γ-干扰素增强RAW264.7巨噬细胞吞噬、杀伤阿萨希毛孢子菌的实验研究

发布时间：2018-06-04 04:29

本文选题：阿萨希毛孢子菌 + 巨噬细胞γ-干扰素　；参考：《安徽医科大学》2012年硕士论文

【摘要】：研究目的 1.研究γ-干扰素对小鼠巨噬细胞株RAW264.7细胞吞噬、杀伤阿萨希毛孢子菌的作用。 2. γ-干扰素对小鼠巨噬细胞株RAW264.7体外抗阿萨希毛孢子菌杀伤机制的影响。研究方法 1.不同浓度的γ-干扰素(IFN-γ)与RAW264.7细胞共孵育18h后，分别与阿萨希毛孢子菌（T.asahii）临床株705共培养45min、4h。45min后在倒置显微镜下观察瑞氏-姬姆萨染色RAW264.7细胞吞噬T.asahii的情况。4h后检测RAW264.7细胞的杀伤活性。 2.不同浓度（10U/ml、100U/ml、1000U/ml）的IFN-γ与RAW264.7共同培养，再加入T.asahii培养后检测菌落形成单位(CFU/ml），计算各浓度组的生长抑制率。在上述体系的上清液中分别检测亚硝酸根离子（NO2-）。将巨噬细胞（Macrophage，Mφ）与不同终浓度的IFN-γ共同孵育后分别检测超氧阴离子(O2-)的OD值。结果 1. IFN-γ呈剂量依赖性地增强RAW264.7细胞吞噬、杀伤T. asahii的活性，各组之间的差异具有显著性意义（P0.01）。 2. IFN-γ浓度分别为10U/ml、100U/ml、1000U/ml时，T.asahii的生长抑制率分别为25.21%、41.95%、55.83%，组间差异具有显著统计学意义（P0.01）。上清液中NO2-的浓度分别为89.16±3.32、108.07±3.81、126.44±3.52，与空白对照组（74.47±1.48）相比差异具有显著性意义(P0.01)。上清液中O2-的OD值分别为0.317±0.006、0.424±0.006、0.518±0.006，，与对照组相比差异具有显著性意义（P0.01）。结论 1.IFN-γ可增强RAW264.7细胞对T.asahii的吞噬、杀伤作用。 2.随着IFN-γ浓度的增加，T.asahii生长抑制率增加。IFN-γ通过产生一氧化氮（NO）及O2-增强巨噬细胞体外抗T.asahii的杀伤活性。
[Abstract]:Research purpose 1. To study the effect of interferon 纬 on phagocytosis of murine macrophage cell line RAW264.7 and the killing of Asaxia sporophyllum. 2. The effect of interferon 纬 on the killing mechanism of murine macrophage cell line RAW264.7. Research method 1. Different concentrations of interferon 纬 (IFN- 纬) were co-incubated with RAW264.7 cells for 18 h. Cultured with T. asahii clinical strain 705 for 45 min, respectively, the killing activity of RAW264.7 cells was observed under inverted microscope. The T.asahii phagocytosis of RAW264.7 cells was detected by Rish-Giemsa staining for 4 h after incubating with T. asahii clinical strain 705 for 45 min. 2. IFN- 纬 was co-cultured with RAW264.7 at different concentrations of 10 U / ml ~ 100 U / ml ~ (-1) U ~ (-1 / ml). Then the colony forming unit (CFU / ml) was detected after T.asahii culture, and the growth inhibition rate of each concentration group was calculated. In the supernatant of the above system, the nitrite ion is determined. Macrophages (M 蠁) were incubated with IFN- 纬 with different final concentrations. The OD values of superoxide anion O _ 2-were determined. Result 1. IFN- 纬 enhanced the phagocytosis of RAW264.7 cells in a dose-dependent manner, killing the activity of T. asahii, and the difference was significant (P 0.01). 2. When the concentration of IFN- 纬 was 10 U / ml, 100 U / ml and 1000 U / ml, respectively, the growth inhibition rate of T. asahii was 25.21% and 41.95%, 55.83%, respectively. The difference between the two groups was statistically significant (P 0.01). The concentration of NO2- in supernatant was 89.16 卤3.32108.07 卤3.81126.44 卤3.52, which was significantly different from that of control group (74.47 卤1.48). The OD values of O _ 2- in supernatant were 0.317 卤0.006, 0.424 卤0.006, 0.518 卤0.006, respectively, which were significantly different from those in the control group (P 0.01). Conclusion 1. IFN- 纬 could enhance the phagocytosis and cytotoxicity of RAW264.7 cells to T.asahii. 2. With the increase of IFN- 纬 concentration, the growth inhibition rate of T. asahii increased. IFN- 纬 increased the killing activity of macrophages against T.asahii by producing nitric oxide (no) and O 2 -.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392

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