川陈皮素对糖尿病肾病大鼠的作用研究

Effect of Nobiletin on Diabetic Nephropathy Rats

目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。

Objective To establish a rat model of diabetic nephropathy,observe the therapeutic effect of Nobiletin on them.Meanwhile to study the protein and mRNA expression of serpin family B member 7(Megsin),platelet-derived growth factor-BB(PDGF-BB),extracellular signal regulated protein kinase(ERK1/2)and type IV collagen(collagen IV).Methods A total of 120 clean male Sprague-Dawley rats were divided into the normal group,diabetic nephropathy model group,benazepril group,Nobiletin low,medium and high dose groups.The diabetic nephropathy model was built by high-fat diet+intraperitoneal injection of streptozotocin(STZ).After the model successfully constructed,the corresponding drug was given for 6 weeks.6 weeks later,the urine,blood and kidney tissue samples were collected to detect the blood glucose,creatinine andβ2-MG in urine and serum,KW/BW,levels of IL-1,IL-6 and TNF-α,the pathology of renal tissue,the proteins and mRNA expression of Megsin,PDGF-BB,pERK1/2 and collagen IV in renal tissues.Results HE staining showed that the slices in the normal group had normal morphology,and those in the model group showed obvious glomerular atrophy and sclerosis.Compared with the model group,the slices in the benazepril group and the Nobiletin low,medium and high dose groups were obvious improved.In addition,those in the benazepril group and the Nobiletin high dose group were basically the same.Compared with the normal group,the blood glucose,KW/BW,UREA andβ2-MG in urine and serum,levels of IL-1,IL-6 and TNF-α,renal tissue Megsin,PDGF-BB,pERK1/2 and collagen IV protein and mRNA in the model group were significantly increased(P<0.05).Compared with the model group,the Nobiletin low,medium and high dose groups’blood glucose,KW/BW,UURE andβ2-MG in urine and serum,levels of IL-1,IL-6 and TNF-α,and the Megsin,PDGF-BB,pERK1/2 and collagen IV protein and mRNA in the benazepril group were decreased(P<0.05).Moreover,the blood glucose in the three Nobiletin groups was significantly lower than that in the benazepril group(P<0.05).The levels of IL-1,IL-6 and TNF-αin the Nobiletin high dose group were significantly lower than those in the benazepril group(P<0.05),but there were no significant differences among three Nobiletin groups and the benazepril group for the KW/BW,urine and serum creatinine andβ2-MG,kidney Megsin,PDGF-BB,pERK1/2 and collagen IV proteins and mRNA expression.Conclusion Nobiletin has a significant therapeutic effect on diabetic nephropathy rats,which is related to improving the expressions of megsin,PDGF-BB,pERK1/2 and collagen IV in the kidney.