摘要
目的 观察益糖康对db/db小鼠肾脏异位脂质沉积及PPAR-α/PGC-1α信号通路的影响,探讨其改善肾脏损伤的作用机制。方法 采用随机数字表法将30只db/db小鼠分为模型组、益糖康组和氯沙坦钾组(每组10只),并设10只db/m小鼠为空白组,分别予相应干预8周。检测小鼠体质量、空腹血糖(FBG)、血肌酐(SCr)、血尿素氮(BUN)、24 h尿蛋白含量,HE、PAS、Masson染色观察肾组织形态及损伤程度,油红O染色观察小鼠肾脏脂质沉积情况,透射电镜观察足细胞超微结构,免疫荧光染色检测肾组织肉碱棕榈酰转移酶1A(CPT1A)、核因子E2相关因子2(Nrf2)表达,实时荧光定量PCR检测过氧化物酶体增殖物激活受体α(PPAR-α)、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)m RNA表达,Western blot检测PPAR-α、CPT1A、PGC-1α、Nrf2、核呼吸因子1(NRF1)、线粒体转录因子A(TFAM)蛋白表达。结果 与空白组比较,模型组小鼠体质量、FBG显著升高(P<0.05),SCr、BUN、24 h尿蛋白含量显著增加(P<0.05);肾小球肥大,系膜细胞与基质增生,基底膜增厚,肾间质胶原纤维异常沉积,肾组织脂质沉积程度增加,足细胞足突广泛融合,足突密度显著降低(P<0.05),线粒体外膜边界模糊,形态肿胀,内嵴模糊不清;肾组织PPAR-α、PGC-1α mRNA表达显著降低(P<0.05),PPAR-α、CPT1A、PGC-1α、Nrf2、NRF1、TFAM蛋白表达显著降低(P<0.05)。与模型组比较,益糖康组小鼠体质量和FBG显著降低(P<0.05),氯沙坦钾组上述指标差异无统计学意义(P>0.05);益糖康组和氯沙坦钾组SCr、BUN、24 h尿蛋白含量显著降低(P<0.05);肾组织病理损伤减轻、脂质沉积减少,足细胞超微结构改善,足突密度显著升高(P<0.05);肾组织PPAR-α、PGC-1α mRNA表达升高,PPAR-α、CPT1A、PGC-1α、Nrf2、NRF1、TFAM蛋白表达升高,差异均有统计学意义(P<0.05)。结论 益糖康能有效改善糖尿病肾病小鼠肾脏损伤,其机制可能与激活PPAR-α/PGC-1α信号通路,减少肾脏脂质沉积和线粒体生物发生有关。
Objective To investigate the effects of Yitangkang on renal ectopic lipid deposition and the PPAR-α/PGC-1αsignaling pathway in db/db mice;To explore its mechanism in improving renal injury.Methods Totally 30 db/db mice were divided into model group,Yitangkang group and losartan potassium group(10 mice in each group)using a random number table method.An additional 10 db/m mice were assigned as the blank group.The mice received corresponding interventions for 8 weeks.Body mass,fasting blood glucose(FBG),serum creatinine(SCr),blood urea nitrogen(BUN),and 24-hour urinary protein content were measured,renal morphology and injury were observed using HE,PAS and Masson staining,lipid deposition in renal tissue was observed by oil red O staining,renal ultrastructure was observed by transmission electron microscopy,immunofluorescence staining was used to detect the expressions of CPT1A and Nrf2 in renal tissue,RT-qPCR was performed to assess the mRNA expressions of PPAR-αand PGC-1α,Western blot was used to detect the protein expressions of PPAR-α,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM.Results Compared with the blank group,the body mass and FBG significantly increased in the model group(P<0.05),and the contents of SCr,BUN and 24-hour urinary protein significantly increased(P<0.05);histopathology revealed glomerular hypertrophy,mesangial cell and matrix proliferation,thickened basement membrane,abnormal deposition of interstitial collagen fibers,increased lipid deposition in renal tissue,widespread foot process effacement,reduced foot process density(P<0.05),blurred mitochondrial outer membranes,swollen morphology,and indistinct cristae;mRNA expressions of PPAR-αand PGC-1αsignificantly decreased(P<0.05),and protein expressions of PPARα,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM significantly decreased(P<0.05).Compared with the model group,the body mass and FBG of mice in Yitangkang group significantly decreased(P<0.05),but there was no significant difference in the above indexes in losartan potassium group(P>0.05);the contents of SCr,BUN and 24-hour urinary protein in Yitangkang group and losartan potassium group significantly decreased(P<0.05);the pathological damage and lipid deposition of renal tissue was alleviated,the ultrastructure of podocytes was improved,and the density of podocytes significantly increased(P<0.05);the mRNA expressions of PPARαand PGC-1αin renal tissue increased,and the protein expressions of PPAR-α,CPT1A,PGC-1α,Nrf2,NRF1 and TFAM increased,with statistical significance(P<0.05).Conclusion Yitangkang can effectively alleviate renal injury in diabetic nephropathy mice.The mechanism may be related to the activation of PPAR-α/PGC-1αsignaling pathway and the reduction of renal lipid deposition and mitochondrial biogenesis.
作者
王诗怡
孙贵炎
张晖
杨宇峰
石岩
WANG Shiyi;SUN Guiyan;ZHANG Hui;YANG Yufeng;SHI Yan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Affiliated Cixi Hospital,Wenzhou Medical University,Ningbo 315300,China)
出处
《中国中医药信息杂志》
2025年第12期89-96,共8页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家中医药管理局青年岐黄学者项目(2022年)
辽宁省应用基础研究计划(2023JH2/101300050)
辽宁省“兴辽英才计划”青年拔尖人才项目(XLYC1807145)
辽宁省科技计划联合计划自然科学基金-博士科研启动项目(2024-BSLH-152)
宁波市卫生健康科技计划项目(2024Y66)。
