| Objective:To observe the effect of Lamps K idney granules on the urinary protein excretion,renal function and renal tissue pathology in diabetic nephropathy rats and to explore Pathological effects and the possible target and mechanism of Lamps K idney granules on diabetic nephropathy,for providing basis that the experimental drug treats diabetic nephropathy.Methods:100 healthy adult male SD rats,Weight(230 + 20)g,8 weeks old.100 rats were randomly divided into normal group(group A),15 rats and streptozotocin model group,85 rats.Streptozotocin model group was treated with 1% citrate buffer solution and streptozotocin solution.The rats were injected by the left side of the abdominal cavity at once,and the A group was injected with the same volume of normal saline.After the model,the standard diet was given,the padding was replaced,and the cage was sterilized regularly.During the STZ model,5 rats died.The level of blood glucose was measured for 3 days after 72 hours of STZ model treatment,and random blood glucose was more than 16.6mmol/L,the success of diabetes model preparation.The rats after the model were fed for 1 weeks,random blood glucose was more than 16.6 mmol/L,urine volume was above of 1.5 times higher than that of control group,24 hours urinary albumin(measured by coomassie brilliant blue)was above of 1.5 times higher than that of control group,for the successful preparation of the model of diabetic nephropathy.A total of 75 rats of streptozotocin model group model successfully.After the rat model was successfully established,the rats were randomlydivided into model group(group B),hydrochloric acid benner pury group(group),and the Lamps K idney granules' s group was divided into low,middle and high dose group(group C,D,E),15 in each group.Each group was treated respectively with drug treatment:Group A and group B were given normal saline 10ml/kg,gavage 1 time a day,C group was given benazepri1 hydroch1 oride tab1ets 1.05mg/kg,gavage 1 time a day,D,E and F group:they were treated with "Lamps K idney granules" 1.89g/kg,9.45g/kg and 18.9g/kg,1 time a day,last for 6 weeks.Determination of random blood glucose and 24 hour urine protein at 1 week before and after model building.Before model building and one day before the end of the experiment,fasting,normal drinking water,collection of urine for examination.On the last day,the rats were weighed,anesthetized with 10% chloral hydrate 4ml/kg,and the blood was collected from the abdominal aorta and centrifuge,for examination.The biochemical indexes of urine protein,urinary creatinine,serum c ystatin C,serum creatinine and serum urea nitrogen were detected by automatic multifunctional biochemical analyzer.The kidneys,weighing,pathology,HE staining,PAS staining and Masson staining were used.The pathological changes of tissue morphology,cell composition and staining characteristics of glomeruli and renal interstitium were observed by light microscopy.Then the score was evaluated according to the extent and the extent of the lesions.The mean values were statistically analyzed.Results: 1 The general condition of the each group of rats:normal group(group A)rat fur is smooth,clean,eyes brimming with radiating vigour,freely,sensitive reaction,water quantity was normal,appetite was good,feces and urine were normal,no special smell,weight increased come naturally(136.60±35.95)g.After modeling,the rats fur was dry and yello w,the lower abdomen and vulva were moist,the spirit was poor,the activity was little,the irritability is easy,food intake decreased,the urine volume is increased evidently,and the weight is reduced(5.34±0.96)g.After durg treatment,the above situation of B group gradually increased,but the C group,D group,E group,Fgroup improved,E group the most obvious.2 Comparison of blood glucose and 24 h urine protein before and after modeling in the each group of rats:The random blood glucose and 24 h urine protein quantity of rats with streptozotocin were significantly different from those before the model(P<0.01).Random blood glucose of all rats in each group was more than 16.6mmol/L,which showed that the model of diabetic model was successful.24 h urine protein of rats in each group was more than 1.5 times that of the normal group,indicating that the diabetic nephropathy model was successfully prepared.3 Comparison of renal coefficient(kidney weight to weight)in the each group of rats: Compared with group A,the renal coefficient increased significantly in group B(P<0.01);the renal coefficient decreased in group C and group D compared with group B(P<0.05),the renal coefficient of group E and group F was significantly lower than that in group B(P<0.01);the decrease o f renal coefficient in group E and group F compared with group C(P<0.05);the renal coefficient decreased in group E and group F compared with group D(P<0.05).4 Comparison of ACR(Urine Microalbumin to Urine creatinine)in the each group of rats:There was a significant difference in the AC R between the rats modelled successfully with STZ after treating and before modelling(P<0.01).Group B was compared with A,and AC R increased observably(P<0.01);the C and D groups were compared with the B group,and the ACR decreased(P<0.05),the E and F groups were compared with the B group,and the AC R decreased observably(P<0.01);the E and F groups were co mpared with the C and D groups,and the ACR decreased(P<0.05).5 Comparison of renal function in the each group of rats:B group BUN,Cys C are observably higher than A group(P<0.01).Compared with the B group,the BUN in the C,D,and F group decreased(P<0.05),BUN observable reduction in group E(P<0.01);Compared with the B group,the Cys C decreased in the C and D group(P<0.05),Cys C observable reduction in group E and F(P<0.01);Group E BUN was lower thanC?D and F groups(P<0.05);Group E and F Cys C were lower than group C and D(P<0.05).6 Comparison of pathological changes of kidney tissue in rats 6.1 Macroscopic observation:normal rat kidney is dark brown,shiny surface,soft texture,round edge sharp;compared with the normal group,the kidneys of model group,hydrochloric acid Benner Pury group group,the Lamps K idney granules' s group was divided into low,middle and high dose group showed no obvious abnormality in the kidney,the volume did not increase or decrease.6.2 Histopathology 6.2.1 Glomerular mesangial,glomerular basement membrane(GBM),renal tubule lumen and renal tubular epithelial cells:Compared with group A,glomerular mesangial matrix hyperplasia,GBM thickening,renal tubular dilatation and vacuolar degeneration of renal tubular epithelial cells in group B were significantly different(P< 0.01);compared with group B,glomerular mesangial matrix hyperplasia,GBM thickening,renal tubular dilatation and vacuolar degeneration of renal tubular epithelial cells were improved in group C and D,the lesion is light(P<0.05),group E and F significantly improved,the lesion is lightest(P<0.01);compared with group C and D,glomerular mesangial matrix hyperplasia,GBM thickening,renal tubular dilatation and vacuolar degeneration of renal tubular epithelial cells were improved in E and F group,the lesion is light,(P<0.05).6.2.2 Inflammatory cell infiltration in renal cortex,renal tubule and kidney tissue : Compared with the A group,the renal cortical necrosis,renal tubular necrosis and inflammatory cell infiltration in the renal tissue were obvious in group B(P<0.01);compared with group B,renal cortical necrosis,renal tubular necrosis,and inflammatory cell infiltration in renal tissue were improved in group D,E and F,the lesion is light(P<0.05);compared with group C,renal cortical necrosis,renal tubular necrosis,and inflammatory cell infiltration in renal tissue were improved in group D,E and F,the lesion is light(P<0.05).Conclusion:The effect of Lamps K idney granules on DN rats:Improve the basic condition of DN rats;reduce ACR excretion in rats with diabetic nephropathy;reduce BUN and C ys C in DN rats and reduce renal function injury;reduce the renal pathological changes:DN can reduce rat mesangial matrix accumulation and glomerular basement membrane hyperplasia,reduce renal tubular lumen expansion and the renal tubular epithelia l cells in vacuolar degeneration,the changes showed a dose-dependent trend;at the same time reduce the renal cortical necrosis,renal tubular necrosis,inflammatory cell infiltration,a dose dependent trend did not show.Benner Pury hydrochloric acid also showed a similar therapeutic effect,which was less effective than the high dose group in Lamps K idney granules. |
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