| ObjectiveIn this study,Nrf2 signaling pathway was taken as the research target to explore the preventive effect of natural Nrf2 activator on DN,so as to provide basic experimental data for DN prevention and control,and further clarify the antioxidant mechanism of GSPE,so as to provide support for the development and utilization of grape resources in xinjiang.MethodsA total of 58 healthy male SD rats were randomly selected as the blank control group,and the remaining 48 SD rats were randomly selected as the control group for diabetic modeling.The modeling method was fed with high-fat and high-sugar diet and intraperitoneal injection of STZ,30mg/kg,once a week,twice in total.After two weeks,random blood glucose ?16.7mmol/L was taken as the standard for successful modeling.Then the model group was stratified and randomly divided into T2 DM group,100mg/kg GSPE intervention group,250mg/kg GSPE intervention group and10mg/kg CDDO-Me intervention group according to blood glucose level.During the experiment,free water and food intake were taken,weight and blood glucose were measured regularly every week,activity,water intake and urine volume were observed and recorded.Pathological changes of kidney were observed by HE staining.The level of insulin was determined by ELISA.Serum creatinine,blood urea nitrogen,24-hour urine albumin and uric acid levels were measured with related kits.The contents of MDA,T-AOC,GSH,gsh-px and SOD were determined with related kits.Western Blot was used to detect the protein expressions of Nrf2,HO-1,GST and NQO1 in the kidney of rats.The m RNA levels of Nrf2,HO-1,GST and NQO1 in rat kidney were detected by q RT-PCR.Results univariate analysis of variance was used to compare the differences among the groups,and Bonferroni method was used for multiple comparisons,with the test level ? =0.05.Results1.Effects of GSPE on blood glucose,insulin level and insulin resistance homeostasis model index in T2 DM ratsFrom the successful modeling to the end of intervention,the blood glucose,insulin level and insulin resistance homeostasis model index of the 100mg/kg and250mg/kg GSPE groups were significantly higher than those of the control group(P<0.05).Compared with the T2 DM group,both the 100mg/kg and 250mg/kg GSPE groups significantly reduced blood glucose levels(P<0.05),significantly increased insulin levels(P<0.05),and significantly reduced the steady-state model index of insulin resistance(P<0.05).Compared with the normal control group,the blood glucose,insulin level and the steady-state model index of insulin resistance in the100mg/kg and 250mg/kg GSPE groups were significantly higher than those in the control group(P<0.05).2.Effects of GSPE on renal pathology in T2 DM ratsPathological results showed that,compared with normal control group,the T2 DM group rat glomerular volume increase,glomerular capillary basement membrane thickening,mesangial proliferation width less than the capillary diameter,the segmental distribution of renal vascular lesions,the hyperplasia of arteriole at visible goal hardening,100 mg/kg of GSPE intervention group in addition to the basement membrane thickening,no other abnormalities,250 mg/kg of GSPE intervention group found no abnormalities.3.Effects of GSPE on renal function in T2 DM ratsCompared with the normal control group,serum creatinine,blood urea nitrogen,24-hour urine albumin and uric acid in the T2 DM group were significantly increased(P<0.05).Compared with the T2 DM group,serum creatinine,blood urea nitrogen and uric acid levels in the 100mg/kg GSPE intervention group decreased significantly(P<0.05),while serum creatinine,blood urea nitrogen and uric acid levels in the250mg/kg GSPE intervention group decreased significantly(P<0.05).Serum creatinine,blood urea nitrogen,24-hour urinary albumin and uric acid in the cddo-me intervention group were significantly lower than those in the T2 DM group(P<0.05).4.Effects of GSPE on oxidative damage and antioxidant indexes of kidney in T2 DM ratsCompared with the normal control group,MDA content in T2 DM group increased significantly(P<0.05),and SOD,T-AOC,GSH,gsh-px levels decreased significantly(P<0.05).Compared with the T2 DM group,MDA level in the 100mg/kg GSPE intervention group was significantly decreased(P<0.05),while GSH-Px and T-AOC levels were significantly increased(P<0.05),but SOD and GSH levels were not significantly different(P>0.05).Compared with the T2 DM group,MDA content in the 250mg/kg GSPE intervention group was significantly decreased(P<0.05),while SOD,T-AOC,GSH and GSH-Px levels were significantly increased(P<0.05).Compared with T2 DM group,MDA content in CDDO-Me group was significantly decreased(P<0.05),while SOD,T-AOC,GSH and GSH-Px levels were significantly increased(P<0.05).5.Regulation of Nrf2 signaling pathway in kidney of T2 DM rats by GSPECompared with the normal control group and the T2 DM group,the GST protein expression of the 100 mg / kg GSPE intervention group was significantly increased(P<0.05),but the protein expression levels of Nrf2,HO-1,NQO1 were not significantly different(P> 0.05);250mg / kg GSPE intervention group Nrf2,HO-1,NQO1,GST protein expression and m RNA expression levels were significantly higher than normal control group and T2 DM group(P <0.05);compared with normal control group and T2 DM group In the CDDO-Me intervention group,the protein expression and m RNA expression of Nrf2,HO-1,NQO1,and GST were significantly up-regulated(P <0.05).Compared with the normal control group and the T2 DM group,the m RNA expression level of NQO1 in the 100 mg / kg GSPE intervention group was significantly increased(P <0.05),but the m RNA expression levels of Nrf2,HO-1,and GST were not significantly different(P> 0.05).ConclusionGrape seed procyanidins can raise Nrf2 and downstream gene m RNA level,and the ability to turn Nrf2 and downstream related protein content is higher,so that SOD and GSH,GSHPX,T-AOC level rise,and thus reduce the MDA content of oxidative damage caused by diabetes,and reduces the blood sugar,reduce insulin resistance and reduced renal function damage,reduce the renal pathological changes. |
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