摘要
目的 观察益肾通络方对糖尿病肾病(DN)小鼠的疗效,并探讨其通过AMPK/mTOR/ULK1通路产生影响的作用机制。方法 将C57BL/6小鼠48只分为对照组8只、造模组40只。造模组STZ腹腔注射建立DN模型,造模成功后随机分为模型组,益肾通络方高、中、低剂量组(18.2,9.1,4.55 g·kg^(-1)),司美格鲁肽组(40μg·kg^(-1)),各8只,连续干预12周。干预后检测血糖、24 h尿蛋白、肾脏系数、血肌酐及尿素氮,进行肾脏HE、PAS、PASM、Masson染色,测定尿β2-MG、NGAL、KIM-1水平;PCR与Western blot检测自噬及AMPK/mTOR/ULK1相关基因与蛋白。结果 模型组血糖、尿蛋白、肾功能损伤指标及肾脏病理改变显著加重,尿β2-MG、NGAL、KIM-1升高,自噬相关基因与蛋白表达紊乱(AMPK/ULK1下调,mTOR/LC3/p62上调)(P<0.05)。与模型组相比,益肾通络方及司美格鲁肽均可降低血糖、尿蛋白及肾损伤指标,改善病理改变,降低尿损伤标志物,促进AMPK、ULK1表达并抑制mTOR、LC3、p62(P<0.05)。结论 益肾通络方可能通过调控AMPK/mTOR/ULK1信号通路改善自噬应激,减轻DN小鼠肾小管间质损伤,从而发挥肾保护作用。
Objective To evaluate the therapeutic effects of Yishen Tongluo Formula(益肾通络方,YSTLF)on diabetic nephropathy(DN)in mice and to investigate its underlying mechanism involving the AMPK/mTOR/ULK1 signaling pathway.Methods Forty-eight C57BL/6 mice were randomly divided into a control group(n=8)and a model group(n=40).DN was induced by intraperitoneal injection of streptozotocin(STZ).Successfully modeled mice were randomly assigned to the following groups(n=8 each):model group,high/medium/low-dose YSTLF(18.2,9.1,and 4.55 g/kg),and semaglutide(40μg/kg)groups.The interventions were administered for 12 weeks.After treatment,blood glucose,24 h urinary protein,kidney index,serum creatinine,and blood urea nitrogen(BUN)were measured.Renal histopathology was assessed by HE,PAS,PASM,and Masson staining.Urinaryβ2-MG,NGAL,and KIM-1 were detected by ELISA.The expression of autophagy-related and AMPK/mTOR/ULK1 pathway-related genes and proteins was analyzed using quantitative PCR and Western blot(WB).Results Compared with controls,DN mice showed significantly increased blood glucose,urinary protein,renal injury indices,and pathological damage,as well as elevated urinaryβ2-MG,NGAL,and KIM-1 levels.Autophagy was dysregulated,characterized by decreased AMPK and ULK1 and increased mTOR,LC3,and p62 expression(P<0.05).Treatment with YSTLF or semaglutide significantly alleviated hyperglycemia,proteinuria,and renal impairment,and ameliorated pathological changes compared with the model group(P<0.05).These treatments also reduced urinary levels of β2-MG,NGAL,and KIM-1.Furthermore,they upregulated AMPK and ULK1 expression while downregulating mTOR,LC3,and p62(P<0.05).Con⁃clusion YSTLF may exert renoprotective effects in DN mice by modulating the AMPK/mTOR/ULK1 signaling pathway,restoring autophagic balance,and alleviating tubulointerstitial injury.
作者
张轶斐
张泽钰
席俊羽
曹梓静
白雪慧
唐靖怡
张术姣
张帅星
谢亦冉
吴宇琪
刘忠杰
刘伟敬
ZHANG Yifei;ZHANG Zeyu;XI Junyu;CAO Zijing;BAI Xuehui;TANG Jingyi;ZHANG Shujiao;ZHANG Shuaixing;XIE Yiran;WU Yuqi;LIU Zhongjie;LIU Weijing(Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Puren Hospital,Beijing 100062,China)
出处
《时珍国医国药》
北大核心
2025年第24期4641-4648,共8页
JOURNAL OF LI-SHIZHEN TRADITIONAL CHINESE MEDICINE
基金
国家自然科学基金面上项目(82374382,82074361,82274293)
中央高校基本科研业务费专项资金资助项目揭榜挂帅项目(2023-JYB-JBZD-037)
国家中医药管理局高水平医院临床研究和成果转化能力提升试点项目揭榜挂帅项目(DZMG-XZYY-23002)
中华中医药学会青年求实项目(ZSL-003-02)。
