| Objective: The rats were induced by high fat and high glucose,left nephrectomy and intraperitoneal injection of STZ combined with SD,,the observation has opened Xuanfu opened Xuanfu on podocytes in diabetic nephropathy rats related protein nephrin,podocin expression,and explore the effects of opening Xuanfu Decoction on24 h proteinuria in diabetic nephropathy rats and the protective effects and mechanism of kidney.Diabetic nephropathy(Diabetic nephropathy DN)was a complication of diabetes in recent years,the incidence of DN was also more and more high,the incidence of diabetic nephropathy in China was about 34.7%,DN has become the leading cause of end-stage renal disease(End Stage Renal,disease,ESRD)the most important single factor [1],and effective in the treatment of DN at present in the field of medicine is a very important issue.Methods: adult male SD rats(Sprague-Dawley),high fat and high sugar feeding plus left nephrectomy plus low dose STZ intraperitoneal injection molding,after the model were divided into three groups: model(n=12),perindopril group(n=12),low dose of Chinese medicine group(n=12),high dose of Chinese medicine group(n=12)and control group(n=12).To the model group and the control group was treated with saline to open Xuanfu,perindopril respectively on high,low dose group and perindopril group intervention.The whole experimental period,body weight of rats was detected,blood glucose and 24 h urinary protein;quantitative measurement of 24 hours urinary protein and biochemical indexes of 8 weeks after the experimental period(serum creatinine(Scr),total cholesterol(Chol),urea nitrogen(BUN),triglyceride(TG)).The podocyte associated protein of nephrin rat glomerular basement of immunohistochemistry podocin staining combined with computer image analysis(Image-proplus)to detect nephrin expression intensity of podocin.The research is based on the traditional Chinese medicine theory innovation,put forward DN "belongs to the category of Xuanfu disease" hypothesis,will open the mechanism of Xuanfu method in treatment of diabetic nephropathy and regulation of podocyte injury related markers of nephrin fabric,link pidocin,contribute to the micro perspective interpretation opened Xuanfu method scientific connotation,combining traditional Chinese and Western medicine for diabetic nephropathy the research for starting point,to develop new ideas.Packet data were analysed by SPSS21.0 software,the standard deviation().Comparative analysis of statistical measurement data between groups using ANOVA.The observation of renal cells under the microscope and oil mirror: the cell membrane of podocyte foot or cytoplasm appeared Brown particles each of which is positive.Slice by microscope randomly collected not repeat 9 glomeruli were photographed image derived and stored.Image analysis system: the positive signal of the immune group of pictures by Image Pro analysis in Plus system.The image input system for photometric conversion,recording light density value.The program to calculate each glomerulus integral optical density of the positive signal area,and taking the average.The integral optical density(IOD)represents the relative expression amount of protein,IOD = average optical density of X positive area,each group selected 9glomeruli were the camera system into the computer,select the image analysis of the immune system module,measurement and analysis by software,relative expression of the mean of the protein.Results: 1,24 h urinary protein: compared with normal control group,urinary protein in diabetic nephropathy rats were increased significantly,the difference was statistically significant(P < 0.05);compared with the model group,opened Xuanfu party and perindopril intervention,high,low dose of Chinese medicine group,perindopril group,urinary protein decreased,the difference was statistically significant(P < 0.05).High,low dose groups of urine protein were lower than the perindopril group,the difference was statistically significant(P < 0.05).Nephrin,podocin distribution location:(x 1000)oil microscope.Normal control group rat kidney tissue by immunohistochemical staining of podocin and nephrin sections are brown,line distribution in the basement membranes of the podocyte.The model group of podocin and nephrin is not uniform in the no rules,glomerular distribution,staining weakened significantly high,low dose of Chinese medicine group and perindopril group staining in the glomerular basement membranedistribution of podocytes is relatively uniform,Tan deposition.Nephrin,the expression of podocin: at the end of the experiment,compared with normal control group,the statistical results showed that the expression of podocin protein in diabetic nephropathy rats strength decreased,the difference was statistically significant(P < 0.05);compared with the model group,the statistical results show that opened Xuanfu and perindopril intervention,high,low dose of Chinese medicine group,perindopril group podocin protein expression intensity increased,the difference was significant(P < 0.05);high,low dose of Chinese medicine group the expression of podocin protein were higher than that of perindopril group,the difference was statistically significant(P < 0.05).The end of the experiment,compared with normal control group,the statistical results showed that the expression of nephrin group protein in diabetic nephropathy rats strength decreased,the difference was statistically significant(P < 0.05);compared with the model group,the statistical results show that opened Xuanfu and perindopril intervention,high,low dose group,perindopril group nephrin egg The expression intensity of white was increased,the difference was statistically significant(P < 0.05).The expression intensity of nephrin protein in high and low dose group was higher than that in perindopril group,the difference was statistically significant(P < 0.05)Conclusion: the opening of Xuanfu can reduce urinary protein excretion and,play a role in protecting the kidney,which might be related to the up regulation of podocyte associated protein nephrin,podocin expression. |
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