The Patho-alteration Of Diabetic Nephropathy In Rosiglitazones Treated Diabetic Rats

With the rapidly development of the global economy,people' s living standard has also been improved,the incidence of Diabetes Mellitus(DM)has been increasing rapidly in China.According to the statistics,the morbidity rate of DM in China is higher than 2.5%,Diabetic nephropathy(DN)is one of the most severe vas capillare complications of DM.In addition,DN related ESRD is a life- threatening disease.Thus,it is very important to control the progress of DN.The main patho-characteristic of DN is thickening of the GBM,and the apposition of the ECM.Rosiglitazone is one of the euglycemic agent,it works by combining and agitating the PPARs and through Peroxisome proliferator-activated receptor gamma(PPARgamma)ligand mostly.It has lots of effects,such as to reduce serum glucose,blood pressure and urinary albumin, to regulate the metabolism of the lipoids and to provide anti-inflammatory effect.ObjectivesTo analyze and identify patho-alteration of Diabetic nephropathy in rosiglitazones treated Diabetic Rats.Materials and MethodsExperimental Animals1.Studies were carried out in 30 Sprague Dawley(SD)male rats,10 weeks old,(from ZheJiang University Animal Laboratories Center).The mean weight was 315g(SD=30.06)and the mean blood sugar was 96 mg/L(SD=4.86).The rats were housed in animal quarters with a 12-hour light/dark cycle and were allowed free access to chow and water throughout the study.The temperature was kept at 20℃to 22℃ 2.SD rats were divided into three groups(the control group,the diabetic group and the ROS-treated diabetic group,each group has 10 rats):Experimental group were given with intraperitoneal injection dosage of STZ 60mg/kg diluted 0.1M pH 4.5 sodium citrate buffer.Control group were treated with a single intraperitoneal injection sodium citrate buffer 0.1M,pH 4.5.10 diabetic rats were given PPARgamma ligands:rosiglitazones(ROS)1mg/kg/d intragastric administration(Corporation)dissolved in normal saline.3.Urine glucose were tested every day and glucose was check every week from tail vein for the measurements of glucose(American Adantage).If urine glucose was less than +++(positive reaction),no treatment was given;if urine glucose was ++++ or above,1～3 u of protamine zinc insulin was injected subcutaneously to prevent them becoming hyperosmolar or ketoacidosis comma or death.4.After 10 weeks,rats were anesthetized by intraperitoneal injection of ketamine(35mg/kg)and phenylethylmalonylurea 50mg/kg.The blood glucose was determined with one Touch Blood Glucouse Monitoring system.To get all the rat' s kidney sample and get them ready for the linght microscope.5.the light microscope sample prepair1.obtaining sample2,fixing 3.dehydration 4.tansparent 5.diping into the wax 6.embedment:to use the iron board 7.slice and paster 8.HE dying steps:1)Apply Dimetbylbenzene for 5-10mins to take off the wax.2)alcoholic dehydration(100%-95%-90%-80%-70%),each step for 2-3mins,to take off dimethylbenzene.3)Wash in the distilled water to take off the alcohol for 10-30mins.4)hematine to dye cell nucleus,2-3mins.5)use 0.5%hydrochloric acid alcohol to differentiate for several seconds.6)apply lotic water washing for 30-40mins. 7)put eosin for 2-3mins,the cell been dyed into pink.8)wash several seconds.9)use alcoholic dehydration(70%-80%-90%-95%-100%),each for 3-5mins.10)apply dimethylbenzene 10mins to get it tanspanrented.11)mounting.Statistical analysis:Data were analyzed using descriptive and inferential statistics with the Statistical Package for the Social Services for Windows(SPSS,Version 11.5). The acceptable level of statistical significance was set at p＜0.05,p＜0.01 was considered very significant.Results were expressed in terms of descriptive statistics such as means,standard deviations and the one-way ANOVA was used to analysis the relationship between variances.ResultsRats weight:The DM rats lost more weight compared to the control group(P＜0.01).The rats weight of group of DM treated with ROS increased compared with DM. Rats glucose concentration:STZ induced Diabetic rates had higher glucose level(P＜0.01)compared with the control.The glucose level of DM group with ROS was up compared with those without ROS,however,it did not have statistical significance(P＞0.05). The patho-alteration of the D group,DR group and the C group under the light microscope.The pathological changes of DR group is small,the glomerular capillary is lightly narrowed,there is a small quantity of lymph-cell infiltrating in the renal tubule-interstitial area.In the C group,the lumen of glomerular capillary is uniformity,non-narrow,and there is no lymph-cell infiltrating in the tubule-interstitial area.The D group has obviously changes.The lumen of glomerular capillary collapsed,obliteration mesangial region widened,basal lamina thickened,and mesenterium base material mult.In addition,the volume of the glomerulum increased and cell population muir.Furthermore, hyalinization;nephric tubule especially the proximal convoluted tubule engorged,degenerated and cavitated,and lots of lymph-cells and mononuclear macrophages can be found in the renal interstitium.ConclusionLingt microscope results suggested that glomerulum,tubule and the interstitium pathological changes of DR group are smaller than those of the D group.It indicated that ROS could protect the kidney of diabetic rats from being damaged by hyperglyceamia.Moreover,the D group have more cells in the glomerulum,and the lumen of blood vessel have become narrower,even to block up.