| Diabetes is the 9th among the 50 kinds causes of death, the prevalence rate increased year by year in China, diabetic nephropathy (diabetic nephropathy,DN) is a common and serious microvascular complication of diabetic, and its incidence rates increased year by year, one third of patients with diabetic may develop into end-stage kidney failure.As the high blood sugar, the advanced glycation end products, the renin-angiotensin system, the oxidative stress, endothelin (ET) and other factors exist in diabetic nephropathy, the secretion of the transforming growth factorβ1 (TGF-β1) increase.Many agents can cause the generation of TGF-β1, TGF-β1 bind with its receptor (RI/RII) and lead to the activation of receptor-type Smad3,TGF-β1/smad signaling pathway can mediate injury of podocytes, the proliferation of mesangial cells, the hypertrophy of tubular cells,and promote the generation of the extracellular matrix production and inhibit the degradation of extracellular matrix, basement membrane thickening, epithelial cells-mesenchymal transformation and apoptosis of the renal cells which lead to glomerular sclerosis and renal interstitial fibrosis. Smad7 is one of the inhibitory Smad family members, it plays a negative role in the regulation TGF-β1/smad pathway. Nuclear factor(nuclear factor,NF)-KappaB(κB) is a important multi-targeted nuclear transcription factor, widely exist in various cells, it is involved in intracellular signal transduction and the regulation of multiple gene expression, It can both inhibit and promote apoptosis. Smad7 can inhibits anti-apoptotic factor NF-κB activity in DN."Consumptive Renal Disease" hypothesis is metioned by Professor Zongjiang Zhao which is based on years of experience in the diagnosis and treatment of kidney disease and he refer to a large number of ancient documents and modern medical knowledge and propose a new kidney theory for Chinese medicine. "Consumptive Renal Disease" refer to the weak and the useless of the kidney. The clinical syndrome of this chronic wasting renal disease is mainly cloudy urine.This disease belong to the generalized "flaccidity" category.The reason of the Atrophy is that long lesions in renal kidney disease,its pathogenesis is mainly hot in the body, Qi, yin and yang Deficiency, the whole renal Deficiency, severe Jing Qi Deficiency. All those resulting in kidney damage.The pathological mechanism are deficiency in root and excess in superficiality. The main treatment is to invigorate the kidney and also treat the superficial diseases.TangShenNing Capsule is a prescription which my teacher made under the "Consumptive Renal Disease" theory to guide the treatment of diabetic nephropathy. It can warm the kidney and spleen, detoxicate,and alleviate water retention.It can give a good treatment for diabetic nephropathy"Consumptive Renal Disease"disease. The experiments used immunohistochemistry, in situ hybridization staining, RT-PCR experimental methods and studied from the different perspective of protein and mRNA in diabetic nephropathy and podocytes which is under high glucose and the express of TGF-β1,Smad7,NF-κB and TangShenNing capsule's function on its expression. To investigate the control mechanism of TangShenNing capsule on DN.Objective:In order to investigate the mechanism of TangShenNing Capsule in relief DN from levels of organ, tissue, cell, protein and gene and improve the DN renal function, reduce oxidative stress,delay the renal pathological damage,we established DN rat model and high glucose cell model then observed the efficiency of TangShenNing Capsule in influence TGF-β1,Smad7,NF-κB gene and protein's expression.Methods:Diabetes induced in 44 SD rats by intraperitoneal injection of streptozotocin(STZ) was used in vivo experiments. The rats were randomly divided into Nomal group, model group, irbesartan group,TangShenNing group, each group were treated accordingly.To observe the mental state of rats, body weight, body hair,water intake, stool and other activities; the body weight of the rats at the 2,4,6,8,10,12 weeks of experimental were mesured,kidneys were removed and weighed at the 12th'week.The kidney weight to body weight ratio were calculated;The urine were collected respectively at the end of the 4,8,12th week, and 24-hour urinary protein were determined by coomassie briliant blue method; serum was used to test the TG, BUN, SCr, NO, MDA, SOD content. HE, methenamine silver,Masson, Mallory staining were used to observe the pathological changes in rats.The Pathological score standard were used to evaluate the renal pathological changes. Immunohistochemistry, In situ hybridization, RT-PCR methods were used to detect the expression of the protein TGF-β1,Smad7, NF-κB protein and mRNA expression of diabetic nephropathy tissue and podocytes under the high glucose.Results:The mechanism of TangShenNing capsule's prevention and cure action forDN maybe lie in:①The body weight:Compared with model group,The body weight of TangShenNing capsule treated rats had increased, in the 12th week, compared with the model group, The body weight of TangShenNing capsule treated rats were different significantly (P<0.01).②kidney weight/body weight ratio:Compared with model group, the kidney weight/kidney weight ratio of TangShenNing capsule treated group decreased significantly, there were significant differences (P<0.05).③24h urine protein:Compared with model group, In the TangShenNing capsule treated group, at the 4th and 8th week, the 24h urine protein excretion decreased significantly (P<0.01),at thel2th week,it decreased (P<0.05).④blood fat: TangShenNing Capsule can reduce the TG content, correct lipid metabolism significantly (P <0.05).⑤Renel function:TangShenNing Capsule can improve the renel function of the DN rats.It can reduce the BUN (P<0.05) and Scr (P<0.01) contents of the serum.⑥oxidative stress:Compared with model group,TangShenNing Capsule can work for the increase of the NO content in the serum of the DN rats (P<0.01), The MDA content was reduced (P<0.01) and the SOD content was increased (P<0.01).⑦TGF-β1-Smad7-NF-KB signal transduction passageway:TangShenNing Capsule can reduce renal TGF-β1 protein and mRNA expression and increase the Smad7 protein expression of diabetic rats and the glomerular visceral epithelial podocytes under high glucose.Conclusion:In short, TangShenNing Capsule has significant protective effect on the kidney of DN"Consumptive Renal Disease" rats. its mechanism may be like this, it can reduce the DN renal tissue TGF-β1, NF-⒘B protein expression, increases the expression of Smad7 protein and inhibit high glucose stimulated podocytes TGF-β1, NF-icBmRNA expression, and increase Smad7 mRNA expression. And it can work for the TGF-β1-Smad7-NF-icB signal transduction passageway, and reduce the podocytes apoptosis and the generation of the albuminuriao... |
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