| Diabetic nephropathy (DN) is one of the chronic microvascular complications of diabetes and a kind for fibrosis to pathological features of renal disease, its pathogenesis is complexity, many mechanisms are still under explored. Recent studies found that connective tissue growth factor (CTGF) in diabetic nephropathy may be the bone morphogenetic-7 (BMP-7) signaling pathway inhibitor, The both play a key role in the kidney fibrosis. Tongxinluo,the components of traditional herb has been widely used in cardiovascular and Cerebrovascular disease prevention and treatment. With Tongxinluo continuous development of new uses, its clinical application are also expanding. This study on Tongxinluo is to explore the the protective effect of Diabetic mellitus (DM) rats and its possible mechanism through the changes of the expression of CTGF and BMP-7 in DN rats'kidney.Objective To study the effects of Tongxinluo on the expression of connective tissue growth factor CTGF) and bone morphogenetic protein-7 (BMP-7) in Diabetic rats,and to explore the protective effects and mechanism on the kidney of Diabetic nephropathy rats.Methods Randomly selected 15 as the normal control group (C group, n= 15) from 70 healthy male Wistar rats.The other 55 rats was established the diabetic (DM) rats model by one-dose introperitoneal injection of streptozotocin(STZ).After 72h,the determination of blood glucose to fasting glucose levels≥16.7mmol/1 (300mg/1) that were diabetes model of success. After animal models were successfully established,they were randomly divided into diabetic groups (DM group), Tongxinluo (500mg-kg-1-d-1) treated groups(TXL group). The ratio of kidney weight to body weight,24h urine microprotein, plasma fasting blood glucose (FPG), glycosylated hemoglobin (HbAlc), advanced glycation end products (AGEs) and kidney function were determined. The renal pathological changes were examined with light microscope PAS staining. The mRNA and protein expression of CTGF and BMP-7 in kidney were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western bloting.Results 1. Weight and blood indices:Compared with C group,DM group weighed significantly less(P<0.01), After treated with Tongxinluo capsules for 24 weeks, rats in TXL group also weighed significantly less than those in C group (P <0.01),but weighed more than those in DM group with statistic significance(P<0.01). As to FPG,HbA1c and AGEs, DM group and C group, FBG, HbA1C, serum AGEs were significantly higher (P<0.05); TXL group was a downward trend than DM group, but the difference was not statistically significant.2. The changes of renal function in rats:DM group than in C group rats with decreased of plasma BUN,Scr and Kw/Bw higher,24h urine protein increased; TXL group than DM group,24h urine protein content decreased, Kw/Bw is also reduced (P<0.05).3. Light microscopic findings of renal morphological changes in rats (PAS×400):C group rats glomerular and tubular interstitial were no significant pathological changes.. In DM group, Most of the glomerular basement membrane were thickened, capillary was narrowed, tubular epithelial cells were vacuolated changes significantly. TXL group compared with DM group, some of glomerular basement membrane is thickened and widened mesangial areas slightly, some of renal tubular epithelial cells were no vacuolar changes.4.The changes of extracellular matrix (ECM) in the rars'kidney: DM group than C group, ECM were significantly higher (P<0.01); TXL group was reduced than DM group, (P<0.05).5.The expression of CTGF and BMP-7mRNA in kidney:RT-PCR results showed, The expression of CTGF mRNA in DM group than C group were significantly increased (P<0.01), TXL group than DM group decreased (P<0.05);The expression of BMP-7mRNA in DM group than C group were significantly decreased (P<0.01), TXL group than DM group increased (P <0.05).6.The expression of CTGF and BMP-7 protein in kidney:Western blotting results were that the expression of CTGF in DM group than C group, were significantly higher (P<0.01), TXL group was lower than the DM group (P<0.05); The expression of BMP-7 protein, DM group than C group, were significantly decreased (P<0.01), TXL group was increased than DM group (P<0.05).Conclusion 1.Tongxinluo can reduced microalbuminuria in diabetic rats and improved the renal pathology changes, and decreased kidney deposition of ECM in diabetic nephropathy rats.2.Its protective effects and mechanism may be related to downregulating expression of CTGF and upregulating expression of BMP-7. |
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