氧环境影响关节软骨细胞表型的相关研究（临床医学范文）.docVIP

氧环境影响关节软骨细胞表型的相关研究（临床医学范文） 目录 TOC \o 1-9 \h \z \u 目录 1 正文 2 文1：氧环境影响关节软骨细胞表型的相关研究 2 1 材料和方法 3 1.1 关节软骨细胞的分离和培养 3 1.2.1 细胞标本收集 4 1.2.2 RT-PCR反应 4 1.3 软骨细胞表型鉴定 4 1.3.1 II型胶原和Aggrecan的免疫化学染色 5 1.3.2 阿利新蓝染色 5 2 结果 5 2.1 软骨细胞形态学观察 5 文2：氧环境影响关节软骨细胞表型的相关研究 5 1 材料和方法 7 1.1 关节软骨细胞的分离和培养 7 1.2.1 细胞标本收集 8 1.2.2 RT-PCR反应 8 1.3 软骨细胞表型鉴定 8 1.3.1 II型胶原和Aggrecan的免疫化学染色 8 1.3.2 阿利新蓝染色 9 2 结果 9 2.1 软骨细胞形态学观察 9 2.2 软骨细胞标志性基因和分化相关基因表达（图1、表2） 9 2.3 细胞免疫化学染色和阿利新蓝染色（图2） 10 2.4 统计分析 10 3 讨论 10 参考文摘引言： 12 原创性声明（模板） 13 文章致谢（模板） 14 正文 氧环境影响关节软骨细胞表型的相关研究（临床医学范文） 文1：氧环境影响关节软骨细胞表型的相关研究 Abstract: ［Objective］To examine the specific gene and protein expression of primary and passaged chondrocytes in normal oxygen teion and hypoxia. ［Methods］Articular chondrocytes were isolated from limb joint cartilage of C57BL/6 mice (3～5 days). Primary chondrocytes (P0) and passaled chondrocytes (P1, P2) were cultured in normal oxygen teion and hypoxia respectively for two days.The mRNA levels of collagen II, aggrecan, sox9, Indian hedgehog (ihh), parathyroid hormone-related protein (PTHrP), bone morphogenetic protein (BMP)-4 and wnt5a were examined with RT-PCR, and protein levels of collagen II,aggrecan were examined with immunohistochemistry and toluidine blue staining.［Results］During the culture of primary chondrocyte, the expression of collagen II and aggrecan increased under hypoxia, mRNA levels of collagen II, wnt5a increased and ihh decreased at the same time.The mRNA levels of special genes and differentiation related genes of passaged chondrocytes were not altered under hypoxia.［Conclusion］Protein and mRNA level of Collagen II of primary chondrocyte under hypoxia increased. It may be regulated by chondrocyte differentiation gene ihh,and wnt5a. The chondrocyte phenotype could not be resumed under hypoxia through short-term monolayer culture in vitro. Key words：chondrocyte; hypoxia; collagen type II; aggrecan 正常生理情况下，关节软骨