铁死亡在鱼藤酮损伤小鼠中脑多巴胺能神经元中的作用研究

The role of ferroptosis in the damage of dopaminergic neurons in rotenone-induced mouse

目的研究铁死亡在农药鱼藤酮损伤小鼠多巴胺能神经元中的作用。方法 8周龄雄性小鼠随机分成对照组(Con)和鱼藤酮组(Rot),每组共4只。通过鱼藤酮对小鼠染毒3周,染毒剂量1.5 mg/(kg·d),通过水合氯醛麻醉小鼠,用磷酸盐缓冲液对小鼠实施心脏灌流至肝脏发白,冰上操作取小鼠中脑。运用Western blot检测和试剂盒检测多巴胺能神经元损伤水平、铁死亡水平。结果对照组酪氨酸羟化酶(TH)表达水平为100.00±1.12,鱼藤酮染毒组TH表达水平为54.47±4.41;对照组游离铁含量为2.18±0.08 ng/mL,鱼藤酮染毒组游离铁含量为3.24±0.05 ng/mL;对照组谷胱甘肽过氧化物酶-4(GPX4)表达水平为100.00±1.16,鱼藤酮染毒组GPX-4表达水平为65.46±3.58;对照组转铁蛋白-1(FPN-1)表达水平为100.00±1.32,鱼藤酮染毒组FPN-1表达水平为62.18±1.18;对照组丙二醛(MDA)表达水平为3.83±0.15μmol/g,鱼藤酮染毒组MDA表达水平为5.73±0.11μmol/g;对照组乳酸脱氢酶(LDH)酶活力(mU/mL)为0.34±0.01,鱼藤酮染毒组LDH酶活力(mU/mL)为0.65±0.01,对照组谷胱甘肽(GSH)表达水平为141.80±4.60μmol/L,鱼藤酮染毒组GSH表达水平为83.44±1.99μmol/L。结论鱼藤酮可能通过诱导铁死亡引发氧化应激损伤小鼠中脑多巴胺能神经元。

Objective To explore the role of ferroptosis in the damage of dopaminergic neurons in mice by the pesticide rotenone.Methods The mice were exposed to rotenone[1.5 mg/(kg·d)] for 3 weeks. The mice were anesthetized with chloral hydrate, and the mice were perfused with phosphate buffer solution until the liver became white. The mice were taken out on ice. Western blot detection and kits were used to detect the level of dopaminergic neuron damage and ferroptosis. Results The TH expression level of the control group was 100.00±1.12, the rotenone-exposed group was 54.47±4.41,the free iron content of the control group was2.18±0.08 and the rotenone-exposed group was 3.24 ±0.05,the expression level of GPX-4 in the control group was 100.00 ±1.16 and the rotenone-exposed group was 65.46 ±3.58. The expression level of FPN-1 in the control group was 100.00 ±1.32,the rotenone-exposed group was 62.18±1.18. The expression level of MDA in the control group was 3.83 ±0.15,the rotenone-exposed group was 5.73±0.11,the expression level of LDH in the control group was 0.34±0.01,the rotenone-exposed group was 0.65± 0.01.The expression level of GSH in the control group was 141.80 ±4.60,and the rotenone-exposed group was 83.44 ±1.99.Conclusion Rotenone may damage dopaminergic neurons in the midbrain of mice through ferroptosis.