Buddecke E, Schmidt A, Tschoerner P
Institut für Physiologische Chemie und Pathobiochemie, Universität Münster.
Klin Monbl Augenheilkd. 1993 Mar;202(3):167-73. doi: 10.1055/s-2008-1045578.
Cultured bovine corneal endothelial cells (CEC) synthesize heparan sulfate and dermatan sulfate containing proteoglycans and distribute them between different compartments.
[35S]sulfate labelled proteoglycans are found associated with the cell layer, secreted into the culture medium and deposited into the underlaying extracellular matrix. In the presence of basic fibroblast growth factor (bFGF)-a strong mitogen for CEC-subconfluent cells incorporate [35S]sulfate into the sulfated proteoglycans at a rate three times higher as compared with the proteoglycans of CEC in the absence of bFGF. The enhanced proteoglycan synthesis is accompanied with a shift in the proteoglycan distribution pattern. While in control cells the cell-associated heparan sulfate accounts for about 30% of the total glycosaminoglycans under the influence of bFGF the HS percentage increases to approximately 60%.
CEC synthesize and deposit endogenous bFGF into the extracellular matrix. Heparitinase treatment of the extracellular matrix releases bFGF activity which is able to stimulate the 35S incorporation into proteoglycans in a comparable manner as exogenous bFGF but does not influence the proteoglycan distribution pattern. Pretreatment of the matrix-bound bFGF activity with polyclonal antibodies against bFGF abolishes its stimulating activity.
培养的牛角膜内皮细胞(CEC)合成含硫酸乙酰肝素和硫酸皮肤素的蛋白聚糖,并将它们分布于不同的区室。
发现[35S]硫酸盐标记的蛋白聚糖与细胞层相关,分泌到培养基中并沉积到下层的细胞外基质中。在碱性成纤维细胞生长因子(bFGF,一种对CEC有强促有丝分裂作用的因子)存在的情况下,亚汇合细胞将[35S]硫酸盐掺入硫酸化蛋白聚糖的速率比不存在bFGF时CEC的蛋白聚糖掺入速率高3倍。蛋白聚糖合成增强伴随着蛋白聚糖分布模式的改变。在对照细胞中,与细胞相关的硫酸乙酰肝素约占总糖胺聚糖的30%,而在bFGF的影响下,硫酸乙酰肝素的百分比增加到约60%。
CEC合成内源性bFGF并将其沉积到细胞外基质中。用肝素酶处理细胞外基质可释放bFGF活性,该活性能够以与外源性bFGF相当的方式刺激[35S]掺入蛋白聚糖,但不影响蛋白聚糖的分布模式。用针对bFGF的多克隆抗体预处理与基质结合的bFGF活性可消除其刺激活性。