中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (12): 1846-1851.doi: 10.3969/j.issn.2095-4344.0202

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

川芎嗪干预早期膝骨关节炎大鼠软骨Ⅱ型胶原纤维α1基因与血管内皮生长因子mRNA及miR20b的表达

谢平金1,2,3,余  翔1,2,4,柴生颋2,3,曹学伟5,孙  赫1,2,5,陈群群2,3,梁桂洪1,2,3   

  1. 1广州中医药大学岭南医学研究中心中医骨伤科学实验室,广东省广州市  510405;2广州中医药大学,广东省广州市  510405;3广州中医药大学第三附属医院骨科,广东省广州市  510240;4广州中医药大学第一附属医院骨科,广东省广州市  510405;5广东省中医院骨科,广东省广州市   510240
  • 收稿日期:2018-01-20 出版日期:2018-04-28 发布日期:2018-04-28
  • 通讯作者: 曹学伟,博士,主任医师,硕士生导师,广东省中医院骨科,广东省广州市 510240
  • 作者简介:谢平金,男,1992年生,广东省普宁人,在读硕士,主要从事骨关节病、骨质疏松症及运动医学临床防治研究。
  • 基金资助:

    广东省自然科学基金课题(2014A030310127);2016年度广州中医药大学中医骨伤重点学科开放基金(YB16);2017年广东省大学生创新训练计划立项项目(201710572084)

Ligustrazine for early-stage knee osteoarthritis in rats: changes in the expression levels of type II collagen fiber alpha 1, vascular endothelial growth factor and miR20b in the cartilage

Xie Ping-jin1, 2, 3, Yu Xiang1, 2, 4, Chai Sheng-ting2, 3, Cao Xue-wei5, Sun He1, 2, 5, Chen Qun-qun2, 3, Liang Gui-hong1, 2, 3   

  1. 1the Lab of Orthopedics and Traumatology of Chinese Medicine of Lingnan Medical Research Center of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 3Department of Orthopedics, the Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240, Guangdong Province, China; 4Department of Orthopedics, the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 5Department of Orthopedics, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510240, Guangdong Province, China
  • Received:2018-01-20 Online:2018-04-28 Published:2018-04-28
  • Contact: Cao Xue-wei, M.D., Chief physician, Master’s supervisor, Department of Orthopedics, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510240, Guangdong Province, China
  • About author:Xie Ping-jin, Master candidate, the Lab of Orthopedics and Traumatology of Chinese Medicine of Lingnan Medical Research Center of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; Department of Orthopedics, the Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240, Guangdong Province, China
  • Supported by:

    the Natural Science Foundation of Guangdong Province, China, No. 2014A030310127; the Open Foundation for the Key Subject of Traumatology and Orthopedics of Traditional Chinese Medicine of Guangzhou University of Chinese Medicine in 2016, No. YB16; the College Student Innovation Training Program of Guangdong Province in 2017, No. 201710572084

摘要:

文章快速阅读:

文题释义:
川芎嗪:是从活血化瘀类中药川芎中提取的一种有效活性生物碱,具有扩张血管、镇静镇痛、改善微循环、增强免疫等作用。课题组前期研究亦发现川芎嗪能明显降低骨关节炎关节液中NO、前列腺素E2水平,对减轻骨性关节炎的炎症有一定作用。
木瓜蛋白酶:是一种蛋白水解酶。木瓜蛋白酶是番木瓜中含有的一种低特异性蛋白水解酶,木瓜蛋白酶对动植物蛋白具有较强的水解能力,其诱导大鼠膝骨性关节炎具有成功率高、时间短、重复性好等特点。已有研究表明,木瓜蛋白酶关节腔注射可作为诱导早期膝骨关节炎大鼠模型的方法。
摘要
背景:
课题组前期研究发现川芎嗪能明显降低骨关节炎关节液中NO、前列腺素E2水平,对减轻骨性关节炎的炎症有一定作用,但具体机制还不清楚。
目的:观察川芎嗪治疗膝骨关节炎模型关节软骨中Ⅱ型胶原纤维α1基因、血管内皮生长因子mRNA及miR20b表达水平,并探讨川芎嗪治疗早期膝骨关节炎大鼠的作用机制。
方法:健康雄性SD大鼠随机分为正常组、模型组、川芎嗪高剂量组、川芎嗪低剂量组、阳性对照组,后4组通过木瓜蛋白酶关节腔注射制作早期膝骨关节炎大鼠模型。造模后川芎嗪高、低剂量组分别灌胃川芎嗪100 mg/kg和50 mg/kg,阳性对照组灌服塞来昔布24 mg/kg;正常组、模型组灌服等量生理盐水。干预6周后取材,分别行鼠软骨大体组织学评分、qRT-PCR检测软骨Ⅱ型胶原纤维α1基因、血管内皮生长因子mRNA及miR20b的表达水平。
结果与结论:①软骨Mankin评分:川芎嗪高、低剂量组和阳性对照组均较正常组高(P < 0.05),较模型组低(P < 0.05),且川芎嗪高剂量组<阳性对照组<川芎嗪低剂量组;②川芎嗪各剂量组和阳性对照组的Ⅱ型胶原纤维α1基因、血管内皮生长因子mRNA表达较正常组上调(P < 0.05),较模型组下调(P < 0.05),且川芎嗪高剂量组<阳性对照组<川芎嗪低剂量组;③川芎嗪各剂量组、阳性对照组及正常组的miR20b表达均较模型组上调(P < 0.05),其中川芎嗪低剂量组<阳性对照组<川芎嗪高剂量组;④提示:川芎嗪对早期膝骨关节炎大鼠治疗具有积极作用,可能通过上调软骨中miR20b从而抑制血管内皮生长因子mRNA的表达,促进软骨的修复。

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: 0000-0002-9554-9033(谢平金)

关键词: 川芎嗪, 膝骨关节炎, miR20b, 血管内皮生长因子, Col2α1, Ⅱ型胶原纤维α1, 大鼠, 软骨

Abstract:

BACKGROUND: Preliminary studies have found that ligustrazine can effectively improve the levels of nitric oxide and prostaglandin E2, and alleviate the inflammatory response of osteoarthritis, but the related mechanism remains unclear.
OBJECTIVE: To observe the effect of ligustrazine on the expression levels of type II collagen fiber α 1, vascular endothelial growth factor (VEGF) mRNA and miR20b in the articular cartilage of knee osteoarthritis model, and to explore the mechanism of ligustrazine for early-stage knee osteoarthritis in rats.
METHODS: Healthy male Sprague-Dawley rats were randomized into normal, model, high- and low-dose ligustrazine, and positive control groups. Rats in the latter four groups were used to establish the model of early-stage knee osteoarthritis by intra-articular injection of papain, and were then intragastrically given the administration of normal saline, 100 and 50 mg/kg ligustrazine, and 24 mg/kg celecoxib, respectively. The normal group was given the same volume of normal saline. The treatment in each group lasted for 6 weeks. Then, the rat cartilage was taken, and changes of cartilage tissues were assessed by Mankin scores. Expression levels of type II collagen fiber α 1, VEGF mRNA and miR20b in the cartilage were detected by qRT-PCR.
RESULTS AND CONCLUSION: The order of Mankin scores was as follows: normal group < high-dose ligustrazine group < positive control group < low-dose ligustrazine group < model group (P < 0.05). The mRNA expression levels of type II collagen fiber α 1 and VEGF were the highest in the normal group, followed by the high-dose ligustrazine group, positive control group, low-dose ligustrazine group, and model group (P < 0.05). The expression level of miR20b was significantly up-regulated except the model group, and its order was follows: high-dose ligustrazine group < positive control group < low-dose ligustrazine group. Our results indicate that ligustrazine has a positive effect on early- stage knee osteoarthritis in rats, and the possible mechanisms by which ligustrazine promotes cartilage repair are to up-regulate miR20b expression and to inhibit VEGF mRNA expression.

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: Osteoarthritis, Knee, Cartilage, Vascular Endothelial Growth Factors, Fibrillar Collagens, Models, Animal, Tissue Engineering

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