| ObjectiveDiabetic nephropathy (DN) is the the frequent extremely complication of type 2 diabetes mellitus,is one of the main cause of death in type 2 diabetic patients aslo. Due to states of an illness delitescencing in early stage , seldom provoke patients attention , it appeared injury in glomeru-lar when clinical symptom of diabetic nephropathy occurred,so it is very important that preveting and curing diabetic disease . PSS is sulfate pro-teoglycans which is half—synthesized with the thing distilled from alga (C5H7O4COOH) , and also is kind of anion electrolyte without side — effect similiar to heparin. PSS can modulate dysfunction of hemodynamics of microcirculation in renal and reduce the excretion of urine albumin;Increase circum—tissue sensitivity to insuline to further improve diabetic glycometabolism. The effects of PSS on glomerular capillary basement membrane and the expression of TGF —β1 ( important causative agent of DN)and type Ⅳ collagen in renal glomerulus , urinary microalbumin excretion rate in diabetes mellitus rats of type 2 were observed, to further explore whether to have preventive and therapeutic effect of PSS on DN .MethodsAnimal ModelWistar rats were randomized into control (n = 8)and diabetic(n=32) groups. Control group had free access to tap water and standard ratchow,throughout the duration of the experiment. Diabetes was fed with high—sugar and high —fat diets , then were injectedO. 4% STZ20mg/kg intravenously, blood glucose>10mmol/Lwith urine glucose + + + rats were served as experimental animals model, divide into groups: experimental animals model wre randomized into diabetic non— therapy(n = 8, normal sodium) group and PSS—treated (n = 8) and positive control(n = 8) group . each group was intragastric administrationed isometric 10ml ? kg"1 ? d-1 for 8 weeks. Therapy group were given 1. 8%pss solution reconstituted with normal sodium, positive control group were given 0. 04% lovastain with normal sodium .specimen collection and processWeigh before and after 8 weeks of treatment , blood glucose levels were determined through by vena caudalis , 0. 2% pentobarbital narcosis (30mg/kg) obtain blood from eye sockets before measure serum creati-nine;collect 12hour urine by metabolic cage for measure umalb and urine creatinine(Ucr). open bdominal cavity in Drugged state on the day before the termination of the experiment. Liberate left side kidney, removal peplos,weigh kidney, calculate kidney weight /body weight(KHI). Left kidney was put in 4% paraformaldehyde to fix. For histology study, im-munohistochemistry obtain many parts of nephridial tissue in renal cortex, then were fixed in 2. 5% glutaraldehyde and postfix in osmic acid, gradient dehydration in acetone plumbi uranium dyeing,observe by transmission electron microscope and photograph.indicator detectionScr^Ucr were measured by nitroxanthic acid ,Ccr was corrected with body weigh. Umalb was observe with immunoturbidimetry, manipulate according to instruction manual of kit. PV—6001 PowerVision. Two — step histostain measure TGF — J3i. Examination of type IV collagen use StreptAvidin—Biotin Complex(SABC). The intensity of glomerular staining of TGF—Pi was evaluated according to the following 0 to 4 scale in coded sections observed at 400 X magnification :0 = no staining;l = weak and spotty intraglomerular staining;2 = moderate and segmental intraglo-merular staining;3 = moderately strong and segmental,or moderate but diffuse (involving more than 50% ) intraglomerular staining;and 4 = strong and diffuse intraglomerular staining. Thickness of GBM measure: At least five sample per group were evaluated, five randomly spots selected in per sample take a photograph and measure, then the means values was calculated.Statistical analysisResults were analyzed using SPSS12. 0 software. The significance of analysis of variance(ANOVA) testing was ascertained using the Turkeys test . A P value urine volume in DM and all therapy groups increased. Blood glucose Aurine volume> KHI in DM higher than normal control group,there was significant differences(p0. 05).After type 2 diabetic animal model was established,umalb hasn't been seen obviously,there is no significant difference in each group, but it was significantly increased in non —control groups at 8 weeks after induction of diabetes,post —treated inl. 8/^oPSS and DM+L group decrease UAER significantly (p0.05>.The localized thickeness of glomerulas basement membranes were seen in diabetic rats at 8 weeks of the course at electromicroscope,endo-thelial cell vacuolar degeneration. GBM was significantly increased in diabetic groups (nondiabeticvs. diabetic mice, 303. 26 + 17. 75vs. 507. 40 + 39. 42,P<0. 01;diabeticvs. DM + PSS, 507.40 +39.42 vs. 422. 58 +31. 18,P<0. 01).ConclusionIn conclusion,PSS can decrease urinary microalbumin excretion rate and thickening of GBM,depress the expression of TGF —^and collagen IV in renal cortex. PSS probably has some protecting effect on diabetic kidney ,delays the onset and progression of diabetic nephropathy, at least in part, through suppression of glomerular expression of TGF—(3iand decreasing the synthesis of collagen IV.
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