摘要
目的:观察运脾和络方减轻Ⅱ型糖尿病大鼠肾损伤的作用并从Sirt1-AMPK-自噬角度探讨其机制。方法:取12周雄性Zucker糖尿病肥胖(ZDF)大鼠24只,根据空腹血糖水平随机分为模型组、Sirt1过表达组和运脾和络方组,高脂高糖喂养10周;另设ZL大鼠作为正常对照组,普食喂养10周。10周后留取尿液和血液检测肾功能指标,处死大鼠并取材留用。Western blot技术测定Sirt1、AMPK、p-AMPK、LC3和P62蛋白水平的变化;real-time PCR技术检测Sirt1的mRNA表达;HE染色和Masson染色法检测肾组织的病理变化。结果:与正常对照组比较,模型组大鼠空腹血糖(FBG)、尿蛋白(UP)、尿白蛋白(U-ALB)和血清肌酐(SCr)均明显升高(P<0.01);Sirt1的mRNA下降(P<0.05),Sirt1、AMPK、p-AMPK和LC3-Ⅱ/-Ⅰ的蛋白水平明显降低(P<0.01),P62的表达明显升高(P<0.01);肾小球局灶性纤维化,局灶性肾小管上皮细胞空泡变、坏死脱落及萎缩,出现管型,肾间质纤维增生。与模型组比较,Sirt1过表达组的FBG明显降低(P<0.01),运脾和络方组的FBG无明显变化(P>0.05),Sirt1过表达组和运脾和络方组的SCr和U-ALB均降低(P<0.05);Sirt1的mRNA表达增加(P<0.05),Sirt1、AMPK、p-AMPK和LC3-Ⅱ/-Ⅰ的蛋白水平明显升高(P<0.01),P62的表达明显降低(P<0.01);Sirt1过表达组和运脾和络方组的肾脏病变均有不同程度减轻。结论:运脾和络方可能通过提高Sirt1表达水平,激活AMPK,调节自噬,从而起到保护肾脏的作用。
AIM:To observe the effect on Yunpiheluo decoction(YPHL)on renal injury in type 2 diabetic rats and to explore the mechanism from the perspective of Sirt1-AMPK-autophagy.METHODS:Male Zucker diabetic fatty(ZDF)rats(n=24)were randomly divided into model group,Sirt1 over-expression group and YPHL group,and fed with high-fat and high-sugar diet for 10 weeks.ZL rats were used as normal control and fed with normal diet for 10 weeks.After 10 weeks,urine and blood were collected for renal function detection.The rats were sacrificed and specimen was submitted.In addition,the mRNA expression of Sirt1 was analyzed by real-time PCR.The protein levels of Sirt1,AMPK,p-AMPK,LC3 and P62 in the renal tissues wene determined by Western blot.The renal pathological changes were observed under light microscope with HE and Masson staining.RESULTS:Compared with control group,fasting blood glucose(FBG),urinary protein(UP),urinary albumin(U-ALB)and serum creatinine(SCr)in model group were obviously increased(P<0.01).The mRNA expression of Sirt1 was decreased(P<0.05).The protein levels of SIRT1,AMPK,p-AMPK and LC3-II/-I were decreased(P<0.01),and P62 was increased(P<0.01).Glomerular focal fibrosis,focal renal tubular epithelial cell vacuolation,necrosis,shedding and atrophy,tubular type,and renal interstitial fibrosis were observed.Compared with model group,FBG was obviously decreased in Sirt1 over-expression group(P<0.01),but it showed no significant change in YPHL group(P>0.05).SCr and U-ALB were decreased(P<0.05),Sirt1 mRNA was increased(P<0.05),the protein levels of SIRT1,AMPK,p-AMPK and LC3-II/-I were increased(P<0.01),and P62 was decreased(P<0.01)in Sirt1 over-expression group and YPHL group.HE and Masson staining showed that the renal damage in Sir1 over-expression group and YPHL group was attenuated.CONCLUSION:Yunpiheluo decoction may protect the kidney by increasing the expression level of Sirt1,activating AMPK,and regulating autophagy.
作者
郭燚
唐比强
赵娜
柴可夫
杜月光
GUO Yi;TANG Bi-qiang;ZHAO Na;CHAI Ke-fu;DU Yue-guang(Pathology Department,Zhejiang Chinese Medical University,Hangzhou 310053,China;National TCM Clinical Research Center,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第12期2156-2162,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81774177)
