【作者】 张凯；

【导师】 雷光华；

【作者基本信息】 中南大学 ， 外科学， 2012， 硕士

【摘要】 目的：探讨骨桥蛋白对体外培养的人膝骨关节炎软骨细胞IL-1β和TNF-a表达的影响,为明确骨关节炎发病机理及其防治提供理论依据。方法：选取接受全膝关节置换手术的原发性膝关节骨关节炎患者关节面截骨软骨标本16份,进行体外培养获得纯化的软骨细胞,取第一代软骨细胞,分为三组：A组(空白对照组)、B组(OPN100ng/ml干预组)和C组(OPN1μg/ml干预组)。在37℃5%C02温箱培养48h后,采用Real-time Q PCR检测软骨细胞IL-1βmRNA和TNF-amRNA表达水平。用SPSS19.0统计软件包分析OPN干预后表达的差异。结果：1、B组与A组相比,人膝OA软骨细胞IL-1β mRNA表达水平(0.562±0.272倍)下降,两者之间差异具有统计学意义(P<0.05)。2、C组与A组相比,人膝OA软骨细胞IL-1β mRNA表达水平(0.305±0.067倍)下降,两者之间差异具有统计学意义(P<0.05)。3、B组与C组人膝OA软骨细胞IL-1β mRNA表达水平相比,两者之间差异具有统计学意义(P<0.05),且C组人膝OA软骨细胞IL-1βmRNA表达水平低于B组。4、人膝OA软骨细胞IL-1β mRNA表达水平与OPN干预浓度呈负相关。(r=-0.557,P<0.01)5、B组与A组相比,人膝OA软骨细胞TNF-a mRNA表达水平(0.330士0.075倍)下降,两者之间差异具有统计学意义(P<0.05)。6、C组与A组相比,人膝OA软骨细胞TNF-a mRNA表达水平(0.286士0.071倍)下降,两者之间差异具有统计学意义(P<0.05)。7、B组与C组人膝OA软骨细胞TNF-a mRNA表达水平相比,两者之间差异无统计学意义(P>0.05)。结论：1、OPN下调人膝OA软骨细胞IL-1β mRNA (?)(?)TNF-amRNA表达。2、人膝OA软骨细胞IL-1β mRNA表达水平与OPN干预浓度呈负相关。更多还原

【Abstract】 Objective:This study is to discuss the effect of osteopontin (OPN) on expression of interleukin1beta and tumor necrosis factor alpha of human knee osteoarthritic chondrocytes in vitro, in order to shed new light on the role of OPN in Osteoarthris and provide a new strategy to cure Osteoarthris.Methods:The study was performed on16specimens of cartilage obtained from osteoarthritic patients who undergo total knee arthoplasty surgery. All the cartilage samples were cultured and purified to obtain chondrocytes in vitro. Taking the firstpassaged cells into three groups:(1) Group A (the control group);(2) Group B (treated with100ng/ml OPN for48hours);(3) Group C (treated with1μg/ml OPN for48hours). Then design and validate the real-time quantitative polymerase chain reaction (Q-PCR) assays with SYBR Green dyes for simultaneous quantification of the expression of the interleukin1beta mRNA and tumor necrosis factor alpha mRNA. All results statistics were analysed by SPSS19.0.Results:1. Compared to controls, mRNA expression of interleukin1beta of Group B decreased (0.562±0.272folds), and there is a statistically significant difference.(P<0.05).2. Compared to controls, Compared to controls, mRNA expression of interleukin1beta of Group C decreased (0.305±0.067folds), and there is a statistically significant difference.(P<0.05).3. Compared to Group B, mRNA expression of interleukin1beta of Group C decreased, and there is a statistically significant difference.(P<0.05).4. There is a negative correlation between the expression of interleukin1beta mRNA and the concentration of OPN in the three groups (r=-0.557,P<0.01).5. Compared to controls, mRNA expression of tumor necrosis factor alpha of Group B decreased (0.330±0.075folds), and there is a statistically significant difference.(P<0.05).6. Compared to controls, mRNA expression of tumor necrosis factor alpha of Group C decreased (0.286±0.071folds), and there is a statistically significant difference.(P<0.05).7. The difference of mRNA expression of tumor necrosis factor alpha between Group B and Group C, showed no difference of statistically significant(P>0.05).Conclusion:1. OPN down-regulate mRNA expression of interleukin1beta and tumor necrosis factor alpha in chondrocytes of patients with osteoarthritis of the knee.2.There may be a negative correlation between the expression of interleukin1beta mRNA and the concentration of OPN in vitro cultured human of knee arthritic cartilage cells in Osteoarthris.更多还原