摘要
目的探究自噬相关蛋白Beclin1、微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)在1型糖尿病肾病(DN)大鼠肾脏组织内的表达量及与肾损伤关系。方法选择健康8~12周龄SD大鼠20只,随机数字表法分成DN组和正常组,各10只。DN组采用链脲佐菌素诱导建立1型糖尿病肾病模型,正常组不做处理。采用全自动生化仪检测2组大鼠体重、血糖、24 h尿蛋白(24 h Upro)、血肌酐、尿素氮,免疫组化检测肾组织中LC3-Ⅱ、Beclin1、肌醇三酸激酶(PI3K)、蛋白激酶B(AKT)表达。另采用高糖刺激人近端肾小管上皮HK-2细胞,分析其对细胞中LC3-Ⅱ、Beclin1、PI3K、AKT蛋白表达的影响。结果DN组大鼠体重低于正常组,血糖、24 h Upro、血肌酐、尿素氮高于正常组,差异均有统计学意义(P<0.05)。病理染色结果显示,DN组大鼠肾小球体积明显变大,肾小管出现局灶性萎缩和细胞坏死,肾小球系膜基出现质增厚。DN组肾组织Beclin1、LC3-Ⅱ、PI3K、AKT蛋白表达积分高于正常组,差异均有统计学意义(P<0.05)。Pearson相关性分析显示,LC3-Ⅱ、Beclin1与24 h Upro、Scr、BUN呈正相关(P<0.05)。高糖组细胞Beclin1、LC3-Ⅱ、PI3K、AKT蛋白相对表达量高于对照组,差异均有统计学意义(P<0.05)。结论自噬相关蛋白Beclin1、LC3-Ⅱ在1型DN中表达上调,且与肾损伤存在一定相关性,而自噬通路激活可能是影响肾细胞损伤的重要因素。
Objective To explore the expression of autophagy-related protein Beclin1 and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in kidney tissues of rats with type 1 diabetic nephropathy(DN)and their relationship with kidney injury.Methods Twenty healthy SD rats aged 8-12 weeks were selected and divided into DN group and normal group according to the random number table method,with 10 rats in each group.The DN group was induced by streptozotocin to establish a type 1 diabetic nephropathy model,and the normal group had no treatment.The body weight,blood glucose,24 h Upro,blood creatinine(Scr),and urea nitrogen(BUN)of the two groups of rats were detected by an automatic biochemical analyzer.Immunohistochemistry was used to detect the expression of LC3-Ⅱ,Beclin1,phytate kinase(PI3K)and protein kinase B(AKT)in kidney tissue.In addition,high glucose was used to stimulate human proximal tubular epithelial HK-2 cells to analyze its effect on the expression of LC3-Ⅱ,Beclin1,PI3K,and AKT protein in the cells.Results The body weight of the rats in the DN group was lower than the normal group,and the blood glucose,24 h Upro,Scr,and BUN were higher than those in the normal group,the differences are statistically significant(P<0.05).The results of pathological staining showed that the glomerulus volume of the rats in the DN group was obviously enlarged,the renal tubules appeared focal atrophy and cell necrosis,and the mesangial base of the glomeruli was thickened.The protein expression scores of Beclin1,LC3-Ⅱ,PI3K and AKT in renal tissues in the DN group were higher than those in the normal group(P<0.05).Pearson correlation analysis showed that,LC3-Ⅱ,Beclin1 was positively correlated with 24 h Upro,Scr,BUN,the differences are statistically significant(P<0.05).The relative expression of Beclin1,LC3-Ⅱ,PI3K,AKT protein in the high glucose group was higher than that of the control group(P<0.05).Conclusion Autophagy-related proteins Beclin1 and LC3-Ⅱare up-regulated in type 1 DN and are related to renal injury.
作者
肖霞
颜雪梅
李慧
黄铄
XIAO Xia;YAN Xue-mei;LI Hui(Department of Internal Medicine,Hunan Maternal and Child Health Hospital,Changsha Hunan 410000,China;Department of Respiratory Medicine,Changsha First Hospital,Changsha Hunan 410000,China)
出处
《临床和实验医学杂志》
2021年第19期2032-2036,共5页
Journal of Clinical and Experimental Medicine
基金
湖南省科技计划项目(编号:2014SK3153)。
关键词
大鼠
自噬相关蛋白
1型糖尿病肾病
肾脏组织
相关性
Rats
Autophagy-related protein
Type 1 diabetic nephropathy
Kidney tissue
Correlation
