成骨细胞旁分泌影响软骨细胞中MMPs与TIMPs的表达  

Paracrine effect of chondrocytes on gene expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in osteoblasts

作　　者：关鹏[1] 赵伟[1] 张全有[2] 谢静[2] 尹利军[1] 赵虎成[2] 许建文[1] 

机构地区：[1]广西中医药大学第一附属医院骨科,广西壮族自治区南宁市530023 [2]清华大学生物力学研究所,北京市100083

出　　处：《中国组织工程研究》2015年第33期5306-5311,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:细胞之间体外共培养能最大限度的模拟体内真实的微环境,细胞划痕实验及炎症因子白细胞介素1β刺激后基质金属蛋白酶及基质金属蛋白酶抑制剂之间的平衡可能破坏,从而导致关节软骨细胞外基质的降解,软骨细胞功能的失调,关节软骨的退变。目的:在成骨细胞上清液与软骨细胞体外共培养下,观察炎症因子白细胞介素1β对体外培养的软骨细胞的迁移、基质金属蛋白酶及组织金属蛋白酶抑制剂表达的影响。方法:实验分为软骨细胞单培养组﹑软骨细胞与成骨细胞上清液共培养组和软骨细胞与成骨细胞上清液共培养+白细胞介素1β组,划痕实验观察3组24 h软骨细胞的迁移变化;半定量PCR实验分析以上3组24 h软骨细胞中基质金属蛋白酶1,2,3,9及组织金属蛋白酶抑制剂1,2,3,4的变化情况。结果与结论:与单培养组比较,共培养组和共培养+白细胞介素1β组细胞迁移率显著增加(P<0.01);与单培养组比较,共培养组中基质金属蛋白酶1,2,3,9基因表达明显增高(P<0.05),共培养+白细胞介素1β组基质金属蛋白酶1,3,9基因表达明显增高(P<0.01);与单培养组比较,共培养组和共培养+白细胞介素1β组中组织金属蛋白酶抑制剂1基因表达明显升高(P<0.01),组织金属蛋白酶抑制剂3,4基因表达明显下降(P<0.05)。提示成骨细胞上清液与软骨细胞共培养促进软骨细胞的迁移,增强软骨细胞中基质金属蛋白酶1,2,3,9的基因表达且调节组织金属蛋白酶抑制剂家族的基因表达。白细胞介素1β抑制共培养的软骨细胞迁移及组织金属蛋白酶抑制剂家族的基因表达。BACKGROUND： Ceil co-culture can maximize the simulation of in vivo microenvironment. Cell scratch test and interleukin-1β can destroy the balance between matrix metalloproteinases （MMPs） and matrix metalloproteinase inhibitors （TIMPs）, resulting in extracellular matrix degradation of the articular cartilage, functional disorders of chondrocytes and articular cartilage degeneration. OBJECTIVE： To study the effect of interleukin-1 β on migration, MMP and TIMP expression of chondrocytes co-cultured with osteoblast supernatant in vitro.METHODS： There were three groups： chondrocyte monoculture group, osteoblast＋chondrocyte group （co-culture group）, osteoblast＋chondrocyte＋interleukin-ll3 group （interleukin-1β group）. Cell scratch test was conducted to observe the migration of chondrocytes within 24 hours. Semi-quantitative PCR test was used to detect the changes in expressions of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, TIMP-3, TIMP-9 in chondrocytes within 24 hours.RESULTS AND CONCLUSION： Compared with the monoculture group, cell migration rate of the other two groups were increased significantly （P 〈 0.01）. Compared with the monoculture group, the gene expressions of MMP-1, MMP-2, MMP-3 and MMP-9 were increased significantly in the coculture group （P 〈 0.05）; the gene expressions of MMP-1, MMP-3, MMP-9 were increased significantly in the interleukin-1 β group （P 〈 0.01 ）. Compared with monoculture group, the gene expression of TIMP-1 was increased significantly （P 〈 0.01）, but the gene expressions of TIMP-3 and TIMP-4 were declined significantly （P 〈 0.05） in the other two groups. These findings indicate that co-culture of chondrocytes with osteoblasts can promote chondrocytes migration, enhance gene expression of chondrocytes MMP-1, MMP-2, MMP-3, MMP-9 and regulate gene expression of TIMPs family. Interleukin-1β inhibits the migration of chondrocytes co-cultured with osteoblasts and gene expression of TIMPs family.

关 键 词：骨关节炎 软骨细胞 成骨细胞 基质金属蛋白酶 组织构建 骨性关节炎 组织金属蛋白酶抑制剂 白细胞介素1β 共培养 

分 类 号：R318[医药卫生—生物医学工程]