Glucose Tolerance Kang Renal Protective Effect And Mechanism Of Diabetic Rats

Diabetic nephropathy(DN) is an important microvascular complication of diabetic mellitus(DM) as well as the common reason resulting in chronic renal failure(CRF),and half of which is caused by DN in some countries or areas. So protecting renal function,reducing the rate of renal damage and improving quality of life becomes to the focus of international researches' attention. On the former study,systematic research on prevention and treatment of DN by Tangnai-Kang concoction has been performed from the whole,organ,cell and molecule levels in this essay which was composed of the reference review and experimental study.1 Reference reviewThe reference review discussed the recent research progresses in prevention and treatment of DN both from the aspects of Chinese medicine and modern medicine. In the theoretical research,we have referred to the related ancient and modern TCM literature in order to explore the source of disease name of TCM for DN, etiology and pathogenesis of TCM,selection of therapeutic methods by different stages or according to the differential syndrome diagnosis,advance on the clinical and experimental research as well as the problems lying in the research of TCM and corresponding strategies.In the meanwhile,we further exploited the pathologic characteristics,diagnosis and treatment of DN.We also have summarized the tutor's academic point of view on the differentiation and treatment of DN,and emphasized that deficiency of qi and yin is the basic pathogenesis of DN,blood stasis and turbid phlegm run through the progress of DN.2 Experimental study2.1 experiments in vivoObjective:To study the affection of Tangnai-Kang concoction for protecting for diabetic kidney in spontaneous diabetic animal model GK mouse,and explaining the mechanism of it.Mwthods:Specific pathogen free(SPF) GK mice 50 were divided randomly according to plasma glucose level into 5 groups:Models,TNK-low dosage,TNK-middle dosage,TNK-high dosage,Pioglitazone and normal wistar mice(normal control group). Each group was tested randomly plasma glucose level every week.60 days later, all animals were tested body weight,renal weight,blood glucose,urinary albumin (ALB),β2 microglobulin(β2-MG) and dying kidney by HE and PAS.At the end, observe the ultrastructure of the kidney through TEM.Results:Compared with Model groups,blood glucose,ALB,β2-MG were all decreased significantly(p＜0.05);the recipe could protected diabetic kidney in some degree and postponed kidney failure,and improve pathological change in diabetic kidney through kidney dying by HE,PAS and TEM.2.2 experiments in vitroObjective:To observe the proliferation of mesangial cells(MCs) in high glucose, and the affection of rat serum containing Tangnai-Kang to it firstly;To study the intervention of the serum in extracellular matrix and extracellular regulated kinases(ERKs) of MCs.Methods:Mesangial cells were cultured,then using 30%high glucose induced it. Divided cells into six groups,such as Models,TNK-low dosage(0.28g/ml) TNK-middle dosage(0.47g/ml),TNK-high dosage(0.80g/ml),Rosiglitazone dosage and normal control group(MCs).Treated the cells 24h,test FN,IV-C,TGF-βin the extracellular matrix,and extracted the total RNA and protein.At the end,tested the expression of cPLA and PAI-l mRNA,and test p-ERK,PKC-α,c-fos protein by reverse transcriptase PCR(RT-PCR) and Western blot.Results:Cultured rat MCs exposed to high glucose environment,MCs proliferation was assessed by MTT colorimetric method.Our studies demonstrated that raising ambient glucose concentration results in transient stimulation of growth in 48 hours,and in inhibition after 48.Biapbasic growth response of MCs to high glucose concentration could be reversed by rat serum containing Tangnai-Kang. Compared with Model groups,FN,IV-C,TGF-βwere downregulated in some digree in the supernatant secreted by MCs following the concentration of Tangnai-Kang Compound,and the expression of everyone were downregulated,such as cPLA and PAI-l mRNA,p-ERK,PKC-αand c-fos protein level.The effecting of Rosiglitazone dosage group were similar with TNK dosage.Conclusion:Tang-nai Kang Concoction have the affection of improving blood glucose and decreasing urine protein excretion,and improving renal function, lessen nephridial tissue damage,prevented the hardening of glomerular,which may realized through these links as follows:①Block ERK signal-transduction effectively,controlled the goal proteins or genes expression level,increasing the activity of the goal points in ERK signal-transduction pathway for which decreasing the FPG urine protein and so on.②Through decreasing the secretion of MCs stimulated by high glucose,such as FN,IV-C,TGF-β,relief the interference factors on ERK signal-transduction pathway.On the other hand,the mechanism of the serum containing Tangnai-Kang Compound improving diabetic nephropathy might be down-regulated the expression of key points.