GSK3β抑制剂对糖尿病肾病大鼠肾组织TGF-β1和血清PAI-1表达的影响

摘要

目的 观察糖原合成酶激酶-3β(GSK3β)抑制剂Licl对糖尿病肾病(DN)大鼠的影响,从而明确GSK-3β抑制剂对DN的作用机制.方法 将60只雄性Wistar大鼠随机分为四组:正常对照组(NC组,n＝10),Licl组(n＝10),DN组(n＝20),DN+Licl组(n＝20).通过一次性腹腔注射链脲佐菌素(STZ)联合长期高脂饮食,建立DN大鼠模型;通过腹腔注射GSK-3β抑制剂Licl进行药物干预.观察各组大鼠一般状态、体重、肾重/体重的变化;检测血糖、尿量、尿蛋白、血肌酐等指标;肾脏PAS染色观察各组大鼠肾脏病理改变;ELISA方法检测各组大鼠血清纤溶酶原激活物抑制物-1(PAI-1)的水平,IHC方法检测肾组织转化生长因子-β1(TGF-β1)的表达,进行组间比较.结果 与NC相比,DN组大鼠一般状态、体重、肾重/体重、血糖、血肌酐、尿量、尿蛋白、肾脏病理均出现了DN特有的变化特征,血清PAI-1明显增高,肾组织TGF-β1的表达明显上调.DN+Licl组较DN组大鼠存活率高,尿蛋白、血清PAI-1表达显著减少,肾脏病理较DN组明显减轻,而肾组织TGF-β1表达明显增加.结论 GSK-3β抑制剂Licl能够降低DN大鼠血清PAI-1水平、蛋白尿及死亡率.DN大鼠较正常大鼠肾组织TGF-β1表达明显上调,GSK-3β抑制剂Licl能进一步增加DN大鼠肾组织TGF-β1蛋白的表达.抑制GSK-3β,对治疗DN有积极的意义.但是GSK-3β抑制剂Licl作为药物治疗DN,应考虑其效应的两面性.%Objective The purpose of this research is to observe the effects of Glycogen synthase kinase-3 betarn(GSK 3-Beta) inhibitor Licl on rats with diabetic nephropathy(DN) in order to determine the effect of GSK-3β inhibitorrnon DN and its mechanism. Methods 60 male Wistar rats were randomly divided into four groups:Normal control grouprn(NC group,n=10),Licl group(n=10),DN group(n=20),DN+Licl group(n=20). The rat model of diabetic nernphropathy was established by single intraperitoneal injection of STZ and feeding on high fat diet. Drug intervention wasrnimplemented by intraperitoneal injection of GSk-3β inhibitor Licl. The changes in general state, weight, kidney weight/rnbody weight of rats in each group were observed; Indexes such as blood glucose, urine volume, urinary protein, serumrncreatinine were detected;Renal pathological changes of rats in each group were observed by PAS staining method. Serrnum PAI-1 of rats in each group was examined by enzyme-linked immunosorbent assay (ELISA) ,and expression of renalrnTGF-β1 was examined by immunohistoehemisty(IHC), and then inter-group comparisons were carried out. ResultsrnCompared with normal control group,general state, weight, kidney weight / body weight, blood glucose, serum creatirnnine, urine volume, urinary protein, renal pathology of rats in DN group appeared specific features of DN, serum PAI-1rnincreased significantly,expression of renal TGF-β1 was up regulated significantly. Compared with DN group, survivalrnrate of rats in DN+Licl group was higher, urinary protein and Serum PAI-1 decreased significantly, renal pathology wasrnalleviated notably,however,expression of renal TGF-β1 increased sharply. Conclusion GSK-3β inhibitor Licl can reducernserum PAI-1, proteinuria and mortality rate of rats with diabetic nephropathy. Compared with normal rats, expression ofrnrenal TGF-β1 of rats with diabetic nephropathy was significantly up regulated. It can be luther increased by GSK-3β inrnhibitor Licl. Inhibition of GSK-3β have a positive significance in the treatment of diabetic nephropathy. However, the two sides of effects of GSK-3β inhibitor Licl should be considered when used as drugs to treat diabetic nephropathy.