舒洛地特对糖尿病大鼠肾脏病变的保护作用与机制探讨

Sulodexide prevents diabetic nephropathy through inhibiting renal NF-κB activation and MCP-1 expression

目的 观察舒洛地特对糖尿病大鼠肾脏组织核因子NF-κB活性及单核细胞趋化蛋白1（MCP-1）表达的影响,探讨舒洛地特对糖尿病肾病的保护机制.方法 高脂高糖喂养联合小剂量链脲佐菌素腹腔注射建立Wistar大鼠糖尿病模型,并按随机数字表法分为非治疗组（DM）及舒洛地特治疗组（ DMS）.非糖尿病大鼠作为正常对照组（NC）.12周后杀检,测定血糖、血肌酐、尿素氮、三酰甘油、胆固醇；免疫比浊法测定24h尿白蛋白；光镜下观察肾小球形态和结构,计算平均肾小球体积；免疫组化法检测肾组织MCP-1表达；Western印迹方法测定NF-κB活性.结果 与NC组比较,DM组及DMS组血糖、三酰甘油、胆固醇均显著升高（均P＜ 0.01）；DMS组与DM组比较,以上指标差异无统计学意义.与NC组相比,DM组及DMS组血肌酐、尿素氮、24 h尿白蛋白显著升高（均P＜0.01）.与DM组比较,DMS组血肌酐[（39.1±0.88） μmol/L比（41.0±2.16） μmol/L,P＜0.05]、尿素氮[（9.12±1.06） mmol/L比（9.87±0.19） mmol/L,P＜0.05]、24h尿白蛋白[（19.92±0.96） mg/24 h比（25.99±0.52） mg/24 h,P＜ 0.01]均显著降低.与NC组比较,DM组平均肾小球体积显著增加[（7.47±1.11）×105 μm3比（4.22±1.09）×105μm3,P＜ 0.01]；DMS组平均肾小球体积[（6.64±0.71 ）×105 μm3,P＜0.05]较DM显著降低,但仍显著高于对照组（P＜0.01）.与NC组相比,DM组肾组织MCP-1表达显著升高[（ 12.17±1.94 ）/HPF比（1.19±0.70）/HPF,P＜0.01]；与DM组比较,DMS组肾组织MCP-1表达[（9.22±1.61 ）/HPF,P＜0.01]显著降低,但仍高于NC组（P＜0.01）.与NC组相比,DM组肾组织NF-κB活性显著升高[（0.89±0.07）比（0.24±0.03）,P＜0.01]；与DM比较,DMS组肾组织NF-κB活性[（0.27±0.01）,P＜0.01]显著降低,与NC组比较,差异无统计学意义.结论 舒洛地特对糖尿病肾病具有防治作用,抑制NF-κB活性及MCP-1表达可能是其作用机制之一.

Objective To study the effects of sulodexide on renal NF-κB activation and monocyte chemotactic protein 1 （MCP-1） expression in diabetic rats and elucidate the possible mechanism of sulodexide in preventing diabetic nephropathy （DN）. Methods Wistar rats were fed with high-sucrose-high-fat diet and injected with a low dose of STZ （streptozotocin,35 mg/kg） into abdominal cavity to induce diabetes.DM rats were randomly divided into non-treated group of treatment,blood glucose （BG）,triglyceride （TG）,cholesterol,serum creatinine （Scr）,urea nitrogen （BUN）,24 h urinary albumin excretion （UAE） were measured.HE staining was performed in renal tissues for light microscopy examination of mean glomerular volume （MGV）.MCP-1 expression was detected by immunohistochemical method.NF-κB activation was determined by Western blot. Results Compared with NC group,DM group and DMS group had significant elevated BG,TG and TC levels （all P〈0.01）.There were no significant differences of BG,TG or TC levels between DM group and DMS group.Compared with NC group,DM group and DMS group had significant increased Scr,BUN,UAE levels （all P〈0.01）.Scr,BUN,UAE levels were significantly lower in DMS group than those in DM group [（39.1±0.88） μmol/L vs （41.0±2.16） μmol/L,（9.12±1.06） mmol/L vs （9.87±0.19） mmol/L; （19.92±0.96） mg/24 h vs （25.99±0.52）mg/24 h,all P〈0.05].Compared with NC group,the MGV of DM group was significantly increased [（7.47±1.11）×105 μm3 vs （4.22±1.09）×105 μm3,P〈0.01].Compared with DM group,the MGA of DMS group was significantly reduced [（6.64±0.71）×105 μm3 vs （7.47±1.11）×105 μm3,P〈0.05],but was still increased compared with that of NC group （P〈0.01）.Compared with NC group,the MCP-1 expression of DM group was significantly higher [（12.17±1.94）/HPF vs （1.19±0.70）/HPF,P〈0.01].MCP-1 expression in DSM group was significantly lower than that of DM group [（9.22± 1.61）/HPF vs （12.17±1.94）/HPF,P〈0.01],but still higher than that of control group （P〈0.01）.Compared with NC group,the NF-κB activity was significantly higher in DM group [（0.89±0.07） vs （0.24±0.03）,P〈0.01].Compared with DM group,NF-κB activity of DMS group was significantly lower [（0.27±0.01） vs （0.89±0.07）,P〈0.01].There was no significant difference of NF-κB activity between DMS group and NC group. Conclusion Sulodexide has protective effects on diabetic nephropathy,and one of the mechanisms may involve the inhibition of NF-κB activation as well as the suppression of MCP-I expression.