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浙江大学学报(医学版)  2016, Vol. 45 Issue (2): 187-194    DOI: 10.3785/j.issn.1008-9292.2016.03.12
原著     
LR-90对骨关节炎模型兔关节软骨的保护效应
黄恺1, 张建方1, 万俊明1, 陆建伟1, 高阳2
1. 浙江省立同德医院骨科, 浙江 杭州 310012;
2. 浙江省磐安中医院骨科, 浙江 金华 322300
Protective effect of LR-90 on articular cartilage in rabbit model of osteoarthritis
HUANG Kai1, ZHANG Jianfang1, WAN Junming1, LU Jianwei1, GAO Yang2
1. Department of Orthopedics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China;
2. Department of Orthopedics, Pan'an County TCM Hospital, Pan'an 322300, China
全文: PDF(1290 KB)  
摘要: 

目的: 体内外观察LR-90对骨关节炎模型兔关节软骨的保护作用,并探讨其作用机制。方法: 体外培养的兔软骨细胞分别用IL-1β(对照组)和IL-1β+LR-90(LR-90组)处理,采用实时定量PCR方法检测各组细胞中MMP-13、ADAMTS-5、Aggrecan和CollagenⅡ的基因表达情况。另取20只雄性新西兰大白兔行双侧前交叉韧带切断术造模,术后4周根据随机原则于每只兔一侧膝关节腔内注射50mg/LLR-90(LR-90组)溶液0.3mL,另一侧关节腔内注射相同剂量的生理盐水作为对照组,每周1次,共注射5周。术后9周取膝关节标本行大体形态及组织病理学评分,用实时定量PCR方法检测各组关节软骨中上述靶基因的表达情况。结果: 大体组织学评分和组织病理学Mankin评分显示,LR-90组软骨病变程度轻于对照组(均P<0.05)。LR-90组软骨细胞MMP-13、ADAMTS-5的mRNA表达低于IL-1β组,而Aggrecan、CollagenⅡ的mRNA表达显著高于对照组(均P<0.01)。体内实验结果与体外实验相符:与对照组比较,关节腔内注射IL-90可下调软骨组织中IL-1β、MMP-13、ADAMTS-5的表达(均P<0.01),减少Aggrecan、CollagenⅡ的丢失(均P<0.01)。结论: LR-90对骨关节炎模型兔关节软骨具有保护作用,可延缓其骨关节炎的进展。下调IL-1β、MMP-13及ADAMTS-5的表达水平,阻止Aggrecan及CollagenⅡ的丢失可能是LR-90对抗骨关节炎骨关节炎,进而保护关节软骨的机制。

关键词 骨关节炎/药物疗法软骨,关节/药物作用模型,动物细胞外基质糖基化终产物,高级/药理学    
Abstract

Objective: To investigate the protective effect of LR-90 on articular cartilage in rabbit model of osteoarthritis. Methods: The cultured rabbits chondrocytes were assigned to be treated with IL-1β (10ng/ml) or IL-1β (10ng/ml)+LR-90 (50 mg/L). The mRNA expression of MMP-13, ADAMTS-5, aggrecan and collagen II in chondrocytes were assessed by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). Twenty male New Zealand white rabbits underwent bilateral anterior cruciate ligament transection (ACLT) to establish a animal model of osteoarthritis. Four weeks after model established, on the basis of randomization one knee of each rabbit was treated with 50 mg/L LR-90 in normal saline (NS) (experimental group) and the other knee was treated with same volume of NS (control group), 1/week × 5. Nine weeks after ACLT all rabbits were sacrificed and the knee joints were evaluated by gross morphology and histology. The mRNA expression of IL-1β, MMP-13, ADAMTS-5, aggrecan and collagen Ⅱ in articular cartilage was analyzed by RT-PCR. Results: Gross morphology and Mankin histological evaluation showed that the extent and grade of cartilage damage in the experimental group were less severe than those in the control group.Compared to IL-1β group, LR-90 treatment suppressed the mRNA expression of MMP-13 and ADAMTS-5, and enhanced aggrecan and collagen Ⅱ mRNA expression. Consistent with the in vitro results, the intraarticular LR-90 administration suppressed the mRNA expression of IL-1β,MMP-13 and ADAMTS-5 (all P<0.01), while enhanced mRNA expression of aggrecan and collagen Ⅱ in cartilage (all P<0.01). Conclusion: LR-90 protects against cartilage degradation and inhibits the progression of osteoarthritis in rabbit mode1 of osteoarthritis, which is associated with the suppressing IL-1β, MMP-13, ADAMTS-5 and promoting aggrecan and collagen Ⅱ mRNA expression in cartilage.

Key wordsOsteoarthritis/drug therapy    Cartilage, articular/drug effects    Models, animal    Extracellular matrix    Glycosylation end products, advanced/pharmacology
收稿日期: 2015-09-30     
CLC:  R684.3  
基金资助:

浙江省科学技术厅公益技术应用研究计划(2013C37025)

通讯作者: 张建方(1967-),男,学士,主任医师,从事骨关节炎人工关节置换研究;E-mail:zhangjf8868@163.com     E-mail: zhangjf8868@163.com
作者简介: 黄恺(1982-),男,硕士,主治医师,从事骨关节炎发病机制研究;E-mail:hzhuangk@163.com
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引用本文:

黄恺 等. LR-90对骨关节炎模型兔关节软骨的保护效应[J]. 浙江大学学报(医学版), 2016, 45(2): 187-194.
HUANG Kai, ZHANG Jianfang, WAN Junming, LU Jianwei, GAO Yang. Protective effect of LR-90 on articular cartilage in rabbit model of osteoarthritis. Journal of ZheJiang University(Medical Science), 2016, 45(2): 187-194.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.03.12      或      http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I2/187

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