CAJ | 학술논문

目的比较不同强度跑台运动对大鼠膝关节软骨MMP-3、COL-Ⅱ、TIMP-3基因及蛋白表达的影响.方法 8周龄雄性SD大鼠24只,随机分为高强度运动组、低强度运动组和对照组.运动组动物按照既定的运动处方在动物跑台上进行运动训练,6周后处死所有实验动物取材.采用COL-Ⅱ免疫组化染色观察膝关节标本COL-Ⅱ染色的平均光密度值,采用RT-RCR技术检测各组大鼠软骨MMP-3 mRNA、COL-ⅡmRNA及TIMP-3 mRNA表达,采用Western blot技术检测各组大鼠软骨MMP-3、COL-Ⅱ及TIMP-3蛋白的表达.结果 COL-Ⅱ免疫组化检测显示高强度运动组光密度值最低,且与低强度运动组和对照组的差异有统计学意义.随着运动强度的增加,MMP-3与COL-Ⅱ的mRNA表达水平上调,而TIMP-3的mRNA表达水平下调.虽然随着运动强度的增加,MMP-3和COL-Ⅱ的蛋白表达水平增高,TIMP-3的蛋白表达水平下降,但在高强度运动组COL-Ⅱ的蛋白表达却较低强度运动组降低,同时MMP-3的蛋白水平明显升高.结论一定程度的运动强度会在基因转录水平调节MMP-3、COL-Ⅱ及TIMP-3的表达,使软骨基质的代谢处于活跃状态,但运动强度过大可能通过MMP-3的降解作用降低COL-Ⅱ蛋白水平的表达.适度的运动有利于关节软骨基质成分的更新,超负荷运动可能通过影响软骨基质的合成导致关节的退变.
목적 비교불동강도포태운동대대서슬관절연골MMP-3、COL-Ⅱ、TIMP-3기인급단백표체적영향.방법 8주령웅성SD대서24지,수궤분위고강도운동조、저강도운동조화대조조.운동조동물안조기정적운동처방재동물포태상진행운동훈련,6주후처사소유실험동물취재.채용COL-Ⅱ면역조화염색관찰슬관절표본COL-Ⅱ염색적평균광밀도치,채용RT-RCR기술검측각조대서연골MMP-3 mRNA、COL-ⅡmRNA급TIMP-3 mRNA표체,채용Western blot기술검측각조대서연골MMP-3、COL-Ⅱ급TIMP-3단백적표체.결과 COL-Ⅱ면역조화검측현시고강도운동조광밀도치최저,차여저강도운동조화대조조적차이유통계학의의.수착운동강도적증가,MMP-3여COL-Ⅱ적mRNA표체수평상조,이TIMP-3적mRNA표체수평하조.수연수착운동강도적증가,MMP-3화COL-Ⅱ적단백표체수평증고,TIMP-3적단백표체수평하강,단재고강도운동조COL-Ⅱ적단백표체각교저강도운동조강저,동시MMP-3적단백수평명현승고.결론 일정정도적운동강도회재기인전록수평조절MMP-3、COL-Ⅱ급TIMP-3적표체,사연골기질적대사처우활약상태,단운동강도과대가능통과MMP-3적강해작용강저COL-Ⅱ단백수평적표체.괄도적운동유리우관절연골기질성분적경신,초부하운동가능통과영향연골기질적합성도치관절적퇴변.
Objective To investigate the effects of different intensities of treadmill running on the expression of matrix metalloproteinase-3 (MMP-3),collagen-Ⅱ (COL-Ⅱ) and tissue inhibitor of metalloproteinase-3 (TIMP-3) in the articular cartilage of the knee joints of rats.Methods Twenty four male SpragueDawley rats were randomly divided into three groups: control group (n=8),low-intensity running (LIR) group (n=8) and high-intensity running (HIR) group (n=8).The rats in LIR and HIR group underwent treadmill running exercise.After 6 weeks,the bilateral knee joints were harvested.The left knee joints were obtained to observe the change of COL-Ⅱ in the cartilage by immunohistochemistry.However,the right knee joints were used to observe the expression of mRNA and protein of MMP-3,COL-Ⅱ and TIMP-3 by RT-PCR or Western bolt.Results The optical density of COL-Ⅱ protein in HIR group was significantly lower than those in control and LIR groups.The MMP-3 and COL-Ⅱ mRNA expression levels in HIR group was significantly higher than those in control and LIR groups.However,the TIMP-3 mRNA expression level in HIR group was lower than those in control and LIR groups.Although MMP-3 and COL-Ⅱ protein expression increased and TIMP-3 protein expression decreased with the increase of exercise intensity,COL-Ⅱ protein expression level in HIR group was lower than that in LIR group.Conclusion Moderate-intensity running could enhance the metabolism of cartilage matrix by regulating the expression of MMP-3,COL-Ⅱ and TIMP-3.Running intensity may be a key factor to regulate cartilage change.