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。
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 Ⅱin chondroe, ytes 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-1 β, 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 model of osteoarthritis, which is associated with the suppressing IL--1β, MMP-13, ADAMTS-5 and promoting aggrecan and collagen Ⅱ mRNA expression in cartilage.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2016年第2期187-194,共8页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省科学技术厅公益技术应用研究计划(2013C37025)
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
Osteoarthritis/drug therapy
Cartilage, articular/drug effects
Models,animal
Extracellular matrix
Glycosylation end products, advanced/pharmacology
