| Objective: To observe the effect of Phytolaccae Radix crude extract on the expression of TGF-β1 、 Smad2 、 Smad3 、 Smad7 protein in kidney tissue of rats with diabetic nephropathy,and to study the efficacy and possible mechanism of Phytolaccae Radix crude extract.Methods: Sixty-five 8-week male SD rats weighing 180～220 g were randomly divided into normal control group(8)and diabetic nephropathy group(57),among which 45 rats were successfully established by intraperitoneal injection of STZ,and were randomly divided into model group,Phytolacca Radix high-dose group,Phytolacca Radix mediumdose group,Phytolacca Radix low-dose group and losartan potassium group,each with 9rats.After the model was successfully established,the corresponding drugs were given by gavage.After continuous administration for 4 weeks,the mice were killed,the specimens were collected and preserved,and the urine protein/creatinine ratio and peripheral fasting blood glucose were detected;the pathological changes of rats kidney were observed and analyzed by HE、Masson;the expression and distribution of Fibronectin(FN)、TGF-β1protein were observed by immunohistochemistry.Western Blot was used to detect the expression of Smad2、Smad 3、Smad 7 and TGF-β1 protein in kidney tissue.Results:(1)The urine protein /creatinine ratio,renal index and blood glucose test results showed that the urine protein/muscle ratio,renal index and blood glucose of rats in the diabetic nephropathy group were significantly higher than that in the negative control group(P<0.05).The indexes after treatment with Phytolaccae Radix and Losartan Potassium showed a decreasing trend compared with the diabetic nephropathy group(P<0.05).(2)The results of renal pathological staining showed that under the light microscope,the rats in the negative group showed complete glomerular structure,clear and neatly arranged renal tubules,and normal tubular interstitial structure.Rats in the diabetic nephropathy group showed different degrees of renal structural changes,such as: extra mesangial matrix increased,glomerular cells proliferation,renal tubule lumen protein cast,renal tubular epithelial cell brush border shedding,cell voids Foaming.The renal pathological manifestations could be improved after treatment with Phytolaccae Radix and Losartan Potassium.(3)Immunohistochemistry results: the intergrated optical density(IOD)of the TGF-β1 and FN protein in the kidney tissue of the diabetic nephropathy group was significantly increased(P<0.05).The IOD values of TGF-β1 and FN protein in the kidney tissue of rats in each drug treatment group were significantly lower than those in the diabetic nephropathy group(P<0.05).(4)WB results: the expression of TGF-β1,Smad2,and Smad3 in the diabetic nephropathy group were significantly higher than the negative control group(P<0.05),and the Smad7 protein expression were significantly lower than the negative control group(P<0.05).The expression of TGF-β1,Smad2,and Smad3 in the kidney tissue of rats in each drug treatment group was significantly lower than that of the diabetic nephropathy group,and the expression of Smad7 protein were significantly higher than that of the diabetic nephropathy group.Among them,the high dose of Pokeweed Compared with the pokeweed low-dose group,the group changed significantly(P < 0.05).Conclusion: Phytolaccae Radix crude extract may participates in regulating the activity of TGF-β1/Smads pathway to improve the condition of diabetic nephropathy rats,which is through inhibit the secretion of TGF-β1,down-regulate the expression of Smad2 and Smad3 protein,and up-regulate the expression of Smad7 protein in kidney tissue. |
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