| Objective: Diabetic nephropathy with increasing mobidity is one of the most important microvascular complications of diabetes,thus it is of great significance to prevent and treat diabetic nephropathy actively and effectively.According to the therapy of JianPiBuShenHuoXue,mentor professor Zhang Gengliang prescribes to protect the kidneys and delay the duration of diabetic nephropathy. After the diabetic rats model set up by the streptozotocin's intraperitoneally injection, the drug intervention was applied. The therapy of JianPiBuShenHuoXue's influences on diabetic rats' general situation, blood glucose, blood lipid, renal function, renal pathological morphology, ratio of kidney weight to body weight, 24-hour urinary albumin excretion rate, hypoxia-inducible factor-1? and heme oxygenase-1 were observed to explore the therapy's function of preventing and treating the diabetic nephropathy.Method:A total of 58 healthy male clean SD rats weighing about 230~250g were selected.After one week's adaptive feeding, ten rats were randomly selected as normal control group(N) and according to 55mg/kg body weight,the other 48 rats received a single intraperitoneal injection of 1% streptozotocin citrate buffer(buffer concentration 0.1mmol/L,pH 4.5) to copy diabetic models.The blood of rats' tail veins were collected after 72 hours.If the blood glucose levels were not less than 16.7mmol/L with the rats' water intake and urine output increasing significantly,the diabetic models were copied successfully.The 48 diabetic rats were randomly divided into model group(M), JianPiBuShenHuoXue's therapy group(H), irbesartan group(W), JianPiBuShenHuoXue's therapy unite irbesartan group(H+W), 12 rats in each group.After one week of the modeling successfully,rats in H,W and H+W groups were given medicine by gavage,in accordance with 10 times as adult dose.Rats in H group were treated by the prescription daily 10ml/kg body weight(equivalent to 35.3g/kg body weight of herbs) while irbesartan tablets suspension(made by acacia gum) were given once a day for rats in W group 10ml/kg body weight(equivalent to 35.3g/kg body weight of original drugs).As for rats in H+W group,the same dose of the prescription were given in the morning, the same dose of irbesartan in the afternoon. Rats in N and M groups were taken the equal volume of saline.The intragastric administration continued 8 weeks.The rats' 24-hour urine was collected by metabolic cages at the last day of the administration to measure the 24-hour urinary albumin excretion rate(UAER).All rats were forced to 12-hour fasting rituals without water deprivation and anesthetized by intraperitoneal injection of 2% pentobarbital sodium 20mg/kg body weight.Taking the rats' left femoral arterial blood for the measurement of blood glucose, blood lipid, renal function hypoxia-inducible factor-1? and heme oxygenase-1,all rats' left kidneys were cut away as samples to test Renal Weight/Body Weight(RW/BW) and observe the effect of renal pathological morphology and the expression of hypoxia-inducible factor-1? and heme oxygenase-1.Results:1 General observationFor rats in N group,the spirit and activity were in a good state while their diet,water intake and urine output were normal and the body weight was increasing significantly with shiny fur. M group's rats had poor spirit and decreased their motion and body weight while their diet,water intake and urine output were increasing with dark yellow fur being lack of luster.For some of the rats diabetic complications emerged such as limbs' edema in the experimental process.The general observation of the other three groups fell in between that of N group and M group. One rat in N group died because of biting each other,while one rat from M group and one rat from W group died for improper gavage. The three rats were rejected.2 Changes of blood glucoseThe blood glucose levels of M,H,W and H+W groups were higher than that of N group,and there were significant differences(P?0.05). The blood glucose levels of H and H+W groups decreased evidently and had significant differences compared with M group.W group's blood glucose levels declined unobvious in comparison with M group and there had no significant differences between the two groups(P?0.05).3 Changes of blood lipidBlood lipid levels increased significantly in M,H,W and H+W groups and there were significant differences compared with N group(P?0.05).Blood lipid levels of H and H+W groups were inferior to that of M group with statistically significant differences(P ? 0.05). There were significant differences between W and M groups(P?0.05).4 Changes of kidney hypertrophy index and 24-hour urinary albumin excretion rateM,H,W and H+W groups exceed N group in kidney hypertrophy index and 24-hour urinary albumin excretion rate,and there were significant differences between the two groups(P?0.05).Kidney hypertrophy indexes and 24-hour urinary albumin excretion rate of H,W and H+W groups were less than that of M group and there were significant differences(P ?0.05).Compared with H and W groups' rats,kidney hypertrophy indexes and 24-hour urinary albumin excretion rate of H+W group's rats were inferior with significant differences(P?0.05).5 Changes of renal functionCompared with N group,M,H,W and H+W groups were superior in rats' BUN and Cr with significant differences(P?0.05).BUN and Cr of H,W and H+W groups were inferior to that of M group and there were significant differences(P?0.05).H+W group's rats' BUN and Cr were inferior to H and W groups' and there were significant differences(P?0.05).6 Effect of renal pathological morphologyN group showed distinct and intact glomerulus structure without glomerulus volume increases,and there had no abnormal changes in glomerular capillary basilar membrane,mesenterium and matrix.For M group,the structure of nephridial tissue was irregular and glomerulus volume increases with expanded mesangial area.Compared with M group,there were different degrees' relief of the renal pathomorphism in the three treatment groups,and H+W group alleviated most.7 Effect of hypoxia-inducible factor-1? and heme oxygenase-1's expressionHypoxia-inducible factor-1? and heme oxygenase-1's antigen immunohistochemical staining appeared to be blown. Deep paint with dense particles expresses strong signal while the opposite of the staining showes the weak mainly in tubular epithelial cells littlely in glomerulus signal.N group's rats showed weak expression of hypoxia-inducible factor-1? and heme oxygenase-1 while M group's rats showed strong positive expression of hypoxia-inducible factor-1? and there were significant differences between the two groups(P ? 0.05).H,W and H+W groups had lower expression of hypoxia-inducible factor-1?than M group and there were significant differences(P?0.05). Among the three groups,H+W group showed the lowest expression(P?0.05).With the method of ELISA to detect hypoxia-inducible factor-1? and heme oxygenase-1,M,H,W and H+W groups were superior compared with N group in rats' HIF-1? and HO-1 with significant differences(P?0.05).HIF-1? and HO-1 of H,W and H+W groups were inferior to that of M group and there were significant differences(P?0.05).H+W group's rats' HIF-1? and HO-1 were inferior to H and W groups' and there were significant differences(P?0.05).Applying the Pearson linear correlation,the correlation coefficient of HIF-1? and HO-1 is 0.554 with significant differences(P?0.05).HIF-1? and HO-1 appears to be positive correlation.Conclusion:1 Diabetic rats expresses the ascension of blood glucose, BUN,Cr and blood lipid,the increase of urinary protein,the enlargement of glomerulus volume,the thickening of glomerular capillary basilar membrane, the accumulation of matrix and high expression of hypoxia-inducible factor-1? and heme oxygenase-1.2 Prescription in accordance with the therapy of JianPiBuShenHuoXue could reduce blood glucose,decrease urinary protein,lower BUN,Cr and blood lipid and suppress the expression of hypoxia-inducible factor-1? and heme oxygenase-1.These indicates that prescription in accordance with the therapy of JianPiBuShenHuoXue makes differences in regulating glucolipid metabolism,improving the state of insulin resistance and oxidative stress.Consequently,it can protect the kidney and achieve the aim of preventing and treating the diabetic nephropathy.A new direction appears for traditional Chinese medicine to delay the course of diabetic nephropathy... |
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