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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5周龄SPF级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处理3 h)和GJIC促进剂——1μmol/L异丙肾上腺素预处理组及蛋白激酶A(PKA)专一抑制剂——10μmol/L H89预处理组与蛋白激酶C(PKC)专一抑制剂——10μmol/L恩扎妥林预处理组(预处理5 min,0.8 mmol/L MP处理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);异丙肾上腺素、H89和恩扎妥林均能缓解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。
Objective To explore the effect of methyl parathion on the gap junctional intercellular communication (GJIC) and progesterone secretion in mouse luteal cells. Methods SPF BALB / c mouse luteal cells were cultured in vitro and were treated with MP exposure group (0,0.05,0.1,0.2,0.4,0.8 and 1.6 mmol / L MP for 3 h) and GJIC promoter - The pretreatment with 1μmol / L isoproterenol and the specific inhibitor of protein kinase A (PKA) --10μmol / L H89 pretreatment with PKC inhibitor - 10μmol / L enzastatone Pretreatment group (pretreatment 5 min, 0.8 mmol / L MP treatment 3 h). The content of progesterone in culture solution was detected by ELISA, and the GJIC of adherent cells was determined by scratch-labeled dye transfer assay. Results MP (0.2 ~ 1.6 mmol / L) could significantly inhibit the secretion of GJIC and progesterone in mouse luteal cells (P <0.05). Both isoproterenol, H89 and enzalutan could alleviate MP in vitro The inhibitory effect of GJIC and progesterone secretion of luteal cells (P <0.05). Conclusion MP can inhibit the secretion of progesterone by inhibiting the GJIC of mouse luteal cells, and PKA and PKC participate in this process.