Effects Of Hedysaryum Polysaccharide On Expre-ssions Of GluT-1、FN And Col-IV In Renal Tissue Of Db/db Mice With Diabetic Nephropathy

Objective To investigate the protective mechanism of Hedysari polysaccharide（HPS）on renal fanction and the effect of expressions on GluT-1, FN, Col-IV mRNA and protein inrenal tissue of db/db mice with diabetic nephropathy, to clarify the possible mechanism.Methods Male db/db mice with diabetic nephropathy and db/m mice were used for thisstudy. The mice were entered experiment after affording early DN by testing urinarymicroalbumin. The db/db mice were randomly divided into5groups: model group, enalaprilgroup and the low、middle and high dose of HPS groups, the db/m mice were taken as thenormal control group, there were10each group. All groups were respectively given0.9﹪physiological saline, enalapril(10mg.kg-1.d-1) and HPS (100,200and400mg.kg-1.d-1) bygavage daily for a period of8weeks. Collecting24h urine of each groups in metabolic cage,then used it to determine the urine albumin (UMALB). The mice′s eyeballs were picked off tocollect blood sample for serum creatinine (SCR) and blood urea nitrogen(BUN). The renaltissues were collected immediately to separate the cortex and medulla after mice death. theleft kidney was saved in-80℃refrigerator for detection of RT-PCR and Western blotting; theright kidney was immediately embedded with paraffin for immunohistochemistry, HE andMasson staining. Picric acid method was used to detected serum creatinine(SCR), enzymecoupling rate method was used to determine blood urea nitrogen (BUN), ELISA method wasused to determine urinary microalbumin (UMALB), RT-PCR method was used to theexpressions of mRNA of GluT-1, FN and Col-IV by. Western blotting andImmunohistochemical method were used to detect the expressions of protein of GluT-1,FNand Col-IV.Results (1) The results of renal function tests: The Enalapril and each HPS groupcompaired with the model group, the levels of SCN, BNM and UMALB after treatment inrenal tissue of db/db mice were lowe. The levels in enalapril and HPS high group was reducedsignificantly (P<0.01).(2) The expressions of GluT-1in renal tissue: The expression of GluT-1in renal tissue ofdb/db mice in Enalaporil group was lower than model group one(P<0.05or P<0.01), Theexpression of GluT-1in renal tissue of db/db mice in HPS high dose group was lower than model group one significantly(P<0.01).(3) The expressions of FN in mice kidney: The expressions of FN in renal tissue of db/dbmice in Enalaporil and PHS high dose group were lower than model group ones significantly(P<0.01); The expressions of mRNA and protein of FN in renal tissue of db/db mice in HPSmiddle ang low dose group were lower than model group ones (P<0.05or P<0.01).(4) The expression of Col-IV in mice kidney: The expressions of C-IV in renal tissue ofdb/db mice in Enalaporil and PHS each group were lower than model group ones. Thedifference was obvious in enalapril group(P<0.01). The area of brown granules dyeinggradually decreases with the increase of HPS dose in kidney tissue.(5) The kidney tissue HE staining test: The degree of glomerular structural disorder, thedistributionm of capillary, the contorts of renal tubular, infiltration of inflammatory cell inrenal interstitial in all treatment groups were significantly better than model group ones. Thedegree of kidney lesions gradually reduced with the increase of HPS dose.(6) Kidney tissue MASSON staining test: The degrees of glomerular structural damagedand capillary disorder, the expression of collagen fiber in all treatment groups weresiginificantly better than model group. With the increase of HPS dose, the expression anddistribution of collagen fiber gradually reduce in kidney tissue.(7) Kidney tissue electron microscopy test: The morphology of glomerular endothelialcells, mesangial cells, podocytes and tubular epithelial cell in all treatment groups were betterthan model group. The microstructure of organelles and wools in all treatment groups werebetter than model group too.There were no significant difference between HPS high-dose andenalapril group. With the increase of HPS dose, lesion level gradually reduce in kidney tissue.Conclusion (1) The expressive trends of GluT, FN and Col-IV are basically identical.(2)HPS could protect renal function and delay the progress of DN by reducing the expressions ofGluT-1, FN and COL-IV with various mechanisms.(3)The pharmacological effects and theresearchful results of HPS could prove that one of the pathogenesis of diabetic nephropathywas " Renal Qi deficiency, change chaotic and stasis", it provides a new theoretical support tochoose drug which benefit Primordial Qi for encouraging Renal Qi, Removing stasis andchanging chaotic to prevent and treat DN.