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Signal transmission of TGF-beta superfamily in chondrocyte

软骨细胞中TGF-β超家族的信号传递

基本信息

批准号：
08407049
负责人：
OKA Masanori
金额：
$ 3.52万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407049/
关键词：
TGF-beta Articular chondrocytes PKC Staurosporine NO Proteoglycan 骨固定軟骨細胞情報伝達形質

项目摘要

From a clinical aspect of joint reconstruction, we have investigated how to repair the joint surface. One of the methods to repair the joint is tissue engineering using cultured chondrocytes which has developed markedly.To repair the joint using cultured chondrocytes, important problem is character of chondrocyte, especially their phenotype as articuler chondrocytes. In other words, it is desirable that the used chondrocytes never ossifies. Considering these character of chondrocytes, we investigated signal transmission of TGF-beta superfamily in chondrocytes. In the present study we examined the effect of phorbol, 12-myristate, 13-acetate (PMA), a potent and specific protein kinase C(PKC) activator and Staurosporine, a potent nonselective PKC inhibitor, on the differentiation of sternal chondrocytes from 13-day-old chick embryo, and whether PKC functions as a mediator of their effects. Sulfated proteoglycan (PG) and type I and type II collagen were used as markers of differentiation. PKC expression was assayd by immunoblotting. PMA inhibited chondrogenesis dose-dependently (1-50 ng/ml), and its effects were possibly mediated by the translocation of PKC from the cytozol to the membrane. Staurosporine enhanced chondrogenesis (1-80nM) and caused the depletion of total PKC in a dose-dependent fashion. A synergistic stimulation of PG synthesis by N-monomethyl, L-arginine (NMA), a specific inhibitor of nitric oxide synthase (NOS), in response to Staurosporine (4 and 80 nM), with a corresponding suppression of NO synthesis, has been observed. The effect PMA on PG synthesis was not modulated by NMA.We suggest that NO may no be involved in the inhibition of chick sternal chondrocyte differentiation by PMA,but that Staurosporine may promote chondrogenic differentiation at least in part through a NO-dependent mechanism

我们从关节重建的临床角度，研究了如何修复关节面。修复关节的方法之一是利用已显着发育的培养软骨细胞进行组织工程。利用培养软骨细胞修复关节，重要的问题是软骨细胞的特性，特别是其作为关节软骨细胞的表型。换句话说，期望所使用的软骨细胞永远不会骨化。考虑到软骨细胞的这些特性，我们研究了软骨细胞中TGF-β超家族的信号传递。在本研究中，我们研究了佛波醇、12-肉豆蔻酸酯、13-乙酸酯 (PMA)（一种有效的特异性蛋白激酶 C(PKC) 激活剂）和星形孢菌素（一种有效的非选择性 PKC 抑制剂）对 13 日龄鸡胚胸骨软骨细胞分化的影响，以及 PKC 是否充当其影响的介质。硫酸化蛋白聚糖（PG）以及I型和II型胶原蛋白被用作分化标记物。通过免疫印迹法测定PKC表达。 PMA 剂量依赖性地抑制软骨形成（1-50 ng/ml），其作用可能是通过 PKC 从胞质到膜的易位介导的。星形孢菌素可增强软骨形成 (1-80nM) 并以剂量依赖性方式引起总 PKC 的消耗。已观察到 N-单甲基、L-精氨酸 (NMA)（一种一氧化氮合酶 (NOS) 的特异性抑制剂）对星形孢菌素（4 和 80 nM）的反应可协同刺激 PG 合成，并相应抑制 NO 合成。 PMA对PG合成的影响不受NMA的调节。我们认为NO可能不参与PMA对鸡胸骨软骨细胞分化的抑制作用，但星形孢菌素可能至少部分通过NO依赖性机制促进软骨细胞分化