摘要
目的探讨熟地黄提取物(RGE)对糖尿病肾病(DN)大鼠蛋白激酶B/糖原合成酶激酶-3β(AKT/GSK-3β)信号通路及足细胞上皮间质转化(EMT)的影响。方法采用高糖高脂、链脲佐菌素(STZ)诱导建立DN大鼠模型,建模成功60只,随机分为模型组、RGE(低、中、高)剂量组、阳性对照组,每组12只;另择12只大鼠作为对照组,对照组基础饲料、0.1 mol/L柠檬酸盐缓冲液对照处理。RGE(低、中、高)剂量组(1、2、4)g/kg RGE灌胃,阳性对照组4 mg/kg贝那普利灌胃,对照组、模型组0.9%氯化钠溶液灌胃,灌胃液体体积均为10 ml/kg,1次/d,连续6周。观察6组大鼠一般情况;血糖仪检测血糖,尿蛋白定量测试盒检测24 h尿蛋白,分光光度计检测肾功能指标尿肌酐、血清肌酐、血清尿素氮水平;HE染色、糖原(PAS)染色、马森三色(MASSON)染色观察肾脏组织形态;qRT-PCR法检测肾脏组织中E-钙黏素(E-Cad)、α-平滑肌肌动蛋白(α-SMA)、纤维粘连蛋白(FN)mRNA表达水平;Western blot法检测大鼠肾脏组织中磷酸化AKT(p-AKT)、AKT、磷酸化GSK-3β(p-GSK-3β)、GSK-3β蛋白表达水平。结果模型组大鼠毛色灰暗、精神萎靡,尿液量明显增多且发臭,肾脏组织肾小管空泡变性,肾小球系膜基质扩张,肾小球硬化和胶原沉积现象明显;RGE(低、中、高)剂量组、阳性对照组较模型组毛色有明显缓和、精神状态缓解,尿液量相对减少,肾脏组织病理变化明显减轻。与对照组相比,模型组血糖、24 h尿蛋白、尿肌酐、血清肌酐、血清尿素氮水平,肾脏组织中α-SMA、FN mRNA表达水平,p-AKT/AKT、p-GSK-3β/GSK-3β蛋白表达水平升高(均P<0.05),E-Cad mRNA表达水平降低(P<0.05)。与模型组相比,RGE低剂量组血糖、24 h尿蛋白、血清肌酐、血清尿素氮,肾脏组织中α-SMA、FN mRNA水平,p-AKT/AKT、p-GSK-3β/GSK-3β蛋白水平降低(均P<0.05);RGE(中、高)剂量组、阳性对照组血糖、24 h尿蛋白、尿肌酐、血清肌酐、血清
Objective To investigate the effects of rehmannia glutinosa extract(RGE)on protein kinase B/glycolgen synthase kinase-3β(AKT/GSK-3β)signaling pathway and podocyte epithelial-mesenchymal transition(EMT)in diabetic nep-hropathy(DN)rats.Methods DN rat models were established by streptozotocin(STZ)injection and feeding with high glucose,high fat diet.Sixty DN rats were randomly divided into 5 groups:model group,low-,medium-and high-dose RGE(1,2,4 g/kg)groups,and positive control group(4 mg/kg benazepril)with 12 rats in each group.Twelve rats fed with normal diet served as control group.The general condition of rats was observed,Blood glucose was measured by blood glucose meter,24 h urine protein was detected by urine protein quantitative test kit,the levels of urinary creatinine,serum creatinine and serum urea nitrogen were measured by spectrophotometer.Hematoxylin eosin(HE),periodic acid-schiff staining(PAS)and MASSON staining were used for histopathological studies.The mRNA expression levels of E-Cadherin(E-Cad),α-smooth muscle actin(α-SMA)and fibronectin(FN)were detected by real-time fluorescence quantitative PCR(qRT-PCR),and the protein levels of phosphorylated-Akt(p-AKT),AKT,phosphorylated-GSK-3β(p-GSK-3β)and GSK-3βwere detected by Western blot.Results In the model group,the hair turned to gray,the activity was reduced,the urine volume was increased and smelly,there were vacuolar degeneration of renal tubules,dilation of glomerular mesangial matrix,obvious glomerulosclerosis and collagen deposition,compared with the model group,the hair color and mental state of the RGE treatment groups and positive control group were significantly alleviated,the amount of urine decreased,the pathological changes were obviously attenuated.Compared with those in the control group,the levels of blood glucose,24-h urinary protein,urinary creatinine,serum creatinine,serum urea nitrogen,the mRNA levels ofα-SMA and FN,the protein levels of p-AKT/AKT and p-GSK-3β/GSK-3βin kidney tissue in model group were higher(all P<0.05),a
作者
管陈安
叶华茂
徐光标
王莎莎
蒋钻红
GUAN Chenan;YE Huamao;XU Guangbiao;WANG Shasha;JIANG Zuanhong(Department of Nephrology,Taizhou Hospital of Zhejiang Province,Taizhou 317000,China)
出处
《浙江医学》
CAS
2021年第4期358-363,I0003,共7页
Zhejiang Medical Journal
基金
台州市科技计划项目(1902ky28)。
