晚期糖化蛋白刺激系膜细胞合成和分泌内皮素的研究  

Advanced glycation of mesangial matrix induces endothelins secretion and gene expression in mesangial cells

作　　者：黄宇峰[1] 林善锬[1] 周江华[1] 顾勇[1] 

机构地区：[1]上海医科大学华山医院肾脏病科,200040

出　　处：《中华肾脏病杂志》1999年第5期273-277,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金!No.3 9470 3 3 5

摘　　要：目的　观察晚期糖化蛋白对系膜细胞合成和分泌内皮素 (ET)的影响。方法　体外正常培养系膜细胞 ,将系膜细胞分泌的基质与 6 磷酸葡萄糖孵育 ,快速制备晚期糖化基质蛋白 ;进而检测该糖化终产物作用后 ,系膜细胞中ET 1和内皮素A受体 (ET AR)mRNA表达的改变 (采用RT PCR) ,以及细胞上清液中内皮素浓度的变化 [用放射免疫法 (放免法 )测定 ]。同时 ,观察高糖对系膜细胞中内皮素表达产生的效应。结果　经晚期糖化蛋白作用的系膜细胞 ,与对照相比 ,其对ET 1和ET AR的mRNA表达明显增加 ,分别为对照组的 1.74倍和 1.47倍。加入氨基胍 (AG)抑制系膜基质糖化和交联后 ,系膜细胞中ET 1和ET AR的mRNA表达随之下降 ,以ET 1mRNA表达下降明显 ,约 5 3% ,ET ARmRNA表达仅下降 18%。系膜细胞对内皮素的分泌亦受糖化基质的作用而显著增加 ,为对照组的 7.15倍 ,氨基胍干预后 ,其分泌明显减少 ,内皮素浓度下降 73.49%。用高糖干预的系膜细胞组 ,却以高糖产生的高渗效应最为明显 ,细胞中ET 1mRNA水平为对照组的 10 .1倍。结论　高糖状态下 ,转变为糖化终产物的细胞外基质对系膜细胞合成和分泌内皮素有直接的上调作用 ,且不依赖于高糖环境。Objective To determine the contribution of advanced glycation end products information to endothelins expression in kidney in diabetes. Methods Mesangial cells were grown in extracellular matrix glycated and crosslinked in vitro for up to 96 hours without passage. The model of in vitro advanced glycation of matrix, as occurs in diabetes, was build up by incubation of mesangial matrix with 200mmol/L glucose 6 phosphate (G6P) for two weeks. Endothelin 1 (ET 1) and its A receptor (ET AR) mRNA expressions were detected by quantitative RT PCR analyses. ET concentration in the media of cultured cells was measured by radioimmunoassay. Results Mesangial cells plated onto matrices glycated with G6P expressed higher levels of ET 1 and ET AR mRNA than did cells plated on control matrices( ET 1: GAPDH mRNA relative ratio was 1.74 and ET AR: GAPDH mRNA relative ratio was 1.47, where the ratio in cells cultured on control matrices were assigned arbitrary values of 1.0). Modification of matrix with G6P in the presence of aminoguanidine (AG) returned the excessive express of ET 1 mRNA towards control values and declined the elevated mRNA level of ET AR, the respective relative ratios of mRNA to that of control state were 1.21 for ET 1 and 1.29 for ET AR. In addition, the stimulated ET 1 mRNA expression induced by advanced glycation of mesangial matrix was accompanied by increases in the translated protein [ vs control: (5 73±0 81)vs (0 65±0 04)pg/ml, P<0 001, by RIA]. AG also largely prevented the glycated matrix induced increase in ET secretion [vs G6P modified matrices:(1 41±0.06)vs(5 73±0 81)pg/ml,P<0.001, by RIA]. However, the effects of nonenzymatic glycation and crosslinking of matrix on the subsequent generation of ET by cultured mesangial cells were different from those of elevated glucose concentration.Conclusion Advanced glycation of mesangial matrix induces directly ET 1 and ET AR mRNA synthesis and ET secretion by mesangial cells.

关 键 词：内皮素 非酶糖化 系膜基质 系膜细胞 

分 类 号：R692.02[医药卫生—泌尿科学]