D-半乳糖诱导大鼠肾脏损害的糖基化机制及药物干预作用  

Glycation Mechanism of D-Galactose-Induced Kidney Injury in Rats and Its Drug Intervention

作　　者：李冰[1] 郑浩[1] 

机构地区：[1]北华大学医学部基础医学院机能实验室,吉林132013

出　　处：《中国实验动物学报》2008年第6期435-440,共6页Acta Laboratorium Animalis Scientia Sinica

摘　　要：目的D-半乳糖(D-galactose)诱导大鼠体内不同糖基化水平,研究其肾脏损伤发生的机理及药物对其干预作用。方法采用不同剂量D-半乳糖[150、75、37.5 mg/(kg.d)]分别腹腔注射(ip)处理大鼠8周,诱导糖基化状态和肾脏损伤,同时D-半乳糖高剂量[150 mg/(kg.d)]分别给与氨基胍[150 mg/(kg.d)]和维生素E[150 mg/(kg.d)]处理8周。采用葡萄糖氧化酶法测定大鼠血糖,硫代巴比妥酸(TBA)比色法测定糖化血红蛋白,硝基四氮唑蓝(NBT)比色法测定血清果糖胺;按文献方法分别测定血红细胞醛糖还原酶活性和晚期糖基化终末产物(AGEs)含量及肾脏组织中AGEs含量,羟胺法和比色法分别测定SOD和GSH-Px活性,硫代巴比妥酸法测定MDA含量;采用CBB法测定尿蛋白含量,二乙酰-肟法测定血尿素氮,苦味酸法测定血肌酐;流式细胞仪检测肾脏细胞凋亡情况。结果D-半乳糖高、中剂量处理8周后,大鼠2 h血糖明显升高,血红细胞醛糖还原酶活性升高,糖化产物形成增多(P<0.01,P<0.05);肾组织中AGEs含量明显升高,SOD及GSH-Px活性下降,MDA含量升高(P<0.01,P<0.05),尿蛋白、血尿素氮和血肌酐量明显增加,肾脏细胞凋亡率明显增加(P<0.01,P<0.05)。而氨基胍和维生素E处理后,可明显抑制高剂量D-半乳糖引起的糖基化反应,减少上述物质的生成,并减轻对肾脏组织的损伤作用,尤其是氨基胍作用更为明显。结论D-半乳糖通过诱导体内蛋白糖基化和肾组织AGEs大量生成,降低抗氧化能力,诱致肾脏细胞凋亡;氨基胍和维生素E对D-半乳糖诱致的肾脏损伤作用具有保护作用。Objective To generate D-galactose-induced glycation at different levels in rats, to understand the role of advanced glycation end-products （AGEs） in the mechanism of kidney injury,and the action of its drug intervention. Methods The rats were administrated intraperitoneally with D-galactose in a dose of 150 mg/kg,75 mg/kg or 37.5 mg/kg,respectively,for 8 weeks to induce the glycation condition and kidney injury. At the same time, aminoguanidine （150 mg/kg） and vitamin E （150 mg/kg） were administrated with D-galactose （150 mg/kg） for 8 weeks. After treatment, fast blood glucose and 2 hours blood glucose were measured by glucose oxidase assay, the serum levels of HbA1C and fructosamine were detected with thiobarbituric acid （TBA） and nitrotetrazolium blue chloride （NBT） photoreduction method, respectively. According to reports in the literature, the activity of aldose reductase and content of AGEs in plasma and in kindey tissue were measured, and the activity of SOD and GSH-Px, the contents of MDA in kindey tissue were assayed by hydroxylamine method, chemical colorimetry and TBA method. 24 h urinary protein was detected by CBB technique, blood urea nitrogen （BUN） was detected by diacetyl oxime method and serum creatinine （Cr） was determined by trinitrophenol assay. The apoptosis in renal cells was detected by flow cytometry. Results After treatment with D-galactose for 8 weeks, the 2 hour glueose level, the activity of aldose reductase and the content of AGEs in blood were increased significantly （P 〈 0.01 or P 〈 0.05）. The levels of AGEs and MDA in renal tissue were also enhanced significantly （P 〈0.01 or P 〈 0.05）. However, the activities of SOD and GSH-Px decreased （P 〈 0.01 or P 〈 0.05）. In addition,the contents of 24 h urine protein, BUN and Cr, and the apoptotie rate of renal cells were significantly increased （P 〈 0.01 or P 〈 0.05）. Aminoguanidine and vitamin E inhibited glycosylation reaction and oxidative stress reaction in body and renal tis

关 键 词：D-半乳糖 糖基化 氧化应激 凋亡 肾脏损伤 

分 类 号：R587.1[医药卫生—内分泌]