School of Public Health/MOE Key Laboratory for Public Health Safety/Collaborative Innovation Center of Social Risks Governance in Health, Fudan University, Shanghai 200032, China.
School of Public Health/MOE Key Laboratory for Public Health Safety/Collaborative Innovation Center of Social Risks Governance in Health, Fudan University, Shanghai 200032, China; Pharmacology and Toxicology Department, Shanghai Institute for Food and Drug Control, Shanghai 201203, China.
Toxicol In Vitro. 2018 Mar;47:228-237. doi: 10.1016/j.tiv.2017.12.006. Epub 2017 Dec 14.
Fluorochloridone (FLC) is a widely used pyrrolidone selective herbicide and reported to induce testis injuries in male rats, but the underlying mechanism is largely unknown. In the present study, primary-cultured Sertoli cells were exposed to FLC at the concentration of 0-10.00μM to study the mechanism of FLC-induced apoptosis. The roles of ROS, intracellular calcium, endoplasmic reticulum (ER), and ERK1/2 were looked at with ROS scavenger N-acetyl-cysteine (NAC), intracellular calcium chelator BAPTA-AM, ER calcium depleting agent thapsigargin (TG), and ERK1/2 inhibitor U0126, respectively. FLC induced dose-dependent apoptosis increase as well as the elevation in levels of ROS, intracellular calcium, and ERK1/2 activation. FLC treatment led to constantly increasing apoptotic rates and ERK1/2 activation over time, while inversed-V shaped change tendencies of ROS and intracellular calcium levels were observed. FLC-induced ROS generation disrupted the intracellular calcium homeostasis by attacking the ER, and the elevated intracellular calcium levels resulted in ERK1/2 over-phosphorylation and consequently promoted Sertoli cell apoptosis. Taken together, ROS and intracellular calcium-mediated ERK1/2 activation led to FLC-induced Sertoli cell apoptosis.
氟氯吡啶（FLC）是一种广泛使用的吡咯烷酮选择性除草剂，据报道会导致雄性大鼠睾丸损伤，但其中的潜在机制尚不清楚。在本研究中，我们用浓度为 0-10.00μM 的 FLC 处理原代培养的支持细胞，以研究 FLC 诱导细胞凋亡的机制。用 ROS 清除剂 N-乙酰半胱氨酸（NAC）、细胞内钙螯合剂 BAPTA-AM、内质网（ER）钙耗竭剂 thapsigargin（TG）和 ERK1/2 抑制剂 U0126 分别观察 ROS、细胞内钙、内质网和 ERK1/2 的作用。FLC 诱导的细胞凋亡呈剂量依赖性增加，同时 ROS、细胞内钙和 ERK1/2 激活水平也升高。FLC 处理导致细胞凋亡率和 ERK1/2 激活随时间持续增加，而 ROS 和细胞内钙水平则呈反“V”形变化趋势。FLC 诱导的 ROS 生成通过攻击内质网破坏了细胞内钙稳态，而升高的细胞内钙水平导致 ERK1/2 过度磷酸化，从而促进支持细胞凋亡。总之，ROS 和细胞内钙介导的 ERK1/2 激活导致 FLC 诱导的支持细胞凋亡。
