We investigated the in vitro effects of pyriproxyfen on ionic balance in the testis of the zebrafish by measuring ⁴⁵Ca²⁺ influx. In vivo pyriproxyfen treatment was carried out to study oxidative stress, and conduct morphological analysis of the testis and liver. Whole testes were incubated in vitro with/without pyriproxyfen (10^-12, 10^-9 or 10^-6 M; 30 min) and ⁴⁵Ca²⁺ influx determined. To study pyriproxyfen’s mechanism of action, inhibitors/activators of ionic channels or pumps/exchangers, protein kinase inhibitors or a calcium chelator were added 15 min before the addition of ⁴⁵Ca²⁺ and pyriproxyfen. We evaluated the in vivo effects of 7 day exposure to waterborne pyriproxyfen (10^-9 M) on reactive oxygen species (ROS) formation, lipid peroxidation, and reduced glutathione content (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) and γ-glutamyltransferase (GGT) activity. Morphological analyses of the testis and liver were carried out after in vivo exposure of D. rerio to pyriproxyfen. Pyriproxyfen increased ⁴⁵Ca²⁺ influx by opening the voltage-dependent T-type channels (T-type VDCC), inhibiting sarco/endoplasmic reticulum ⁴⁵Ca²⁺-ATPase (SERCA) and the NCX exchanger (forward mode) and by mobilizing calcium from stores. The involvement of potassium channels and protein kinase C (PKC) was also demonstrated in pyriproxyfen-induced intracellular calcium elevation. In vivo pyriproxyfen treatment of D. rerio increased lipid peroxidation, decreased GSH content and increased GST activity in testes, in addition to increasing the number and size of spermatogonia cysts and inducing hepatocyte basophilia and dilation of blood vessels in the liver. The toxicity of pyriproxyfen is mediated by calcium overload, increased lipid peroxidation, and a diminished antioxidant capacity in the testis, due to GSH depletion, and altered spermatogenesis. The development of high basophilia in the liver suggests that pyriproxyfen may have estrogenic activity, possibly acting as an endocrine-disruptor. These findings indicate that these alterations may contribute to pyriproxyfen toxicity and spermatogenesis disruption.

我们通过测量⁴⁵ Ca ^2+的流入量，研究了吡吡洛芬对斑马鱼睾丸中离子平衡的体外影响。进行体内吡吡氧芬治疗以研究氧化应激，并对睾丸和肝脏进行形态分析。整个睾丸孵育在体外有/无蚊蝇醚（10 ^-12，10 ^-9或10 ^-6米; 30分钟）和⁴⁵的Ca ²⁺内流来确定。为了研究吡ip昔芬的作用机理，在加入盐酸之前15分钟，应加入离子通道或泵/交换剂的抑制剂/活化剂，蛋白激酶抑制剂或钙螯合剂。⁴⁵ Ca ²⁺和吡吡^氧芬。我们评估了7天暴露于水性吡咯氧芬（10 ^-9 M）对体内活性氧（ROS）形成，脂质过氧化和降低的谷胱甘肽含量（GSH），谷胱甘肽S-转移酶（GST），超氧化物歧化酶（ SOD），过氧化氢酶（CAT）和γ-谷氨酰转移酶（GGT）活性。在体内将雷氏梭状芽胞杆菌暴露于吡ip昔芬后，对睾丸和肝脏进行形态学分析。吡咯氧芬通过打开电压依赖性T型通道（T型VDCC），增加肌钙蛋白/内质网⁴⁵ Ca ²⁺，从而增加了⁴⁵ Ca ^2+的流入量。-ATPase（SERCA）和NCX交换剂（正向模式），并从商店中调集钙。钾通道和蛋白激酶C（PKC）的参与也被证明在吡ip昔芬诱导的细胞内钙升高。吡吡洛芬的体内治疗瑞氏梭菌除了增加精子囊肿的数量和大小，并诱导肝细胞嗜碱性和肝血管扩张外，还增加了睾丸的脂质过氧化作用，降低了GSH含量并增加了GST活性。由于谷胱甘肽（GSH）耗竭和精子发生改变，吡pyr昔芬的毒性由钙超载，脂质过氧化增加和睾丸的抗氧化能力下降介导。肝脏中高度嗜碱性粒细胞的发展表明吡吡洛芬可能具有雌激素活性，可能充当内分泌干扰物。这些发现表明，这些改变可能导致吡吡氧芬毒性和精子发生破坏。