中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (16): 2495-2501.doi: 10.3969/j.issn.2095-4344.2017.16.007

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

六味地黄丸对椎间盘体外退变模型椎间盘细胞外基质组分的影响

徐无忌1,杨  康2,刘思华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岳阳市中医院骨伤科,湖南省岳阳市  414000;3桃江县人民医院骨二科,湖南省桃江县  413400
  • 修回日期:2017-01-06 出版日期:2017-06-08 发布日期:2017-07-06
  • 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,主要从事脊柱疾患中医药防治研究。
  • 基金资助:

    国家自然青年科学基金课题(81302993);湖南中医药科技计划重点项目(201317)

Effect of Liuwei Dihuang Pills on the extracellular matrix components of intervertebral disc degeneration models in vitro

Xu Wu-ji1, Yang Kang2, Liu Si-hua3   

  1. 1Department of Spine Surgery, the Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410005, Hunan Province, China; 2Department of Traumatology and Orthopaedics, Yueyang Hospital of Traditional Chinese Medicine, Yueyang 414000, Hunan Province, China; 3Second Department of Orthopaedics, Taojiang People’s Hospital, Taojiang 413400, Hunan Province, China
  • Revised:2017-01-06 Online:2017-06-08 Published:2017-07-06
  • About author:Xu Wu-ji, M.D., Associate chief physician, Lecturer, Department of Spine Surgery, 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 Key Science and Technology Program for Traditional Chinese Medicine in Hunan Province, No. 201317

摘要:

文章快速阅读:

文题释义:
肿瘤坏死因子α:
肿瘤坏死因子有几十种之多,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。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,且向兔椎间盘内注射外源性肿瘤坏死因子α时可加速影响蛋白聚糖含量的变化而导致椎间盘退变。

 

摘要
背景:
六味地黄丸作为补益肝肾名方,临床应用其治疗腰痛已取得良好的效果,而有关六味地黄丸治疗肾虚型腰痛的作用机制尚不明确。
目的:观察六味地黄丸对兔椎间盘体外退变模型细胞外基质组分的影响,探讨六味地黄丸防治椎间盘退变的疗效。
方法:将20只新西兰兔带终板的L1-L6椎间盘100个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+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,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+六味地黄丸组含10 mg/L 肿瘤坏死因子α 2 μL和体积分数为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:在培养基中加入肿瘤坏死因子α能明显上调Ⅰ型胶原mRNA和蛋白的表达,下调糖胺多糖含量、硫酸软骨素/硫酸角质素比值、透明质酸含量,蛋白多糖mRNA和蛋白的表达、Ⅱ型胶原mRNA和蛋白的表达(P < 0.05),六味地黄丸含药血清可部分对抗肿瘤坏死因子α所导致的改变,提示六味地黄丸可在一定程度上延缓了椎间盘退变。

 

 

 

 

关键词: 组织构建, 组织工程, 六味地黄丸, 椎间盘退变, 细胞外基质, 蛋白多糖, 糖胺多糖, 硫酸软骨素, 硫酸角质素, 透明质酸, Ⅰ型胶原, Ⅱ型胶原, 国家自然科学基金

Abstract:

BACKGROUND: Liuwei Dihuang Pills (LWPs), a famous Chinese prescription for replenishing the liver and kidney, have achieved good effect in the treatment of backache. However, the mechanism underlying LWPs treating backache with kidney deficiency is unclear. 
OBJECTIVE: To observe the effect of LWPs on extracellular matrix components of rabbit intervertebral disc degeneration models in vitro, and to explore the effect of LWPs in the prevention and treatment of intervertebral disc degeneration. 
METHODS: Totally 100 L1-6 intervertebral discs with endplate removed from 20 New Zealand rabbits were randomly divided into control, tumor necrosis factor α (TNF-α), p38-JNK/SAPK blocked, LWPs, and combination groups (n=20 per group). 2 μL of 10 mg/L TNF-α, 10 μL of 20 μmol/L SB203580 (specific inhibitor of p38MAPK) and SP600125 (inhibitor of JNK), 10% LWPs serum, 10% LWPs combined with 2 μL of 10 mg/L TNF-α were added into the medium of the corresponding group. Samples were collected at 2, 4, 6 and 14 days of culture.
RESULTS AND CONCLUSION: The addition of TNF-α significantly upregulated the mRNA and protein expression levels of collagen type I, while significantly downregulated the levels of glycosaminoglycan and hyaluronic acid, chondroitin sulfate/keratan sulfate ratio, as well as the protein and mRNA expression levels of proteoglycan and collagen type II (P < 0.05). LWPs could partly reverse the changes caused by TNF-α, indicating that LWPs can alleviate intervertebral disc degeneration to a certain extent.

 

 

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

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