培养的人脐静脉内皮细胞合成的蛋白聚糖

以~(35)S]-Na_2SO_4为示踪物,观察培养的人脐静脉内皮细胞(EC)合成及分泌的蛋白聚糖(PG),经DEAE-Sephacel离子交换及Sepharose6B凝胶滤柱层析分析发现细胞层及培养液均含有三种PG单体,即硫酸乙酰肝素蛋白聚糖(HS-PG)、硫酸软骨素蛋白聚糖(CS-PG)及硫酸皮肤素蛋白聚糖(DS-PG)HS-PG又可分为大小两种,前者(HS-PG_L)位于V_o处,后者(HS-PG_s)Kd=0.53(sepharose6B);CS-PG/DS-PG分为三个峰,峰Ⅰ位于V_0处,峰Ⅱ、峰Ⅲ的Kd值分别为0.26及0.52(sepharose6B)。汇合前后细胞层及培养液中各种PG的含量不同。细胞层PG总量汇合前低于汇合后,无论是细胞层还是培养液汇合前HS-PG_L均低于汇合后,HS-PG_L与HS-PG_s比值亦为汇合前低于汇合后,而CS-PG/DS-PG含量则高于汇合后。汇合前后EC合成及分泌PG的差异与文献报道的EC损伤及正常者类似。

PROTEOGLYCANS SYNTHESIZED BY CULTURED HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS.

Proteoglycans (PGs) synthesized and secreted by cultured human umbilical vein endothelial cells (EC) were studied with (35S)-Na2SO4 as labelling precursors.PGs in the medium and the cell layer were fractionated by DEAE-Sephacel ion exchange and Sepharose 6B gel chromatography.Three species of PG monomers,heparin sul-fate-PG (HS-PG), chondroitin sulfate-PG (CS-PG) and dermatan sulfate-PG(DS-PG),were identified. HS-PG in the cell layer and the medium could be further fractionated into a larger excluded peak (HS-PGL)and a smaller included one(HS-PGs, Kd = 0.53),and CS-PG/DS-PG into three peaks one at Vo the others with Kd = 0.26 and 0.53, respectively. The relative percentages of HS-PG (especially HS-PGL) in both the cell and the medium of nonconfluent cultures were lower than those of confluent cultures, while CS-PG/DS-PG were just the reverse, i.e.higher in nonconfluent cultures. In the synthesis of PGs, nonconfluent EC shared certain characteristics with injured EC and confluent EC with normal EC.