Nrf2在镧活化MMP9致脉络丛上皮细胞紧密连接蛋白表达改变中的作用  

The role of Nrf2 in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated MMP9

作　　者：孙静 徐兴博 苏宏悦 闫立成 张艳淑 张丽锦 Sun Jing;Xu Xingbo;Su Hongyue;Yan Licheng;Zhang Yanshu;Zhang Lijin(Public Health and Preventive Medicine,School of Public Health,North China University of Science and Technology,Tangshan 063210,China;Preventive Medicine,School of Public Health,North China University of Science and Technology,Tangshan 063210,China;Department of Health Toxicology,School of Public Health,North China University of Science and Technology,Tangshan 063210,China;Laboratory Animal Center,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学公共卫生学院公共卫生与预防医学专业,唐山063210 [2]华北理工大学公共卫生学院预防医学专业,唐山063210 [3]华北理工大学公共卫生学院卫生毒理学教研室,唐山063210 [4]华北理工大学实验动物中心,唐山063210

出　　处：《中华劳动卫生职业病杂志》2023年第1期2-7,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：河北省自然科学基金(青年科学基金,H2020209177);河北省教育厅河北省高等学校科学技术研究项目(QN2020215)。

摘　　要：目的探讨核因子E2相关因子2(Nrf2)在镧活化基质金属蛋白酶9(MMP9)致脉络丛上皮细胞紧密连接蛋白表达改变中的作用研究。方法于2020年10月,采用永生化大鼠脉络丛上皮细胞(Z310细胞)体外模拟血脑脊液屏障,分为对照组和0.125、0.25、0.5 mmol/L氯化镧处理组。不同浓度氯化镧处理Z310细胞24 h后,倒置显微镜观察细胞形态学变化,细胞免疫荧光法观察MMP9、闭合蛋白(occludin)和胞质附着蛋白1(ZO-1)蛋白表达水平,蛋白免疫印迹法检测MMP9、组织金属蛋白酶抑制剂1(TIMP1)、occludin、ZO-1和Nrf2蛋白表达水平,流式细胞仪检测细胞内活性氧(ROS)水平。结果与对照组比较,氯化镧处理组Z310细胞体积变小,细胞间连接变少,死亡细胞、细胞碎片逐渐增多。0.25、0.5 mmol/L氯化镧处理组细胞内MMP9蛋白表达水平明显高于对照组(P<0.05),且TIMP1及紧密连接蛋白occludin、ZO-1蛋白表达水平较对照组明显降低(P<0.05);而与对照组比较,0.25、0.5 mmol/L氯化镧处理组细胞内ROS产生水平明显增加(P<0.05),0.125、0.25、0.5 mmol/L氯化镧处理组细胞内Nrf2蛋白表达水平明显下降(P<0.05)。结论镧可能通过下调Nrf2表达造成细胞内ROS水平增加,从而活化MMP9致细胞间紧密连接蛋白occludin、ZO-1表达水平下降。Objective To investigate the effect of nuclear factor erythroid 2-related factor 2(Nrf2)in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated matrix metalloproteinase 9(MMP9).Methods In October 2020,immortalized rat choroid plexus epithelial cell line(Z310)cells were used as the blood-cerebrospinal fluid barrier in vitro,and were divided into control group and 0.125,0.25,0.5 mmol/L lanthanum chloride(LaCl3)treatment group.After treating Z310 cells with different concentrations of LaCl3 for 24 hours,the morphological changes of Z310 cells were observed under inverted microscope,the protein expression levels of MMP9,occludin and zonula occludens-1(ZO-1)were observed by cellular immunofluorescence method,and the protein expression levels of MMP9,tissue inhibitors of metalloproteinase1(TIMP1),occludin,ZO-1 and Nrf2 were detected by Western blotting.The level of reactive oxygen species(ROS)in cells was detected by flow cytometry.Results Compared with the control group,Z310 cells in the LaCl3 treatment group were smaller in size,with fewer intercellular junctions,and more dead cells and cell fragments.The expression level of MMP9 protein in cells treated with 0.25 and 0.5 mmol/L LaCl3 was significantly higher than that in the control group(P<0.05),and the expression level of TIMP1 and tight junction proteins occudin and ZO-1 was significantly lower than that in the control group(P<0.05).Compared with the control group,the ROS production level in the 0.25,0.5 mmol/L LaCl3 treatment group was significantly increased(P<0.05),and the Nrf2 protein expression level in the 0.125,0.25,0.5 mmol/L LaCl3 treatment group was significantly decreased(P<0.05).Conclusion Lanthanum may increase the level of ROS in cells by down regulating the expression of Nrf2,thus activating MMP9 to reduce the expression level of intercellular tight junction proteins occludin and ZO-1.

关 键 词：镧 氯化镧 核因子E2相关因子2 活性氧 基质金属蛋白酶9 细胞间紧密连接 

分 类 号：R114[医药卫生—卫生毒理学]