兔关节软骨损伤生物标志物的蛋白质组学检测

背景:采用蛋白质组学的方法可以检测出关节、血液、尿液等体液中一些能反映关节软骨损伤程度的特异性标志物的水平。目的:进一步验证采用生物芯片技术发现兔关节软骨损伤生物标志物。方法:采用改良的Hulth方法建立兔关节软骨损伤模型,建模后自由活动,不固定伤肢。30min/d分2次驱赶,连续12周。以不做任何处理兔膝关节为正常对照。并在造模后0,4,8,12周时采取部分标本(血清、关节液),验证观察关节软骨的损伤程度;各个时间点的动物关节液、血清样本放入采用PBSⅡ-C型蛋白质芯片阅读机,采用CM10芯片检测。结果与结论:改良的Hulth造模方法建立膝关节软骨损伤模型,比较全面地反映了关节软骨损伤从早、中、晚、失代偿各期的变化。与正常对照组相比,血清学样品:模型组3475蛋白表达下调,7558,15475,33665蛋白表达上调;关节液样品:模型组7558,33278蛋白表达下调,3950,16055蛋白表达上调;质核比3475,7558,16884为血清学和关节液样本共有差异蛋白质谱波峰显著蛋白。结果提示样本中出现的部分差异蛋白质可能为关节软骨损伤的生物标志物。

Proteomic detection of biological markers for rabbit articular cartilage damage

BACKGROUND:The specific markers in joints,blood,urine and other body fluids that can reflect the degree of articular cartilage damage can be detected by proteomics.OBJECTIVE:To further verify the application of biological chip technology to find rabbit articular cartilage damage biological markers.METHODS:The improved Hulth method was used to establish the rabbit articular cartilage damage model.Postoperative free activities and the injury limb were not fixed,30 min/d points off twice for 12 weeks.The rabbit knee without any treatment was as normal control.At 4,8 and 12 weeks after modeling,part of the specimens (such as serum and joint liquid) were collected,and the articular cartilage damage degree was observed and verified;the joint fluid and serum sample of the animals at each time points were put into the PBS Ⅱ-C protein microarray reading machine and detected with CM10 chip.RESULTS AND CONCLUSION:The articular cartilage damage model established by improved Hulth method could quite comprehensively reflect the change of articular cartilage damage from the early,middle,and late and decompensate periods.Compared with normal control group,the expression of 3 475 protein in serum specimen was decreased and the expression of 7 558,15 475 and 33 665 protein was increased in model group;the expression of 7 558 and 33 278 protein in joint fluid specimen was decreased and the expression of 3950 and 16 055 protein was increased in model group.The significant differences protein peak spectrum with the nuclear mass ratio of 3 475,7 558,16 884 was expressed in both serum specimen and joint fluid specimen.Part of the differential protein appeared in the specimen may be the articular cartilage damage biological markers.