Abstract：
Beauvericin is an emerging Fusariotoxin naturally occurring in cereal grains throughout the world whereas glyphosate (N-phosphonomethyl-glycine) is a non-selective systemic herbicide used worldwide. The purpose of this study is to evaluate a newly developed ovarian cell culture system (that includes both granulosa and theca cells) as an in vitro model for toxicological studies. Specifically, the effects of beauvericin and glyphosate in formulation with Roundup on ovarian cell numbers and steroid production were evaluated. Ovaries collected from cattle without luteal structures were sliced into 30-70 pieces each, and granulosa and theca cells were collected. Harvested cells were cultured for 48 h in 10% fetal bovine serum-containing medium followed by 48 h in serum-free medium containing testosterone (500 ng/mL; as an estrogen precursor) with the following eight treatments: (1) controls, (2) FSH (30 ng/mL) alone, (3) FSH plus insulin-like growth factor-1 (IGF1; 30 ng/mL), (4) FSH plus IGF1 plus beauvericin (3 µM), (5) FSH plus IGF1 plus glyphosate in Roundup (10 µg/mL), (6) FSH plus IGF1 plus fibroblast growth factor 9 (FGF9, 30 ng/mL), (7) a negative control without added testosterone, and (8) IGF1 plus LH (30 ng/mL) with basal medium without added testosterone. In the presence of FSH, IGF1 significantly increased cell numbers, estradiol and progesterone production by severalfold. Glyphosate in Roundup formulation significantly inhibited IGF1-induced cell numbers and estradiol and progesterone production by 89-94%. Beauvericin inhibited IGF1-induced cell numbers and estradiol and progesterone by 50-97% production. LH plus IGF1 significantly increased androstenedione secretion compared with controls without added testosterone indicating the presence of theca cells. In conclusion, the present study demonstrates that toxicological effects of beauvericin and glyphosate in Roundup formulation are observed in a newly developed ovarian cell model system and further confirms that both glyphosate and beauvericin may have the potential to impair reproductive function in cattle.
摘要：
Beauvericin是一种新兴的镰刀菌毒素，天然存在于世界各地的谷物中，而草甘膦（N-膦酰基甲基-甘氨酸）是全球使用的非选择性系统性除草剂。这项研究的目的是评估新开发的卵巢细胞培养系统（包括颗粒细胞和卵泡膜细胞）作为毒理学研究的体外模型。具体来说,评价了与农达制剂中的波维菌素和草甘膦对卵巢细胞数量和类固醇产生的影响。从没有黄体结构的牛身上采集的卵巢被切成30-70块，收集颗粒细胞和卵泡膜细胞。将收获的细胞在含10％胎牛血清的培养基中培养48小时，然后在含睾酮（500ng/mL；作为雌激素前体）的无血清培养基中培养48小时，进行以下八种处理：（1）对照，（2）单独使用FSH（30ng/mL），（3）FSH加胰岛素样生长因子-1（IGF1；30ng/mL），（4）FSH+IGF1+白僵素（3µM），（5）FSH加IGF1加草甘膦在农达（10µg/mL），（6）FSH+IGF1+成纤维细胞生长因子9（FGF9，30ng/mL），（7）不添加睾酮的阴性对照，和(8)IGF1加LH(30ng/mL),具有不添加睾酮的基础培养基。在FSH存在的情况下,IGF1显著增加细胞数量，雌二醇和孕酮的产量提高了几倍。农达制剂中的草甘膦显着抑制IGF1诱导的细胞数量以及雌二醇和孕酮的产生89-94％。Beauvericin抑制IGF1诱导的细胞数量以及雌二醇和孕酮产生50-97％。与未添加睾丸激素的对照相比，LH加IGF1显着增加雄烯二酮的分泌，表明存在卵泡膜细胞。总之,本研究表明，在新开发的卵巢细胞模型系统中观察到了农达制剂中的博维菌素和草甘膦的毒理学效应，并进一步证实了草甘膦和博维菌素可能具有损害牛生殖功能的潜力。
