安徽医科大学学报Issue(4)：446-451,6.
环巴胺对佐剂性关节炎大鼠关节软骨细胞增殖凋亡的影响及其抗凋亡机制
Effects of cyclopamine on the proliferation and apoptosis of chondrocytes in adjuvant-induced arthritis rats and its anti-apoptotic mechanisms
摘要
Abstract
Objective To explore the effects of cyclopamine, a hedgehog ( Hh) signal pathway inhibitor, on the proliferation and apoptosis of articular chondrocytes in adjuvant-induced arthritis ( AIA) rats and its anti-apoptotic mechanisms. Methods Freund’ s complete adjuvant was used to induce AIA. Articular chondrocytes were isolated and cultured by trypsin and collagenase digestion method. AIA articular chondrocytes were treated by cyclopamine in vitro. The cell proliferation was detected by MTT assay. The cell apoptosis was evaluated with DNA agarose gel electrophoresis, hoechst staining and flow cytometry analysis. The levels of Shh, Ptch1, Gli1, Bcl-2, Bax and Caspase-3 mRNA were detected by RT-PCR. Results Cyclopamine (0. 08, 0. 4, 2, 10, 50 μmol/L) could in-crease AIA articular chondrocytes proliferation in a dose-dependent. DNA ladder pattern was formed typically in AIA articular chondrocytes, while cyclopamine (0. 4, 2, 10μmol/L) treatment could reduce the formation of DNA ladder . AIA articular chondrocytes displayed nuclei fragmentation with condensed chromatin and cyclopamine could promote the number of uniform blue fluorescent cells. Flow cytometry analysis also indicated cyclopamine could vis-ibly reduce the rate of apoptotic cells in AIA articular chondrocytes. Compared with normal group, the levels of Hh pathway-related genes ( Shh, Ptch-1 and Gli1 ) mRNA apparently increased in AIA articular chondrocytes. Anti-apoptotic gene Bcl-2 mRNA level significantly decreased, while pro-apoptotic genes Bax and Caspase-3 mRNA lev-els were significantly increased. Cyclopamine treatment could inhibit the excessive activation of Hh pathway, up-regulate Bcl-2 mRNA expression, and reduce Bax and Caspase-3 mRNA expressions. Conclusion Cyclopamine could promote the proliferation and inhibit excessive apoptosis of AIA articular chondrocytes, which might be relat-ed to regulation of Bcl-2, Bax, and Caspase-3 expressions. Our results suggest that intervention of Hh signal in ar-ticular chondrocytes has potential therapeutic significance for articular cartilage protection in rheumatoid arthritis.关键词
Hedgehog信号通路/环巴胺/佐剂性关节炎/关节软骨细胞/增殖/凋亡Key words
hedgehog signal pathway/cyclopamine/adjuvant-induced arthritis/articular chondrocytes/prolifera-tion/apoptosis分类
医药卫生引用本文复制引用
丁婧,李荣,胡向阳,蔡莉,张晓亮,汪巧..环巴胺对佐剂性关节炎大鼠关节软骨细胞增殖凋亡的影响及其抗凋亡机制[J].安徽医科大学学报,2015,(4):446-451,6.基金项目
国家自然科学基金(编号:81102273) （编号:81102273）
教育部博士学科点专项科研基金(编号:20113420120005) （编号:20113420120005）
