MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, China.
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, China.
Food Chem Toxicol. 2024 May;187:114629. doi: 10.1016/j.fct.2024.114629. Epub 2024 Mar 31.
Chlorpyrifos (CPF), a widely used organophosphorus pesticide (OP) to control pests has been verified reproductive toxicity on mammalian oocytes. However, limited information exists on its correlation with the dysfunction of the intercellular communication in cumulus-oocyte complexes (COCs). Herein, our study utilized porcine COCs as models to directly address the latent impact of CPF on the communication between cumulus cells (CCs) and oocytes during in vitro maturation. The results demonstrated that CPF exposure decreased the rate of the first polar body (PB1) extrusion and blocked meiosis progression. Notably, the cumulus expansion of CPF-exposed COCs was suppressed significantly, accompanied by the down-regulated mRNA levels of cumulus expansion-related genes. Furthermore, the early apoptotic level was raised and the expression of BAX/BCL2 and cleaved caspase 3 was up-regulated in the CCs of CPF-exposed COCs (p < 0.05). Moreover, CPF exposure impaired mRNA levels of antioxidant enzyme-related genes, induced higher levels of reactive oxygen species (ROS) and reduced the levels of mitochondrial membrane potential (MMP) in CCs (p < 0.05). Additionally, the integrated optical density (IOD) rate (cumulus/oocyte) of calcein and the expression of connexin 43 (CX43) was increased in CPF treatment groups (p < 0.05). As well, CPF exposure reduced the expression levels of FSCN1, DAAM1 and MYO10, which resulted in a significant decrease in the number and fluorescence intensity of transzonal projections (TZPs). In conclusion, CPF inhibited the expansion of cumulus and caused oxidative stress and apoptosis as well as disturbed the function of gap junctions (GJs) and TZPs, which eventually resulted in the failure of oocyte maturation.
毒死蜱(CPF)是一种广泛用于防治害虫的有机磷农药(OP),已被证实对哺乳动物卵母细胞具有生殖毒性。然而,关于其与卵丘-卵母细胞复合物(COCs)细胞间通讯功能障碍的相关性的信息有限。在此,我们的研究利用猪 COCs 作为模型,直接研究 CPF 对体外成熟过程中卵丘细胞(CCs)和卵母细胞之间通讯的潜在影响。结果表明,CPF 暴露降低了第一极体(PB1)排出率并阻断了减数分裂进程。值得注意的是,CPF 暴露的 COCs 的卵丘扩展受到显著抑制,伴随着卵丘扩展相关基因的 mRNA 水平下调。此外,CPF 暴露的 COCs 中 CCs 的早期凋亡水平升高,BAX/BCL2 和 cleaved caspase 3 的表达上调(p<0.05)。此外,CPF 暴露会损害抗氧化酶相关基因的 mRNA 水平,诱导更高水平的活性氧(ROS)并降低 CCs 中线粒体膜电位(MMP)水平(p<0.05)。此外,CPF 处理组的 calcein 整合光密度(IOD)率(卵丘/卵母细胞)和连接蛋白 43(CX43)的表达增加(p<0.05)。同样,CPF 暴露降低了 FSCN1、DAAM1 和 MYO10 的表达水平,导致跨带突(TZP)的数量和荧光强度显著减少。总之,CPF 抑制了卵丘的扩展,并导致氧化应激和细胞凋亡,同时扰乱了缝隙连接(GJs)和 TZPs 的功能,最终导致卵母细胞成熟失败。