| Objective:Diabetic nephropathy is one of the most common microvascular complications of diabetes,is a major cause of ESRD,seriously endangering the health of diabetics,is the leading cause of death in patients with diabetes.Therefore,active and effective prevention and treatment of diabetic nephropathy is of great significance.According to the therapy of Jian Pi Bu Shen Huo Xue,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 Jian Pi Bu Shen Huo Xue's influences on diabetic rats' general situation,blood glucose,blood lipid,renal function,renal pathological morphology,kidney hypertrophy index,24-hour urinary albumin excretion rate,tumor necrosis factor-? and monocyte chemotactic protein-1 were observed to explore the renal protection in diabetic nephropathy.Methods: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 other48 rats received a single intraperitoneal injection of 1% streptozotocin citrate buffer(buffer concentration 0.1mmol/L,p H 4.5)to copy diabetic models,normol control group received the same volume of single intraperitoneal injection of 1% citric acid buffer.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),Jian Pi Bu Shen Huo Xue's therapy group(H),irbesartan group(W),Jian Pi Bu Shen Huo Xue'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 group10ml/kg body weight(equivalent to original drug 2.5mg/ml).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 tumor necrosis factor-? and monocyte chemotactic protein-1,all rats' bilateral kidneys were cut away as samples to test kidney hypertrophy index,and taken for light microscopy morphological changes of renal pathological and immunohistochemical determination of the expression of tumor necrosis factor-? and monocyte chemotactic protein-1.Results:1 General observationThe rats in the normal control group were generally in good condition,with bright color,sensitive reaction,free movement,normal diet,water intake and urine output,and weight gain.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.Part of the rats with the progress of the experiment,appear a series of diabetes complications,foot ulcers,limb edema.After drug treatment,the generalobservation of the other three groups fell in between that of N group and M group.During the experiment,the rats of group N induced skin ulcers due to bite each other only 1 died of infection;1 rats of M group died,considered as slow,due to reduced diet;W group died of suffocation caused by intragastric administration of 1 mistakes.2 Changes of blood glucose and blood lipidCompared with N group,M,H,W and H+W groups of blood glucose and blood lipid were significantly increased,there was a significant difference(P<0.05).The blood glucose and blood lipid levels of H and H+W groups decreased evidently and had significant differences compared with M group(P<0.05).Compared with the M group,W group decreased blood glucose and blood lipid is not obvious,no significant difference(P>0.05).3 Changes of kidney hypertrophy index and 24-hour urinary albumin excretion rateCompared with N group,M,H,W and H+W groups of kidney hypertrophy index and 24-hour urinary albumin excretion rate were increased,and there were significant differences(P<0.05).Kidney hypertrophy indexes and 24-hour urinary albumin excretion rate of H,W and H+W groups were inferior to that of M group and there were significant differences(P<0.05).Compared with H and W groups' rats,kidney hypertrophy indexes and24-hour urinary albumin excretion rate of H+W group's rats were inferior with significant differences(P<0.05).4 Changes of renal functionCompared with N group,M,H,W and H+W groups were superior in rats' urea nitrogen and creatinine with significant differences(P<0.05).BUN and Cr 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' s rats,the urea nitrogen and creatinine in H+W group's rats were lower and there were significant differences(P<0.05).5 Changes of renal pathological morphologyN group showed distinct and intact renal unit structure withoutglomerulus volume increases,and there had no abnormal changes in glomerular capillary basilar membrane,mesenterium and matrix,no inflammatory cell infiltration.In the M group,the glomerular volume was significantly increased,the basement membrane was thickened and the mesangial area widened.Some tubular epithelial cells showed vacuolar or granular degeneration,inflammatory cell infiltration was observed.Compared with the M group,the renal pathological damage of the three groups were reduced to different degrees,the most obvious in the joint group.6 Effect of tumor necrosis factor-? and monocyte chemotactic protein-1's expressionImmunohistochemical staining of tumor necrosis factor-? and monocyte chemotactic protein-1 antigens showed brown granular positive reaction,mainly in tubular epithelial cells and interstitial cells of renal interstitium.N group's rats showed weak expression of tumor necrosis factor-? and monocyte chemotactic protein-1 while M group's rats showed strong positive expression of tumor necrosis factor-? and monocyte chemotactic protein-1,and there were significant differences between the two groups(P<0.05).H,W and H+W groups had lower expression of tumor necrosis factor-? and monocyte chemotactic protein-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 tumor necrosis factor-? and monocyte chemotactic protein-1,M,H,W and H+W groups were superior compared with N group in rats' TNF-? and MCP-1 with significant differences(P<0.05).TNF-? and MCP-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' TNF-? and MCP-1 were inferior to H and W groups' and there were significant differences(P<0.05).Conclusions:1 The therapy of Jian Pi Bu Shen Huo Xue can improve the general status of diabetic rats,lower blood glucose,reduce total cholesterol,triglyceride,bloodurea nitrogen,creatinine,reducing urinary protein excretion,restoring hypertrophy of kidney,improve its pathological form.2 The therapy of Jian Pi Bu Shen Huo Xue could suppress the expression of TNF-? and MCP-1 in renal tissue and serum of diabetic rats,reduce the renal inflammatory reaction,and has a certain renal protective effect,so as to play a role in the prevention and treatment of diabetic nephtopathy. |
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