摘要
目的利用家兔膝关节前交叉韧带切断(ACLT)及内侧半月板前1/3切除的方法制作骨关节炎(OA)模型,测定关节炎发病的不同时期,关节软骨中基质金属蛋白酶-1(MMP-1),基质金属蛋白酶-13(MMP-13)以及细胞外信号调节激酶1/2(ERK1/2)蛋白表达水平的变化情况,探索它们之间的变化规律及关系。方法实验组家兔30只行ACLT及内侧半月板前1/3切除术造模,分别于术后4w、8w、12w处死10只,假手术对照组家兔10只于术后12w处死。进行股骨髁关节软骨退变的大体评分;取退变的软骨组织,用Westernblot印记杂交方法测定软骨组织中MMP-1,MMP-13和ERK1/2的蛋白表达水平。结果ACLT及内侧半月板前1/3切除术后4w即可见软骨退变,8w和12w时退变进一步加重,对照组无明显软骨退变,不同组动物关节软骨评分差异有统计学意义。MMP-1和ERK1/2的蛋白表达在正常对照组水平均较低,造模4w开始升高,到第8w及第12w持续升高;而MMP-13在造模4w时表达开始升高,造模第8w时持续升高,但在造模第12w时MMP-13的蛋白表达下降。结论家兔膝关节ACLT及内侧半月板前1/3切除制作OA模型的方法简便可行,MMP-1和MMP-13在骨关节炎的发病过程中有不同程度的升高,可以作为OA研究的评价指标。本实验结果提示,在OA发展的过程中,MMP-1和MMP-13的表达不平行,两者上游信号调控通路可能存在差异。蛋白激酶ERK1/2的表达水平从造模第4w开始持续升高,说明ERK1/2信号转导通路在骨关节炎发病过程中持续发挥作用。
Objective To detect protein expression of matrix metalloproteinase-1 （MMP-1）, matrix metalloproteinase-13 （ MMP-13 ） and exlracellular regulated kinase 1/2 （ ERK1/2 ） in the joint cartilage of osteoarthritie rabbit knee at different time points after ACLT and partial excision of the medial meniscus. Methods Thirty rabbits in the experimental goup received unilateral ACLT and partial excision of the medial meniscus, and another 10 rahhits received unilateral arthrotomy as pseudo-operation control. Ten of the 30 experimental rabbits were sacrificed at week 4, 8 and 12 after surgery respectively, and 10 pseudo-operation rabbits were sacrificed at 12 weeks after surgery. The knees were harvested to observe maeropathologie changes of the joint cartilage. MMP-1, MMP-13 and ERK1/2 proteins near the cartilage lesions were detected by Western Blot. Results Cartilage degeneration was obvious at week 4 and beeame more severe at week 8 and 12 after operation, while the cartilage remained normal at week 12 after pseudo-operation. The expression of MMP-1, MMP-13 and ERK1/2 protein was constantly low in the normal cartilage of the pseudo-operation group. In the experimental group, the expression of MMP-1 and ERK1/2 protein began increasing at week 4, and remained at a high level at week 8 and 12 after operation, while the expression of MMP-13 protein began increasing at week 4, remained increased at week 8, and hegan decreasing at week 12. Conclusions MMP-1 and MMP-13, especially MMP-1, seem to he good markers for study of osteoarthrltis. MMP-13 expression was different from MMP-l expression at week 12 after operation, suggesting that their upstream signaling pathways may be different. The ERK1/2 signal transduction pathway seems to play a constant role in the pathogenesis of osteoarthritis.
出处
《老年医学与保健》
CAS
2007年第6期338-342,共5页
Geriatrics & Health Care
基金
国家自然科学基金资助项目(编号:30471745)
关键词
骨关节炎
基质金属蛋白酶
动力传导
细胞
软骨
关节
前交叉韧带
蛋白激酶类
Osteoarthritis
Matrix metalloproteinases
Meehanotransduetion, cellular
Cartilage, articular
Anterior cruciate ligament
Protein kinases
