| Background and ObjectiveThe appearing of microalbuminuric in the patients of diabetic nephropathy means the coming of diabetic nephropathy. It is not only the clinical of the diabetic nephropathy,but also the reason of evolutional. The electrostatic and machinery barrier of the glomerular basement membrane,the multihole of the endotheliocyte and the slit diaphragm of foot cell is very important in the coming of the albuminuric. Heparan sulfate proteoglycan is one of the glycosaminoglycans,is also the biomacromolecules material in the structure of glomerular basement membrane and mesangial region extracellular matrix. Heparanase is one of the glucuronic acid incision enzyme in the degradation of HSPGs.It can modify the express of the GBM and ECM.It also destroys the electrostatic barrier of the GBM. The high of the fat,glucose and hemodynamics, the activation of the renin-angiotensin systemRAS, the damage of the oxygen radicals,the effect of sharpen is the pathogenesy of the DN. In the few years,in the urine of the appearing of microalbuminuric in the patients of diabetic nephropathy detect the HPA and HS.The further research also discover that the expressions of HPA were significantly high in the kidney tissue of the diabetic nephropathy patients,at the same time the expressions of HS grow downwards.The effect of the the expressions of HPA in the kidney tissue of the albuminuric become important for the past few years. Therefore we decide to investigate the expressions of HPA,b-FGF in rats with STZ diabetic nephropathy, explore the mechanism of proteinuria occurrence. In addition, we use sulodexide to treat the rats and observe the changes of biochemical indicators,glomerulus immunohistochemistry, reverse transcription PCR in order to investigate its kidney protective effect and provide firm experimental basis and theoretic evidence.Method70 normal four-week male SD rats were randomly divided into following two groups after one week adaptive breeding:normal control group (n=20)and DN group (n=50).Diabetic nephropathy group were induced by intraperitoneal injection of 60mg/kg 0.1mmol/L streptozotocin(STZ).The levels of blood glucose and the 24hour urine were measured after 72 hours,we considered that the model were successful if the blood glucose of the rat was greater than or equal to 16.6mmol/L and the 24hour urine was 1.5 greater than initial urine. The rats of normal group were intraperitoneal injection partes aequales STZ buffer as control.Rats in sulodexide group received intragastric administration of (10mg/kg.d) of sulodexide for 8 weeks, while the rats in both diabetic group and control group received intragastric administration normal saline in same amount for 8 weeks. During the period of the experiment, free food and water were given to these rats. We don't use insulin and other hypoglycemic agents. Ten rats were sacrificed after four and eight weeks.24h urinary protein excretion, ALB, Cys-C were measured,the kidneys were removed and renal morphology was observed, the methods of immunohistochemistry and reverse transcription PCR were used to detect the expression of HPA and b-FGF of kidney of all rats.Result1.24UPE in group DN and group DNS were significantly higher than that in normal control group either at the 4th week or 8th week (P<0.05), meanwhile,24UPE in group DNS is significantly lower than that in group DN (P<0.05). ALB in group DN were significantly lower than that in normal control group and group DNS either at the 4th week or 8th week(P<0.05). Compared to normal control group, ALB in group DNS have not statistic difference (P>0.05). Shenbizhong in group DN and group DNS were significantly higher than that in normal control group either at the 4th week or 8th week(P<0.05). Compared to the group DN, Shenbizhong in group DNS is significantly lower (P<0.05). Cys-C in group DN is significantly higher than that in normal control group and group DNS. Cys-C in group DNS and normal control group have not statistic difference (P>0.05).2. The organization of kidney in normal control group is approximately normal.After four weeks, shenbizhong, volum of the glomerulus become augentation,and extracellular matrix increase,basilar membrane become increased thickness, till eight weeks glomerulosclerosis and interstitial fibrosis appear. However, compared to the group DN, the pathological changes in group DNS are obviously alleviated.3. The reverse transcription PCR shows that the expressions of HPA mRNA in glomeruli and kidney tubules in normal control group are significantly wild while in group DN at the 4th week are significantly up-regulated(P<0.05), and these continue to exist into the 8th week.The expressions of HPA mRNA in group DNS is significantly lower than those in group DN (P<0.05).4. The immunohistochemistry shows that the expressions of HPA,b-FGF in glomeruli and kidney tubules in normal control group are significantly wild while in group DN at the 4th week are significantly up-regulated(P<0.05), and these continue to exist into the 8th week.The expressions of HPA and b-FGF in group DNS is significantly lower than those in group DN(P<0.05).Conclusion1. With progressing of STZ nephropathy, the expressions of HPA and b-FGF were significantly increased in glomeruli and kidney tubules,which may be important factors of the proteinuria in rats with STZ nephropathy.2. Sulodexide confers a protective effect through down-regulation of HPA and b-FGF in adriamycin nephropathy. |
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