摘要
目的通过肾小球系膜细胞培养及2型糖尿病动物模型来研究糖尿病对葡萄糖转运蛋白(GLUT)1的影响。方法体外培养大鼠肾小球系膜细胞,分别给予高糖(HG)刺激12 h、24 h后提取RNA及蛋白。高脂饮食结合小剂量链脲佐菌素诱导2型糖尿病肾病(DN)大鼠模型,随机分成对照(Control)组、DN组,记录大鼠体重、血糖,留取尿液、血清、肾脏组织进行生化指标、12脂氧化酶(12-LO)、GLUT1测定及组织病理等检查。结果在体外,HG刺激下肾小球系膜细胞内12-LO转录及蛋白水平均升高,而GLUT1转录及表达在HG刺激12 h后升高,24 h后下降。在体内,PAS染色显示DN组肾小球体积明显增大,系膜基质增生,囊腔裂隙变窄,肾小管管腔狭窄,肾小管肿胀。与Control组相比,DN组肾组织内12-LO表达明显升高,GLUT1表达明显下降(P<0.05)。免疫组化示DN组肾小球内12-LO表达显著增加,GLUT1表达显著降低(P<0.05)。结论DN条件下不同阶段肾小球内12-LO和GLUT1表达不完全同步,提示12-LO和GLUT1在DN肾小球损伤过程中可能既有致病作用,又有保护性作用。
Objective To study the effect of diabetes on glucose transporter protein(GLUT)1 by renal mesangial cell culture and type 2 diabetes animal model.Methods Rats glomerular mesangial cells were cultured in vitro, RNA and protein were extracted after high glucose(HG) stimulation for 12 and 24 h, respectively. High-fat diet combined with low-dose streptozotocin was used to induce type 2 diabetic nephropathy(DN) rats model, the rats were randomly divided into control(Control) group and DN group, their body weight and blood glucose were recorded, urine, serum and kidney tissues were collected for biochemical indexes, 12 lipid oxidase(12-LO), GLUT1 determination and histopathological examination. Results In vitro, 12-LO level of transcript and protein level were elevated in mesangial cells under HG stimulation, while GLUT1 transcript and expression were increased after 12 h of HG stimulation and decreased after 24 h of HG stimulation. In vivo, PAS staining showed that the glomeruli of DN group were significantly enlarged, with hyperplasia of the mesangial matrix, narrowing of the cystic fissure, narrowing of the tubular lumen and swelling of the tubules. Compared with Control group, the expression of 12-LO in renal tissue of DN group was significantly increased and the expression of GLUT1 was significantly decreased(P<0.05).Immunohistochemistry showed the expression of 12-LO was significantly increased and the expression of GLUT1 was significantly decreased in glomeruli of DN group(P<0.05).Conclusions The expression of 12-LO and GLUT1 in glomeruli at different stages of DN is not completely synchronous, suggesting that 12-LO and GLUT1 may have both pathogenic and protective effects during the process of DN glomeruli injury.
作者
张丽
吴美延
许颖
许钟镐
ZHANG Li;WU Mei-Yan;XU Ying(Department of Nephrology,The First Hospital of Jilin University,Changchun 130061,Jilin,China)
出处
《中国老年学杂志》
CAS
北大核心
2022年第23期5749-5753,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(81974094)
吉林省自然科学基金(20200201362JC)。
