| Objective:Through experimental studies,it was observed that vascular endothelial injury in DN rat kidney was associated with NO-ONOO-pathway-mediated oxidative stress injury and affected the expression of SUR2B/Kir6.1 subtype KATP channel in renal vascular endothelial cells.The stilbene glycosides extracted from Polygonum multiflorum can protect the kidney damage of DN rats by intervening in the above two processes.Methods:A total of 40 male Wister rats aged 8 weeks and weighing 200±20g were randomly divided into 2 groups:8 normal controls?NC group?and 32 rats with high glucose and high fat for 8 weeks.A rat model of diabetic nephropathy was injected into the tail vein of 35 mg/kg streptozotocin?STZ?.72 hours after the injection of STZ,the blood was collected from the tail vein of the rats to measure blood glucose.The randomized blood glucose?16.7mmol/L was successfully established for diabetes?DM?,and STZ 30mg/Kg was added to the tail vein of the rat whose blood glucose did not reach the target value.Thirty-two rats were randomly divided into diabetic nephropathy model group?DN group?12;benazepril group?DN+BE group?8;stilbene group?DN+TSG group?12 rats.0.1 g of TSG was added to 10 ml of NaCl physiological saline to prepare a solution of1%concentration,and the rats in the stilbene glucoside group were intraperitoneally injected for 8 weeks at a dose of 30 mg/kg/d;The method was to calculate the equivalent dose of 10mg/kg/d of benazepril into a solution,and the rats in the benazepril group were treated with intragastric intervention for 8 weeks.At the same time,the rats in the blank group and the model group were given daily peritoneal cavity.Inject an equal dose of normal saline.The general living state of the rats was observed every day,and the body weight,blood sugar,24-hour urine protein and urine volume of the test rats were periodically measured.At the end of the 8th week of the experimental administration,20%of the urethane anesthetized rats were anesthetized by anesthesia,and the abdominal aorta was harvested by phlebotomy and centrifuged at the same time.Morphological changes of kidney tissues in each group were observed by HE staining and electron microscopy.The expression levels of ET-1 and vWF in rat kidney were determined by immunohistochemical staining.Western-blot protein quantitative analysis The expression level of NT protein in rat kidney tissue was detected by Real-Time PCR.The expression of SUR2B/Kir6.1 mRNA in rat kidney was detected by automatic biochemical analyzer.The whole body biochemical analyzer was used to detect rat urine ALBU/Ucr.Automated biochemical analyzer was used to detect total serum cholesterol in rats.Results:Part I:At 8 weeks after the experimental administration,the ALBU/Ucr and serum total cholesterol were significantly increased in the model group compared with the blank group.The ALBU/Ucr and the model of the benazepril group and the stilbene group were compared.The group comparison index decreased;the serum total cholesterol of the stilbene group decreased compared with the model group.HE:In the experimental rats for 8weeks,the renal tubular epithelial cells in the experimental rats were shed and vacuolar degeneration;the renal interstitial lymphocytes and monocytes infiltrated,the arterial wall thickened,and occasionally the renal interstitial fibrosis The glomerulus showed infiltration of inflammatory cells and mesangial hyperplasia.Electron microscopy:partial glomerular focal podocyte fusion,mitochondrial degeneration,glomerular basement membrane heterogeneous heterogeneous thickening,a large number of immune complex deposition;visible proximal convoluted villi,epithelial cell lysosome comparison More appearances.The performance of the stilbene glycoside group was significantly reduced compared to the model group and the benazepril group.Part II:After 8 weeks of experimental administration,it was found by immunohistochemistry that ET-1 and vWF were widely present in the kidney tissues of diabetic nephropathy rats.The experimental group found that compared with the blank group rats,the model group rats and The expression of ET-1 and vWF in renal tissue was significantly increased in the stilbene glycoside group,and the expression of ET-1 and vWF in the stilbene glycoside group was less than that in the model group.Statistically significant?P<0.05?.The Western blotting method showed that the gray value of NT protein expression in the kidney tissue of the diabetic nephropathy model group was significantly higher than that in the blank resistance rats,while the NT protein expression in the kidney tissue of the stilbene glycoside group was higher than that of the model group.Rats were reduced and had statistical differences.Real-Time PCR method was used to detect the expression of SUR2B/Kri6.1 mRNA in the kidney of rats with diabetic nephropathy.The expression of SUR2B/Kir6.1 mRNA in the kidney of rats in the model group and the stilbene glycoside group was lower than that in the blank group.Compared with the model group,the expression of SUR2B/Kir6.1 mRNA in the kidney tissue of the stilbene glycoside group increased,and there was a statistical difference?P<0.05?.Conclusion:1.The main active ingredient of Polygonum multiflorum TSG can better control the weight loss,blood sugar,urine volume,and proteinuria and serum total cholesterol in rats.2.TSG can reduce the expression of total NT protein?ONOO-marker?and reduce the expression of von Willebrand factor?vWF?and endothelin?ET-1?in renal tissues.3.The SUR2B/Kir6.1 subtype KATP channel exists in the kidney tissue of DN rats,which can regulate the balance of potassium ions and the release of NO,and affect the damage of renal vascular endothelium in DN rats.4.TSG has a good protective effect on kidney injury in early DN rats,its mechanism and the SUR2B/Kir6.1 subtype KATP channel affecting the vascular endothelial cells of DN rats,and through the intervention of NO-ONOO-pathway-mediated oxidation.Stress vascular endothelial injury,thereby delaying the progression of DN... |
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