黄花倒水莲通过激活SIRT1/FoxO1通路抑制糖尿病肾病大鼠氧化应激

The Inhibition Effect of Polygala Fallax Hemsl on Oxidative Stress in Diabetic Kidney Disease Rats by the Activating SIRT1/FoxO1 Pathway

研究黄花倒水莲通过激活SIRT1/FoxO1通路对糖尿病肾病大鼠氧化应激的影响.采用链脲霉素联合高脂高糖饮食建立糖尿病肾病(DKD)大鼠模型.将36只SD大鼠随机分为对照组,模型组,黄花倒水莲低、中、高剂量组,EX527组.黄花倒水莲组灌胃50、100、200 mg/kg黄花倒水莲酒精性提取物,EX527组腹腔注射5 mg/kg EX527,每天1次,连续8周.测量各组大鼠体重、血糖、24 h尿蛋白(24h-UP)、尿素氮(BUN)、血肌酐(Scr)水平;H&E染色观察肾脏组织的病变;ELISA检测肾组织内MDA、SOD、GSH-Px水平;Masson染色观察肾组织胶原沉积,免疫组化检测纤连蛋白(fibronectin)、胶原Ⅳ蛋白(collagenⅣ)的表达;western blot检测SIRT1、FoxO1、CAT、MnSOD、p-FoxO1蛋白表达.与模型组比较,黄花倒水莲治疗后,大鼠体重增加(P<0.05),血糖和24 h尿蛋白水平降低,BUN、Scr水平降低(P<0.01或P<0.001);肾组织病变明显减轻;肾组织ROS和MDA水平降低(P<0.05或P<0.001),SOD和GSH-Px活性升高(P<0.01或P<0.001);胶原沉积显著减少,fibronectin和collagenⅣ阳性表达水平明显降低(P<0.05或P<0.001);CAT、SIRT1和MnSOD的表达明显增加(P<0.001),p-FoxO1/FoxO1的比值显著降低(P<0.001).EX527干预显著减弱黄花倒水莲的治疗效果.黄花倒水莲可改善DKD模型大鼠肾组织氧化应激状态,其机制可能与激活SIRT1/FoxO1信号通路有关.

This study was to investigate the effect of Polygala fallax Hemsl on oxidative stress in rats with diabetic kidney disease(DKD).A rat model of DKD was established by using streptozotocin(STZ)combined with high-fat and high-sugar diet.Thirty-six SD rats were randomly divided into control group,model group,and low,medium and high dose groups of Polygala fallax Hemsl,and EX527 group.The Polygala fallax Hemsl groups was intragastric with 50,100 and 200 mg/kg Alcoholic extract of Polygala fallax Hemsl,the EX527 group was intraperitoneally injected with 5 mg/kg EX527,once a day for 8 weeks.The body weight,blood glucose,24 h urine protein(24h-UP),serum BUN and Scr were measured in each group;H&E staining was performed to observe the lesions in kidney tissues;ELISA was performed to detect levels of MDA,SOD and GSH-Px in kidney tissues;Masson staining was performed to observe collagen deposition in kidney tissues;the expression levels of fibronectin andcollagen IV were detected by immunohistochemistry;the protein expression of SIRT1,FoxO1,CAT,MnSOD and p-FoxO1 were detected by western blot.Compared with model group,the rats in Polygala falla xHemsl group had increased body weight(P<0.05),decreased blood glucose and 24h-UP levels,decreased serum BUN and Scr levels(P<0.01 or P<0.001);significantly reduced renal tissue lesions;decreased renal tissue ROS and MDA levels(P<0.05 or P<0.001),increased SOD and GSH-Px activities(P<0.01 or P<0.001);collagen deposition was significantly reduced,and positive expression levels of fibronectin and Collagen IV were significantly reduced(P<0.05 or P<0.001);The expressions of CAT,SIRT1 and MnSOD were significantly increased(P<0.001),and the ratio of P-FoxO1 to FoxO1 was significantly decreased(P<0.001).The EX527 intervention significantly reduced the therapeutic effect of Polygala fallax Hemsl.The oxidative stress state of kidney tissue of DKD model rats can be improved by Polygala fallax Hemsl,and the mechanism may be related to activation of SIRT1/FoxO1 signaling pathway.