铁死亡参与高蛋氨酸饮食所致的ApoE^(-/-)小鼠肾脏损伤  

Ferroptosis participates in renal damage induced by high methionine diet in ApoE^(-/-) mice

作　　者：畅思容 孙岳 杨安宁[2,3] 李媛媛 王梦 宝瑞 余梦雪 刘志宏 CHANG Si-rong;SUN Yue;YANG An-ning;LI Yuan-yuan;WANG Meng;BAO Rui;YU Meng-xue;LIU Zhi-hong(*School of Public Health and Management,Ningxia Medical University,Yinchuan,Ningxia 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院,宁夏银川750004 [2]宁夏医科大学基础医学院 [3]国家卫生健康委员会代谢性心血管疾病研究重点实验室,宁夏银川750004

出　　处：《现代预防医学》2023年第17期3133-3138,3150,共7页Modern Preventive Medicine

基　　金：国家自然科学基金项目(81960018,82160088,82060264);宁夏回族自治区科学技术厅重点研发计划项目(2020BEG03008,2021BEG02030,2020BFH02003)

摘　　要：目的探讨铁死亡是否参与高蛋氨酸饮食所致的ApoE^(-/-)小鼠肾脏损伤。方法C57BL/6J普通饲料喂养小鼠作为正常对照组、将ApoE^(-/-)小鼠随机分为模型对照组(普通饲料喂养)和高蛋氨酸组(高蛋氨酸饲料喂养),每组6只小鼠。PAS和Masson染色观察小鼠肾脏损伤以及纤维化情况;免疫荧光染色检测肾组织纤维连接蛋白(fibronectin)含量;比色法检测肾组织二价铁离子含量;免疫荧光染色和硫代巴比妥酸(thiobarbituric acid,TBA)法检测肾组织丙二醛(malondialdehyde,MDA)含量;Western blotting和qRT-PCR检测p53、SLC7A11和GPX4的蛋白以及mRNA表达水平,多组数据比较采用单因素方差分析,多重比较采用Tukey检验。结果染色结果显示,载脂蛋白E(apolipoprotein E,ApoE)基因缺失引起肾脏细胞外基质沉积,出现肾损伤,而高蛋氨酸饮食则进一步加重了肾脏细胞外基质沉积,肾损伤加重;与正常对照组相比,模型对照组和高蛋氨酸组Fibronectin含量增加(F=87.330,P<0.001),铁离子(F=111.200,P<0.001)和MDA(F=205.200,P<0.001)含量增加,p53的蛋白水平增加(F=32.380,P<0.001)、mRNA水平增加(F=24.840,P<0.001),SLC7A11和GPX4的蛋白水平降低(F_(SLC7A11)=18.620,P<0.001;F_(GPX4)=20.830,P<0.001)、mRNA水平降低(F_(SLC7A11)=11.200,P<0.001;F_(GPX4)=23.530,P<0.001);与模型对照组相比,以上指标在高蛋氨酸组变化更显著(P<0.05)。结论铁死亡参与了高蛋氨酸饮食所致的ApoE^(-/-)小鼠肾脏损伤。Objective To investigate whether ferroptosis is involved in renal damage in Apo E^(-/-)mice induced by high methionine diet.Methods C57BL/6J mice fed with normal diet were used as normal control group.ApoE^(-/-)mice were randomly divided into model control group(fed with normal diet)and high methionine group(fed with high methionine diet),with 6 mice in each group.Renal injury and fibrosis in mice were observed by PAS and Masson staining.Detection of Fibronectin content by immunofluorescence staining and iron content was measured by colorimetry in renal tissue.The content of malondialdehyde(MDA)in renal tissue was detected by immunofluorescence staining and Thiobarbituric acid(TBA)method.Protein and mRNA expression of p53,SLC7A11,and GPX4 were determined by Western blotting and qRT-PCR.One-way ANOVA was used to compare multiple groups,and Tukey test was used for multiple comparisons.Results The staining results showed that apolipoprotein E(ApoE)gene deletion caused renal extracellular matrix deposition and renal injury,while high methionine diet further aggravated renal extracellular matrix deposition and renal injury.Compared with the normal control group,the content of Fibronectin in the model control group and high methionine group increased(F=87.330,P<0.001),the content of iron ion(F=111.200,P<0.001)and MDA(F=205.200,P<0.001)increased,the protein level of p53 increased(F=32.380,P<0.001),the level of mRNA increased(F=24.840,P<0.001),the protein level of SLC7A11 and GPX4 decreased(F_(SLC7A11)=18.620,P<0.001;F_(GPX4)=20.830,P<0.001),and mRNA level decreased(F_(SLC7A11)=11.200,P<0.001;F_(GPX4)=23.530,P<0.001).Compared with the model control group,the above indexes changed more significantly in the high methionine group(P<0.05).Conclusion Ferroptosis is involved in renal damage in Apo E^(-/-)mice induced by high methionine diet.

关 键 词：肾脏损伤 铁死亡 HHCY GPX4 脂质过氧化 

分 类 号：R692[医药卫生—泌尿科学] R595[医药卫生—外科学]