中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (12): 1861-1866.doi: 10.3969/j.issn.2095-4344.2017.12.010

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

六味地黄丸对椎间盘体外退变模型髓核细胞MAPK信号级联的影响

徐无忌1,刘思华2,杨  康3   

  1. 1湖南中医药大学第二附属医院脊柱科,湖南省长沙市  410005;2桃江县人民医院骨二科,湖南省桃江县  413400;3岳阳市中医医院骨伤科,湖南省岳阳市  414000
  • 收稿日期:2016-12-18 出版日期:2017-04-28 发布日期:2017-05-16
  • 作者简介:徐无忌,男,1978 年生,湖南省祁东县人,土家族,2008 年广州中医药大学毕业,博士,副主任医师,讲师,主要从事脊柱疾患的中医药防治研究。
  • 基金资助:

    国家自然科学基金青年项目(81302993);湖南中医药科技计划重点项目(201317)

Liuwei Dihuang Pill inhibits mitogen-activated protein kinase signaling cascade of nucleus pulposus cells in a model of intervertebral disc degeneration

Xu Wu-ji1, Liu Si-hua2, Yang Kang3   

  1. 1Department of Spine, the Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410005, Hunan Province, China; 2Second Department of Orthopaedics, Taojiang People’s Hospital, Taojiang 413400, Hunan Province, China; 3Department of Orthopaedics and Traumatology,  Yueyang Chinese Medicine Hospital, Yueyang 414000, Hunan Province, China
  • Received:2016-12-18 Online:2017-04-28 Published:2017-05-16
  • About author:Xu Wu-ji, M.D., Associate chief physician, Lecturer, Department of Spine, the Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410005, Hunan Province, China
  • Supported by:

    the National Natural Science Foundation for the Youth of China, No. 81302993; the Scientific Research Plan for Traditional Chinese Medicine in Hunan Province, No. 201317

摘要:

文章快速阅读:

文题释义:
丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)级联:是信号从细胞表面转导到细胞核内部的重要传递者之一,参与细胞生长、发育、分化和凋亡等一系列生理病理过程。到目前为止,在哺乳动物细胞中已至少发现有4条MAPK通路:细胞外信号调节蛋白激酶(extracellular signa1-regulated protein kinase,ERK)通路,c-jun氨基末端激酶(c-jun N-terminal kinase,JNK)通路,ERK5通路和p38MAPK通路。JNK与p38MAPK主要被应激反应如渗透压的改变、热休克、炎症反应因子白细胞介素1α和肿瘤坏死因子α、DNA损伤等因素所激活,引起细胞凋亡。 
肿瘤坏死因子α:是肿瘤坏死因子家族中发现最早,研究较多的一种。一般认为,它由单核细胞和巨噬细胞产生,不仅具有选择性杀伤某些肿瘤细胞的功能,而且有多种免疫调节作用,尤其是参与炎症反应。目前许多研究结果显示,椎间盘退变时椎间盘组织中肿瘤坏死因子α表达增多。
摘要
背景:
JNKs和p38作为丝裂原活化蛋白激酶(MAPKs)家族中重要的成员,对细胞增殖、分化、转化及凋亡起着至关重要的作用。
目的:观察六味地黄丸对兔椎间盘体外退变模型髓核细胞MAPK信号级联的影响。
方法:将100个带终板软骨的椎间盘随机分为空白对照组、肿瘤坏死因子α组、p38-JNK/SAPK阻断组、六味地黄丸组、肿瘤坏死因子α+含六味地黄丸组,每组20个。肿瘤坏死因子α组培养液中含10 mg/L 肿瘤坏死因子α 2 μL;p38-JNK/SAPK阻断组培养液中含p38MAPK特异性阻断剂SB203580,JNK特异性阻断剂SP600125各20 μmol/L 10 μL,10 mg/L 肿瘤坏死因子α 2 μL;六味地黄丸组培养液中含体积分数为10%六味地黄丸血清;肿瘤坏死因子α+六味地黄丸组培养液中含10 mg/L 肿瘤坏死因子α 2 μL,体积分数为10%六味地黄丸血清,分别于培养的第2,4,8,14天收集标本待测。
结果与结论:RT-PCR、Western Blot检测显示,随着培养时间的延长,髓核细胞JNK及P38通路的mRNA含量、蛋白表达量明显逐步升高,而含六味地黄丸血清可以延缓这一进程,提示六味地黄丸可以通过一定程度阻滞JNK及P38通路的信号表达,延缓椎间盘退变进程,起到保护椎间盘的作用。

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: 0000-0001-9509-6874(徐无忌)

关键词: 组织构建, 组织工程, 六味地黄丸, 椎间盘退变, 肿瘤坏死因子α, 丝裂原活化蛋白激酶(MAPK), JNK通路, p38MAPK通路, 国家自然科学基金

Abstract:

BACKGROUND: JNKs and p38 MAPK, as important members of mitogen-activated protein kinase (MAPK), play critical roles in the cell proliferation, differentiation, transformation and apoptosis.
OBJECTIVE: To observe the effects of Liuwei Dihuang Pill on the MAPK signaling cascade of nucleus pulposus cells in a rabbit model of intervertebral disc degeneration.
METHODS: 100 discs samples with endplate cartilages were equivalently randomized into five groups and cultured in the medium containing 10 mg/L tumor necrosis factor-α (TNF-α, 2 μL), p38-JNK/SAPK blockers (10 μL of 20 μmol/L SB203580 and SP600125, 2 μL of 10 μmol/L TNF-α), 10% Liuwei Dihuang Pill serum, 10 mg/L TNF-α (2 μL) plus 10% Liuwei Dihuang Pill serum, respectively. Those cells received no intervention as controls. Samples were collected and detected at 2, 4, 8 and 14 days of culture.
RESULTS AND CONCLUSION: Real time-PCR and western blot assay showed that mRNA and protein expressions of JNK and P38 pathways by the nucleus pulposus cells were upregulated with time, while all above indicators were significantly decreased by the Liuwei Dihuang Pill serum. These findings reveal that Liuwei Dihuang Pill can delay the process of disc degeneration by blocking JNK and P38 signaling pathways, thus exerting protective effect on the intervertebral discs.

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: Intervertebral Disk Degeneration, Tumor Necrosis Factor-alpha, JNK Mitogen-Activated Protein Kinases, p38 Mitogen-Activated Protein Kinases, Drugs, Chinese Herbal

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