李翠珍, 霍倩, 张晨, 周志俊, 吴庆. 苯并(a)芘对大鼠睾丸支持细胞连接蛋白基因表达的影响[J]. 环境与职业医学, 2013, 30(3): 161-166.
引用本文: 李翠珍, 霍倩, 张晨, 周志俊, 吴庆. 苯并(a)芘对大鼠睾丸支持细胞连接蛋白基因表达的影响[J]. 环境与职业医学, 2013, 30(3): 161-166.
LI Cui-zhen , HUO Qian , ZHANG Chen , ZHOU Zhi-jun , WU Qing . Effects of Benzo(a)pyrene on mRNA Expression of Junction Protein Genes in Primary Cultured Sertoli Cells[J]. Journal of Environmental and Occupational Medicine, 2013, 30(3): 161-166.
Citation: LI Cui-zhen , HUO Qian , ZHANG Chen , ZHOU Zhi-jun , WU Qing . Effects of Benzo(a)pyrene on mRNA Expression of Junction Protein Genes in Primary Cultured Sertoli Cells[J]. Journal of Environmental and Occupational Medicine, 2013, 30(3): 161-166.

苯并(a)芘对大鼠睾丸支持细胞连接蛋白基因表达的影响

Effects of Benzo(a)pyrene on mRNA Expression of Junction Protein Genes in Primary Cultured Sertoli Cells

  • 摘要: 目的 探讨苯并(a)芘(BaP)对大鼠原代培养睾丸支持细胞的连接蛋白(connexin 43、occludin、ZO-1、N-cadherin)基因mRNA表达的影响及可能机制。

    方法 取出生21 d SD大鼠睾丸组织, 分离支持细胞, 细胞分离后, 均按照1.5& #215;106 个/mL 浓度接种于细胞培养板中。培养48 h 后, 以0、1、10、50、100 μmol/L 浓度的BaP 同时染毒于原代培养的大鼠支持细胞12 h, 每个剂量设置6 个平行孔。检测支持细胞活力、凋亡蛋白caspase-3 活性, 以及连接蛋白基因的mRNA表达。

    结果 在BaP 50 μmol/L 时, 支持细胞活力降低(和对照组相比, P < 0.01);在BaP 10 μmol/L 时caspase-3活性升高, 连接蛋白connexin 43、occludin、和ZO-1 基因的mRNA表达降低(和对照组相比, P < 0.05)。在BaP 100 μmol/L时连接蛋白N-cadherin 基因的mRNA表达降低(和对照组相比, P < 0.05)。加入caspase-3 抑制剂后, BaP 所诱导的连接蛋白基因表达下降, 且随caspase-3 活性变化而发生改变。

    结论 BaP 能诱导大鼠支持细胞连接蛋白mRNA表达下降, BaP 诱导的caspase-3 活性上升是调节连接蛋白mRNA表达下降的可能机制。

     

    Abstract: Objective To investigate the effect of benzo(a)pyrene (BaP) on mRNA expression of junction protein genes in sertoli cells.

    Methods Sertoli cells were isolated from 21-day-old male rats and incubated at a density of 1.5& #215;106 cells/mL. After incubation for 48 h, the sertoli cells were exposed to varied concentrations of BaP (0, 1, 10, 50, and 100 μmol/L) for 12 h. Every group contained 6 samples. Cell viability, caspase-3 activity, and mRNA expression levels of junction protein genes were measured.

    Results The cell viability was significantly decreased by the BaP treatment at 50 μmol/L compared with the control group (P < 0.01). The caspase-3 activity was increased and the mRNA expression levels of connexin 43, occludin, and ZO-1 were decreased after BaP exposure at 10 μmol/L compared with the control group (P < 0.05). A down-regulated mRNA expression level of N-cadherin was observed after BaP administration at 100 μmol/L compared with control group (P < 0.05). The mRNA expression levels of junction protein genes were changed along with the alteration of caspase-3 activity after caspase-3 inhibitor was added.

    Conclusion BaP exposure could decrease mRNA expression levels of junction protein genes in primary sertoli cells, in which an up-regulated caspase-3 activity by BaP interference may play an important role.

     

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