中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (42): 6244-6251.doi: 10.3969/j.issn.2095-4344.2016.42.002

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

骨桥蛋白对人膝骨关节炎软骨细胞透明质酸表达的影响

罗  伟1,张方杰1,李宇晟1,雷光华1,2   

  1. 1中南大学湘雅医院骨科,湖南省长沙市  410008;2中南大学骨科研究所,湖南省长沙市  410008
  • 收稿日期:2016-07-20 出版日期:2016-10-14 发布日期:2016-10-14
  • 通讯作者: 雷光华,博士,教授,中南大学湘雅医院骨科,湖南省长沙市 410008;中南大学骨科研究所,湖南省长沙市 410008
  • 作者简介:罗伟,男,1983年生,湖北省广水市人,汉族,2009年北京大学毕业,博士,主治医师,主要从事骨关节外科,关节镜,骨肿瘤的研究。
  • 基金资助:

    国家自然科学基金资助项目(81401838);中南大学自由探索计划(2012QNZT095)

Effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes

Luo Wei1, Zhang Fang-jie1, Li Yu-sheng1, Lei Guang-hua1, 2   

  1. 1Department of Orthopedics, Xiangya Hospital of Central South University, Changsha 410008, Hunan Province, China; 2Institute of Orthopedics, Central South University, Changsha 410008, Hunan Province, China
  • Received:2016-07-20 Online:2016-10-14 Published:2016-10-14
  • Contact: Lei Guang-hua, M.D., Professor, Department of Orthopedics, Xiangya Hospital of Central South University, Changsha 410008, Hunan Province, China; Instiute of Orthopedics, Central South University, Changsha 410008, Hunan Province, China
  • About author:Luo Wei, M.D., Attending physician, Department of Orthopedics, Xiangya Hospital of Central South University, Changsha 410008, Hunan Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81401838; the Free Exploration Program of Central South University, No. 2012QNZT095

摘要:

文章快速阅读:


文题释义:
骨桥蛋白:1979年Senger等首次报道与恶性转化有关的一种包含RGD整合素结合区的磷酸化糖蛋白与肿瘤的关系,称之为转化相关性磷酸蛋白。后来Franzen等从骨基质和牙齿中分离出一种磷酸蛋白,特性与转化相关性磷酸蛋白相似,人们将其命名为OPN。后来一些学者在不同的组织细胞中发现了骨桥蛋白,因此骨桥蛋白曾被称为44kD骨酸蛋白,PP69、骨延蛋白1、尿蛋白、分泌的磷蛋白、骨唾液酸蛋白、44kD骨磷蛋白和2aR。自1986年来相继克隆了大、小鼠骨桥蛋白,人骨桥蛋白,猪骨桥蛋白,牛骨桥蛋白,鸡骨桥蛋白和兔骨桥蛋白cDNA。近年来,骨桥蛋白在早期细胞免疫应答、肉芽肿炎症、肿瘤发生及转移的作用中倍受关注。
反转录酶:是一种存在于部分RNA病毒中具有反转录活性、能以单链 RNA作为模板合成DNA的酶。由反转录酶催化合成的DNA称为互补DNA(cDNA)。反转录酶可用于RT-PCR,将RNA转变为DNA后扩增,以获得 RNA的序列。
摘要
背景:
在骨关节炎病程中,骨桥蛋白水平和透明质酸水平的改变可以使一系列细胞因子和酶表达水平改变,但是软骨细胞中骨桥蛋白高表达是否与透明质酸高表达相关目前尚不清楚。
目的:通过调节体外培养人膝骨关节炎软骨细胞的骨桥蛋白水平,观察其对软骨细胞透明质酸表达水平的影响。
方法:从人膝骨关节炎软骨组织标本中获取并体外培养软骨细胞,实验分为3组:空白对照组不进行任何干预,骨桥蛋白干预组以1 mg/L重组人骨桥蛋白干预,骨桥蛋白siRNA组以骨桥蛋白siRNA进行干预。采用real time PCR法检测各组骨桥蛋白,透明质酸合成酶1,透明质酸合成酶2和透明质酸合成酶3 mRNA表达,ELISA法检测透明质酸水平。
结果与结论:①与空白对照组相比,重组人骨桥蛋白干预组透明质酸合成酶1,2,3表达升高,而骨桥蛋白siRNA干预组透明质酸合成酶1,2,3表达下降,3组间透明质酸合成酶1,2,3表达比较差异均有显著性意义(P < 0.05);②重组人骨桥蛋白干预组软骨细胞透明质酸表达显著高于空白对照组和骨桥蛋白siRNA干预组(P < 0.05)。③结果说明,提示骨桥蛋白可以通过刺激透明质酸合成酶分泌进而诱导骨关节炎软骨细胞透明质酸表达。

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
ORCID: 0000-0003-1818-3469(罗伟)

关键词: 组织构建, 软骨细胞, 骨桥蛋白, 透明质酸, 透明质酸酶, 骨关节炎, 国家自然科学基金

Abstract:

BACKGROUND: Both osteopontin and hyaluronic acid involve in the pathological process of osteoarthritis, resulting in the abnormal expression levels of various cytokines and enzymes. However, the relationship between the high expression of osteopontin and hyaluronic acid in chondrocytes remains unclear.
OBJECTIVE: To investigate the effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes in vitro by modulating the level of osteopontin.
METHODS: Chondrocytes from human knee osteoarthritic cartilage were cultured in vitro, and were then divided into three groups: blank control group without any treatment; osteopontin group and and pontin siRNA group were treated with 1 mg/L recombinant human osteopontin and osteopontin siRNA, respectively. Expression levels of osteopontin, hyaluronic acid synthase 1, 2 and 3 mRNA were detected by real-time PCR, and the levels of hyaluronic acid were measured using ELISA.
RESULTS AND CONCLUSION: Compared with the blank control group, the mRNA expressions of hyaluronic acid synthase 1, 2 and 3 were remarkably increased in the osteopontin group, while siRNA made the significantly inhibitory effects on the hyaluronic acid synthase 1, 2 and 3 mRNA expressions (P < 0.05). The level of hyaluronic acid in chondrocytes in the osteopontin group was significantly higher than that in the other two groups (P < 0.05). Our results suggest that osteopontin induces the synthesis of hyaluronic acid in osteoarthritic chondrocytes through upregulating the hyaluronic acid synthases expression levels.

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: Osteopontin, Chondrocytes, Hyaluronic Acid, Osteoarthritis, Tissue Engineering

中图分类号: