| 游晓星，马小华，刘良专，曾焱华，朱翠明，何军，蒋传好，余敏君，吴移谋.支原体巨噬细胞活化脂肽-2经PI3K/Nrf2诱导单核细胞表达HO-1和NQO1.[J].中南医学科学杂志.,2014,42(1):6-10. |
| 支原体巨噬细胞活化脂肽-2经PI3K/Nrf2诱导单核细胞表达HO-1和NQO1 |
| Macrophage-activating Lipopeptide-2 Induces the Expression ofHO-1 and NQO1 via PI3K/Nrf2 Pathways in Monocytes |
| 投稿时间：2013-11-06 |
| DOI： |
| 中文关键词: 支原体巨噬细胞活化脂肽-2 血红素氧合酶-1 NAD(P)H∶醌氧化还原酶-1 NF-E2相关因子2 |
| 英文关键词:macrophage-activating lipopeptide-2 hemoxygenase-1 NAD(P)H∶quinone oxidoreductase 1 NF-E2-related factor 2 |
| 基金项目:国家自然科学基金（31000091，81072418）,湖南省科研条件创新专项（2013TT2013）. |
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| 中文摘要: |
| 目的观察支原体巨噬细胞活化脂肽-2（MALP-2）对THP-1单核细胞血红素氧合酶-1（HO-1）和NAD(P)H∶醌氧化还原酶-1（NQO1）表达的影响，以明确机体抵抗支原体感染所致炎性损伤的自我防御机制。方法体外培养THP-1细胞并分为对照组和实验组。其中对照组加入等体积培养基，实验组根据不同的实验目的加入不同浓度的MALP-2作用5～120 min或12 h，Western blot 检测HO-1和NQO1表达以及Akt磷酸化水平。同时，采用PI3K抑制剂LY294002处理细胞，以证实PI3K参与HO-1表达。提取细胞核蛋白，凝胶迁移率实验和免疫荧光观察NF-E2相关因子2（Nrf2）的DNA结合活性和核转位情况，并采用特异性siRNA沉默Nrf2后，Western blot观察其对HO-1和NQO1表达的影响。结果Western blot结果显示，MALP-2能诱导THP-1细胞表达HO-1和NQO1蛋白，且呈一定的剂量依赖性。此外，MALP-2能激活PI3K，且PI3K抑制剂能抑制HO-1和NQO1的表达；凝胶迁移率实验和激光共聚焦结果显示，MALP-2能增强Nrf2的DNA结合活性及核转位，而PI3K抑制剂处理后，Nrf2的DNA结合活性以及核转位水平进一步降低。RNA干扰Nrf2后，HO-1和NQO1表达显著降低。结论MALP-2能诱导THP-1细胞表达HO-1和NQO1，其机制可能受PI3K/Nrf2调控。 |
| 英文摘要: |
| ObjectiveThis study aimed to determine the role of macrophage-activating lipopeptide-2 (MALP-2) on the expression of hemoxygenase-1 (HO-1) and NAD(P)H∶quinone oxidoreductase 1 (NQO1),and then investigate its regulatory mechanism to better understand the self-protecting mechanism upon mycoplasma infection.MethodsTHP-1 cells were cultured in vitro and divided into control group and experimental group.In the control group,equal volume of culture medium was added.For the experimental group,different concentrations of MALP-2 were added for 5～120 min or 12 h,and the expression of HO-1 and NQO1,phosphorylation of Akt were detected by Western blot.Also,PI3K inhibitor LY294002 was used to investigate the role of PI3K in HO-1 expression.Immunofluorescence and electrophoretic mobility shift assay were applied to examine the nuclear translocation and DNA-binding activity of NF-E2-related factor 2 (Nrf2).Silencing of Nrf2 was achieved by RNA interference and expression of HO-1 and NQO1 were detected by Western blot.ResultsMALP-2 induced THP-1 cells production of HO-1 and NQO1 in a dose-dependent manner,as demonstrated by Western blot.In addition,MALP-2 could also activate PI3K,and LY294002 inhibited HO-1 and NQO1 expression.MALP-2 could also induce the translocation of Nrf2 to the nucleus,and increase its DNA-binding activity;but this activity could be inhibited by PI3K inhibitor.Silencing of Nrf2 expression significantly inhibited HO-1 and NQO1 expression.Conclusion |
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