The Protecting Effect Of Melatonin Combine Gliquidone In Diabetic Nephropathy Rat On Kidney

Objective:Through copying the model of rat diabetic nephropathy, to observe the protecting effect of Melatonin combine Gliquidone in diabetic nephropathy rat on kidney. Methods:①rats were randomly divided into normal control (NC) and diabetic nephropathy (DN) model group. DN model group were replicated by intraperitoneal injection of STZ 60mg/kg,NC group were injected with an equal volume citrate buffer. After 8 weeks, NC group and the DN model group were randomly selected six rats to detect fasting blood glucose (FBG), serum creatinine (Scr), blood urea nitrogen (BUN) and kidney morphology, to determine the models was replicated successfully.②DN model rats were randomly sub-divided into diabetic nephropathy control (DN) group, melatonin treatment (Mel) group, Gliquidone (G) group and Melatonin associate with Gliquidone (MG) group. Since nine weeks of the experiments,Mel groups were treated with melatonin 10mg/kg/d by gavage; G group was treated with Gliquidone 15mg/d by gavage, MG Group were treated with melatonin 10mg/kg/d and Gliquidone 15mg/d by gavage.NC group and the DN group were treated with normal equal Saline wich containing 2% ethanol by gavage,Four weeks later,we detected FBG by glucometer; the method of diacetyl monoxime to measuring BUN, the method of picric acid assay to measuring Scr, CBB assay to detected 24h urine protein (24hUP); the expression of cysteine-aspartic protease 3 (caspase-3) protein to detect with Western blot; RT-PCR method to measur renal cortex bax, bcl-2mRNA; the methods thibbarbituric acid to measuring malondialdehyde (MDA) in renal tissue, use the hydroxylamine to Measuring superoxide dismutase (SOD), using dithio-2 nitrobenzoic acid assay to detected the activity of glutathione peroxidase (GSH-Px); the renal cell Apoptosis Index using in situ end labeling (terminal deoxynucleotidyl transferasemediated dUTP nick endlabeling, TUNEL) to detected; histological changes was using HE and Masson staining in the kidney rat. Results:①Compared with the normal control group, The FBG, BUN, Scr and 24hUP were significantly increased in diabetic nephropathy model group (P<0.01); the renal mesangial cell was proliferated, mesangial region broadening,tubular dilation, dropsy in epithelial cells, the basement membrane renal tubular was thickening irregularly,Renal interstitial fibrosis and most of glomerulus of the kidney moderate sclerosis, this all show that the renal had obvious functional damage and morphological changes,,it is indicat that the model of diabetic nephropathy rats were successfully reproduced. Compared with diabetic nephropathy control group,FBG was insignificant decrease in melatonin group (P>0.05), and BUN, Scr,24hUP were significantly lower (P＜0.01, P<0.05);the activity of SOD, GSH-Px was significantly higher in renal tissue (P<0.05, P <0.01), and the content of MDA was decreased significantly (P<0.05), the apoptosis index was significantly lower (P<0.01); its show that the renal function and renal pathological changes has significantly improved in the role of melatonin on diabetic nephropathy,while the blood glucose were not significant lower; the FBG,24hUP were significantly lower in gliquidone group (P<0.05), and Scr, BUN decreased (P> 0.05); the activity SOD, GSH-Px in renal tissue were higher (P> 0.05), and MDA content was lower (P> 0.05);the expression Bcl-2mRNA was significantly increased (P<0.05),and expression of BaxmRNA was significantly lower (P<0.05), protein Caspase-3 was lower (P>0.05), and apoptosis index of the kidney was significantly decreased (P<0.05); show that the Gliquidone have obviously effect on hypoglycemic in diabetic nephropathy rats and also on the DN of renal function and renal pathological changes in the role has some improvements,FBG was significantly decreased in MG group (P <0.01), BUN, Scr and 24hUP significantly decreased (P<0.01); the expression of Bcl-2mRNA was significantly increased (P<0.01), BaxmRNA,protein Cas-pase-3 and apoptosis index were significantly decreased (P<0.01), The results showed that Melatonin combine Gliquidone had obvious hypoglycemic effect,also on the diabetic nephropathy of renal function and renal pathological changes in the role has significantly improvements.③renal mesangial cells and mesangial matrix to reduce and epithelial cell edema was alleviated, Renal interstitial fibrosis and glomerular sclerosis was reduced,especially group MG. Conclusion:Melatonin,Gliquidone and melatonin combine Gliquidone have all protecting effect on kidneys in diabetic nephropathy rats;The effect of protecting in MG group was better than melatonin or gliquidone when it used alone; which may be related to lower blood sugar, to reduce the damage of oxidative stress and apoptosis in the kidney and so on.