淫羊藿苷影响软骨细胞基质分泌和基因表达的体内研究  

The in vivo study of the effect of icariin on ECM secretion and gene expression of chondrocytes

作　　者：郭筵[2] 吴瑕[1] 张璇[2] 李东晓[1] 肖玉梅[2] 樊渝江[2] 张兴栋[2] 

机构地区：[1]亚健康中医药防治实验室,四川省中医药科学院,成都610041 [2]四川大学,成都610064

出　　处：《中药药理与临床》2012年第2期42-45,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家重点基础研究发展计划项目(No.G2005cb623901);国家自然科学基金项目(No.51173113);四川省青年科技基金项目(No.090ZQ026-057,2011JQ0060)

摘　　要：目的:研究淫羊藿苷(icariin,Ica)对体内异位组织工程化软骨细胞基质分泌和特异性基因表达的影响,以评估Ica作为软骨组织工程促进剂的可能性。方法:分离、培养新生家兔关节软骨细胞;制备封装在包埋盒中含Ica终浓度为1×10-5M的软骨细胞-胶原水凝胶构建物,体外培养1周后植入家兔背部皮下,分期取材。激光共聚焦显微镜观察Ica对构建物中软骨细胞生长活性的影响;生化方法检测糖胺聚糖(gLycosaminogLycans,GAG)的分泌量;ELISA测定II型胶原(CollagenⅡ,ColⅡ)的合成情况;实时荧光定量聚合物酶联反应检测软骨特有基因表达情况。结果 :在凝胶中载入1×10-5M浓度范围内的Ica可使细胞-凝胶构建物外观更为致密,类似于软骨组织;维持软骨细胞表型及生长活性;促进GAG和ColⅡ的分泌;提高Aggrecan、CollagenⅡ和Sox9等软骨细胞特有基因的表达。结论:Ica可维持体内异位环境下组织工程化软骨细胞表型,促进细胞外基质分泌和软骨特有基因的表达,有望成为软骨组织工程中一种安全有效的促进剂。In order to evaluate the possibility of icarrin （Ica） as accelerator for cartilage tissue engineering, the effect of Ica on the ex- tracellular matrix （ECM） secretion and gene expressions of tissue engineered chondrocytes in vivo was investigated. Methods： Rabbit chon-drocytes were isolated from articular cartilage and cultured in medium containing ct-MEM. The stock solution of Ica was added into the neutral collagen （Col） solution and the final concentrations of Ica was 1×10-5 M. The second passage of chondrocytes was mixed gently with the pre- pared neutral Ica/Col solution, and then the cells-lea/Col mixture was then injected into the sealed boxes and formed hydrogel constructs. These sealed boxes were cultured in vitro for 1 week, implanted into subcutaneous tissue of rabbit＇s back and taken out at predetermined time intervals. The effect of Ica on chondrocyte morphology and viability in vivo was observed by confocal laser scanning microscope. The secretion of Col Ⅱ and glycosaminoglycans （GAG） were analyzed by biomedical assay and enzyme-linked immunosorbent （ELISA） assay, respective- ly. The expressions of cartilage-specified genes were also investigated by real time polymerase chain reaction （Real time PCR）. Results： The results found that the cell-Ica/Col hydrogel constructs were more similar in appearance to cartilage tissue compared with cell-Col hydrogel con- structs. The presence of Ica in Iea/Col hydrogel constructs could better maintain chondrocyte phenotype and viability. Moreover, the Iea might promote the secretion of ECM （GAG and Col Ⅱ） and enhance the expressions of cartilage-specified genes （Aggrecan, Col Ⅱ and Sox9） of chondrocytes. Conclusion： Ica could maintain the phenotype of tissue engineered chondrocytes in vivo and promote the secretion of ECM and the expressions of cartilage-specified genes of chondrocytes in vivo. Thus, Ica could be a potential promoting compound for cartilage tis- sue engineering.

关 键 词：淫羊藿苷 胶原 水凝胶 软骨组织工程 

分 类 号：R285[医药卫生—中药学]