中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (24): 3568-3573.doi: 10.3969/j.issn.2095-4344.2016.24.011

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

透骨消痛胶囊及其拆方对退变软骨细胞Wnt/β-catenin信号通路的影响

吴追乐1,陈星强1,陈春蓉1,于  超1,朱晓勤1,庄志强2   

  1. 1福建中医药大学,福建省福州市  350122;2福建中医药大学附属漳州市中医院,福建省漳州市  363000
  • 出版日期:2016-06-10 发布日期:2016-06-10
  • 通讯作者: 庄志强,主任医师,硕士生导师,福建中医药大学附属漳州市中医院,福建省漳州市,363000
  • 作者简介:吴追乐,男,1982年生,汉族,福建省石狮市人,2011年福建中医药大学毕业,博士,讲师,主要从事中西医结合骨伤科研究。
  • 基金资助:

    国家自然科学基金资助项目(81202713);陈可冀中西医结合发展基金资助项目(CKJ2015010);福建省自然科学基金项目(2015J05161)

Effect of Tougu Xiaotong Capsule and its disassembled recipes on chondrocyte degeneration of rats via Wnt/beta-catenin signaling pathway

Wu Zhui-le1, Chen Xing-qiang1, Chen Chun-rong1, Yu Chao1, Zhu Xiao-qin1, Zhuang Zhi-qiang2   

  1. 1Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China; 2Affiliated Zhangzhou Chinese Medicine Hospital of Fujian University of Traditional Chinese Medicine, Zhangzhou 363000, Fujian Province, China
  • Online:2016-06-10 Published:2016-06-10
  • Contact: Zhuang Zhi-qiang, Chief physician, Master’s supervisor, Affiliated Zhangzhou Chinese Medicine Hospital of Fujian University of Traditional Chinese Medicine, Zhangzhou 363000, Fujian Province, China
  • About author:Wu Zhui-le, M.D., Lecturer, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81202713; the Chen Keji Integrative Chinese and Western Medicine Development Foundation, No. CKJ2015010; the Natural Science Foundation of Fujian, No. 2015J05161

摘要:

文章快速阅读:

文题释义:
Wnt信号通路:1982年在小鼠乳腺癌发现了Wnt基因,由于此基因激活依赖小鼠乳腺癌相关病毒基因的插入,因此,当时被命名为Int1基因,之后的研究表明,Int1基因在小鼠正常胚胎发育中起重要作用,相当于果蝇的无翅(Wingless)基因,可控制胚胎的轴向发育。此后大量研究提示了Int1基因在神经系统胚胎发育中的重要性,因此将Wingless与Int1结合,称为Wnt基因。人Wnt基因定位于12q13.在胚胎发育中,Wnt基因调控的重要信号传导系统即为Wnt通路。
透骨消痛胶囊:为福建中医药大学附属第二人民医院研发的院内制剂(闽制字Z20100006),主要成分为巴戟天、白芍、川芎和肿节风。前期研究表明其对骨性关节炎有治疗作用。
软骨细胞与信号通路:当细胞里要发生某种反应时,信号从细胞外到细胞内传递了一种信息,细胞要根据这种信息来做出反应的现象,叫做信号通路。软骨细胞是关节软骨中唯一的细胞成分,其老化和凋亡在软骨退变中起到重要作用。而Wnt 信号通路在软骨退变和软骨损伤后的修复反应中起重要作用。
摘要
背景
:前期研究表明,透骨消痛胶囊能调控软骨细胞Rho/Rock信号通路、抑制软骨细胞线粒体凋亡通路的信号转导、改变软骨下骨重塑的速率和模式、改善软骨下骨胶原纤维和钙磷结晶的排列等对骨性关节炎发挥治疗作用。
目的:进一步观察透骨消痛胶囊及其拆方含药血清对大鼠退变软骨细胞Wnt/β-catenin信号通路的影响,揭示方中治疗骨性关节炎的主要治法。
方法:①取SD大鼠40只随机分为透骨消痛胶囊组、补肾柔肝组、活血祛风组和生理盐水组,按照药理试验中动物间和动物与人体间的等效剂量换算,前3组分别灌服等效剂量的透骨消痛胶囊、补肾柔肝药、活血祛风药,生理盐水组灌服生理盐水,制备含药血清用于细胞实验。②体外培养SD大鼠软骨细胞并分为5组:空白对照组、模型组、透骨消痛胶囊组、补肾柔肝组和活血祛风组。模型组及各给药组用白细胞介素1β干预软骨细胞24 h后,加入各组血清干预软骨细胞72 h,空白对照组用盐水血清培养,同一时间收集空白对照组软骨细胞。③采用实时定量反转录-聚合酶链反应和Western blot法分析Wnt 4、β-catenin、基质金属蛋白酶13的表达。
结果与结论:①与空白对照组相比,模型组Wnt 4、β-catenin、基质金属蛋白酶13表达均明显增加。②与模型组相比,透骨消痛胶囊组、补肾柔肝组和活血祛风组的Wnt 4、β-catenin、基质金属蛋白酶13表达均明显降低,且降低程度为透骨消痛胶囊组> 补肾柔肝组> 活血祛风组。③结果提示透骨消痛胶囊对白细胞介素1β诱导的软骨细胞具有协同保护作用,补肾柔肝为透骨消痛胶囊治疗骨性关节炎的主要治法。

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: 0000-0001-7816-9453(吴追乐)

关键词: 组织构建, 软骨细胞, 骨关节炎, 退变模型, Wnt/β-catenin信号通路, 透骨消痛胶囊, 补肾柔肝, 活血祛风, 国家自然科学基金

Abstract:

BACKGROUND: Previous studies have showed that Tougu Xiaotong Capsule (TGXTC) exerts better effects on osteoarthritis, by regulating Rho/Rock signaling pathway, inhibiting signal transduction of chondrocyte mitochondrial apoptosis pathway, varying the rate and pattern of subchondral bone remodeling and improving the arrangement of subchondral bone collagen fibers and calcium-phosphate crystallization.
OBJECTIVE: To observe the effects of the serum containing TGXTC and its disassembled recipes on chondrocyte degeneration of rats via Wnt/β-catenin signal pathway, and to explore the main therapeutic method for osteoarthritis in the TGXTC.
METHODS: Forty Sprague-Dawley rats were randomly assigned to receive the treatment of TGXTC, Bushen Rougan (BSRG), Huoxue Qufeng (HXQF) and normal saline, respectively, according to the dose conversion methods of animal to animal and animal to human. Then various drug-containing serums were prepared for the following cellular experiment. After culture and passage, chondrocytes from Sprague-Dawley rats at passage 3 were divided into five groups: blank control, model, TGXTC, BSRG, HXQF groups. Cells in the latter four groups were cultured in appropriate drug-containing serums (normal saline serum for the model group) for 72 hours, following intervention with interleukin-1β for 24 hours. Cells in the blank control group were cultured in normal saline serum. Afterwards, cells in all the five groups were collected for detecting expression of Wnt 4, β-catenin and matrix metalloproteinase 13 at mRNA and protein levels using real-time PCR and western blot assay, respectively.
RESULTS AND CONCLUSION: Compared with the blank control group, the expression of Wnt 4, β-catenin, matrix metalloproteinase 13 was significantly increased in the model group. Compared with the model group, the expression of Wnt 4, β-catenin, matrix metalloproteinase 13 in the TGXTC, BSRG and HXQF groups were decreased significantly, sequenced as TGXTC group < BSRG group < HXQF group. These results indicate that TGXTC plays a synergistic protection against interleukin-1β induced degeneration of chondrocytes. In addition, BSRG as a disassembled recipe of TGXTC is the main therapeutic method for osteoarthritis.

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: Medicine, Chinese Traditional, Osteoarthritis, Cartilage, Tissue Engineering

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