甲基对硫磷对小鼠黄体细胞的细胞间隙连接通讯和孕酮分泌的影响
首发时间:2013-04-15
摘要:目的 探索甲基对硫磷(methyl parathion,MP)对小鼠黄体细胞的细胞间隙连接通讯(gap junctional intercellular communication,GJIC)和孕酮分泌的影响。方法 体外培养BALB/c小鼠黄体细胞,分别设MP暴露组(0、0.05、0.1、0.2、0.4、0.8、1.6 mmol/L的MP培养3 h)和异丙肾上腺素预处理组[1 μmol/L异丙肾上腺素(GJIC促进剂)处理5 min后,0.8 mmol/L的MP培养3 h]及H89与Enzastaurin预处理组[用蛋白激A(PKA)与蛋激酶C(PKC)专一抑制剂H89与Enzastaurin(10 μmol/L)处理5 min后,用0.8 mmol/L的MP培养3 h]。采用ELISA法检测培养液中孕酮含量,划痕标记染料转移法测定贴壁细胞的GJIC。结果 高浓度(0.2~1.6 mmol/L)MP可明显抑制小鼠黄体细胞的GJIC和孕酮分泌(P<0.05);GJIC促进剂异丙肾上腺素、PKA专一抑制剂H89和PKC专一抑制剂Enzastaurin均能显著降低MP对体外小鼠黄体细胞的GJIC和孕酮分泌的抑制作用(P<0.05)。结论 MP可通过抑制小鼠黄体细胞的GJIC导致孕酮分泌受抑制,而PKA与PKC参与该过程。
关键词: 环境卫生学 甲基对硫磷 细胞间隙连接通讯 生殖毒性 孕酮
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Effect of Methyl Parathion on Progesterone Secretion and Gap Junctional Intercellular Communication of Mouse Luteal Cells
Abstract:Objective The present study aims at exploration of the effect of methyl parathion (MP) on progesterone secretion and gap junctional intercellular communication (GJIC) of mouse luteal cells. Methods Mouse luteal cells were treated by (1) MP with various concentrations (0,0.05,0.1,0.2,0.4,0.8,1.6 mmol/L) for 3 h;(2) Isoprenaline (1 μmol/L, a promoter of GJIC) for 5 min followed by treatment of 0.8 mmol/L MP for 3 h; (3) H89 and Enzastaurin (10 μmol/L, single or combined) for 5 min followed by 0.8 mmol/L MP for 3 h. Progesterone content in medium was detected by ELISA kit specific for mouse progesterone and GJIC of mouse luteal cells was detected using the scrape loading and dye transfer technique. Results The data showed that MP with higher concentrations (0.2-1.6 mmol/L) inhibited significantly (P < 0.05) progesterone secretion and GJIC of mouse luteal cells. Isoprenaline,H89 and Enzastaurin could significantly (P < 0.05) alleviate the inhibition effect of MP on progesterone secretion and GJIC of mouse luteal cells. Conclusion MP inhibited progesterone secretion of luteal cells through inhibition of GJIC,involving PKA and PKC in the process.
Keywords: Methyl parathion Gap junctional intercellular communication Reproductive toxicity Progesterone
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