SOCS3在骨关节炎中的作用.DOC

SOCS在骨关节炎的表达和含义 孙晓辉新乡医学院第一附属医院骨科(IL-1β)和肿瘤坏死因子α(TNFa)，在关节损伤中起作用。目的：在本研究中，我们研究了正常和关节炎患者的软骨细胞表达SOCS1，2,3和CIS-1的基因表达，以及分析IL-1β和TNFα对上述SOCS家族成员mRNA的表达水平的影响。方法：在晚期OA和因车祸需要置换的关节中通过胰蛋白酶、透明质酸酶和胶原蛋白酶B分离软骨细胞。从软骨细胞提取总RNA，用定量RT-PCR检测SOCS1,2,3和CIS-1 mRNA。此外，健康的软骨细胞用或不用 IL-1β和 oncostatin M (OSM, both 2.5 ng/ml) 或TNFa (10 ng/ml)培养。用单个细胞因子处理24和72小时后收获细胞，和长期培养周期性用细胞因子刺激长期培养维持4-5周。总ＲＮＡ被提取和qRT－PCR检测ｍＲＮＡ水平。结果：与对照相比，ＯＡ样本中ＳＯＣＳ２和ＣＩＳ－１ｍＲＮＡ减少8倍，而在ＯＡ和健康人中显示类似的ＳＯＣＳ１和ＳＯＣＳ３的表达模式。用IL-1b和联合 OSM 或TNFa长期处理后ＳＯＣＳ２和ＣＩＳ－１ｍＲＮＡ减少5和３倍。结论：本研究证明了ＯＡ中ＳＯＣＳ２和ＣＩＳ－１减少了表达，而ＳＯＣＳ１和ＳＯＣＳ３的表达未受影响。用炎性细胞因子长期处理软骨细胞模拟ＯＡ减少ＳＯＣＳ２和ＣＩＳ－１的表达，意味着潜在的正反馈机制。 关键词：细胞因子信号抑制因子；骨关节炎 The expression and implication of SOCS in osteoarthritis (OA) SUN Xiao-hui Department of Orthopedics, The First Affiliated Hospital of Xinxiang Medical College, Henan province China Weihui 453100 Abstract：Suppressor of cytokine signalling (SOCS) proteins are inhibitors of cytokine signalling that function via the JAK/STAT pathway. SOCS1, 2, and 3 and CIS-1 belong to SOCS family. Proinflammatory cytokines, such as interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNFa) secrected by articular chondrocytes may be induced by mechanotransduction and contribute to cartilage destruction. Objectives: In this study, we have studied the gene expression of SOCS1, 2, 3 and CIS-1 in healthy and OA human chondrocytes, and also analyzed the effects of IL-1βand TNFa on the levels of mRNA encoding these SOCS family members. Methods: Chondrocytes from patients of OA and traffic accident were isolated by sequential treatment with trypsin, hyaluronidase and collagenase B. Total RNA were extracted from the same chondrocytes, and the levels of SOCS1, 2, 3 and CIS-1 mRNA were determined by qRT-PCR. In addition, healthy chondrocytes were cultured with and without a mixture of IL-1b and oncostatin M (OSM, both 2.5 ng/ml) or TNFa (10 ng/ml). The short-term cultures with single cytokine treatment were harvested 24 and 72 h after treatment, and the long-term cultures were main