土曲霉对小鼠肝氧化损伤及铁死亡相关指标的影响

Effects of Aspergillus terreus on Oxidative Damage and Ferroptosis Related Indicators in Mice Liver

旨在探究土曲霉引起小鼠肝损伤的机制。将20只昆明小鼠随机分为对照组和试验组,试验组腹腔注射0.3 mL(5×10^(7) CFU·mL^(-1))土曲霉MSF2菌株孢子悬液,试验周期为7 d。采集小鼠肝用丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化酶(GSH-PX)、总超氧化物歧化酶(T-SOD)试剂盒检测肝氧化损伤相关指标;铁检测试剂盒检测肝铁离子含量;制备肝石蜡切片,进行HE染色和普鲁士蓝染色,观察其病理变化及铁沉积;透射电镜观察肝细胞超微结构;qPCR检测肝铁死亡相关基因mRNA相对转录水平。结果表明:土曲霉可引起试验组小鼠肝MDA含量极显著升高(P<0.01),GSH含量、GSH-PX活力和T-SOD活力极显著降低(P<0.01),铁离子含量极显著升高(P<0.01),肝细胞肿胀、坏死,炎性细胞浸润,肝细胞中可见蓝染的铁离子沉积,肝细胞线粒体萎缩、嵴减少和膜密度增加,转铁蛋白1(TFR 1)、二价金属离子转运体1(DMT 1)、铁蛋白重链1(FTH 1)和电压依赖性阴离子通道3(VDAC 3)基因mRNA相对转录水平显著升高(P<0.05),过氧化物酶4(GPX 4)和胱氨酸/谷氨酸转运受体11(SLC 7 A 11)基因mRNA相对转录水平显著降低(P<0.05)。综上表明,土曲霉MSF2菌株致小鼠肝损伤的机制是铁死亡,为深入研究土曲霉的致病机制提供参考资料。

In order to explore the mechanism of Aspergillus terreus causing liver injury of mice,20 Kunming mice were divided into a control group and a test group.The test group was intraperitoneally injected with 0.3 mL(5×10^(7) CFU·mL^(-1))spore suspension.The trail was 7 days.The indicators of oxidative damage in mouse liver tissue were evaluated by malondialdehyde(MDA),glutathione(GSH),glutathione peroxidase(GSH-PX)and superoxide dismutase(T-SOD)kits.The iron content of liver tissue was determined by iron kit.Prepare liver paraffin sections to observed the pathological changes and iron accumulation by HE and Perls stain.The ultrastructure of hepatocytes was observed by transmission electron microscope.The mRNA relative transcription level of ferroptosis-related genes in liver tissue were detected by qPCR.The results were as follows:compared with control group,the content of MDA in test group was significantly increased,but the content of GSH,the activities of GSH-PX and T-SOD in test group were significantly decreased(P<0.01).Meanwhile,the content of iron ions in test group was significantly increased(P<0.01).Some hepatocytes were swollen and necrotic.A few inflammatory cells were infiltrated.Iron ions accumulated in the center of hepatic loules.The mitochondria atrophied,cristae reduced,and membrane density increased in hepatocytes.The relative mRNA transcription level of transferrin receptor protein 1(TFR 1),iron uptake-related proteins divalent metal transporter-1(DMT 1),ferritin heavy chain 1(FTH 1),and voltage-dependent anion channel 3(VDAC 3)genes were significantly up-regulated(P<0.05),while,the relative mRNA transcription level of glutathione peroxidase 4(GPX 4)and solute carrier family-7 member-11(SLC7A 11)genes were significantly down-regulated(P<0.05).These results indicated that Aspergillus terreus MSF2 could cause ferroptosis in the liver of mice.It benefits for further study of the pathogenic mechanism of Aspergillus terreus.