Intracellular signal transaction of osteoblastic cells cultured in a low calcium environments

低钙环境中培养的成骨细胞的细胞内信号传输

• 批准号: 02670822
• 负责人: HISADA Yoh
• 金额: $ 1.41万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670822/
• 关键词: Osteoblastic cells; Low calcium environment; [Ca^<2+>]i; Protein kinase C (PKC); PKC activity; PKC localization; PKC isozyme; 細胞内遊離カルシウム; プロテインキナ-ゼC活性; アルカリフォスファタ-ゼ活性

It is widely known that low calcium(Ca)conditions in body fluids cause various malformations of hard tissue. Our previous studies have shown a corelation between the malfortation and the elnvironment for the rat-femur in vivo and in vitro. The results clearly showed that disturbances to the calcification mechanism occur with cells exposed to the low-Ca environment. Osteoblasts play a central role in the Ca regulation of bone formation. In the present study, we examined the effect of a lowCa environment on intracellular sggnal trunsduction ofosteoblastic cells isolated from the embryonic calvaria of Wistar rats. The cells were cultured up to 10 days in BGJ_b medlum. The Ca concentration in the medium was 1.87 mM for control cells and 0.34 mM for low-Ca cells. The changes in the low-Ca cells were as follows : The Ca^<2+> contents decreased in both total the cellular fractions. Intracellular free Ca([Ca^<2+>]i)and'inositol 1, 4, 5-triphosphate(IP_3)contents decreased markedly. The[Ca^2+]i increased rapidly after stimulation with concanavalin A or the addition of exogenous Ca. The activity of protein kinase C(PKC)decreased in both cytosol and membrane fractions. When the medium was changed to the control medium, the increase of PKC activity in the membrane fraction was much higher than that in cytosol one. The immunocytochemical stinning of PKC(subspecies : TypeI, II andIII)was observed by similar in the both cells. TypeI was cleary observed in the cytoplasm. Especially, the staining was more markedly in a part of cytoplasm. Also, it was presented at the inside of the plasmamembrane. TypeII was the strongestisozne. Typel showed almost no staining or only a faint staining. These findings suggest that the cells under non-physiological conditions such as low-Ca, may perform a functional replacement to maintain normal calcium metabolism in hard tissue. Supported by Grant-in-Aid for Scientific Research(Nos. 02670822, 63480408)of Japan.

众所周知，体液中的低钙（Ca）条件导致硬组织的各种畸形。我们以前的研究表明，在体内和体外的大鼠股骨的畸形和消除之间的相关性。结果清楚地表明，细胞暴露于低钙环境中会发生钙化机制的紊乱。成骨细胞在骨形成的钙调节中起核心作用。本研究观察了低钙环境对Wistar大鼠胚胎颅骨成骨细胞胞内信号转导的影响。细胞在BGJ_B培养基中培养10天。培养基中的Ca浓度对于对照细胞为1.87 mM，对于低Ca细胞为0.34 mM。低钙细胞的变化如下：两种细胞组分中Ca^<2+>含量均降低。细胞内游离Ca（[Ca^<2+]i）和1，4，5-三磷酸肌醇（IP_3）含量显著降低。在伴刀豆球蛋白A刺激或加入外源性Ca后，[Ca^2+]i迅速增加。胞浆和胞膜组分中蛋白激酶C（PKC）活性均降低。当培养基改为对照培养基时，细胞膜组分中PKC活性的增加明显高于胞浆组分。两种细胞中PKC（Ⅰ、Ⅱ、Ⅲ型）的免疫细胞化学染色结果相似。Ⅰ型细胞胞浆内可见。尤其是部分细胞质染色更为明显。而且，它还存在于质膜的内部。Ⅱ型为强雌激素。Typel几乎没有染色或仅显示微弱的染色。这些发现表明，在非生理条件下，如低钙，细胞可能执行功能替代，以维持正常的钙代谢在硬组织。科学研究补助金资助（编号：02670822，63480408）。

项目成果

Matsumoto, A., Hisada, Y. and Yoshimura, Y.: "Changes in alkaline and acid phosphatase activites in bone cells cultured in a low calcium environment." The 64th Annual Meeting of Japanese Pharmacological Society, 1991. Jpn. J. Pharmacol. 55 (Suppl I). 239

Matsumoto, A.、Hisada, Y. 和 Yoshimura, Y.：“在低钙环境中培养的骨细胞中碱性和酸性磷酸酶活性的变化。”

Hisada, Y., Matsumoto, A. and Yoshimura, Y.: "Changes in protein kinase C activity in rat calvarial bone cells cultured in a low calcium environment." Archs oral Biol.(1992)

Hisada, Y.、Matsumoto, A. 和 Yoshimura, Y.：“在低钙环境中培养的大鼠颅骨骨细胞中蛋白激酶 C 活性的变化。”

久田 洋・吉村 善隆・松本 章: "培養骨系細胞の細胞内遊離カルシウムに及ぼす環境のカルシウム濃度の影響" 歯科基礎医学会雑誌. 32. 65 (1990)

Hiroshi Hisada、Yoshitaka Yoshimura 和 Akira Matsumoto：“环境钙浓度对培养骨细胞内游离钙的影响”日本基础牙科医学会杂志 32. 65 (1990)。

Hisada, Y., Yoshimura, Y. and Matsumoto, A.: "Changes in intracellular-free calcium concentrations after stimulation with concanavalin A or phospholipase C of bone cells cultured in low-calcium environments." Dentisty in Japan. 28. (1991)

Hisada, Y.、Yoshimura, Y. 和 Matsumoto, A.：“用伴刀豆球蛋白 A 或磷脂酶 C 刺激低钙环境中培养的骨细胞后，细胞内游离钙浓度发生变化。”

Hisada,Y.,Matsumoto,A.: "The changes of intracellular free calcium concentration after the stimulation of concanavalin A or phosrholipase C in bone cells cultured in a lowーcalcium environment" Archs.Oral Biol.

Hisada, Y., Matsumoto, A.：“低钙环境中培养的骨细胞中伴刀豆球蛋白 A 或磷脂酶 C 刺激后细胞内游离钙浓度的变化”Archs.Oral Biol。