| Objective:This study made diabetic rat mode by using high sugar,high fat feed and streptozotocin, which was then treated by traditional Chinese medicine or irbesartan. We explore traditional Chinese medicine’ s protective effect on DN rat renal and possible mechanism, through observing urinary protein,biochemical criterion,pathomorphological changes of kidney and expression of protein kinase C(PKC) and transforming growth factor-β1(TGF-β1) in rats.Methods: Selected 85 heathy male SD rats, weighing 160-180 g, adaptive feed one week, 15 were randomly divided into the normal group(NG), and the other 70 into model making group after measurement of blood sugar and urinary protein were negative. The normal group were given ordinary feed, while another group were on the high sugar and high fat diet. After 12 weeks the rats of model making group were given intraperitoneal injection STZ35mg/kg, and the normal group were injected by the equal dose of citrate buffer. The rats whose tail vein blood sugar were greater than or equal to 16.7mmol/l after 72 hours were as successful diabetic rat mode, death and blood sugar not reaching the standard were rejected. The model rats were randomly divided into four groups: 17 in model group(MG), 17 in traditional Chinese medicine group(ZG), 17 in irbesartan group(XG), and 16 in traditional Chinese medicine and irbesartan group(JG), and then the treatment group were given corresponding drug intervention for 16 weeks, while the others were administered equal volume of saline. The tail vein blood glucose and 24-hour urine protein were measured on week 4,8,12 respectively during the treatment. At the end of 16 weeks, rats’ 24-hour urine were collected for the quantitative determination of 24-hour urinary protein, blood were drew under anesthesia condition for testing biochemical criterion, and the right kidney was weighed. What’s more,specimens from renal tissue were taken for observation by light microscopy, electron microscopy, immunohistochemistry and Real-Time PCR.Results: 1 The body weight, right kidney weight and hypertrophy index of each groupCompared with normal group,the body weight of model group and treatment group was obviously lighter(P<0.05), there was no significant difference between model group and treatment group(P>0.05); The right kidney weight was no difference in different groups(P>0.05); The hypertrophy index of model group and treatment group was significantly higher than normal group(P<0.05), there was no statistical difference between model group and treatment group(P>0.05). 2 The 24-hour urinary total protein(UTP) of each groupThe UTP of model group and treatment group was significantly higher than that of normal group(P<0.05).Compared with model group, the quantity of treatment group was obviously lower(P<0.05). Compared with ZG and XG, the UTP of JG was distinctly lower(P<0.05). There was no statistical difference between ZG and XG(P>0.05). 3 The serum total protein(TP) and albumin(ALB) of each groupThe TP and ALB of model group and treatment group was significantly lower than that of normal group(P<0.05). Compared with model group, the quantity of treatment group was obviously higher(P<0.05).Compared with ZG and XG, the TP and ALB of JG was distinctly higher(P<0.05). There was no statistical difference between ZG and XG(P>0.05). 4 The total cholesterol(TC) and triglyceride(TG) of each groupThe TC and TG of model group and treatment group was significantly higher than that of normal group(P<0.05). Compared with model group, the quantity of treatment group was obviously lower(P<0.05). Compared with ZG and XG, the TC and TG of JG was distinctly lower(P<0.05). There was no statistical difference between ZG and XG(P>0.05). 5 The serum creatinine and urea nitrogen of each groupThere was no statistical difference among different groups(P>0.05). 6 The glycated hemoglobin(Hb A1c) of each groupThe Hb A1 c of model group and treatment group was significantly higher than that of normal group(P<0.05). Compared with model group and XG, the Hb A1 c of ZG and JG was obviously lower(P<0.05). There was no statistical difference between ZG and JG(P>0.05). Model group and XG was no significant difference as well(P>0.05). 7 The observation of renal pathomorphological changes 7.1 Light microscopy observationThe structure of rats’ renal glomerular of normal group was clear and the size of that was suitable. The basement membrane was not thickening and the mesangial matrix was normal. The size of model group rats’ renal glomerular was hypertrophy, basement membrane thickening and mesangial matrix increasing significantly. The pathological changes of treatment group were lighter than model group, and there was no significant difference between each treatment group. 7.2 Electron microscopy observationThe structure of glomerular basement membrane of normal group was clear and suitable. Foot process did not show abnormal change. In model group,the basement membrane incrassation,endothelial cell proliferation and podocyte fusion widely were observed. The treatment group showed the similar pathological changes, but the degree was lighter than model group. There was no significant difference between each treatment group. 8 Real-Time PCR result of the expression of TGF-β1m RNA in each groupThe expression of model group and treatment group was significantly higher than normal group(P<0.05). Compared with model group, the expression of treatment group was obviously lower(P<0.05). Compared with ZG and XG, the expression of JG was lower(P<0.05). There was no statistical difference between ZG and XG(P>0.05). 9 Immunohistochemical result of the expression of PKC and TGF-β1 in renal cortexPKC and TGF-β1 was mainly expressed in nephric tubule of rats in five groups, but only few in glomerulus. The expression of model group and treatment group was significantly higher than normal group(P<0.05). Compared with model group, the expression of treatment group was obviously lower(P<0.05). Compared with ZG and XG, the expression of JG was lower(P<0.05). There was no statistical difference between ZG and XG(P>0.05).Conclusions:1 Xiezhuojiedu Huoxuetongluo traditional Chinese medicine can reduce urinary protein quantitative, blood sugar,total cholesterol and triglyceride, increase the serum total protein and albumin, relieve renal pathological damage of diabetic rat, and delay the progress of DN.2 Xiezhuojiedu Huoxuetongluo traditional Chinese medicine can protect kidney by inhibiting the basement membrane thickening and mesangial matrix increasing, possibly because of inhibiting overexpression of PKC and TGF-β1 in renal tissues of diabetic rat.3 The study shows that traditional Chinese medicine has similar therapeutic effect to irbesartan, and combined application is better. |
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