糖肾宁对糖尿病肾病小鼠肾小管损伤及铁蛋白自噬介导的铁死亡的影响  

Effects of Tangshenning on Renal Tubular Injury and Ferritinophagy-mediated Ferrop⁃tosis of Diabetic Kidney Disease Mice

作　　者：陈纯苇 单晓萌 邹大威[1,2] 高彦彬 朱智耀[1,2] 郑亚琳 侯伟欣[1,2] 崔家霖 CHEN Chun-wei;SHAN Xiao-meng;ZOU Da-wei;GAO Yan-bin;ZHU Zhi-yao;ZHENG Ya-lin;HOU Wei-xin;CUI Jia-lin(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China;Beijing Key Laboratory of TCM Collateral Disease Theory Research,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院,北京100069 [2]中医络病研究北京市重点实验室,北京100069

出　　处：《时珍国医国药》2023年第8期1871-1875,共5页Lishizhen Medicine and Materia Medica Research

基　　金：北京市自然科学基金(7172032);首都医科大学科研培育基金(PYZ21027);国家中医药管理局全国名老中医药专家传承工作室建设项目。

摘　　要：目的研究糖肾宁(TSN)对糖尿病肾病(DKD)小鼠肾小管损伤及铁蛋白自噬介导的铁死亡的影响。方法采用高脂饲料诱导KK-Ay小鼠建立DKD模型,与正常组相比24小时尿微量白蛋白排泄率(UAER)明显升高(P<0.05)且随机血糖≥16.7 mmol·L^(-1)视为模型建立成功。模型建立成功后,将KK-Ay小鼠随机分为模型组和糖肾宁组,连续给药10周。实验期间,动态监测血糖和UAER;给药10周后,检测血肌酐(Scr)、血尿素氮(BUN)水平;取肾组织进行HE、PAS和普鲁士蓝病理染色,电镜观察肾小管线粒体的超微结构,Western blot检测肾组织中LC3、P62、FTH1、NCOA4、KIM-1的蛋白表达水平,并检测MDA和Fe^(2+)含量。结果与正常组相比,模型组小鼠血糖、UAER、Scr、BUN明显升高(P<0.05),与模型组相比,糖肾宁组小鼠血糖没有明显变化(P>0.05),UAER、Scr、BUN明显降低(P<0.05);与正常组相比,模型组小鼠LC3Ⅱ/LC3Ⅰ、FTH1、KIM-1蛋白表达水平明显升高(P<0.05);P62和NCOA4蛋白表达水平明显降低(P<0.05);MDA和Fe^(2+)含量均明显升高(P<0.05)。与模型组相比,糖肾宁组小鼠LC3Ⅱ/LC3Ⅰ、FTH1、KIM-1的蛋白表达水平显著降低(P<0.05),P62和NCOA4蛋白表达水平显著升高(P<0.05),MDA和Fe^(2+)含量均明显降低(P<0.05)。光镜观察显示:模型组小鼠出现肾小管扩张且伴有空泡变性的病理损伤以及肾小管的铁离子沉积增加。电镜观察显示:模型组小鼠肾小管的线粒体体积明显缩小,线粒体嵴减少,线粒体双层膜密度增加,具备铁死亡的典型形态学特征。经中药TSN治疗后,上述病变均有所改善。结论糖肾宁可降低KK-Ay小鼠蛋白尿,保护肾功能,减轻肾小管损伤,其机制可能与抑制铁蛋白自噬介导的铁死亡有关。Objective To study the effect of Tangshenning(TSN)on renal tubular injury and ferritinophagy-mediated ferropto⁃sis in diabetic kidney disease(DKD).Methods KK-Ay mice were induced by high-fat diet to establish the DKD model.Com⁃pared with the normal group,when the 24-hour urinary microalbumin excretion rate(UAER)was significantly increased(P<0.05)and the random blood glucose was≥16.7 mmol·L^(-1),the KK-Ay mice were considered as successful models.After the models were successfully established,the KK-Ay mice were randomly divided into the model group and the Tangshenning group,And they were administered continuously for 10 weeks.During the experiment,blood glucose and UAER were monitored dynami⁃cally.After 10 weeks of administration,serum creatinine(Scr)and blood urea nitrogen(BUN)levels were detected.Kidney tis⁃sues were taken for HE,PAS and Prussian blue pathological staining and using electron microscopy to observe the ultrastructure of renal tubular mitochondria.the protein expressions of LC3,P62,FTH1,NCOA4,KIM-1 were assessed by Western blot.MDA and Fe^(2+)were examined.Results Compared with the normal group,the blood glucose,UAER,Scr and BUN in the model group were significantly increased(P<0.05);Compared with the model group,the blood glucose of the Tangshenning group did not change significantly(P>0.05),while UAER,Scr,and BUN were significantly decreased(P<0.05).Compared with the normal group,the protein expressions of LC3II/LC3I,FTH1 and KIM-1 were significantly increased(P<0.05),the protein expressions of P62 and NCOA4 were significantly decreased(P<0.05),and the contents of MDA and Fe^(2+)were significantly in⁃creased in the model group(P<0.05).Compared with the model group,the protein expressions of LC3II/LC3I,FTH1 and KIM-1 were significantly decreased(P<0.05),and the protein expressions of P62 and NCOA4 were significantly increased(P<0.05),the contents of MDA and Fe^(2+)were significantly decreased in the Tangshenning group(P<0.05).The pathological stai⁃ning of kidney tissue showed that

关 键 词：糖尿病肾病 糖肾宁 肾小管损伤 铁蛋白自噬 铁死亡 

分 类 号：R285.5[医药卫生—中药学]