中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (9): 1410-1415.doi: 10.3969/j.issn.2095-4344.1594

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

姜黄素和硫酸镁联合应用协同促进炎性软骨细胞的逆分化

王 洋,宋卓悦,李广恒   

  1. 郑州大学第一附属医院,河南省郑州市 450000
  • 修回日期:2018-11-29 出版日期:2019-03-28 发布日期:2019-03-28
  • 通讯作者: 李广恒,博士,教授,博士生导师,郑州大学第一附属医院骨科,河南省郑州市 450000
  • 作者简介:王洋,男,1991年生,河南省洛阳市人,汉族,郑州大学在读硕士,主要从事关节外科基础及临床研究。
  • 基金资助:

    国家自然科学基金资助项目(81472136),项目负责人:李广恒

Synergistic promotion of inflammatory chondrocyte reverse differentiation by the combination of curcumin and magnesium sulfate

Wang Yang, Song Zhuoyue, Li Guangheng   

  1. the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
  • Revised:2018-11-29 Online:2019-03-28 Published:2019-03-28
  • Contact: Li Guangheng, MD, Professor, Doctoral supervisor, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
  • About author:Wang Yang, Master candidate, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81472136 (to LGH)

摘要:

文章快速阅读:

文题释义:
姜黄素:
即二十二烷基甲烷,是姜黄的活性成分,是从姜科植物的根茎中提取的,它是抗氧化剂,具有抗炎、抗肿瘤以及清除自由基等多方面药理作用。
3D培养:在细胞培养过程中为细胞提供一个更加接近体内生存条件的微环境的培养技术,其主要利用磁性微球载体和磁悬浮技术使贴有细胞的微球载体悬浮在培养液中,确保了高质量、高密度的细胞繁殖,突破了传统有盖培养皿、培养瓶或微孔板细胞培养耗时繁琐,细胞产量微小等局限性。

 

摘要
背景:
骨性关节炎是老年人疼痛和致残的常见原因,其根本原因在于不适当的机械应力、炎症递质和生化因子的共同作用。姜黄素已被证明具有抗氧化、抗炎、抗菌、抗肿瘤、神经保护、心脏保护、辐射防护以及治疗骨关节炎等作用,硫酸镁(MgSO4)同样具有缓解关节疼痛,促进关节破坏的作用。
目的:探究姜黄素和MgSO4联合应用协同促进炎性软骨细胞逆分化的作用机制。
方法:原代培养炎性软骨细胞,在体外单层培养条件下,细胞增殖实验检测炎性软骨细胞的增殖情况。实验细胞分为4组,进行体外3D诱导逆分化培养,炎性软骨细胞组为炎性软骨细胞单独培养,姜黄素组采用姜黄素干预培养,MgSO4组为采用MgSO4干预培养,联合治疗组为姜黄素+MgSO4联合干预培养,共培养18 d。
结果与结论:①细胞增殖实验显示,第3代炎性软骨细胞早期增殖速率较高,分化程度较低;②定量PCR检测显示,联合治疗组Ⅱ型胶原、蛋白聚糖和SOX9 mRNA表达量均高于姜黄素组或MgSO4组(P < 0.01);③体外3D诱导分化培养中联合治疗组的大体标本的尺寸和软骨逆分化的阿尔新蓝染色、番红O染色的阳性区域均明显高于其他3组(P < 0.01);④TUNEL染色显示,联合治疗组及MgSO4组凋亡特异性染色的阳性区域均明显小于另外2组(P < 0.01);⑤结果证实,联合应用姜黄素和MgSO4具有协同促进炎性软骨细胞逆分化的作用。


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程
ORCID: 0000-0002-6475-0359(李广恒)

关键词: 骨性关节炎, 姜黄素, 硫酸镁, 3D培养, 软骨细胞, 炎性软骨细胞, 软骨细胞逆分化, 软骨退变, 细胞增殖

Abstract:

BACKGROUND: Osteoarthritis is a common cause of pain and disability in the elderly. The underlying cause is the combination of inappropriate mechanical stress, inflammatory mediators and biochemical factors. Curcumin has been shown to have anti-oxidant, anti-inflammatory, anti-bacterial, anti-tumor, neuroprotective, and cardioprotective effects, radiation protection and therapeutic effects on osteoarthritis. Similarly, magnesium sulfate can relieve joint pain and inhibit joint destruction.
OBJECTIVE: To investigate the mechanism by which the combined use of curcumin and magnesium sulfate exerts synergistic effect to promote inflammatory chondrocyte reverse differentiation.
METHODS: Primary cultured inflammatory chondrocytes were subjected to monolayer culture in vivo. Cell proliferation assay (MTS) was used to detect the proliferation of inflammatory chondrocytes under in vitro monolayer culture conditions. The experimental cells were divided into four groups and underwent 3D induced reverse differentiation culture for 18 days: single culture of chondrocytes (chondrocyte group), inflammatory chondrocyte cultured with curcumin (curcumin group), inflammatory chondrocytes cultured with magnesium sulfate (magnesium sulfate group), and inflammatory chondrocytes cultured with curcumin and magnesium sulfate (combination group). 
RESULTS AND CONCLUSION: MTS proliferation experiments showed that inflammatory chondrocytes at passage 3 had a higher rate of early proliferation and a lower degree of differentiation. Quantitative PCR results showed that the mRNA levels of type II collagen, proteoglycan and SOX9 were significantly higher in the combination group than in the curcumin group or magnesium sulfate group (P < 0.01). The size of the gross specimens and the positive area of chondrocyte reverse differentiation for alcian blue staining and safranin O staining in the combination group were significantly larger than those in the other three groups (P < 0.01). TUNEL staining results indicated that the positive area of apoptosis-specific staining in the combination group and magnesium sulfate group was significantly smaller than that in the other two groups (P < 0.01). Therefore, the combined use of curcumin and magnesium sulfate has the synergistic effect to promote the reverse differentiation of inflammatory chondrocytes.


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Curcumin, Magnesium, Chondrocytes, Tissue Engineering

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