解脲脲原体LAMPs通过激活NF-κB诱导小鼠巨噬细胞表达iNOS和凋亡
发布时间：2019-02-22 15:31
【摘要】：目的 研究从解脲脲原体(Uu)提取的脂质相关膜蛋白(LAMPs)在体外诱导小鼠巨噬细胞表达诱导性一氧化氮合酶(iNOS)及产生一氧化氮(NO)的水平,研究该膜蛋白能否激活小鼠巨噬细胞中核因子kappa B(NF-κB)及诱导细胞发生凋亡的情况,以便了解Uu潜在的致病性,为进一步探讨Uu LAMPs诱导小鼠巨噬细胞表达iNOS及诱导巨噬细胞发生凋亡的分子机制提供实验依据。 方法 用从Uu提取的LAMPs刺激小鼠巨噬细胞,用Griess试剂测定经刺激后的小鼠巨噬细胞产生的NO水平,以RT-PCR和Western blot方法分析iNOS的表达水平,用细胞免疫组化、间接免疫荧光检测NF-κB的激活,用Western blot检测核提取物中NF-κB蛋白的表达,且利用RT-PCR和Western blot方法检测NF-κB的特异性抑制剂二硫代氨基甲酸吡咯烷(PDTC)和蛋白酶抑制剂放线菌酮(CHX)对iNOS的表达及对NF-κB激活的影响。用细胞凋亡检测试剂盒检测经Uu LAMPs处理的小鼠巨噬细胞的凋亡情况。 结果 Uu LAMPs能诱导小鼠巨噬细胞表达iNOS,且能以时间和剂量依赖方式刺激小鼠巨噬细胞产生NO,当LAMPs的浓度为3μg/mL时产生的NO量最多,为13.56±0.5μmol/L;但当LAMPs从3μg/mL增加到5μg/mL时,NO产生的量反而减少。另外当LAMPs刺激细胞4h后即可从培养基中检测到NO,而刺激32h后产生的NO量则达到高峰。在用该膜蛋白与NF-κB的抑制剂PDTC或蛋白酶抑制剂
[Abstract]:Objective To study the expression of nitric oxide synthase (iNOS) and the level of nitric oxide (NO) in mouse macrophages induced by Uu in vitro. whether the membrane protein can activate the nuclear factor kappab (NF-B) in the mouse macrophages and induce the apoptosis of the cells in order to understand the potential pathogenicity of the Uu, In order to further study the mechanism of Uu LMPs to induce the expression of iNOS in mouse macrophages and induce the apoptosis of macrophages, the experimental basis was provided. Methods The mouse macrophages were stimulated with LAMs extracted from Uu, and the level of NO produced by the stimulated mouse macrophages was determined by Griess reagent. The expression level of iNOS was analyzed by RT-PCR and Western blot, and the expression level of iNOS was detected by immunohistochemistry and indirect immunofluorescence. The expression of NF-EMAB in the nuclear extract was detected by Western blot and the expression of the specific inhibitor of NF-EMAB by RT-PCR and Western blot was detected by Western blot.-The effect of the activation of the antigen-B. The mice treated with the Uu LAMPs were tested with a cell apoptosis assay kit. Results Uu LMPs can induce the expression of iNOS in mouse macrophages, and can stimulate the macrophage to produce NO in time and dose-dependent manner. When the concentration of LAMs is 3. m from 3 & mu; g/ mL to 5 in mu. g/ ml, the amount of NO was reduced, and no NO was detected from the medium after the LAMs stimulated the cell for 4 h. The amount of NO produced after stimulation of 32h reached a peak. The membrane protein and NF were used.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R375
本文编号：2428358
[Abstract]:Objective To study the expression of nitric oxide synthase (iNOS) and the level of nitric oxide (NO) in mouse macrophages induced by Uu in vitro. whether the membrane protein can activate the nuclear factor kappab (NF-B) in the mouse macrophages and induce the apoptosis of the cells in order to understand the potential pathogenicity of the Uu, In order to further study the mechanism of Uu LMPs to induce the expression of iNOS in mouse macrophages and induce the apoptosis of macrophages, the experimental basis was provided. Methods The mouse macrophages were stimulated with LAMs extracted from Uu, and the level of NO produced by the stimulated mouse macrophages was determined by Griess reagent. The expression level of iNOS was analyzed by RT-PCR and Western blot, and the expression level of iNOS was detected by immunohistochemistry and indirect immunofluorescence. The expression of NF-EMAB in the nuclear extract was detected by Western blot and the expression of the specific inhibitor of NF-EMAB by RT-PCR and Western blot was detected by Western blot.-The effect of the activation of the antigen-B. The mice treated with the Uu LAMPs were tested with a cell apoptosis assay kit. Results Uu LMPs can induce the expression of iNOS in mouse macrophages, and can stimulate the macrophage to produce NO in time and dose-dependent manner. When the concentration of LAMs is 3. m from 3 & mu; g/ mL to 5 in mu. g/ ml, the amount of NO was reduced, and no NO was detected from the medium after the LAMs stimulated the cell for 4 h. The amount of NO produced after stimulation of 32h reached a peak. The membrane protein and NF were used.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R375
【参考文献】
相关期刊论文 前4条
1 范延红,赵连友,郑强荪,薛玉生,杨学东,田建伟,徐琳;核因子κB在精氨酸血管升压素诱导大鼠心肌成纤维细胞一氧化氮合成中的作用[J];生理学报;2003年04期
2 林红,蔡英年;一氧化氮合酶的研究进展[J];生物学通报;1997年05期
3 黄培林,金月玲,朱世能,陆世伦,邓江平;脂肪酸合酶抑制剂对大肠癌细胞NF-κB活性的抑制作用[J];中国肿瘤临床与康复;2000年05期
4 黎庶,汪正清;LPS诱导巨噬细胞凋亡的研究[J];中华微生物学和免疫学杂志;2003年07期
，本文编号：2428358
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2428358.html
最近更新
教材专著
