健脾升清降浊方干预糖尿病肾病小鼠蛋白尿的作用及机制研究  

Study on the Effect and Mechanisms of Jianpi Shengqing Jiangzhuo Recipe on Proteinuria in Mice with Diabetic Kidney Disease

作　　者：冯慧 凌云[1] 夏子琪 朱晓云[3] 朱鹏飞 FENG Hui;LING Yun;XIA Ziqi;ZHU Xiaoyun;ZHU Pengfei(School of Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Integrated Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Guang anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;Department of Endocrinology,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210004,China)

机构地区：[1]南京中医药大学中医学院,江苏南京210023 [2]南京中医药大学中西医结合学院,江苏南京210023 [3]中国中医科学院广安门医院,北京100053 [4]江苏省中医院内分泌科,江苏南京210004

出　　处：《南京中医药大学学报》2025年第2期213-222,共10页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省科技发展计划青年人才项目(QN202102);江苏省高等学校基础科学(自然科学)研究项目(23KJB360007);国家中医药管理局高水平中医药重点学科建设项目(国中医药人教函〔2023〕85号);江苏省中医院优秀青年博士培养计划(2024QB005)。

摘　　要：目的探讨健脾升清降浊方对糖尿病肾病(DKD)db/db小鼠的保护作用及可能的机制。方法32只8周龄雄性db/db小鼠随机分为模型组,西药组[达格列净(1.0 mg·kg^(-1)·d^(-1))],健脾升清降浊低、高剂量组(19.63、58.89 g·kg^(-1)·d^(-1)),每组8只。8只db/m小鼠作为正常组。每日灌胃给药1次,连续10周。观察小鼠一般生存状况,动态监测小鼠体质量、空腹血糖(FBG)、24 h尿量;检测尿肌酐(Ucr)、尿微量白蛋白,计算尿微量白蛋白排泄(UAE)及蛋白肌酐比(ACR)。末次干预结束后,禁食12 h,麻醉取血,分离肾脏组织;检测血肌酐(Scr)、尿素氮(BUN)、甘油三脂(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)水平;HE、Masson染色观察肾脏组织病理变化;油红O染色观察肾脏内脂滴沉积情况,采用Image J进行定量分析;ELISA检测肾组织TNF-α、IL-1β水平;Western blot检测肾组织SIRT1、SREBP-1、PPAR-α蛋白表达。结果与正常组小鼠相比,模型组小鼠体质量、FBG、Scr、TG、TC、HDL-C、LDL-C、尿量、UAE及ACR显著升高(P<0.05,P<0.01),肾小球体积明显增大,肾脏纤维化改变,肾脏脂滴沉积增多(P<0.01),TNF-α、IL-1β、SREBP-1表达增加(P<0.01),SIRT1、PPAR-α表达减少(P<0.01)。与模型组相比,干预第5、10周,西药组FBG水平显著降低(P<0.05,P<0.01);干预结束后,西药组与健脾升清降浊高剂量组Scr、UAE与ACR均显著降低(P<0.05,P<0.01),而健脾升清降浊高剂量组血清TG水平亦显著降低(P<0.01);西药组与健脾升清降浊高剂量组肾脏病理变化、脂质沉积改善(P<0.05,P<0.01),TNF-α、IL-1β水平降低(P<0.05,P<0.01),SIRT1、PPAR-α蛋白表达增加(P<0.05,P<0.01);健脾升清降浊高剂量组SREBP-1表达降低(P<0.01)。结论健脾升清降浊方能改善DKD蛋白尿、肾损伤、肾脏脂质沉积与炎症反应,其机制可能与激活SIRT1/SREBP-1/PPAR-α通路而调节脂稳态有关。OBJECTIVE To explore the protective effect of Jianpi Shengqing Jiangzhuo Recipe on db/db mice with diabetic kidney disease(DKD)and its possible mechanism.METHODS Thirty-two 8-week-old male db/db mice were randomly divided into four groups(n=8),including the model group,the Western medicine group[Dapagliflozin(1.0 mg·kg^(-1)·d^(-1))],low-dose Jianpi Shengqing Jiangzhuo group(19.63 g·kg^(-1)·d^(-1)),and high-dose Jianpi Shengqing Jiangzhuo group(58.89 g·kg^(-1)·d^(-1)).Additionally,8 db/m mice were used as the normal group.The mice were orally administered once a day for 10 consecutive weeks.The general survival status of the mice was observed,and the body weight,fasting blood glucose(FBG),and urine volume of the mice were dynamically monitored;urine creatinine and urine microalbumin were detected,and urine microalbumin excretion(UAE)and protein-creatinine ratio(ACR)were calculated.After the last intervention,mice were fasted for 12 h,blood was collected under anesthesia,and kidney tissue was separated;blood creatinine(Scr),serum urea nitrogen(BUN),triglycerides(TG),total cholesterol(TC),low-density lipoprotein(LDL-C),and high-density lipoprotein(HDL-C)were tested;HE and Masson staining were used to observe pathological changes in renal tissue;Oil Red O staining was used to observe the deposition of lipid droplets in the kidneys,and Image J was used for quantitative analysis;ELISA was used to detect the levels of TNF-αand IL-1βin renal tissue;Western blot was used to detect the expression of SIRT1,SREBP-1,and PPAR-αproteins in renal tissue.RESULTS Compared with the normal group mice,the body weight,FBG,Scr,TG,TC,HDL-C,LDL-C,urine volume,UAE,and ACR of the model group mice were significantly increased(P<0.05,P<0.01);the glomerular volume was significantly increased,renal fibrosis was altered,and renal lipid droplet deposition increased(P<0.01);renal TNF-α,IL-1β,SREBP-1 expression increased(P<0.01),and SIRT1 and PPAR-αexpression decreased(P<0.01).Compared with the model group,the FBG levels in the Western

关 键 词：糖尿病肾病 健脾升清降浊方 异位脂质沉积 脂稳态 炎症反应 

分 类 号：R285.5[医药卫生—中药学]