不同硫酸化程度的糖胺聚糖成软骨作用的体外研究

doi:10.7518/hxkq.2023.2023055

华西口腔医学杂志 ›› 2023, Vol. 41 ›› Issue (4): 395-404.doi: 10.7518/hxkq.2023.2023055

不同硫酸化程度的糖胺聚糖成软骨作用的体外研究

郑雯¹^,²^,³(), 蔡明详¹^,²^,³, 彭荟桢¹^,²^,³, 刘敏义¹^,²^,³, 刘湘宁¹^,²^,³()

^1.暨南大学口腔医学院，广州 510632
^2.暨南大学附属第一医院口腔医疗中心，广州 510630
^3.暨南大学口腔交叉学科临床研究平台，广州 510630

收稿日期:2023-02-17 修回日期:2023-06-15 出版日期:2023-08-01 发布日期:2023-07-21
通讯作者: 刘湘宁 E-mail:zhengwen.dds@outlook.com;liuxiangning2003@126.com
作者简介:郑雯，硕士，E-mail：zhengwen.dds@outlook.com
基金资助:
国家自然科学基金资助项目(82201001);广东省自然基金项目(2021A1515010882);广州市科技计划项目(2021020-10040);暨南大学附属第一医院临床前沿新技术立项项目(a01210)

Effect of glycosaminoglycans with different degrees of sulfation on chondrogenesis

Zheng Wen¹^,²^,³(), Cai Ming-xiang¹^,²^,³, Peng Huizhen¹^,²^,³, Liu Minyi¹^,²^,³, Liu Xiangning¹^,²^,³()

^1.School of Stomatology, Jinan University, Guangzhou 510632, China
^2.Hospital of Stomatology, The First Affiliated Hospital of Jinan University, Guangzhou 510630, China
^3.Clinical Research Platform for Interdiscipline of Stomatology, Jinan University, Guangzhou 510630, China

Received:2023-02-17 Revised:2023-06-15 Online:2023-08-01 Published:2023-07-21
Contact: Liu Xiangning E-mail:zhengwen.dds@outlook.com;liuxiangning2003@126.com
Supported by:
The National Natural Science Foundation of China(82201001);The Natural Science Foundation Projects of Guangdong(2021A1515010882);The Science and Technology Projects in Guangzhou(202102010040);The Clinical Frontier Technology Program of the First Affiliated Hospital of Jinan University(a01210);Correspondence: Liu Xiangning, E-mail: liuxiangning2003@126.com

摘要/Abstract

摘要：

目的本研究旨在探讨硫酸软骨素（CS）、硫酸皮肤素（DS）与肝素（HEP）对软骨形成细胞成软骨分化和小鼠关节软骨状态维持的作用及其机制。方法小鼠软骨发生细胞系（ATDC5）和小鼠关节软骨组织块在含不同硫酸化程度糖胺聚糖的培养基中培养后，使用细胞增殖试验、阿利新蓝染色、实时荧光定量聚合酶链反应（RT-qPCR）和蛋白质印迹（Western blot）分析来观察细胞增殖、成软骨分化、软骨形成、软骨组织维持的作用，并进一步探讨其潜在机制。结果 HEP和DS主要通过激活骨形态发生蛋白（BMP）信号通路，CS主要通过激活蛋白激酶B（AKT）信号通路促进软骨形成细胞的增殖能力、提高基质蛋白多糖的生成、增加Sox9、Ⅱ型胶原蛋白（Col2a1）和聚集蛋白聚糖（Aggrecan）的表达水平。结论本研究探讨了不同硫酸化程度的糖胺聚糖对细胞成软骨与软骨稳态的维持作用差异及其机制，HEP有助于促进软骨形成和维持软骨组织正常状态，CS在损伤软骨组织再生方面效果更好。

关键词: 软骨形成, 软骨损伤修复, 硫酸化糖胺聚糖, 硫酸化程度

Abstract:

Objective This study aims to investigate the effects and mechanisms of chondroitin sulfate (CS), dermatan sulfate (DS), and heparin (HEP) on chondrogenesis of murine chondrogenic cell line (ATDC5) cells and the maintenance of murine articular cartilage in vitro. Methods ATDC5 and articular cartilage tissue explant were cultured in the medium containing different sulfated glycosaminoglycans. Cell proliferation, differentiation, cartilage formation, and mechanism were observed using cell proliferation assay, Alcian blue staining, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blot, respectively. Results Results showed that HEP and DS primarily activated the bone morphogenetic protein (BMP) signal pathway, while CS primarily activated the protein kinase B (AKT) signal pathway, further promoted ATDC5 cell proliferation and matrix production, and increased Sox9, Col2a1, and Aggrecan expression. Conclusion This study investigated the differences and mechanisms of different sulfated glycosaminoglycans in chondrogenesis and cartilage homeostasis maintenance. HEP promotes cartilage formation and maintains the normal state of cartilage tissue in vitro, while CS plays a more effective role in the regeneration of damaged cartilage tissue.

Key words: chondrogenesis, cartilage repair, sulfated glycosaminoglycan, degree of sulfation

中图分类号:

Q 249

郑雯, 蔡明详, 彭荟桢, 刘敏义, 刘湘宁. 不同硫酸化程度的糖胺聚糖成软骨作用的体外研究[J]. 华西口腔医学杂志, 2023, 41(4): 395-404.

Zheng Wen, Cai Ming-xiang, Peng Huizhen, Liu Minyi, Liu Xiangning. Effect of glycosaminoglycans with different degrees of sulfation on chondrogenesis[J]. West China Journal of Stomatology, 2023, 41(4): 395-404.

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不同硫酸化程度的糖胺聚糖成软骨作用的体外研究

Effect of glycosaminoglycans with different degrees of sulfation on chondrogenesis

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