水通道蛋白4调控1,2-二氯乙烷致小鼠血脑屏障通透性机制  

Mechanism of aquaporin 4 in regulating 1,2-dichloroethane-induced blood-brain barrier permeability in mice

作　　者：江俊莹 汪波 刘俊 林莉 黄煜基 林茜 梁博萱 钟怡洲 谢植伟[3] 黄振烈[2] 余日安 JIANG Junying;WANG Bo;LIU Jun;LIN Li;HUANG Yuji;LIN Xi;LIANG Boxuan;ZHONG Yizhou;XIE Zhiwei;HUANG Zhenlie;YU Rian(School of Public Health,Guangdong Pharmaceutical University,Guangzhou,Guangdong 510315,China;不详)

机构地区：[1]广东药科大学公共卫生学院,广东广州510315 [2]南方医科大学公共卫生学院,广东广州510515 [3]广东省职业病防治院毒理实验所广东省职业病防治重点实验室,广东广州510300

出　　处：《中国职业医学》2020年第5期519-525,共7页China Occupational Medicine

基　　金：国家自然科学基金(82073519,81872601);广东省自然科学基金(2018A030313068);广东省医学科学技术研究基金(A2017193,B2019056,A2020394);广东省热带病重点实验室(2017B030314035)。

摘　　要：目的探讨水通道蛋白4(AQP4)在1,2-二氯乙烷(1,2-DCE)亚急性吸入暴露对血脑屏障(BBB)通透性的影响。方法将无特定病原体级健康Aqp4基因工程雄性CD-1小鼠[Aqp4野生型(Aqp4+/+)和Aqp4基因敲除(Aqp4-/-)]随机分为对照组和低、中、高剂量组,分别予剂量为0.00、100.00、350.00和700.00 mg/m3的1,2-DCE进行全身动式吸入暴露,6 h/d,连续28 d。染毒结束后,以伊文思蓝染色法检测小鼠脑组织BBB通透性;以实时荧光定量聚合酶链反应法检测小鼠大脑BBB紧密连接蛋白(TJP)相关基因Tjp1、Tjp2、Tjp3、闭合蛋白(Cldn)3、Cldn5、Cldn11、咬合蛋白(Ocln)、基质金属蛋白酶(Mmp)2、Mmp9和钠-钾-氯共转运体1(Nkcc1)的mRNA相对表达水平。结果小鼠BBB通透性在1,2-DCE染毒剂量与Aqp4基因型的交互效应上有统计学意义(P<0.01);其中,低、中、高剂量组Aqp4+/+基因型小鼠BBB通透性均高于同基因型对照组小鼠(P值均<0.05);对照组Aqp4+/+基因型小鼠BBB通透性低于同组Aqp4-/-基因型小鼠(P<0.05),但3个剂量组Aqp4+/+基因型小鼠BBB通透性分别高于同组Aqp4-/-基因型小鼠(P值均<0.05)。小鼠大脑皮质中Cldn3和Ocln的mRNA相对表达水平仅在基因型的主效应上差异有统计学意义(P值均<0.01);其中,Aqp4-/-基因型小鼠大脑皮质中Cldn3和Ocln的mRNA相对表达水平均高于Aqp4+/+基因型小鼠(P值均<0.05)。小鼠大脑皮质中Tjp1、Tjp2、Tjp3、Cldn5、Cldn11、Mmp2、Mmp9和Nkcc1的mRNA相对表达水平在染毒剂量与基因型的主效应及两者交互效应上均无统计学意义(P值均>0.05)。结论 1,2-DCE染毒可导致小鼠BBB通透性增加;其机制可能与1,2-DCE通过下调Aqp4继而引起大脑皮质TJP相关分子Cldn3和Ocln的mRNA表达上调有关。Objective To study the effect of aquaporin 4(AQP4) in regulating the permeability of blood-brain barrier(BBB) induced by subacute 1,2-dichloroethane(1,2-DCE) inhalation. Methods Specific pathogen free healthy CD-1 male Aqp4 genetically engineered mice(Aqp4+/+and Aqp4-/-) were randomly divided into control and low-, medium-and high-dose groups. The mice were exposed to 1,2-DCE at the dosages of 0.00, 100.00, 350.00 and 700.00 mg/m3 for 6 hours per day for consecutive 28 days by systemic dynamic inhalation. After the end of 1,2-DCE exposure, the BBB permeability was evaluated by Evans blue staining. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of genes related to BBB tight junction protein(Tjp)1, Tjp2, Tjp3, claudin(Cldn)3, Cldn5, Cldn11, occludin(Ocln), matrix metalloproteinase(Mmp)2, Mmp9 and Na-K-Cl cotransporter-1(Nkcc1). Results The BBB permeability in mice showed significant change with 1,2-DCE dose and Aqp4 genotype(P<0.01). The BBB permeability of Aqp4+/+ genotype mice was higher in low-, medium-and high-dose groups than that of control group(all P values were <0.05). The permeability of BBB was lower in Aqp4+/+ genotype mice in the control group than that of Aqp4-/- genotype mice in the same group(P<0.05), but BBB permeability was higher in Aqp4+/+ genotype mice in the three dose groups than that of Aqp4-/- genotype mice in the same group(all P values were <0.05). The Cldn3 and Olcn mRNA relative expression in the brain cortex had statistical difference in mice with different genotype(all P values were <0.01). The mRNA relative expressions of Cldn3 and Olcn in the brain cortex were higher in Aqp4-/- genotype mice than that of Aqp4+/+ genotype mice(all P values were <0.01). The relative mRNA expression levels of Tjp1, Tjp2, Tjp3, Cldn5, Cldn11, Mmp2, Mmp9 and Nkcc1 in the cerebral cortex of mice were not statistically significant in aspect of 1,2-DCE exposure dose and genotype(all P values were >0.05). Conclusion Exposure to 1,2-DCE can increase

关 键 词：1 2-二氯乙烷 水通道蛋白4 血脑屏障 通透性 紧密连接蛋白 小鼠 

分 类 号：R114[医药卫生—卫生毒理学] R135.1[医药卫生—公共卫生与预防医学]