氯化钴对小鼠肝氧化损伤及铁死亡相关指标的影响

Effects of cobalt chloride on liver oxidative damage and ferroptosis in mice

目的观察氯化钴暴露对小鼠肝氧化损伤及铁死亡相关指标的影响。方法将24只雄性SPF级C57BL/6小鼠随机分为对照组和实验组,每组6只,实验组分别腹腔注射4、8和12 mg/kg氯化钴溶液,对照组腹腔注射等量生理盐水,每天染毒一次,共染毒30 d。染毒结束,计算动物肝脏脏器系数;采用试剂盒测定动物肝组织匀浆上清中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG);Western blot检测动物肝组织谷胱甘肽过氧化物酶4(GPX4)、铁死亡抑制蛋白1(FSP1)和长链酯酰辅酶A合成酶4(ACSL4)蛋白表达水平。结果与对照组比较,4 mg/kg染毒组动物肝脏脏器系数显著升高(F=6.15,P=0.027),12 mg/kg氯化钴暴露组SOD降低(F=6.56,P=0.028);MDA随着氯化钴染毒剂量增加而增加(F=30.93,P<0.001);GSH和GSH/GSSG随着氯化钴染毒剂量增加而降低(F值分别为22.78和24.57,P<0.01);GPX4和FSP1表达水平随着氯化钴染毒剂量增加而降低(F值分别为41.47和19.397,P<0.01);ACSL4表达水平随着氯化钴染毒剂量增加而增加(F=16.01,P=0.004)。结论氯化钴暴露可引起小鼠肝脏氧化损伤,通过降低GPX4和FSP1表达,升高ACSL4表达促发肝脏铁死亡。

Objective To explore the effects of cobalt chloride exposure on the indicators of oxidative damage and ferroptosis in the liver of mice.Methods A total of 24 male C57BL/6 specific pathogen free(SPF)grade mice were randomly divided into three exposure groups and one control group to receive intraperitoneal injection of cobalt chloride solution at doses of 4,8,and 12 mg/kg and an equal volume of saline,respectively,once daily,for a total of 30 days.Then the mice were sacrificed to obtain the liver to calculate the organ coefficient.The liver homogenate supernatant was prepared to determine the content of malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH)and oxidized glutathione(GSSG)by kits.Western blot was performed to measure the protein expression levels of glutathione peroxidase 4(GPX4),ferroptosis suppressor protein 1(FSP1)and long-chain acyl coenzyme A synthetase 4(ACSL4)in the liver tissue.Results Compared with that of the control group,the liver coefficient was significantly increased in the 4 mg/kg exposure group(F=6.15,P=0.027).The content of SOD was significantly decreased in the 12 mg/kg exposure group than in the control group(F=6.56,P=0.028).As the dose of cobalt chloride increased,the content of MDA was significantly increased(F=30.93,P<0.001),the GSH content and GSH/GSSG ratio were significantly decreased(F=22.78,24.57,both P<0.01),the levels of GPX4 and FSP1 were significantly decreased(F=41.47,19.397,both P<0.01),and the level of ACSL4 was significantly increased(F=16.01,P=0.004).Conclusion Cobalt chloride exposure can cause hepatic oxidative damage in mice,and promote hepatic ferroptosis by downregulating GPX4 and FSP1 and upregulating ACSL4..