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Basic fibroblast growth factor (bFGF) requires heparan sulfate (HS) that contains bFGF-binding domains for regulating gllomerular cell apoptosis (AP)

碱性成纤维细胞生长因子 (bFGF) 需要含有 bFGF 结合域的硫酸乙酰肝素 (HS) 来调节肾小球细胞凋亡 (AP)

基本信息

批准号：
07671247
负责人：
SHINZATO Toru
金额：
$ 1.28万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671247/
关键词：
heparansulfate apoptosis mesangial cells basic fibroblast growth factor ヘパリン

项目摘要

Basic FGF can function as a survival factor in certain cell lineages. It requires a cofactor, HS or heparin (HP), for the activation of signal-transducing bFGF receptors. HS/HP are heterogeneous carbohydrates. We hypothesized that specific interaction between bFGF and bFGF-binding domains of HS might optimize bFGF's ability of regulating AP induced in glomerular cells. To test this hypothesis, we compared the effects of various forms of glycosaminoglycans (GAG). Two types of Engelbreth-Holm-Swarm tumor HS with respect to the affinity for bFGF were fractionated : the bound and unbound HS.The bound HS were enriched in IdoA(2SO4)-GlcNSO3 units. In order to trigger AP,a) isolated glomeruli from Wistar rats were incubated with Trimeresurus flavoviridis vebom, or b) rat mesangial cells in culture were deprived growth factors. The bound HS,te unbound HS,or either one of the other forms of GAG (HP,hyaluronate, chondroitin sulfate, dermatan sulfate and keratan sulfate) was added to the culture media at various concentrations (4ng/ml-40mug/ml) with bFGF (0.5-5.0ng/ml) in the presence or absence of cycloheximide (250muM). To detect DNA cleavage and subsequent nuclear changes typical of AP,DNA electrophoresis, light microscopic examination and TdT-mediated dUTP-biotin nick and labeling (TUNEL) were conducted. AP of the isolated glomeruli and cultured mesangial cells were successfully introduced. To rescue AP,5.0ng/ml of bFGF required HS or HS (5.0mug/ml or more as uronate) that contains bFGF-binding portions. Cycloheximide treatment inhibited this activity of bFGF.Other GAG did not give such regulatory effects at the same concentration.Conclusion : The results suggest that the bFGF-binding portions help regulate AP.These studies may contribute toward establishing a framework for the molecular design of carbohydrates capable of optimizing bFGF activity.

碱性成纤维细胞生长因子在某些细胞系中可以作为一种生存因子。它需要辅因子HS或肝素（HP）来激活信号转导bFGF受体。HS/HP是异质碳水化合物。我们推测bFGF和bFGF结合域之间的特异性相互作用可能优化bFGF调节肾小球细胞中诱导的AP的能力。为了验证这一假设，我们比较了各种形式的糖胺聚糖（GAG）的影响。Engelbreth-Holm-Swarm肿瘤HS对bFGF的亲和力分为结合型和非结合型两类，结合型HS富含IdoA（2SO 4）-GlcNSO 3单位。为了触发AP，a）从Wistar大鼠分离的肾小球与竹叶青蛇（Trimeresurus flavoviridis vebom）孵育，或B）培养的大鼠肾小球系膜细胞被剥夺生长因子。在存在或不存在放线菌酮（250 μ M）的情况下，将结合的HS、未结合的HS或其它形式的GAG（HP、透明质酸盐、硫酸软骨素、硫酸皮肤素和硫酸角质素）中的任一种以各种浓度（4 ng/ml-40 μ g/ml）与bFGF（0.5-5.0ng/ml）一起加入到培养基中。为了检测AP的典型DNA裂解和随后的核变化，进行了DNA电泳、光学显微镜检查和TdT介导的dUTP-生物素缺口标记（TUNEL）。成功地将AP导入分离的肾小球和培养的系膜细胞。为了挽救AP，5.0ng/ml的bFGF需要含有bFGF结合部分的HS或HS（5.0ug/ml或更高，以糖醛酸计）。放线菌酮处理抑制这种活性的bFGF。其他GAG没有在相同concentration.Conclusion：结果表明，bFGF结合部分有助于调节AP。这些研究可能有助于建立一个框架，能够优化bFGF活性的碳水化合物的分子设计。