Sun J, Xu X B, Su H Y, Yan L C, Zhang Y S, Zhang L J
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.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2023 Jan 20;41(1):2-7. doi: 10.3760/cma.j.cn121094-20220121-00039.
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) . In October 2020, immortalized rat choroid plexus epithelial cell line (Z310) cells were used as the blood-cerebrospinal fluid barrier , and were divided into control group and 0.125, 0.25, 0.5 mmol/L lanthanum chloride (LaCl(3)) treatment group. After treating Z310 cells with different concentrations of LaCl(3) 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. Compared with the control group, Z310 cells in the LaCl(3) 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 LaCl(3) was significantly higher than that in the control group (<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 (<0.05) . Compared with the control group, the ROS production level in the 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly increased (<0.05) , and the Nrf2 protein expression level in the 0.125, 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly decreased (<0.05) . 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.
探讨核因子红细胞2相关因子2(Nrf2)在镧激活基质金属蛋白酶9(MMP9)所致脉络丛上皮细胞紧密连接蛋白表达改变中的作用。2020年10月,将永生化大鼠脉络丛上皮细胞系(Z310)细胞作为血脑屏障模型,分为对照组和0.125、0.25、0.5 mmol/L氯化镧(LaCl₃)处理组。用不同浓度的LaCl₃处理Z310细胞24小时后,在倒置显微镜下观察Z310细胞的形态变化,采用细胞免疫荧光法观察MMP9、闭合蛋白和紧密连接蛋白1(ZO-1)的蛋白表达水平,并用蛋白质免疫印迹法检测MMP9、金属蛋白酶组织抑制剂1(TIMP1)、闭合蛋白、ZO-1和Nrf2的蛋白表达水平。通过流式细胞术检测细胞内活性氧(ROS)水平。与对照组相比,LaCl₃处理组的Z310细胞体积变小,细胞间连接减少,死细胞和细胞碎片增多。0.25和0.5 mmol/L LaCl₃处理组细胞中MMP9蛋白表达水平显著高于对照组(<0.05),TIMP1及紧密连接蛋白闭合蛋白和ZO-1的表达水平显著低于对照组(<0.05)。与对照组相比,0.25、0.5 mmol/L LaCl₃处理组细胞内ROS生成水平显著升高(<0.05),0.125、0.25、0.5 mmol/L LaCl₃处理组细胞中Nrf2蛋白表达水平显著降低(<0.05)。镧可能通过下调Nrf2表达增加细胞内ROS水平,从而激活MMP9,降低细胞间紧密连接蛋白闭合蛋白和ZO-1的表达水平。