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- 2017
IL-1β和MMP-13在兔骨关节炎模型软骨和滑液中的表达
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Abstract:
摘要：目的 评价兔膝关节内侧半月板损伤制作骨关节炎(OA)模型的方法的可行性，探讨白细胞介素-1β(IL-1β)和金属蛋白酶-13(MMP-13)在兔骨关节炎模型软骨和滑液病变中的作用机制。方法 新西兰大白兔40只，随机分为实验组(n=30)和对照组(n=10)，于术后2、6、12周观察股骨髁关节软骨的病理变化并进行评分；用免疫组化方法检测关节软骨中IL-1β和MMP-13的表达情况；用酶联免疫吸附法测定关节液中IL-1和MMP-13的含量。结果 实验组和对照组在大体和病理观察不同时间点关节软骨评分及HE染色差异均有统计学意义(P<0.05)。免疫组织化学检测结果显示IL-1β在两组中均有表达，实验组和对照组2周时细胞分数差异无统计学意义，6、12周差异有统计学意义(P<0.05)。MMP-13在对照组关节软骨细胞中未见表达，实验组的关节软骨细胞中可见MMP-13蛋白的表达，差异有统计学意义(P<0.05)。关节滑液中IL-1β表达和在软骨中表达一致。结论 采用损伤兔膝关节半月板的方法可建立合理的动物OA模型；IL-1β和MMP-13在OA发病过程中表达水平有明显变化，需要进一步的临床研究来探讨其是否可以作为早期诊断OA的指标之一。
ABSTRACT: Objective To investigate the feasibility of osteoarthritis (OA) model in rabbits by injuring medial meniscus so as to understand the role of interleukin-1 beta (IL-1β) and matrix metalloproteinase-13 (MMP-13) in pathological mechanism of articular cartilage and synovial fluid of OA model in rabbits. Methods We randomly divided 40 New Zealand white rabbits into experimental group (n=30) and control group (n=10). Pathological changes in articular cartilage of the femoral condyle were scored at weeks 2, 6 and 12 after surgery. We detected the expressions of IL-1β and MMP-13 by immunohistochemistry. The cell fractions of IL-1β and MMP-13 were recorded by ELISA. Results The articular cartilage score and HE staining significantly differed at various time points of gross and pathological observation between control group and experimental group (P<0.05). Immunohistochemistry showed that IL-1β was expressed in both groups and that the cell fraction differed significantly at weeks 6 and 12 ？？(P<0.05),？？ but not at week 2 between the two groups. MMP-13 protein expression was not detected in articular condrocytes in control group, but was detected in experiment group with a significant difference (P<0.05). IL-1β expression was consistent in articular cartilage and synovial fluid. Conclusion The reasonable rabbit animal OA model could be established by knee meniscus injury caused by surgical method. Expressions of IL-1β and MMP-13 change obviously in the pathomechanism of OA. Further clinical studies are needed to determine whether they can be used as markers in early diagnosis of OA
| [1] | KEKKEY WN. Textbook of Rheumatology 6th ed[M]. Philadelphis: Saunders, 2001:18-24. |
| [2] | PELLTER JP, JOVANOVIC D, FERNANDES JC, et al. Reduced progression of experimental osteoarthritis in vivo by selective inhibition of Inducible nitric oxidesynthase[J]. Arthritis Rheum,1998, 41(7):1275-1286. |
| [3] | ASSIRELLI E, PULSATELLI L, DOLZANI P, et al. Human osteoarthritic cartilage shows reduced in vivo expression of IL-4, achondroprotective cytokine that differentially modulates IL-1β stimulated production of chemokines and matrix-degrading enzymes in vitro[J]. PLoS One, 2014, 9(5):e96925. |
| [4] | BREYHOLZ HJ, WAGNER S, LEVKAU B, et al. A18F-radiolabeled analogue of CGS27023A as a potential agent for assessment of matrix-metalloproteinase activity in vivo[J]. QJ Nucl Med Mol Imaging, 2007. 51(1):24-32. |
| [5] | MESZAROS E, MALEMUD CJ. Prospects for treating osteoarthritis: enzyme-protein interactions regulating matrix metalloproteinase activity[J]. Therapeutic Advances in Chronic Disease, 2012, 3(5):219-229. |
| [6] | ATTUR M, STATNIKOV A, SAMUELS J, et al. Plasma levels of interleukin-1 receptor antagonist (IL1Ra) predict radiographic progression of symptomatic knee osteoarthritis[J]. Osteoarthr Cartilage, 2015, 23(11):1915-1924. |
| [7] | 王欣，秦宇. 白细胞介素-1β对大鼠软骨细胞MMP-13表达的影响及miR-27b的调控作用[J]. 天津医药, 2015, 43(8):871-875. |
| [8] | YANG L, ZHANG J, WANG G. The effect of sodium hyaluronate treating knee osteoarthritis on synovial fluid interleukin-1β and clinical treatment mechanism[J]. Pak J Pharm Sci, 2015, 28(1):407-410. |
| [9] | GHIVIZZANI SC, KANG B, GEORGESEU HI, et al. Constitutive Intra-articular expression of human IL-1 beta following gene transfer to rabbit synovium produces all major pathologies of human rheumatoid arthritis[J]. Immunol, 1997, 159(7):3604-3612. |
| [10] | COOK JL, ANDERSON CC, KREEGER JM, et al. Effects of human recombinant interleukin-1β on canine articular chondroocytes in three dimensional culture[J]. AJVR, 2000, 61(7):766-770. |
| [11] | 宋朋飞，阚卫兵，袁琴，等. 基质金属蛋白酶与骨关节炎的关系[J]. 广东医学, 2011, 32(20):2736-2738. |
| [12] | MITEHELL PG, MAGNA HA, REEVES LM, et al. Cloning, expression and type-Ⅱ collagenolytic activity of matrimetalloProteinase-13 from human osteoarthritic cartilage[J]. J Clin Invest, 1996, 97(3):761-768. |
| [13] | SALMINEN H, SAAMANEN AM, VANKEMMELBEKE M, et al. Differential expression patterns of matrix metalloproteinases and their inhibitors during the development of osteoarthritis in a transgenic mouse model[J]. Ann Rheum Dis, 2002，61(7):591-597. |
| [14] | 吴宏斌，杜靖远，郑启新，等. 基质金属蛋白酶-1在创伤性骨关节炎软骨及滑膜中的表达[J]. 中华创伤杂志, 2003, 19(1):42-46. |
| [15] | PELLETIER JP, MCCOLLUM R, CLOUTIER JM, et al. Synthesis of metalloproteases and interleukin-6 (IL-6) in human osteoarthritic synovial membrance is an IL-1 mediated process[J]. J Rheumatol Suppl, 1995, 43:109-114. |
| [16] | MATHIEU P. Interleukin-I: Its role, its dosage the difficulties in advance in arthritis[J]. Rev Prat, 1999, 13:515-518. |
| [17] | 周晓莉.IL-1 β在骨关节炎发病及治疗应用中的研究现状[J]. 海南医学，2010, 21(3):101-104. |
| [18] | PELLETIER JP, FFAURE MP, DIBATTISTA JA, et al. Coordinate synthesis of stromelysin, interleukin-1, and oncogene proteins in experimental osteoarthrisis.An immunohistochemical study[J]. Am J Pathol, 1993, 142(1):95-105. |
| [19] | SHINMEI M, MASUDA K, KIKUCHI T, et al. Interleukin-l, tumor necrosis factor, and interleukirl-6 as mediators of cartilage destruction[J]. Semin Arthritis Rheum, 1989, 18(suppl 1):27-32. |
| [20] | 李敏，梁翼，吴晓惠，等. 伴骨髓水肿的膝骨性关节炎患者白细胞介素-16、白细胞介素-6、肿瘤坏死因子-α的相关性研究[J]. 中华风湿病杂志, 2012, 16(2):107-110. |
| [21] | HERAUD F, HERAUD A, HARMAND MF. Apoptosis in normal and osteoarthritis human artcular cartilage[J]. Ann Rheum Dis, 2000, 59(12):959-965. |
