| Objectives To observe effects of Shizidaiping formula on the renal function and renal tissue of diabetic rats and the expression of FN TGF-?1 and PTEN protein,Further discussion on its effect on mi RNA-21 in plasma.This experiment is aimed at clearing the effect of Shizidaiping formula on the T2 DM rats' s renal fibrosis,and exploring whether it is related to the regulation of TGF-?1 and PTEN protein.Methods Selected 60 SPF male SD rats,and adaptive feed for a week.Choose 50 of them to established diabetic disease by feeding with high-fat diet for 4 weeks combined with intraperitoneal injection of Streptozotocin.Other 10 rats were selected as the normal control group feeding with normal diet.72 hours later,take the tail vein blood to make a IPGTT tests.Detected the FBG ? 7.0 mmol/L,PG2 h ? 11.1 mmol/L was considered succeed T2 DM model.The T2 DM rats were randomly divided into model group,metformin group,Shizidaiping formula groups with high,medium and low dose respectively.Detected the rats' random plamsa glucose on 1,3,5 and 7 weeks.Collect urine for testing CCR and 24 H UPro after 8 weeks of treatment.Fasting for 12 hours,the rats are anesthesia by intraperitoneal injection of Chloral hydrate and charged the abdominal aortic blood.Some of the blood collected by common blood collection,centrifugal to get the red blood cells for determination of GHB in 4 hours.Taken for determination of GSP and serum biochemical indexes such as CRE and UREA levels.Some of the blood collected using EDTA anticoagulant tubes,centrifuge the blood plasma and mi RNA-21 expression level were observed by RT-PCR.Weighing the left kidney to calculate kw/bw,cut it into 10mm×10mm size,and saved in 10% formaldehyde for HE staining and Masson staining.FN,TGF-?1 and PTEN protein expression was detected by immunohistochemical method.Results 1 At the end of the experiment,the random blood glucose,GHB and GSP of model was higher than that of normal Group(P<0.05).Accompanied by more food,polydipsia,and other symptoms,it means that the rats are succeed to induction with type 2diabetes.The treatment group can significantly decreased the rats' random blood glucose,GHB and GSP(P<0.05)compared with model group.2 At the end of the experiment,the CRE,UARE,CCR,kw/bw and 24 h UPro of model group was higher than that of normal Group(P<0.05).This means that the rats of model Group rats kidney damage has occurred.The treatment group can significantly decreased the rats ' CRE,UARE,CCR,kw/bw and 24 h UPro(P<0.05)compared with model group.3 The pathological changes of renal tissue of T2 DM SD rats in each group:The normal renal tissue structure are arrange in the complete rules without infiltration of inflammatory.And a small amount of collagen deposition in the glomerular mesangial and epithelial cells of renal tubules.While in the diabetic model Group,the renal glomeruli dilated significantly.Thickening of the glomerular mesangial;renal cystic dilatation and adhesion of renal glomerulus;Some part of renal tubules are damaged with less smooth and shrink,Tubular vacuolar change are visible;tubulointerstitial obviously a large number of inflammatory cell accumulation;renal interstitial zone is widened.But we have not found K-W nodules yet.There were collagen accumulation in renal tissue and fibrosis is more serious by Masson staining.The pathological changes above are significantly mitigated after 8 weeks treatment.4 The level of FN,TGF-?1,PTEN in rat kidney tissue by IHC:FN and TGF-?1 expression in renal tubular epithelial and mesangial are stronger in model group than any other groups.Semi-quantitative statistical result shows : Compared with model Group,the treatment groups TGF-?1 was significantly reduced(P<0.05);PTEN in the glomeruli of normal group were negative,in the renal tubular epithelial cells were strongly positive,PTEN protein stainingshowed that in model Group,the glomeruli were negative,in the renal tubule was slightly positive.Semi-quantitative statistical result shows : Compared with model Group,the treatment groups PTEN are increased(P<0.05).5 Detection of mi RNA-21 expression in rat plasma by RT-PCR: Compared with normal group,the model Group significantly increased mi RNA-21 in plasma(P<0.05),compared with model Group,the treatment groups mi RNA-21 was significantly reduced(P<0.05).Conclusions 1 Shizidaiping Formula can reduce the random blood glucose,GHB and GSP and kidney function,decreased kidney weight ratio of T2 DM SD rats.2 Shizidaiping formula can alleviate the pathological changes of kidney in type 2 diabetic rats,reduce the FN gathering,protect the diabetic kidney fromrenal fibrosis.3 Shizidaiping formula protect the kidney of type 2 diabetic rats by decreased mi RNA-21 expression in plasma,reduced the expression of TGF-?1,increased of PTEN or other related pathways regulating. |
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