【摘要】 目的:检测基质金属蛋白酶(MMPs)在兔膝关节软骨细胞损伤后不同时间点的表达变化。方法:无菌条件下获取兔膝关节软骨细胞,原代体外培养软骨细胞,6孔板内制备细胞损伤模型。显微镜观察正常细胞和划伤后1、3和7 d的软骨细胞的增殖情况。实时荧光定量PCR检测正常软骨细胞和损伤后1、3和7 d MMPs的mRNA表达水平。结果:成功分离关节软骨细胞,原代培养后成功建立细胞损伤模型。MMP-2 mRNA表达水平在损伤后1、3、7 d比正常组均升高,其中损伤后7 d最高(P<0.05～P<0.01)。MMP-3 mRNA表达水平在细胞损伤后1、3、7 d与正常组均明显下降,损伤后7 d最低(P<0.01)。MMP-9 mRNA表达水平在损伤后1、3、7 d均比正常组升高,划伤后3 d最高,随后开始下降(P<0.01)。结论:MMPs在细胞损伤后的不同时间点表达不同,可为调节细胞外基质基因治疗关节软骨损伤提供实验依据。更多还原

【Abstract】 Objective: To explore the mRNA expressions of matrix metalloproteinases( MMPs) in rabbit articular chondrocytes at different time-points after injury. Methods: The rabbit knee joint cartilage cells were obtained under sterile condition,primarily cultured in vitro,and established the cell scratch model in six-well plates. The proliferation of chondrocytes after 1,3 and 7 days of scratching and normal cells were observed under the microscope. The MMPs mRNA expression levels in normal cells and different time-points after injury were detected using real-time PCR. Results: The articular chordrocyte was successfully isolated,the scratch model of chondrocytes was established after primary culture. Compared with normal cells,the mRNA expression levels of MMP-2 gene in chondrocytes increased at 1,3 and 7 days aftere injury,and which was the highest on the seventh day( P < 0. 05 to P < 0. 01). Compared with normal cells,the mRNA expression levels of MMP-3 gene in chondrocytes decreased significantly at 1,3 and 7 days after injury,and which was the lowest at on the seventh day( P < 0. 01). Compared with normal cells,the mRNA expression levels of MMP-9 gene in chondrocytes increased at 1,3 and 7 days after injury,and which was the highest on the third day and then began to decrease( P < 0. 01).Conclusions: The mRNA expression levels of MMPs are different after injury,which can provide the experimental evidence in gene therapy of articular cartilage injury.更多还原