| Purpose:Observing the change,effect and the clinical trial result of urinary albumin excretion(UAE)of diabetic nephropathy(DN)and transforming growth factor(TGF-β) in streptozotocin(STZ)which caused by A compound prescription of traditional Chinese medicine called Tang Ke Decoction,to discuss its effect and mechanism in precaution and treatment of diabetic nephropathy(DN)Material and method:1.Trial Study1.1 Methods of modeling With high fat,high sugar diet combined with intraperitoneal injection of streptozotocin(STZ) to duplicate Diabetis model the method is one-month high-sugar high-fat feed to induce obesity and insulin resistance on Wistar rats,and then give STZ(40mg/kg) injection through abdomen to lead hyperglycemia,and miner albuminuria.1.2 Method of grouping and oral medication administering Deride all the rats into normal group(10 cases)and trial group(70 cases)randomly by weight of rats.and then devide 61 rats which has stable blood sugar after one week modeling into model group(12 cases),Chinese medicine precautious group(12 cases),low dozed chinese medicine group(12 cases),high dozed chinese medicine group(13 cases) and western medication compared group(12 cases).Giving Tang ke decoction 8-9 am q.d after injecting STZ to build DM model in chiense medicine precautious group and giving meds in other groups after building DM model 4 weeks.Giving same doze physiological saline in model group and normal group and giving Lotensin 1mg/kg·d in western medication compared group.In Chinese medicine precautious group,the medicine giving lasts 12 weeks.And in Chinese medicine high doze group and low doze group and western medication compared group,the medicine giving lasts 8 weeks.All the rats allow drinking unlimitedly during this trial.The medicine giving in Chinese medicine precautious group and lowdoze Chinese medicine group is 6.3 times as human adult doze.Triple the low doze medicine group doze in High doze medicine group.1.3 Observating index and measuring methodObserving the effect index of Tang ke Dedoction including the effection of general situation,weight,kidney weight and comparatively kidney weight on DN rats.Testing blood sugar once in a week with GT-1810 blood sugar testing mechine made by Aike itd,Japan.Testing blood lipids after trial with enzymating method.Testing glycosylated hemoglobin(GHB) after the test with. ion-exchange chromatography.Testging 24h urinary protein and urinary albumin excretion rate(UAER)after the test with Lmmulite method.He staining and Masson staining the renal biopsy and observing under optical ordinary microscope,then uranyl acetate-lead citrate staining the renal biopsy and observing under electronic microscope to find the change in pathomorphology on all the DN rats.Measuring transforming growth factor(TGF-β1) and its internal transduction molecules of the protein expression of smad4 with immunohistochemical method.Meanwhile, testing the antioxidant enzyme activity of renal tissue glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD) After the above testing,analyzing and discussing the effect and mechanism of Tang ke decoction on DN rats.2.Clinical research, With randomly compared method,selecting 90 early stage DN patients which have Qi-deficiency and blood stasis,therapy group(45 cases) with Tang ke decoction(Astragalus 30g,cyrtonema 30g,Chinese yam 20g,Poria 20g, Eupatorium 30g,Kun Grass 20g,Arctii 30g),positive drug control group(45 cases) with Lotensin(Beijing Novartis Pharma Ltd.,10mg/d),treating 8 weeks,to make a comprehensive clinical ovservation on early stage DN patients which have qi-deficiency and blood stasis in urinary protein excretion rate (VAER),plasma glucose(FPG),glycosylated hemoglobin(GHb),lipids,blood flow changes,and symptom score3.Statistical methods:Analyzing the group differentia in cariance analysis and T test with SPSS (16.0 vorsion).Measuring date with(?)±s.P＜0.05 is significant.Results:1.Results of animal experiments1.1 The affection on general situation of ratsThe weight of normal compared group rats increase obviously,they are well, sensitive,agile and shiny colour.The experimental groups are over-drinked, over-ate,over-urinated,fatigued,insensitive,slow and tarnished.In graphs of weight,water-taking,urine volume,it showed the weight of rats increase from 5th week and increase rapid at 12th week.It also showed water-taking became normal at 8th week.Urine volume has not difference from normal groups(P＞0.05).Compared the effect on model group,it works more obviously on precautious group(p＜0.01). it manifested that Tang ke Decoction can improve the symptoms of DN rats and prevent the DN get worse.1.2 The affection to the rate of urinary protein excretion on rats:The rate of urinary protein excretion in compared group is lower than it is in model group.Moreover,it works very well in precautious group and high dozed medicine group,compared with model group(P＜0.01).1.3 The affection on blood sugar:The blood sugar in experimental groups are all higher than it is in normal group,Chinese medicine group are obvious different from model group,the glycosylated hemoglobin in Chinese medicine group is lower than it is in model group.However,the effect in precautious group is significant(P＜0.01).1.4 The affection on blood lipids:The total cholesterol and triglyceride levels in precautious group were lower than they are in model group,and the effects of decrease cholesterol in precautious group is obviously different from model group(P＜0.01)1.5.The affection on kidney weight and relative kidney weight:Compared with normal group,both the kidney weight and KW/BW increased in model group.After 8 weeks therapies of Tnag ke Decoction and Lotensin.the kidney weight and KW/BW are both decreased.Moreover,it decreased more in Chinese medicine group.1.6 The affection on renal pathological changes1.6.1 the naked eye and optical microscope observation:The kidney volume increased in most of rats in model group.There are three 2x3mm nodules on the surface of a kindey in 1 case.Under optical microscope, it could be seen as glomerular swelling,congestion,markedly increased volume and the narrowed renal capsule,some thickened glomerular GBM,widened mesangial,mesangial cell proliferation,proximal tubule epithelial swelling, there also could be seen as small transparent tube,widen distal convoluted tubule cytosolic space,balloon-like degeneration.Some kidney tube appears narrow portion,edema in renal interstitial and minor fibrosis.All the groups except normal group are all have volume decrease but still has volume increase compared with normal group.Under optical microscope, glomerular swelled slightly,congested GBM thickened slightly,mesangial widened slightly and kidney tube got swelled.The extent of pathological change were small and slight which has no glomerular sclerosis and renal interstitial fibrosis.There was no big difference between all the therapy groups.1.6.2 electron microscope observation:The GBM was thickened equally in model group.The formation of foot process fusion is sufficient to reduce intracellular organelle,the expansion of mesangial areas,increased mesangial matrix,the cytoplasm of mitochondrial swelling,decreased matrix density,and some ridge reduced,the increase in the number of lysosomes,and larger.The based membrane of segmental tubular thinning or thickening in some sections;some renal tubular epithelial has cell loss,reduced microvilli, shorter,or even falling visible capillary erythrocyte aggregation,and vacuolar degeneration.In all the therapy groups,compared with model group,it can be seen glomerular GBM under electron microscope and tubular glomerular basement membrane get thicken,the epithelial cell foot process get broaden,renal tubular epithelial cells and microvilli fell off slightly. There was no big difference between all the therapy groups.1.7 The affection on kidney tissue TGF-β1 and smad4:Normal group kidney has little volume and slight colour TGF-β1 protein expression in both glomerular and tubular epithelial cells.Compared with normal group,the model group has more volume of TGF-β1 protein expression which showed extremely positive expression and deep colour in both places. The TGF-β1 in all therapy groups have week positive expression which goes down compared with model groups(P＜0.05).It manifested that after the therapies of Tang ke decoction and Lotensin,the TGF-β1 decreased in DN rats. Moreover,the testing result of smad4 is similar to TGFβ1.1.8.The affection on GSH-Px and SOD in kidney issue.Model groups has lower Renal tissue GSH-Px and SOD vitality than normal group. And it is also lower than all the other groups.Comparing precautious group and western medication group,It is significant in statistics(P＜0.05).It manifested that Tang ke decoction could improve Renal tissue GSH-Px and SOD vitality.And it works better than Lotensin.2.result of clinical observation Mainly test indexes:24h urinary albumin excretion rate(UAER) in treatment group decrease from 74.36±47.90μg/min to 13.25±1.26μg/min which has obvious dirrerence (P＜0.01),compared with compared group P＜0.05.The other indexes including fasting blood glucose(FBG),glycosylated hemoglobin(GHb),lipids,blood flow change are all have obvious change.Syndrome points decrease from 19.58±2.55 to 4.42±1.65 after therapy which has obvious difference(P＜0.01). It shoes that,the rate of treating early stage of DN with Tang ke Decoction is 44%.Totally effect is 95.56%,which are all better than hotensin compared group,P＜0.01.Conclusion:1.Tang ke decoction has obvious effect on Qi-deficiency and blood stasis DN patients.2.Tang ke decoction can improve the general state of rats and reduce diabetic symptoms.3.Tang ke decoction can significantly reduce the rate of 24h urinary albumin excretion in early stage of DN.Therefore,it can reduce the kidney damage caused by urinary protein and provide the protection to the kidney.4.Tang ke decoction can lower blood sugar,regulate lipid metabolism disorders,improve the blood flow changes,thereby improve renal microcirculation,increased renal blood flow,increase antioxidant capacity and vascular endothelial protection.5.Tang ke decoction can inhibit kidney hypertrophy,protect glomerular and tubular structure and improve kidney function. 6.Tang ke decoction can inhibit TGF-β1/smad4 protein expression to inhibit mesangial cell ECM synthesis and reduce the abnormal accumulation of ECM. Moreover,it can abate glomerular sclerosis.7.Tnag ke decoction can increase GSH-Px and SOD vitality of kidney tissue of DN rats,prevent kidney tissue damage caused by free radical and protect renal function.8.Enough doze of Tang ke decoction in early stage can alleviate DN symptoms, delay the disease progression.It is worth to be used and recommended.
|
PDF Full Text Request