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EMAP-Ⅱ通过LKB1/AMPK/mTOR信号通路增加BTB的通透性

         

摘要

Aim To investigate the effects of Endothe-lial-monocyte-activating polypeptide Ⅱ ( EMAP-Ⅱ) on the permeability of BTB model in vitro and the pos-sible mechanisms. Methods The BTB model in vitro was established and treated with EMAP-Ⅱ. After that, millipore resistance measurement system was used to n detect the transendothelial resistance ( TEER ) value. The expressing levels of pLKB1/LKB1,pAMPK/MAPK and p-mTORC1/ mTORC1 were assessed by Western blot assay. After pretreatment with LKB1 inhibitor radicicol, AMPK inhibitor compound C and mTORC1 activator IGF 1 respectively , the BTB model in vitro was n treated with EMAPⅡ. The TEER value of BTB model was detected. Western blot assay was used to measure the expressing levels of tight junction associated protein ZO-1 and occludin. Results Compared with EMAP-Ⅱ0 h group, EMAP-Ⅱ decreased the TEER value of BTB model in vitro, increased the expressing levels of p-LKB1/ LKB1 and p-AMPK/AMPK and decreased the levels of p-mTORC1/mTORC1 significantly, and the most obvious change of above indicators appeared at EMAP Ⅱ 1 h. Radicicol, compound C and n IGF1 could partially block the role of EMAP-Ⅱ, caused the increase of the TEER value and the express-ing levels of tight junction associated proteins ZO-1 and occludin. Conclusion EMAP-Ⅱ might increase the permeability of BTB model in vitro, and the activation of LKB1/AMPK/mTOR signal pathway is involved in this process.%目的:研究内皮-单核细胞激活多肽( EMAP-Ⅱ)对体外血肿瘤屏障( BTB)通透性的影响及可能机制。方法建立体外BTB模型, EMAP-Ⅱ处理后, millipore电阻测量系统检测跨内皮细胞阻抗( TEER)值,采用Western blot方法检测pLKB1/LKB1、pAMPK/MAPK 和 p-mTORC1/ mTORC1的蛋白表达水平;分别应用LKB1抑制剂radicicol、AMPK抑制剂compound C和mTORC1激活剂IGF1进行预处理后再给予EMAPⅡ,检测体外BTB模型的TEER值,Western blot方法检测紧密连接相关蛋白ZO-1和occludin的蛋白表达水平。结果与 EMAP-Ⅱ0 h组相比, EMAP-Ⅱ可明显降低体外BTB 模型的 TEER 值,增加 p-LKB1/ LKB1和 p-AMPK/AMPK的表达水平,降低 p-mTORC1/mTORC1的表达水平,以上指标在EMAP-Ⅱ作用1 h时效果最明显;radicicol、com-pound C和IGF1预处理能够部分阻断EMAP-Ⅱ的作用,使体外BTB模型的TEER值及紧密连接相关蛋白ZO-1和occlu-din的表达水平明显增加。结论 EMAP-Ⅱ能明显增加体外BTB模型通透性,其机制可能与激活LKB1/AMPK/mTOR信号通路相关。

著录项

  • 来源
    《中国药理学通报》 |2015年第3期|351-355356|共6页
  • 作者单位

    中国医科大学96期七年制;

    辽宁 沈阳 110001;

    中国医科大学基础医学院神经生物教研室;

    辽宁 沈阳 110001;

    中国医科大学附属盛京医院神经外科;

    辽宁 沈阳 110004;

    中国医科大学附属盛京医院神经外科;

    辽宁 沈阳 110004;

    中国医科大学基础医学院神经生物教研室;

    辽宁 沈阳 110001;

    中国医科大学附属盛京医院神经外科;

    辽宁 沈阳 110004;

    中国医科大学基础医学院神经生物教研室;

    辽宁 沈阳 110001;

    中国医科大学附属一院神经内科;

    辽宁 沈阳 110001;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类
  • 关键词

    内皮 -单核细胞激活多肽; 血肿瘤屏障; 胶质瘤; LKB1; AMPK; mTORC1;

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