跑台运动对小鼠骨性关节炎的影响机制

Mechanism Research of Effects of Exercise on Osteoarthritis

研究运动对软骨细胞基质降解酶MMP13、Adamts4、Adamts5及软骨标记蛋白Col2、Col10、Runx2、Sox9的影响,为骨性关节炎运动医学角度提供科学依据。方法:对36只8周雄性小鼠,随机分成不手术对照组(C),手术自由活动组(F)和手术跑台组,手术为DMM关节炎手术,对照组不手术自由喂养,手术自由组进行DMM手术后自由喂养,手术跑台组进行DMM手术后5天后进行跑台,每天进行20m跑台训练,6周后处死取样。对3组小鼠膝关节进行石蜡包埋切片,通过fast green-Safranin O染色对关节炎进行评级,然后在通过Q-PCR和Western Blotting检测相关基因mRNA和蛋白水平的表达。结果:1)与C组比较,F组和R组关节均有不同级别损伤,而R关节受损程度更加严重。2)与C组和F组相比,R组MMP13mRNA表达均有非常显著性升高(P<0.01),F组与C组无显著性差异;与C组和F组相比,R组Adamts4mRNA表达有均有非常显著性升高(P<0.01),与C组相比,F组Adamts4mRNA也有显著性升高(P<0.05);与C组和F组相比,R组Adamts5mRNA表达均有非常显著性升高(P<0.01),F组与C组相比也有非常显著性升高(P<0.01);与C组和F组相比,R组Col2mRNA表达有均有非常显著性降低(P<0.01),F组与C组相比有非常显著性降低(P<0.01)。3)与C组和F组相比,R组col2蛋白表达量均有非常显著性下降(P<0.01),F组比C组的Col2蛋白表达量也有非常显著性下降(P<0.01);与C组和F组相比,R组Sox9蛋白表达均有非常显著性下降(P<0.01),F组比C组Sox9蛋白表达量也有非常显著性下降(P<0.01);与C组和F组相比,R组Col10蛋白表达都有非常显著性增加(P<0.01),F组比C组Col10蛋白表达量也有非常显著性增加(P<0.01);与C组和F组相比,R组Runx2蛋白表达都有非常显著性增加(P<0.01),F组比C组Runx2蛋白表达量也有非常显著性增加(P<0.01)。结论:跑台运动加快骨性关节炎小鼠关节软骨的降解和钙化,加重了关节软骨损伤。

The purpose of this research was to study the effects of exercise on cartilage matrix degradation enzymes MMP13,Adamts4,Adamts5,and cartilage marker protein Col2,Col10,Runx2,Sox9 in order to provide a scientific basis for osteoarthritis sports medicine.36 adult male mice of 8weeks were randomly divided into 3groups：no surgery group（C）,free surgery group（F）and surgical treadmill group,surgery for DMM osteoarthritis surgery,the control group without surgery free feeding,free surgery group after DMM surgery free feeding,treadmill group into 5operation days after DMM operation treadmill,20 minutes of treadmill training in everyday,after 6weeks of sampling.The paraffin embedded sections of the 3groups of mice were evaluated by fast green-Safranin O staining,and then the expression of mRNA and protein was detected by QPCR and Western.Results showed that：1）Compared with Group C,Group F and Group R had different levels of jointdamage,and R joint damage was more serious.2）Compared with Group C and Group F,MMP13 mRNA expressions in Group R significantly increased（P〈0.01）,no significant differences were seen between Group F and Group C.Compared with Group C and Group F,Adamts4 mRNA expressions of Group R significantly increased（P〈0.01）.Compared with the Group C,Adamts4 mRNA of Group F also significantly increased（P〈0.05）.Compared with Group C and Group F,Group R,Adamts5 mRNA expression significantly increased（P〈0.01）,Group F and Group C significantly increased（P〈0.01）.Compared with Group C and Group F,there was a significant decrease（P〈0.01）in Col2 mRNA expression in Group R.Group F and Group C significantly decreased（P〈0.01）.3）Compared with Group C and Group F,col2 protein expressions in Group R significantly decreased（P〈0.01）.Col2 protein expression in Group F significantly decreased（P〈0.01）more than that of Group C.Compared with Group C and Group F,Group R,Sox9 protein expression significantly decreased（P〈0.01）,Group F Sox9 protein expression significantly decreased than Group C（P〈0.01）.Compared with Group C and Group F,the expression of R protein in Col10 group significantly increased（P〈0.01）,Col10 protein expression in Group F significantly increased than that in Group C（P〈0.01）.Compared with Group C and Group F,the expression of R protein in Runx2 group significantly increased（P〈0.01）,Runx2 protein expression in Group F significantly increased than that in Group C（P〈0.01）.It could be concluded that exercise could accelerate the degradation and calcification of articular cartilage and aggravate the injury of cartilage.