鼠暴露于海拔4 200 m、2 200 m和400 m的不同环境30 d后:(1)流式细胞仪检测小鼠腹腔M_φ对FITC标记的金黄色葡萄球菌的吞噬能力;(2)双乙酰基二氯荧光素(DCFH-DA)荧光探针法检测M_φ的呼吸爆发功能;(3)ELISA法检测小鼠M_φ培养上清中NO稳定氧化代谢产物亚硝酸盐(NO_2^-)的水平;(4)ELISA法检测小鼠M_φ培养上清中的IL-6、TNF-α的释放水平。结果:低氧暴露30 d后,海拔4 200 m和2 200 m的低氧状态下,小鼠腹腔M_φ的吞噬能力、呼吸爆发水平、NO的释放水平明显低于平原对照组(P<0.05),培养细胞上清中的IL-6、TNF-α的浓度明显升高(P<0.05)。结论:低氧暴露30 d可降低M_φ的吞噬能力和氧依赖杀伤功能,升高IL-6、TNF-α的分泌水平,进而影响机体M_φ的固有免疫应答能力。Objective:To explore the influence of high altitude hypoxia on the phagocytosis and killing functions of peritoneal macrophages in mice by establishing mouse model in high altitude hypoxic environment.Methods:①After exposure of mice to an altitude of 4 200 m,2 200 m and 400 m for 30 d respectively,flow cytometry was used to detect the phagocytosis and killing functions of peritoneal macrophages on staphylococcus aureus labeled with FITC.②The respiratory burst level of the cultured macrophage in mice was detected in 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescent probe method.③The concentration of NO2-as a stable oxidative metabolite of NO in the supernatant of the cultured macrophages was measured with ELISA kits;④The release level of IL-6 and TNF-αin the cultured mice macrophage supernatant was also determined with ELISA kits.Results:After exposed under high altitude hypoxia for 30 d,compared with the control group(400 m),the phagocytosis,respiratory burst level and NO release in high altitude groups(4 200 m and 2 200 m)were all than those in the control group(400 m)(P<0.05).While the concentration of IL-6 and TNF-αin the Mφcultured supernatant showed an obvious increase(P<0.05).Conclusion:Exposure under high altitude hypoxia(at altitude of 4 200 m and 2 200 m)for 30 d compromised the phagocytosis and oxygen dependent cytolyticactivity functions,and also raised the cytokines secretion level of IL-6 and TNF-αin Mφ,thereby affecting the innate immune response ability of Mφin body.

关 键 词：低氧 巨噬细胞 吞噬能力 呼吸爆发 一氧化氮 

分 类 号：R392.11[医药卫生—免疫学]