【作者】 马婵娟；

【导师】 孙子林；

【作者基本信息】 东南大学 ， 内科学， 2006， 硕士

【摘要】 目的:观察糖基化终产物(advanced glycation end products,AGEs)对人肾系膜细胞(human renal mesangial cells,HRMCs)galectin-3表达和分泌的影响以及罗格列酮的干预作用。方法:1、运用体外制备的糖基化修饰的牛血清白蛋白(AGE-bovine serum albumin,AGE-BSA)干预HRMCs,并进一步用罗格列酮与AGE-BSA共同干预HRMCs;2、用逆转录聚合酶链式反应检测galectin-3 mRNA表达,蛋白免疫印迹法检测galectin-3蛋白表达,收集细胞上清浓缩后用蛋白免疫印迹法检测上清中galectin-3的含量。结果:1、与DMEM及BSA组相比,AGE-BSA可使HRMCs的galectin-3 mRNA和蛋白表达及细胞上清中galectin-3的含量增加(P<0.05),其表达量及上清中的含量随着AGE-BSA浓度的增加和干预时间的延长而增加(P<0.05);2、与DMEM组及单独AGE-BSA组相比,罗格列酮与AGE-BSA共同干预HRMCs,细胞的galectin-3 mRNA和蛋白表达及细胞上清中galectin-3的含量增加(P<0.05),且表达量及上清中的含量随着罗格列酮浓度的升高而增加(P<0.05);而单独罗格列酮干预HRMCs,galectin-3 mRNA和蛋白表达及细胞上清中galectin-3的含量与DMEM组相比无差异(P>0.05)。结论:1、AGEs可以浓度和时间依赖的方式增加HRMCs galectin-3的表达和分泌。Galectin-3可能通过增加AGEs的清除在糖尿病肾病的发生中起保护作用。2、罗格列酮与AGEs共同干预HRMCs,可进一步增加HRMCs galectin-3的表达和分泌,推测罗格列酮增加细胞galectin-3的表达和分泌可能是其发挥肾保护作用的机制之一。更多还原

【Abstract】 Objectives:To investigate the effects of advanced glycation end products(AGEs)and rosiglitazone on the expression and secretion of galectin-3 in cultured human renal mesangial cells (HRMCs).Methods:1. HRMCs were incubated with different concentrations of AGE-bovine serum albumin (AGE-BSA) for different time, and exposed to AGE-BSA (200 mg·l-1) in presence of different concentrations of rosiglitazone. 2. The mRNA and protein expression of galectin-3 in HRMCs were analyzed by use of RT-PCR and Westernblot respectively. 3. The supernatant of HRMCs was collected and concentrated. Then the content of galectin-3 in the supernatant was detected by Westernblot.Results:1. Both RT-PCR and Westernblot revealed that AGE-BSA upregulated the expression of galectin-3 in HRMCs in a concentration- and time- dependent manner compared with DMEM and BSA (P<0.05). 2. Compared with DMEM and BSA, AGE-BSA elevated the content of galectin-3 in the supernatant of HRMCs (P<0.05) time- and concentration- dependently. 3. Exposure of HRMCs to different concentrations of rosiglitazone in presence of AGE-BSA but not to rosiglitazone alone and, to a lesser extent, AGE-BSA alone, increased both protein and mRNA expression of galectin-3, and its secretion in the medium (P<0.05). Rosiglitazone in presence of AGE-BSA increased the expression and secretion of galectin-3 dose-dependently (P<0.05).Conclusions:1. AGEs upregulates the expression and secretion of galectin-3 in HRMCs. These results indicate that galectin-3 might serve as a protective mechanism toward AGE-induced injury by favoring the removal of AGEs. 2. Rosiglitazone further upregulates the expression and secretion of galectin-3 in HRMCs. The data suggests rosiglitazone plays their role of reno-protection via upregulation of galectin-3.更多还原