兔前交叉韧带切断骨关节炎模型中MMP-1 MMP-13及TIMP-1的mRNA表达研究

Expression of MMP-1 MMP-13 and TlMP-1 mRNA in cartilage and synovium of experimentally induced rabbit ACLT osteoarthritis

目的 研究兔膝关节前交叉韧带切断 (ACLT)骨关节炎模型关节软骨及滑膜中基质金属蛋白酶 (MMP) 1,MMP 13及组织源性基质金属蛋白酶抑制剂 (TIMP) 1在不同造模时期的表达情况 ,探讨上述因子在骨关节炎 (OA)发病过程中的作用。为该模型作OA防治研究提供理论依据。方法 实验组ACLT模型兔 2 0只 ,分别于造模 4周及 8周时各处死 10只 ,假手术对照组大白兔10只于术后 8周处死。解剖显微镜下行股骨髁关节软骨退变的大体评分 ,取股骨内髁内侧退变软骨及邻近滑膜 ,用反转录 多聚酶链反应 (RT PCR)的方法检测MMP 1,MMP 13及TIMP 1的mRNA表达。结果 ACLT术后 4周即可见软骨退变 ,8周时退变进一步加重 (P <0 0 2 ) ,对照组无明显软骨退变。对照组软骨及滑膜中MMP 1及TIMP 1的检出率较低 ,造模 4周时检出率明显增高 (P <0 0 5 ) ,部分阳性标本表达量有一定增高 ,造模 8周时MMP 1仍有很高的检出率 ,表达量持续增高 ,而TIMP 1检出率较 4周时下降 ;MMP 13在对照组滑膜中没有检测出 ,造模 4周时有一定的检出率 ,8周时检出率明显增加 (P <0 0 0 2 ) ,软骨中对照组MMP 13的表达率及表达量都很低 ,造模 4周时表达率明显增高 (P <0 0 5 ) ,8周时表达率却明显下降。结论 MMP 1,MMP 13及TIMP

Objective To explore the roles of MMP 1,MMP 13 and TIMP 1 in the development of osteoarthritis and to select proper gene for the prophylaxis and treatment research on osteoarthritis,and mRNA expression of the above genes in cartilage and synovium of rabbit ACLT knee osteoasrthritis was determined at different time after ACLT operation.Methods Twenty white rabbits received unilateral anterior cruciate ligament transection (ACLT),another 10 rabbits received unilateral knee arthrotomy as pseudo operation controls.Ten experimental rabbits were killed at 4 and 8 weeks post surgery respectively,and 10 pseudo operation rabbits were killed at 8 weeks post surgery.Knees were harvested to observe the macro pathologic changes of the femoral condyle.Cartilage at the margine of lesion and synovium nearby were obtained and mRNA expression of MMP 1,MMP 13 and TIMP 1 was detected by RT PCR.Results Cartilage degeneration was obvious at 4 weeks and became more severe at 8 weeks after ACLT operation,while cartilage still remained normal at 8 weeks after pseudo operation.Expression rates of MMP 1 and TIMP 1 in control cartilage and synovium were low,and the rates increased significantly at 4 weeks after ACLT with some positive samples showing increased expression amount.At 8 weeks the rates of MMP 1 still remained high and the amount remained increasing,but the rates of TIMP 1 decreased.MMP 13 was not detectable in control synovium.Its expression rates were low at 4 weeks and increased significantly at 8 weeks after ACLT. The expression rate of MMP 13 was very low in control cartilage,it increased significantly at 4 weeks but decreased at 8 weeks after ACLT.Conclusion MMP 1,MMP 13 and TIMP 1 play an important role in the development of osteoarthritis,especially in its early stage.But only MMP 1 remained highly detectable both at 4 and 8 weeks,and its amount increased with time going,indicating MMP 1 could be a good marker in the prophylaxis and treatment research of osteoarthritis.