铅暴露对体外血脑脊液屏障功能及ZO-1和闭合蛋白表达的影响

发布时间：2018-12-18 00:52

【摘要】：目的通过建立体外血脑脊液屏障(BCB)模型,研究铅诱导BCB损伤的可能机制。方法使用Z310细胞建立BCB体外细胞模型,分别加入Pb(AC)22.5,5.0和10.0 mmol·L-1,暴露24,48和72 h。采用跨内皮电阻(TEER)检测法及葡聚糖法检测BCB模型通透性;Western蛋白印迹法和免疫荧光法检测BCB中紧密连接蛋白ZO-1和闭合蛋白表达及分布。结果与细胞对照组相比,Pb(AC)22.5,5.0和10.0 mmol·L-1暴露48 h对Z310细胞存活率和密度均无明显影响。暴露后第12天,与细胞对照组比较,Pb(AC)25.0和10.0 mmol·L-1组TEER值明显降低(P0.05),葡聚糖通过率显著增高(P0.05)。Western蛋白印迹法和免疫荧光法检测结果显示,与细胞对照组相比,Pb(AC)2各浓度暴露48 h后,ZO-1和闭合蛋白表达水平明显降低(P0.05)。结论铅暴露能够破坏BCB模型的通透性,其机制可能与抑制紧密连接蛋白的表达有关。
[Abstract]:Objective to study the possible mechanism of BCB damage induced by lead by establishing (BCB) model of blood-cerebrospinal fluid barrier in vitro. Methods BCB cell model was established by using Z310 cells in vitro. The cells were exposed to Pb (AC) 22.5 mmol 5.0 and 10.0 mmol L -1 for 24 h and 72 h, respectively. Transendothelial resistance (TEER) and dextran were used to detect the permeability of BCB model, and Western blot and immunofluorescence were used to detect the expression and distribution of close-junction protein ZO-1 and closed protein in BCB. Results compared with the control group, Pb (AC) 22.5 and 10.0 mmol L-1 exposure for 48 h had no significant effect on the survival rate and density of Z310 cells. On the 12th day after exposure, the TEER values of, Pb (AC) 25.0 and 10.0 mmol L-1 groups were significantly lower than those of the control group (P0.05). The pass-through rate of dextran was significantly increased (P0.05). Western blot and immunofluorescence assay showed that compared with the control group, Pb (AC) _ 2 was exposed to different concentrations for 48 h. The expression level of ZO-1 and closed protein was significantly decreased (P0.05). Conclusion lead exposure can damage the permeability of BCB model, and its mechanism may be related to the inhibition of tight junction protein expression.
【作者单位】：中国人民解放军第四军医大学军事预防医学系军队劳动与环境卫生学教研室;
【基金】：国家自然科学基金(81472942) 第四军医大学青年英才培育计划(4139Z3B6BC)~~
【分类号】：R114

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