淫羊藿总黄酮调控骨性关节炎p38MAPK信号转导通路的研究

项目名称： 淫羊藿总黄酮调控骨性关节炎p38MAPK信号转导通路的研究

项目编号： No.81001529

项目类型： 青年科学基金项目

立项/批准年度： 2011

项目学科： 金属学与金属工艺

项目作者： 于波

作者单位： 山东中医药大学

项目金额： 10万元

中文摘要： 分离培养人OA软骨细胞，MTT检测淫羊藿对软骨细胞的增殖影响。观察淫羊藿对人OA软骨组织GAG和II型胶原降解的作用。检测淫羊藿对软骨细胞aggrecanases、MMPs的作用；甲苯胺蓝、番红O染色及Real-time PCR检测淫羊藿对软骨细胞成软骨分化的作用；TUNEL、Hoechst 33342染色检测淫羊藿对软骨细胞凋亡的作用；western blot检测淫羊藿经MAPK信号通路保护软骨细胞去分化的机制。 研究结论如下：1.淫羊藿可显著抑制OA软骨组织内GAG及Ⅱ#22411;胶原降解；2.淫羊藿可降低aggrecanases-1/-2和MMP-1/-3/-13mRNA表达，提高TIMP-1mRNA表达；3.淫羊藿可抑制软骨细胞凋亡；4.淫羊藿可降低软骨细胞MAPK（ERK、JNK和p38 MAPK）磷酸化。 本课题阐明了淫羊藿经MAPK信号通路减缓软骨组织基质降解、抑制软骨细胞凋亡，从而保护OA软骨及软骨细胞的去分化。

中文关键词： 淫羊藿；MAPK信号通路；软骨细胞；骨性关节炎

英文摘要： Chondrocytes were isolated from OA patients, while MTT assay was performed for the detection of cell viability. Treating cartilage explants with icariin, Type Ⅱcollagen and GAG levels were determined; We tested whether icariin regulated mRNA expression of aggrecanases and MMPs from the chondrocytes. Chondrocytes were stained with toluidine blue and safranin O for glycosaminoglycan distribution; For the detection of in situ DNA breaks, the TUNEL reaction and Hoechst 33342 Staining were applied; MAPK pathway activation, as measured by western blotting, was analyzed to understand the molecular mechanism by which icariin inhibits OA induced de-differentiation. Results: 1.Icariin effectively blocks TNF-αnduced degradation of GAG and collagen II from human osteoarthritic cartilage explants; 2.The expression of aggrecanase-1, aggrecanase-2, MMP-1, MMP-3, and MMP-13 mRNA decreased significantly while TIMP-1 mRNA increased in the icariin treated group; 3.Icariin dramatically decreased TNF-αnduced apoptosis, as indicated by a decreased red fluorescent signal in the nucleus; 4. Icariin suppressed ERK, JNK and p38 phosphorylation and activation. In conclusion, our studies demonstrate that icariin has a positive role in protecting OA cartilage and chondrocytes. In addition, the mechanism for the potency of icariin was probably accomplished by suppressing MAPK pathway phosphorylation.

英文关键词： icariin;MAPK;chondrocyte;osteoarthritis