慢病毒介导siRNA沉默ERK2对创伤性骨性关节炎大鼠软骨退变的影响及其机制  

Effect of siRNA mediated by lentivirus silencing ERK2 on cartilage degeneration in rat models of post traumatic osteoarthritis and its mechanism

作　　者：宁仁德 孔令超 周业金 王祥[2] 

机构地区：[1]安徽医科大学第三附属医院,合肥230031 [2]上海交通大学医学院附属第三人民医院

出　　处：《山东医药》2016年第16期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81101380)

摘　　要：目的观察慢病毒介导siRNA沉默细胞外信号调节激酶2(ERK2)对创伤性骨性关节炎大鼠软骨退变的影响,并探讨其机制。方法将30只右膝创伤性骨性关节炎大鼠随机分为三组,每组10只,分别于右膝关节腔注射无菌PBS(模型组)、ERK2 siRNA阴性慢病毒溶液(对照组)及ERK2 siRNA慢病毒溶液(观察组)。8周后处死大鼠,取其右膝关节观察膝关节软骨形态并进行评分,行HE、甲苯胺蓝及番红"O"染色后观察软骨组织病理形态,采用Mankin半定量法进行关节软骨评分。采用Real-time PCR法检测各组软骨组织MMP3、MMP13及Col2a mRNA相对表达量。结果观察组软骨面溃疡、缺损程度均轻于模型组和对照组。HE、甲苯胺蓝、番红"O"染色均显示,观察组软骨面较光滑,裂隙出现、表面组织丢失、基质染色减轻、软骨细胞增生和排列紊乱程度均轻于模型组和对照组。模型组、对照组软骨形态评分、关节软骨评分均高于观察组(P均<0.05),模型组和对照组比较无统计学差异(P均>0.05)。观察组MMP3、MMP13 mRNA相对表达量均低于模型组和对照组,Col2a mRNA相对表达量均高于模型组和对照组(P均<0.01);模型组和对照组MMP3、MMP13及Col2a mRNA相对表达量比较无统计学差异(P均>0.05)。结论慢病毒介导siRNA沉默ERK2能够减轻创伤性骨性关节炎大鼠的软骨退变,机制可能与降低MMP3、MMP13表达及增加Col2a表达有关。Objective To observe the effect of siRNA mediated by lentivirus silencing extracellular signal-regulated kinase 2（ ERK2） on cartilage degeneration in rat models of post-traumatic osteoarthritis and to investigate its mechanism.Methods Thirty light knees of traumatic osteoarthritis model rats were randomly divided into 3 groups with 10 rats in each group,then the right knee joint cavities of theirs were injected with sterile PBS（ PBS group）,negative ERK2 siRNA lentivirus solution（ negative control group） and ERK2 siRNA lentivirus solution（ observation group）,respectively. After 8weeks,the rats were sacrificed and the general morphology of the articular cartilage was observed and scored,then the articular cartilage pathological changes were observed under the optical microscope with HE,toluidine blue and safranin ＂ O＂ staining,respectively,and the degree of cartilage degeneration was analyzed by Mankin semi-quantitative method. Meanwhile,the relative mRNA expression of matrix metalloproteinase 3（ MMP3）,MMP13 and Col2 a of cartilage tissues was analyzed by real-time fluorescence quantitative PCR. Results The articular cartilage surface ulcers and defects in the observation group were significantly lighter than those of the PBS group and negative control group,the extent of smooth articular surface of cartilage,crack area,loss of tissue,matrix staining,the proliferation of chondrocytes and the disorder of arrangement in the cartilage tissues of the observation group were lighter than those of the PBS group and negative control group; and the normal morphology and articular cartilage score in the observation group were significantly lower than those in the PBS group and negative control group（ all P 0. 05）,there was no significant difference between the PBS and negative control groups（ P 0. 05）. In addition,the relative mRNA expression levels of MMP3 and MMP13 in the observation group were lower than those in the PBS group and negative control group,while the relative mRNA expression le

关 键 词：骨性关节炎 软骨退变 慢病毒 小干扰RNA 细胞外信号调节激酶2 大鼠 

分 类 号：R684.3[医药卫生—骨科学]