摘要
背景:在慢性肾脏病中常常伴有同型半胱氨酸水平升高,导致足细胞凋亡,但是其具体机制还尚未清楚。目的:探讨高同型半胱氨酸血症诱导Cbs^+/-小鼠肾损伤的作用机制。方法:选取体质量相近的Cbs^+/+小鼠(对照组)和Cbs^+/-小鼠(模型组),每组10只,两组小鼠均饲以高蛋氨酸饮食,8周后处死,分离血清,留取肾脏组织。采用全自动生化分析仪检测血清同型半胱氨酸、尿素氮、肌酐水平;糖原染色法及透射电镜观察肾脏损伤情况;TUNEL染色观察肾小球中细胞凋亡情况;Western blot检测Bax、Bcl-2和caspase12的蛋白表达水平。结果与结论:①与Cbs^+/+小鼠比较,Cbs^+/-小鼠血清同型半胱氨酸、尿素氮及肌酐水平均明显升高(P<0.01);②糖原染色结果显示,Cbs^+/+小鼠肾小球基底膜清楚,粗细均匀,而Cbs^+/-小鼠肾小球基底膜则呈现不同程度的粗细不均,膜区增宽、基质增厚;③透射电镜显示,Cbs^+/+小鼠肾小球基底膜清晰,足突规则,而Cbs^+/-小鼠肾小球基底膜局灶性增厚,足突不规则融合;④TUNEL染色结果显示,与Cbs^+/+小鼠比较,Cbs^+/-小鼠肾小球内凋亡细胞数量明显增加;⑤Western blot检测结果显示,Bax/Bcl-2和caspase12的蛋白水平均升高(P<0.05);⑥结果表明:足细胞凋亡在高同型半胱氨酸血症诱导Cbs^+/-小鼠肾损伤中发挥着重要的作用。
BACKGROUND:In chronic kidney disease,there is often an increase in the level of homocysteine,which can lead to podocyte apoptosis,but the specific mechanism is not clear.OBJECTIVE:To investigate the mechanism of hyperhomocysteinemia-induced renal injury in Cbs^+/-mice.METHODS:Cbs^+/+mice(control group)and Cbs^+/-mice(model group)with similar body weight were selected,with 10 mice in each group,and were fed with high methionine diet.After 8 weeks,the mice were killed,the serum was separated and the kidney tissue was obtained.The levels of serum homocysteine,urea nitrogen and creatinine were measured by automatic biochemical analyzer.The renal injury was observed by Periodic Acid-Schiff staining and transmission electron microscope.TUNEL staining was used to observe the apoptosis of glomeruli.The protein expression levels of Bax,Bcl-2 and caspase12 were detected by western blot.RESULTS AND CONCLUSION:Compared with Cbs^+/+mice,the level of serum homocysteine,urea nitrogen and creatinine in Cbs^+/-mice were significantly increased(P<0.01).The Periodic Acid-Schiff staining results showed that the glomerular basement membrane of Cbs^+/+mice was clear and the thickness was uniform,while the Cbs^+/-mouse glomerular basement membrane showed varying degrees of uneven thickness,widening of membrane area and thickening of matrix.Under the transmission electron microscope,the glomerular basement membrane of Cbs^+/+mice was clear and the foot process was regular,while the glomerular basement membrane of Cbs^+/-mice was locally thickened and the foot process was irregular fusion.TUNEL staining showed that the number of apoptotic cells in glomeruli of Cbs^+/-mice was significantly increased compared with Cbs^+/+mice;meanwhile,western blot detection showed that the protein levels of Bax/Bcl-2 and caspase12 were significantly increased(P<0.05).To conclude,podocyte apoptosis plays an important role in hyperhomocysteinemiainduced renal injury in Cbs^+/-mice.
作者
吴凯
刘昆
谢琳
卢冠军
马胜超
李桂忠
曹军
揭育祯
姜怡邓
郝银菊
Wu Kai;Liu Kun;Xie Lin;Lu Guanjun;Ma Shengchao;Li Guizhong;Cao Jun;Jie Yuzhen;Jiang Yideng;Hao Yinju(Clinical Medical College of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Key Laboratory of Metabolic Cardiovascular Diseases Research of National Health Commission,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Ningxia Key Laboratory of Vascular Injury and Repair Research,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Basic Medical College of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2021年第11期1728-1732,共5页
Chinese Journal of Tissue Engineering Research
基金
宁夏高等学校科学研究项目(NZY2017122),项目负责人:郝银菊。
