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[伴刀豆球蛋白A对静止软骨细胞培养物形态及DNA合成的影响]

[Effect of concanavalin A on morphology and DNA synthesis of resting chondrocyte cultures].

作者信息

Yan W Q, Yang T S, Hou L Z, Suzuki F, Kato Y

机构信息

Institute of Perclinical Sciences, Norman Bethune University of Medical Sciences, Changchun.

出版信息

Shi Yan Sheng Wu Xue Bao. 1994 Sep;27(3):300-5.

PMID:7801723
Abstract

Rabbit chondrocytes were isolated from resting cartilage of ribs of 4-week-old New Zealand rabbits. Chondrocytes were seeded at low density and grown to confluency in medium (DME) with 10% fetal bovine serum, 50 micrograms/ml ascorbic acid, and antibiotics, at 37 degrees C under 5% CO2 in air, and then serum concentration was reduced to 0.3%. At the low serum concentration, chondrocytes adopted fibroblastic morphology. Addition of concanavalin A to the culture medium induced a morphologic alteration of the fibroblastic cells to polygonal or spherical chondrocytes that were surrounded by refractile matrix. Wheat germ agglutinin and garden pea lectin induced similar cell shape changes without any increase in matrix synthesis. Concanavalin A decreased [3H]thymidine incorporation into DNA in a dose-dependent manner with an ED50 of 0.4-1 micrograms/ml in the presence of various concentrations of serum ranging from 0.3 to 20%. The concanavalin A inhibition of DNA synthesis was abolished by 10 mmol/L methyl-D-mannopyranoside. The inhibition of chondrocyte proliferation does not seem to be specific to concanavalin A. Wheat germ agglutinin, lentil lectin, phytohemagglutinin, Ulex europeaus agglutinin, and garden pea lectin also decreased, dose-dependently, [3H]thymidine incorporation into DNA in chondrocytes. Other lectins at 0.01-20 micrograms/ml had little effect on [3H] thymidine incorporation. Because the molecular structure of concanavalin A and its mode of actions have been extensively characterized, chondrocytes exposed to this lectin will be useful as a novel model in studying of the control of cellular differentiation.

摘要

兔软骨细胞从4周龄新西兰兔肋骨的静止软骨中分离得到。将软骨细胞以低密度接种,在含有10%胎牛血清、50微克/毫升抗坏血酸和抗生素的培养基(DME)中,于37℃、5%二氧化碳的空气中培养至汇合,然后将血清浓度降至0.3%。在低血清浓度下,软骨细胞呈现成纤维细胞形态。向培养基中添加伴刀豆球蛋白A可诱导成纤维细胞形态改变为多边形或球形软骨细胞,周围有折光性基质。小麦胚凝集素和豌豆凝集素诱导类似的细胞形态变化,但基质合成没有增加。伴刀豆球蛋白A以剂量依赖方式降低[3H]胸腺嘧啶核苷掺入DNA,在0.3%至20%的各种血清浓度存在下,其半数有效剂量(ED50)为0.4 - 1微克/毫升。10毫摩尔/升甲基 - D - 甘露吡喃糖苷可消除伴刀豆球蛋白A对DNA合成的抑制作用。软骨细胞增殖的抑制似乎并非伴刀豆球蛋白A所特有。小麦胚凝集素、扁豆凝集素、植物血凝素、欧洲荆豆凝集素和豌豆凝集素也均以剂量依赖方式降低软骨细胞中[3H]胸腺嘧啶核苷掺入DNA。其他浓度为0.01 - 20微克/毫升的凝集素对[3H]胸腺嘧啶核苷掺入影响很小。由于伴刀豆球蛋白A的分子结构及其作用方式已得到广泛研究,接触该凝集素的软骨细胞将作为研究细胞分化调控的新型模型具有重要作用。

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