Cechowska-Pasko Marzanna, Surazyński Arkadiusz, Bańkowski Edward
Department of Pharmaceutical Biochemistry, Medical University of Białystok, Mickiewicza 2A, 15-089 Białystok, Poland.
Mol Cell Biochem. 2009 Jul;327(1-2):211-8. doi: 10.1007/s11010-009-0059-8. Epub 2009 Feb 19.
It was decided to study the effect of glucose deprivation on collagen synthesis and degradation in fibroblast cultures and a correlation of these processes with the expression of oxygen/glucose regulated proteins (ORP150/GRP170). The incorporation of radiolabeled proline into collagenase-sensitive and hydroxyproline-containing proteins was used as an index of collagen synthesis, whereas pulse-chase technique was employed to evaluate the degradation of newly synthesised proteins. We demonstrated that fibroblasts incubated in high-glucose medium synthesised detectable amounts of collagenous proteins. Most of them were secreted into the culture medium. The shortage of glucose resulted in about 30% reduction in synthesis of collagenous proteins, both those secreted into culture medium and remaining in the cell layer. The pulse-chase experiments demonstrated that the reduced amount of newly synthesised collagen was protected against intracellular degradation. Proportionally less collagen was degraded in cultures incubated in low-glucose than in high-glucose media. These phenomena were accompanied by an increase in the expression of chaperon-ORP150 in cultures growing in low-glucose medium. We suggest that the increased expression of ORP150 is a factor which protects collagen against intracellular degradation induced by glucose deprivation.
决定研究葡萄糖剥夺对成纤维细胞培养物中胶原蛋白合成与降解的影响,以及这些过程与氧/葡萄糖调节蛋白(ORP150/GRP170)表达的相关性。将放射性标记的脯氨酸掺入胶原酶敏感且含羟脯氨酸的蛋白质中,以此作为胶原蛋白合成的指标,而采用脉冲追踪技术评估新合成蛋白质的降解。我们证明,在高糖培养基中孵育的成纤维细胞合成了可检测量的胶原蛋白。其中大部分分泌到培养基中。葡萄糖短缺导致胶原蛋白合成减少约30%,无论是分泌到培养基中的还是留在细胞层中的。脉冲追踪实验表明,新合成的胶原蛋白量减少,但免受细胞内降解。在低糖培养基中孵育的培养物中,胶原蛋白降解的比例比在高糖培养基中少。这些现象伴随着低糖培养基中生长的培养物中伴侣蛋白ORP150表达的增加。我们认为,ORP150表达的增加是保护胶原蛋白免受葡萄糖剥夺诱导的细胞内降解的一个因素。