小分子药物kartogenin保护终板软骨退变的实验研究

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (8): 846-850.

小分子药物kartogenin保护终板软骨退变的实验研究

郑权¹, 徐宏光¹, 汪景², 熊寿良¹, 王弘¹, 张晓玲²

¹ 皖南医学院第一附属医院弋矶山医院脊柱外科,芜湖 241001,安徽;
² 中国科学院上海生命科学院骨科细胞与分子生物学实验室, 上海 200025

收稿日期:2014-04-02 修回日期:2014-07-21 出版日期:2014-08-26 发布日期:2014-08-26
通讯作者: 徐宏光,男,主任医师、教授,研究方向:脊柱外科及老年骨病。 Tel: 13855356303 E-mail: pumchxuhg@126.com
作者简介:郑权,男,硕士研究生, 研究方向:脊柱外科及老年骨病。 Tel: 15855986712 E-mail: zhquan2817@163.com
基金资助:
国家自然科学基金(81272048,30973025,81311130314); 卫生部公益性行业专项基金(201002018); 安徽省自然科学基金(1308085MH152)

Experimental study on small molecule drug kartogenin protecting endplate cartilage degeneration

ZHENG Quan¹, XU Hong-guang¹, WANG Jing², XIONG Shou-liang¹, WANG Hong¹, ZHANG Xiao-ling²

¹ Department of Orthopedic Surgery,Yijishan Hosptial ,Wannan Medical College,Wuhu 241001,Anhui,China;
² Key Laboratory of Stem Cell Biology, Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS) & Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025, China;

Received:2014-04-02 Revised:2014-07-21 Online:2014-08-26 Published:2014-08-26

摘要/Abstract

摘要： 目的研究小分子药物kartogenin(KGN)对终板软骨组织细胞外基质分泌的影响,探讨其在终板软骨以及椎间盘退变中的作用。方法建立大鼠椎间盘体外器官退变模型,分为对照组、退变组(穿刺注射生理盐水)、KGN处理组(穿刺注射 100 nmol/L KGN),利用HE染色观察终板软骨及椎间盘形态,番红-固绿染色观察终板软骨细胞外基质的分泌。通过Real time-PCR评价三组终板软骨组织中Ⅱ型胶原、蛋白多糖、SOX-9的表达变化,Western blot检测Ⅱ型胶原、SOX-9蛋白表达变化。结果 HE染色结果显示与对照组比较,退变组椎间盘髓核消失,终板软骨减少,发生明显退变,KGN处理组椎间盘结构较完整;番红-固绿染色可见实验组较对照组终板软骨细胞外基质分泌增多。Realtime-PCR结果显示退变组终板软骨组织Ⅱ型胶原、蛋白聚糖及SOX-9表达明显下调,KGN处理组中终板软骨组织Ⅱ型胶原、蛋白聚糖及SOX-9表达上调;Western blot结果显示实验组软骨细胞中Ⅱ型胶原、SOX-9蛋白表达明显增高。结论体外穿刺及培养能够诱导椎间盘及终板软骨的退变,小分子药物KGN能够促进终板软骨细胞外基质的分泌从而保护终板软骨及椎间盘的退变。

关键词: 椎间盘, 终板软骨, 退变, kartogenin

Abstract: AIM: To observe the effect of small molecule drug kartogenin (KGN) on promoting extracellular matrix secretion in endplate cartilage, and explore its protective role in the endplate cartilage or intervertebral disc degeneration. METHODS: Established an in vitro organ culture model of rat intervertebral disc, punctured the annulus and cultured in vitro. The rats were divided into three groups: control group, degeneration group and KGN treated group. The change of histomorphology in intervertebral disc tissue was assessed by HE staining, the extracellular matrix of endplate cartilage was observed by safranin O-fast green staining. The expressions of type Ⅱ collagen, proteoglycans and SOX-9 in the three groups were detected by Real time-PCR. The expressions of typeⅡCollagen and SOX-9 protein were measured by western blot.RESULTS: HE staining indicated that the nucleus disappeared and the endplate cartilage tended to thin in the degeneration group, and the disc structure is more complete after KGN treated; safranin O- fast green staining showed that the secretion of extracellular matrix were increased in the KGN treated group. Realtime-PCR showed type Ⅱcollagen, aggrecan, and SOX-9 mRNA levels were significantly reduced in the degeneration group, and the cartilage-related gene increased after KGN treated; Western blot results showed similar changes. CONCLUSION: Small molecule drug KGN can promote the extracellular matrix secretion in the endplate cartilage in order to protect the endplate cartilage and intervertebral disc degeneration.

Key words: intervertebral disc, endplate chondrocytes, degeneration, kartogenin

中图分类号:

郑权, 徐宏光, 汪景, 熊寿良, 王弘, 张晓玲. 小分子药物kartogenin保护终板软骨退变的实验研究[J]. 中国临床药理学与治疗学, 2014, 19(8): 846-850.

ZHENG Quan, XU Hong-guang, WANG Jing, XIONG Shou-liang, WANG Hong, ZHANG Xiao-ling. Experimental study on small molecule drug kartogenin protecting endplate cartilage degeneration[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(8): 846-850.

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