Effects of cell proliferation, protein synthesis and intra-cellular signal transduction

细胞增殖、蛋白质合成和细胞内信号转导的影响

• 批准号: 07670768
• 负责人: TAKATA Masanobu
• 金额: $ 1.41万
• 依托单位: TOYAMA MEDICAL AND PHAMACEUTICAL 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-07670768/
• 关键词: erythropoietin; cell proliferation; protein synthesis; Intra cellular calcium ion; 慢性腎不全; 高血圧症; 血管平滑筋細胞; 細胞増殖作用

In patients with chronic renal failure, recombinant human erythropoietin (rHuEpo) improves renal anemia. However, rHuEpo induced-hypertension occurs commonly (about 30%). The aim of this study was to assess the relation of rHuEpo to cell proliferation, protein synthesis and cytosolic free calcium ([Ca^<2+>] i).We investigated whether rHuEpo induce cell proliferation and protein synthesis in cultured smooth muscle cells or not. Smooth muscle cells were incubated with various concentration of rHuEpo (0,0.1,5,50U/ml). RHuEpo induced DNA and protein synthesis measured by ^3H-thymidine and ^3H-leucine in dose dependent manner.We tried to measure the erythropoietin receptor by binding assay using ^<125>I-rHuEpo, but failed. In 1996, Ammarguellat reported that northern blot analysis revealed the expression of Epo receptor messenger RNA in smooth muscle cells of rats.We also investigated [Ca^<2+>] i in platelets using fluorescent indicator quin 2 in patients with chronic renal failure. Resting [Ca^<2+>] i was higher in patients with chronic renal failure than in controls. RHuEpo (40U/ml) increased [Ca^<2+>] i in platelets in both chronic renal failure and controls. However, the increase in [Ca^<2+>] i in platelets was higher in chronic renal failure than that in controls. There was a positive relationship between changes in [Ca^<2+>] i in platelets and mean blood pressure ((r=0.56, p<0.05).It is suggested that rHuEpo induce cell proliferation and protein synthesis and increase intracellar free calcium, which may attribute to rHuEpo-induced hypertension in part.

在慢性肾衰竭患者中，重组人促红细胞生成素（rHuEpo）可改善肾性贫血。然而，rHuEpo诱导的高血压发生率很高（约30%）。本研究旨在探讨重组人促红细胞生成素（rHuEpo）对平滑肌细胞增殖、蛋白质合成及胞浆游离钙（[Ca^2+] i）的影响。用不同浓度的rHuEpo（0、0.1、5、50 U/ml）孵育平滑肌细胞。用^3 H-胸腺嘧啶核苷和^3 H-亮氨酸测定RHuEpo诱导的DNA和蛋白质合成呈剂量依赖性，我们尝试用^ I-rHuEpo结合试验测定促红细胞生成素受体<125>，但没有成功。1996年，Ammarguellat等报道了北方印迹法检测大鼠血管平滑肌细胞Epo受体mRNA的表达，并应用荧光指示剂quin 2检测了慢性肾功能衰竭患者血小板[Ca^&lt;2+&gt;] i。慢性肾功能衰竭患者的静息[Ca^&lt;2+&gt;] i高于对照组。rhuEpo（40 U/ml）可增加慢性肾功能衰竭患者和对照组血小板[Ca^&lt;2+&gt;] i。慢性肾功能不全组血小板[Ca^&lt;2+&gt;] i较正常对照组增高。血小板[Ca^&lt;2+&gt;] i变化与平均血压呈正相关（r=0.56，p&lt;0.05），提示rHuEpo可诱导细胞增殖和蛋白质合成，增加细胞内游离钙，这可能是rHuEpo所致高血压的部分原因。

项目成果

上野 均ほか7名: "エリスロポエチンの血管平滑筋細胞に及ぼす細胞増殖作用" 日本腎臓学会誌. 36suppl. 252 (1994)

Hitoshi Ueno 等 7 人：“促红细胞生成素对血管平滑肌细胞的细胞增殖作用”，日本肾病学会杂志 36suppl.252（1994 年）。

大橋信也: "腎不全患者におけるエリスロポエチンの血小板内遊離Ca濃度への影響" 第5回腎とエリスロポエチン研究会Proceeding. (印刷中).

Shinya Ohashi：“促红细胞生成素对肾功能衰竭患者血小板中游离 Ca 浓度的影响”第五届肾脏和促红细胞生成素研究组论文集（正在出版）。

Ueno H et al: "Time course of regression of vascular structural changes and its relation to cytosolic free calcium in hypertensives after nilvadipine treatment" J Human Hypertens. 9. 735-739 (1995)

Ueno H 等人：“尼伐地平治疗后高血压患者血管结构变化消退的时间过程及其与胞质游离钙的关系”J Human Hypertens。

Ueno H et al: "Effects of erythropoietin on cell proliferation in cultured smooth muscle cells" Jpn J Nephrol. 36 Suppl.252 (1994)

Ueno H 等人：“促红细胞生成素对培养平滑肌细胞增殖的影响”Jpn J Nephrol。

Oh-hashi S et al.: "Effects of erythropoietin intra-cellular free calcium in platelets in patients with chronic renal failure." (In press).

Oh-hashi S 等人：“促红细胞生成素对慢性肾功能衰竭患者血小板中细胞内游离钙的影响。”