中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (11): 1901-1904.doi: 10.3969/j.issn.1673-8225.2011.11.001

• 骨组织构建 bone tissue construction •    下一篇

重组人胰岛素样生长因子Ⅰ对成骨细胞的影响

徐  萍1，张克勤2，刘  超2，陈军建1，袁国跃1，俞  力1，钱方方1   

  1. 1江苏大学附属医院内分泌科，江苏省镇江市 212001
    2南京医科大学第一附属医院内分泌科，江苏省南京市 210029
  • 收稿日期:2010-09-07 修回日期:2010-10-29 出版日期:2011-03-12 发布日期:2011-03-12
  • 通讯作者: 张克勤，博士，主任医师，研究方向：内分泌基础和临床。南京医科大学第一附属医院内分泌科，江苏省南京市 210029 keqzhang@hotmail.com
  • 作者简介:徐萍★，女，1975年生，江苏省镇江市人，汉族，2005年南京医科大学毕业，硕士，主治医师，研究方向：内分泌基础和临床。 jennyxu1234@yahoo.com.cn
  • 基金资助:

    江苏省自然科学基金(BK2009208)，C反应蛋白调控脂联素表达的分子机制研究。

Effects of recombinant human insulin-like growth factor on rat osteoblasts

Xu Ping1, Zhang Ke-qin2, Liu Chao2, Chen Jun-jian1, Yuan Guo-yue1, Yu Li1, Qian Fang-fang1   

  1. 1Department of Endocrinology, the Affiliated Hospital of Jiangsu University, Zhenjiang  212001, Jiangsu Province, China
    2Department of Endocrinology, the First Affiliated Hospital of Nanjing Medical University, Nanjing  210029, Jiangsu Province, China
  • Received:2010-09-07 Revised:2010-10-29 Online:2011-03-12 Published:2011-03-12
  • Contact: Zhang Ke-qin, Doctor, Chief physician, Department of Endocrinology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China keqzhang@hotmail.com
  • About author:Xu Ping★, Master, Attending physician, Department of Endocrinology, the Affiliated Hospital of Jiangsu University, Zhenjiang 212001, Jiangsu Province, China jennyxu1234@yahoo.com.cn
  • Supported by:

    the Natural Science Foundation of Jiangsu Province, No. BK2009208*

PDF

421

被引次数

12

摘要：

背景：人胰岛素样生长因子Ⅰ在成骨细胞中含量丰富，与骨密度有密切关系。
目的：观察重组人胰岛素样生长因子Ⅰ对体外培养大鼠成骨细胞的增殖、凋亡及碱性磷酸酶合成的影响。
方法：不同质量浓度重组人胰岛素样生长因子Ⅰ刺激体外培养的大鼠成骨细胞，噻唑蓝法测定活细胞数量；肿瘤坏死因子α单独或与重组人胰岛素样生长因子Ⅰ共同刺激成骨细胞，流式细胞仪检测细胞周期和凋亡率，采用对硝基酚磷酸盐法测定细胞裂解液中碱性磷酸酶活性。
结果与结论：与对照组相比，一定剂量的重组人胰岛素样生长因子Ⅰ能明显增加大鼠成骨细胞数量(P< 0.05)，在质量浓度为0.1~100 μg/L时，成骨细胞数量的增加与质量浓度呈正相关；肿瘤坏死因子α在0.1~100 μg/L范围内呈剂量依赖性地促进成骨细胞凋亡(P< 0.05)，并使S期细胞减少(P < 0.05)，而重组人胰岛素样生长因子Ⅰ能抑制肿瘤坏死因子α对成骨细胞的促凋亡作用(P < 0.05)；与对照组相比，重组人胰岛素样生长因子Ⅰ刺激的成骨细胞碱性磷酸酶的活性明显增高(P< 0.05)。重组人胰岛素样生长因子Ⅰ对大鼠成骨细胞有明显的促增殖作用，且能明显抑制肿瘤坏死因子α诱导的大鼠成骨细胞凋亡，提示增加成骨细胞数量可能是重组人胰岛素样生长因子Ⅰ促进骨形成的机制之一；重组人胰岛素样生长因子Ⅰ能促进大鼠成骨细胞碱性磷酸酶的合成，提示重组人胰岛素样生长因子Ⅰ有可能促进骨基质的合成和钙化。

关键词: 重组人胰岛素样生长因子Ⅰ, 成骨细胞, 增殖, 凋亡, 碱性磷酸酶, 肿瘤坏死因子α, 骨组织工程

Abstract:

BACKGROUND: Recombinant human insulin-like growth factor (rhIGF-Ⅰ) is rich in osteoblasts, which has close contact with bone mineral density. 
OBJECTIVE: To investigate the effects of rhIGF-Ⅰ on proliferation, apoptosis and synthesis of alkaline phosphatase (ALP) of rat osteoblasts in vitro.
METHODS: Rat calvaria cells were cultured in medium with or without rhIGF-Ⅰ. The living cell number was assessed by MTT colorimetric assay. Tumor necrosis factor α (TNF-α) was added in the medium to induce apoptosis. The cell cycle and apoptosis were analyzed by flow cytometry after the cells were incubated with medium containing TNF-α or TNF-α plus rhIGF-Ⅰ. The ALP content within osteoblasts was measured by PNPP method.
RESULTS AND CONCLUSION: The cell number of osteoblasts exposed to rhIGF-Ⅰ at 102-105 ng/mL significantly increased compared with control group (P < 0.05). TNF-α (102-105 ng/L) caused a dose-dependent increase of apoptosis (P < 0.05) and decreased the cell number in S phase (P < 0.05). rhIGF-Ⅰ inhibited the inducement of TNF-α for apoptosis. The ALP activity in rhIGF-Ⅰ-treated cells was higher than that in vehicle-treated cells (P  < 0.05). IGF-Ⅰ can stimulate proliferation of osteoblasts and inhibit apoptosis induced by TNF-α in vitro, these results suggested that increasing the cell number of osteoblasts might be one of the mechanisms for IGF-Ⅰ to promote bone formation. IGF-Ⅰ can increase ALP synthesis in osteoblasts in vitro, this result suggested that IGF-Ⅰ could probably promote the synthesis of organic matrix and mineralize action of bone.

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