EMAP-Ⅱ通过LKB1/AMPK/mTOR信号通路增加BTB的通透性  

EMAP-Ⅱ increases the permeability of BTB by LKB1/AMPK/mTOR signal pathway

作　　者：刘啸白[1] 刘丽波[2] 陈方周[3] 刘云会[3] 薛一雪[2] 李振[3] 李志清[2] 商秀丽[4] 

机构地区：[1]中国医科大学,辽宁沈阳110001 [2]中国医科大学基础医学院神经生物教研室,辽宁沈阳110001 [3]中国医科大学附属盛京医院神经外科,辽宁沈阳110004 [4]中国医科大学附属一院神经内科,辽宁沈阳110001

出　　处：《中国药理学通报》2015年第3期351-356,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金(No 81172197,81171131,81100893,81272564,81272795,81372484,81372682,81402573);辽宁省科技计划项目(No 2012225016);沈阳市科学技术计划项目(No F11-264-1-15,F12-277-1-05,F13-318-1-16,F13-318-1-19,F13-220-9-15)

摘　　要：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p LKB1/LKB1、p AMPK/MAPK和p-m TORC1/m TORC1的蛋白表达水平;分别应用LKB1抑制剂radicicol、AMPK抑制剂compound C和m TORC1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-m TORC1/m TORC1的表达水平,以上指标在EMAP-Ⅱ作用1 h时效果最明显;radicicol、compound 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occludin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/m TOR信号通路相关。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 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 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 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.

关 键 词：内皮-单核细胞激活多肽 血肿瘤屏障 胶质瘤 LKB1 AMPK mTORC1 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R341.6[医药卫生—基础医学]