[阿特拉津对培养的大鼠支持细胞凋亡的影响]

[Effect of atrazine on apoptosis in cultured rat Sertoli cells].

作者信息

Song Yang, Zhang Yu, Ma Mengting, Zhang Lishi

机构信息

Sichuan Center for Disease Control and Prevention, Chengdu 610041, China.

出版信息

Wei Sheng Yan Jiu. 2017 Jan;46(1):103-108.

PMID:29903161

Abstract

OBJECTIVE

To elucidate the effects of atrazine( ATZ) on apoptosis in cultured rat Sertoli cells.

METHODS

The Sertoli cells isolated from male SD rats( 18-21day) were used to set up Sertoli cells primary culture model in vitro, and these cells were treated with 0, 10, 50, 100 and 200 μmol / L ATZ for 24 h. Cell viability was assessed by MTT assay, and the vimentin expression of Sertoli cells was analyzed by immunofluorescence assay, and apoptosis were evaluated by flow cytometry. Real timePCR was used to analyze the mRNA levels of Fas L, caspase-3 and caspase-8, and the protein contents of Fas L, caspase-3 and caspase-8 were determined by ELISA.

RESULTS

The viability of Sertoli cells of 50, 100 and 200 μmol / L ATZ groups were 82. 47%, 75. 23% and 53. 76%. Compared with the control group, the viability of Sertoli cells decreased by 17. 53%( P < 0. 05), 24. 77%( P < 0. 05) and 46. 24%( P < 0. 01), respectively. The apoptosis rates of Sertoli cell of 50, 100 and 200 μmol / L ATZ groups were 8. 53%, 13. 07% and 14. 45%. Compared with the control group, the apoptosisrates of Sertoli cells were increased by 86. 86%( P < 0. 01), 186. 13%( P < 0. 01), 216. 06%( P < 0. 01), respectively. the mRNA and protein expressions of Fas L, caspase-3 and caspase-8 were significantly increased at 50, 100 and 200 μmol / L ATZ groups( P < 0. 05, P < 0. 01). The Vimentin expression of Sertoli cells was inhibited in treated groups.

CONCLUSION

ATZ can affect the viability of Sertoli cells, and induce apoptosis of Sertoli cells through a Fas L dependent pathway.

摘要

目的

阐明莠去津（ATZ）对培养的大鼠支持细胞凋亡的影响。

方法

采用从18 - 21日龄雄性SD大鼠分离的支持细胞建立体外支持细胞原代培养模型，将这些细胞分别用0、10、50、100和200 μmol/L的ATZ处理24小时。通过MTT法评估细胞活力，通过免疫荧光法分析支持细胞波形蛋白的表达，并通过流式细胞术评估细胞凋亡。采用实时荧光定量PCR分析Fas L、caspase - 3和caspase - 8的mRNA水平，通过ELISA法测定Fas L、caspase - 3和caspase - 8的蛋白含量。

结果

50、100和200 μmol/L ATZ组支持细胞的活力分别为82.47%、75.23%和53.76%。与对照组相比，50、!00和200 μmol/L ATZ组支持细胞活力分别下降了17.53%（P < 0.05）、24.77%（P < 0.05）和46.24%（P < 0.01）。50、100和200 μmol/L ATZ组支持细胞的凋亡率分别为8.53%、13.07%和14.45%。与对照组相比，支持细胞的凋亡率分别增加了86.86%（P < 0.01）、186.13%（P < 0.01）、216.06%（P < 0.01）。在50、100和200 μmol/L ATZ组中，Fas L、caspase - 3和caspase - 8的mRNA和蛋白表达均显著增加（P < 0.05，P < 0.01）。处理组中支持细胞波形蛋白的表达受到抑制。