| 佟娜仁其木格,邹菊,陈聪,申海艳,刘彦.日本血吸虫外源性短肽干预小鼠巨噬细胞极化.[J].中南医学科学杂志.,2019,(2):120-124, 138. |
| 日本血吸虫外源性短肽干预小鼠巨噬细胞极化 |
| Schistosoma japonicum exogenous short peptide on macrophage polarization in mice |
| 投稿时间：2018-12-20 修订日期：2019-02-26 |
| DOI：10.15972/j.cnki.43-1509/r.2019.02.002 |
| 中文关键词: 日本血吸虫 多肽 巨噬细胞 碱性磷酸酶 |
| 英文关键词:Schistosoma japonicum peptide macrophage alkaline phosphatase |
| 基金项目: |
| 作者 | 单位 | | 佟娜仁其木格 | 南华大学衡阳医学院病原生物学研究所,湖南省特殊病原体防治重点实验室,湖南省分子靶向新药研究合作创新中心,湖南 衡阳 421001 | | 邹菊 | 南华大学衡阳医学院病原生物学研究所,湖南省特殊病原体防治重点实验室,湖南省分子靶向新药研究合作创新中心,湖南 衡阳 421001 | | 陈聪 | 南华大学衡阳医学院病原生物学研究所,湖南省特殊病原体防治重点实验室,湖南省分子靶向新药研究合作创新中心,湖南 衡阳 421001 | | 申海艳 | 南华大学衡阳医学院病原生物学研究所,湖南省特殊病原体防治重点实验室,湖南省分子靶向新药研究合作创新中心,湖南 衡阳 421001 | | 刘彦 | 南华大学衡阳医学院病原生物学研究所,湖南省特殊病原体防治重点实验室,湖南省分子靶向新药研究合作创新中心,湖南 衡阳 421001 |
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| 中文摘要: |
| 日本血吸虫肝硬化与宿主巨噬细胞极化密切相关,本研究为验证多肽干预宿主巨噬细胞极化的分子机制,人工培养小鼠巨噬细胞,分别用单磷酸腺苷、日本血吸虫碱性磷酸酶、外源性短肽处理,ELISA检测各试剂最佳浓度；免疫印迹法检测短肽作用后细胞内一氧化氮合酶以及精氨酸酶1蛋白含量变化；酶联免疫吸附法检测细胞上清液中腺苷浓度的改变,对硝基苯磷酸二钠法检测日本碱性磷酸酶酶活,实时荧光定量PCR法检测细胞上清液中一氧化氮合酶、精氨酸酶1 mRNA的表达变化,硝酸还原酶法测定一氧化氮的浓度,免疫荧光技术鉴别M1和M2型巨噬细胞。结果显示,单磷酸腺苷、碱性磷酸酶及短肽最佳浓度分别为2.5 mmol/L、1.1 mmol/L及0.6 mmol/L；经多肽刺激小鼠巨噬细胞后分泌的一氧化氮合酶的蛋白及mRNA表达均上调,精氨酸酶1表达下调,碱性磷酸酶酶活降低(P<0.01),腺苷受体激活率下降(P<0.01),M1型巨噬细胞增多,M2型巨噬细胞减少。表明多肽通过降低碱性磷酸酶酶活,减少腺苷A合成,从而促进巨噬细胞经典途径激活极化,抑制巨噬细胞替代途径激活极化。 |
| 英文摘要: |
| Schistosoma japonicum cirrhosis is closely related to macrophage polarization. In order to verify the molecular mechanism of ZLW intervention polarization, RAW264.7 cells was artificially cultured, and treated with adenosine monophosphate, Schistosoma japonicum alkaline phosphatase, exogenous short peptide, ELISA to determine the optimal concentration of each reagent；Western Blot was used to detect changes of nitric iNOS and Arg1 protein in cells after treatment with ZLW. ELISA was used to detect changes in adenosine concentration in cell supernatants. The activity of Japanese alkaline phosphatase was detected, and the expression of iNOS and Arg1 mRNA in the supernatant was detected by real-time PCR. The concentration of nitric oxide was determined by nitrate reductase method, immunofluorescence identified M1 and M2 macrophages.The results showed that the optimal concentrations of AMP, SmAP and ZLW were 2.5mmol/L, 1.1mmol/L and 0.6mmol/L, respectively.The expression of iNOS protein and mRNA secreted by ZLW-stimulated mouse macrophages was up-regulated, and the expression of Arg1 was down-regulated, the activity of alkaline phosphatase is decreased(P<0.01), and the activation rate of adenosine receptor is decreased(P<0.01).The classical pathway activates macrophage, and alternative pathways activate macrophage reduction.It is indicated that the ZLW can reduce the activation of adenosine A by decreasing the activity of alkaline phosphatase, thereby promoting the activation of polarization of the macrophage classical pathway and inhibiting the activation of polarization by the macrophage alternative pathway. |
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