中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (46): 7391-7375.doi: 10.3969/j.issn.2095-4344.2015.46.003

• 骨组织构建 bone tissue construction • 上一篇    下一篇

氟化钠致人成骨细胞凋亡相关基因分析

邓 强1，张亚楼2，盛伟斌1   

  1. 1新疆医科大学一附院脊柱外科，新疆维吾尔自治区乌鲁木齐市 830011；2新疆医科大学基础医学院组织胚胎学教研室，新疆维吾尔自治区乌鲁木齐市 830011
  • 收稿日期:2015-09-22 出版日期:2015-11-12 发布日期:2015-11-12
  • 通讯作者: 张亚楼，博士，副教授，新疆医科大学基础医学院组织胚胎学教研室，新疆维吾尔自治区乌鲁木齐市 830011
  • 作者简介:邓强，男，1974年生，湖南省益阳市人，汉族， 2004年新疆医科大学毕业，硕士，副主任医师，主要从事脊柱外科的研究。
  • 基金资助:

    国家自然科学基金资助项目(81102084，81460481)；新疆自治区自然科学基金项目(2015211C040)

Apoptosis-related genes in human osteoblasts induced by sodium fluoride

Deng Qiang1, Zhang Ya-lou2, Sheng Wei-bin1   

  1. 1Department of Spinal Surgery, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China; 2Department of Histology and Embryology, Preclinical Institute of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
  • Received:2015-09-22 Online:2015-11-12 Published:2015-11-12
  • Contact: Zhang Ya-lou, Ph.D., Associate professor, Department of Histology and Embryology, Preclinical Institute of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
  • About author:Deng Qiang, Master, Associate chief physician, Department of Spinal Surgery, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81102084, 81460481; the Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2015211C040

PDF

294

被引次数

4

摘要：

背景：目前氟致成骨细胞凋亡对线粒体凋亡通路研究不系统，缺乏系统连贯性，不能明确氟引起成骨细胞凋亡具体传导路径。

目的：分析氟致成骨细胞凋亡的可能途径及分子特征。
方法：以人成骨肉瘤细胞系Saos-2建立体外染氟模型，体外培养细胞后经不同质量浓度NaF(0，5，10，20，40，80 mg/L)处理。用流式细胞仪检测干预24 h后的线粒体膜电位；用PCR功能芯片检测84个与凋亡相关基因；对部分差异表达基因用免疫印记法予以验证。
结果与结论：当氟化钠质量浓度为20，40，80 mg/L时，成骨细胞线粒体膜电位分别为27.0%，28.8%，38.6%      (P均< 0.05)；PCR芯片检测发现13个基因表达上调，15个基因表达下调。免疫印记显示Bim、Caspase 9、Caspase 14、BCL2、BAX表达随氟化钠剂量增高而增强；Caspase 3在5 mg/L时表达减弱，10 mg/L以上表达逐渐增强。Caspase 7在各组的表达未见明显差异。Caspase 10表达随氟化钠剂量增高而减弱。结果提示，氟致成骨细胞凋亡可能是通过线粒体途径(包括内质网应激途径)及死亡受体途径。
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 组织构建, 骨细胞, 成骨细胞, 凋亡, 氟中毒, PCR芯片, 国家自然科学基金

Abstract:

BACKGROUND: There are no systematic and coherent studies on the mitochondrial apoptotic pathway of fluoride-induced osteoblast apoptosis, and the specific pathways to induce apoptosis in osteoblasts by fluorine are still unclear.
OBJECTIVE: To explore the possible pathways of apoptosis in osteoblasts induced by fluoride and its molecular characteristics.
METHODS: A fluorosis model of human osteosarcoma cell line Saos-2 was established in vitro. After in vitro culture, the cells were treated with sodium fluoride at different concentrations (0, 5, 10, 20, 40, 80 mg/L). Flow cytometry was used to inspect the mitochondrial membrane potential at 24 hours after intervention; 84 apoptosis-related genes were detected by PCR Array; parts of the differentially expressed genes were verified by western blot method.
RESULTS AND CONCLUSION: When the concentrations of sodium fluoride were 20, 40 and 80 mg/L, the mitochondrial membrane potentials in osteoblasts were 27.0%, 28.8%, 38.6%, respectively (all P < 0.05). PCR  
array found 13 genes upregulated and 15 genes down-regulated. Immunoblotting results showed Bim, Caspase 9, Caspase 14, BCL2, BAX expressions enhanced with increasing doses of sodium fluoride; Caspase 3 expression was decreased at the concentration of 5 mg/L, but increased gradually at over 10 mg/L. Caspase 7 expression had no significant difference. The expression of Caspase 10 decreased with the increasing doses of sodium fluoride. These findings indicate that fluoride may induce apoptosis in osteoblasts through the mitochondrial pathway (including the endoplasmic reticulum stress pathway) and death receptor pathway.
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoblasts, Apoptosis, Fluorine, Tissue Engineering