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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 observe the effect of advanced glycated protein on synthesis and secretion of endothelin (ET) in mesangial cells. Methods Normal mesangial cells were cultured in vitro, and the matrix secreted by mesangial cells was incubated with glucose 6 phosphate to prepare advanced glycated matrix protein. Then the effect of endogenous glycated end products on ET 1 and endothelin A receptor ET AR) mRNA expression (using RT PCR), and changes in endothelin concentrations in the cell supernatant [measured by radioimmunoassay (RIA)]. At the same time, observe the effect of high glucose on the expression of endothelin in mesangial cells. Results Compared with the control, the mesangial cells treated with advanced glycosylated protein had significantly increased mRNA expression of ET 1 and ET AR, which were 1.74 and 1.47 times of that of the control group, respectively. Addition of aminoguanidine (AG) inhibited mesangial cell glycosylation and crosslinking, mesangial cells ET 1 and ET AR mRNA expression decreased, ET 1mRNA expression decreased significantly, about 53%, ET ARmRNA expression decreased only 18 %. The secretion of endothelin by mesangial cells was also significantly increased by the effect of glycosylation matrix, which was 7.15 times that of control group. After aminoguanidine intervention, the secretion of endothelin significantly decreased, and the concentration of endothelin decreased by 73.49%. The mesangial cells treated with high glucose had the most obvious hypertonic effect with high glucose, the ET 1 mRNA level in cells was 10.1 times that of the control group. Conclusion In high glucose, the extracellular matrix that is converted to final glycation end products has a direct up-regulation of the synthesis and secretion of endothelin by mesangial cells, and does not depend on the high glucose environment.