治疗型关节炎护膝对实验性骨关节炎软骨细胞凋亡基因Bcl-2及p53的影响

目的：通过检测OA护膝对日本大耳白兔膝骨性关节炎软骨细胞凋亡基因Bcl-2、p53mRNA表达的影响，探讨OA护膝防治兔膝骨性关节炎软骨细胞凋亡的分子生物学机制。方法：健康6月龄日本大耳白兔54只，雌雄各半，空腹体重2～2．2kg，采用改良Huhh法复制膝骨性关节炎模型，随机分为6组，即正常组、模型组、对照组（微波组）、实验1组（电组）、实验2组（热组）、实验3组（护膝组）．正常组10只，常规饲养；模型组9只，造模后常规饲养；对照组9只，微波仪治疗30min，每日1次；实验1组9只，电（疏密波）治疗30min，每日1次；实验2组8只，热（热软膜）治疗30min，每日1次；实验3组9只，电热（OA护膝）治疗30min，每日1次，连续治疗16周时处死。采用荧光定量RT—PCR法检测各组膝关节软骨细胞Bcl-2、p53mRNA的表达水平。结果：16周时，各组所有抽提的兔关节软骨组织总RNA的OD260／OD280值均在1．80～2．00范围内，表明RNA纯度高：模型组、对照组、实验1组、实验2纽、实验3组关节软骨细胞p53的mRNA相对呈高表达，而关节软骨细胞Bcl-2的mRNA相对低表达，与正常组差异有统计学意义（P〈0．01）：关节软骨细胞Bcl-2、p53的mRNA相对表达水平，对照组、实验1组、实验2组、实验3组与模型组差异有统计学意义（P〈0．01）；对照组、实验1组、实验2组与实验3组差异有统计学意义（P〈0．01）。结论：OA护膝能提高关节软骨细胞Bcl-2 mRNA表达，减弱软骨细胞p53mRNA表达，从而抑制软骨细胞凋亡，延缓膝关节软骨的退变：

Effect of OA kneepad on Apoptosis Genes Bcl-2 and p53 expression in articular cartilage cells of experimental knee osteoarthritis

Objective: To study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis,so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree. Methods: Forty-four Japanese healthy 6-month-old rabbits(equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups:normal group,model group,control group(microwave),experimental group 1 (electricity),experimental group 2(thermal),experimental group 3(kneepad). Ten rabbits in the normal group were breed with conventional method;9 rabbits in the model group were breed with conventional method after model made;9 rabbits in the control group were treated with microwave for 30 minutes,one time daily;9 rabbits in the experimental group 1 were treated with electricity(density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes,one time daily;9 rabbits in the experiment group 3 were treated with electrothermal(OA knee pad) for 30 minutes,one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method. Results: At the 16 th week,the OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,which indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,the control group,the experimental group 1,group 2,group 3 were overexpressed,and Bcl-2 mRNA expression levels of articular cartilage cells were low expression,and compared with the normal group there were significant differences (P<0.01). Bcl-2,p53 mRNA expression in articular cartilage cells,there were significant differences(P<0.01) between the control group,experimental group 1,group 2,group 3 and model group. The results between the control group,experimental group 1,group 2 and group 3 had significant differences(P<0.01). Conclusion: OA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53,thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.