| ObjectiveTo investigate the expression of TSP-1 in diabetic rats and the effect of TSP-1 on the activation of TGF-β1,and whether inhibiting TSP-1-mediated activation of TGF-β1 could delay the progression of renal fibrosis in diabetic nephropathy,so that provide new ideas for Prevention of diabetic nephropathy.MethodsHealthy 42 male Wistar rats were randomly divided into model group and normal control group (A group), with streptozotocin (STZ) by intraperitoneal injection of 60mg/kg one-time-induced diabetes model, the diabetic model group of diabetic rats were randomly divided into control group (B group) and diabetic rats treated group (C group), Drug treatment group from the STZ diabetic model injection immediately after the establishment of a successful beginning to the TSP-1-specific antagonist, namely: synthetic peptide (LSKL) 100μg / d Subcutaneous injection until the end of the experiment。Group B untreated rats and normal control group A group were given the same dose of normal saline. Different times were measured in diabetic rats treated group C serum creatinine, creatinine clearance rate, blood lipids, fasting blood glucose, glycosylated hemoglobin, 24 h urinary protein, urinary albumin excretion rate (UAER) and Group A Group B at the same time period comparison of results. The first 24 weeks with HE and PAS staining in each group the morphological changes of renal pathology, determination of kidney weight, index of kidney hypertrophy (KW / BW) and immunohistochemical detection of TSP-1 and TGF-β1, both for the regression analysis, Pearson linear correlation.Results(1)Survival of rats: normal control group (A) all survived; untreated diabetic nephropathy group (B group) 20,51,93,108 days of the death of the first one; diabetic nephropathy treatment group (C group) 98 the death of a day; (2) General condition: the normal control group (A group) of rats in good condition, color gloss, response, and weight gain. DM model group (B group) in rats appeared typical food intake, water intake and urine output increased, significant weight loss in diabetes, "a little more than three" symptoms, dark matte color, activity less, there are two cataract rats , a rat repeatedly diarrhea, four rats died during the experiment. Compared with B group, C group, "a little more than three" significant improvement in symptoms, shiny coat, weight gain; LSKL prompted to a certain extent, symptoms of DM control;(3)Treatment of diabetic nephropathy were determined during different time 24h urinary protein, urinary albumin excretion rate, serum creatinine and renal function compared with non-treatment group decreased significantly(P<0.01); but better than the same period in the normal control group slightly significant difference (P <0.05);(4)Blood glucose, glycosylated hemoglobin and blood lipids situation:B group and C group of diabetic rats blood glucose, glycosylated hemoglobin and blood lipids were significantly higher in normal rats(P<0.01);And between the diabetic group there was no significant difference in blood glucose(P>0.05);C group glycosylated hemoglobin and blood lipids different than the B group, the extent of the decline (P<0.05);(5)The 24th week diabetic nephropathy renal pathological changes(Glomerulosclerosis, interstitial fibrosis, tubular atrophy, vascular intimal lesions), Determination of renal weight index of kidney hypertrophy (KW / BW), glomerular volume, mesangial area, the treatment group was significantly lighter compared with non-treatment group, significant differences(P<0.05); However, these changes of diabetic nephropathy group than those in normal control group, significant differences (P<0.05);(6)The 24th week when the use of immunohistochemistry in renal tissue observed TSP-l in the normal control group, no positive expression, only a small amount TGF-β1 expressed. And diabetic nephropathy group (B group, C group) TSP-l in the cytoplasm of tubular epithelial cells, glomerular parietal epithelial cells positive for the expression of inflammatory infiltration and fibrosis in the interstitial expression of the more obvious, Compared with normal control group (A group), the significant difference(P< 0.01). The positive expression of TGF-β1 also significantly enhance the area, can be found in glomerular mesangial cells, glomerular parietal epithelial cells, renal tubular epithelial cells, renal interstitial cells, as compared with normal control group, significant differences(P<0.01); Diabetes treatment group, diabetic control group, TSP-l, TGF-β1-positive area and intensity of expression significantly different from(P<0.05).Conclusion(1)Renal tissue of diabetic rats with TSP-1, TGF-β1 expression in the presence of high;(2)TSP-1-mediated activation of TGF-β1 in diabetic rats is an important mechanism of renal fibrosis;(3)LSKL able to block specific TSP-1-mediated activation of TGF-β1, TGF-β1 content of the overall significantly reduced, indicating that: LSKL can significantly slow down the process of renal fibrosis in diabetic rats and improve the long-term survival rate of diabetic nephropathy. |
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