摘要
为探讨中药丹参的主要活性成份丹酚酸B对人软骨细胞系C28112细胞的增殖作用及其调节作用机制中相关因子的初步辨识采用MTS法检测丹酚酸B的对软骨细胞作用的有效浓度;吖碇橙荧光标记软骨细胞的DNA及RNA,观察细胞分裂及形态学改变;Western Blotting(WB)检测β-Catenin及新型软骨生长因子类细胞介素1的蛋白质表达。结果显示MTS法检测加入丹酚酸B后的所有剂量实验组与对照组比较吸光度值均有不同程度的增高,差异均有统计学意义(P<0.01);丹酚酸B实验组可见明显核分裂及较多的双核现象,细胞增生较活跃;半定量WB分析结果显示丹酚酸B实验组比对照组的类细胞介素1蛋白表达量显著升高,β-Catenin蛋白质表达量亦显上调趋势。表明丹酚酸B能促进人软骨细胞的增殖并上调了相关因子的表达。其作用机制可能与丹酚酸B作用于软骨细胞中Wnt信号调节通路,上调信号调节因子β-Catenin表达量,激活靶基因类细胞介素1的转录并促进其表达释放有关。
To investigate the proliferative function of the major component of Salvia, the salvianolic acid B, on human chondrocyte cell line C28112 and to identify associated factors in regulatory signaling path-ways, the study investigated the effective concentration of salvianolic acid B for chondrocyte growth in culture using MTS method and observe the cell division and the morphological changes by using Acridine orange fluorescent labeling of DNA and RNA to as well as detect protein expression levels of 15 - Catenin and the new types of cartilage growth factor cytokine-like 1 （ CYTL - 1 ） by Western Blotting （ WB ） method. The results showed that the absorbance values （A） in experimental group detected by the MTS methods were higher than the control group in all doses with statistically significance （p 〈 0. 01 ） ; In salvianolic acid B experimental group, splitting cell nucleus with more dual-nucleus cells were obviously seen, indicating more active cells in growth process; the semi-quantitative Western Blot analysis showed that the CYTL - 1 expression in salvianolic acid B experimental group was significantly up-regulated com- pared to the control group, the expression of 15 - Catenin in experimental group showed a trend of up-reg- ulation. So, salvianolic acid B promoted the growth of human chondrocytes. The functional mechanism could be that the salvianolic acid B acted on the Wnt signal transduction pathway in chondrocytes, up- regulated the expression level of 15 - Catenin, activated the transcription of the target gene CYTL - 1, and then stimulated its expression and release.
出处
《中山大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第4期110-115,共6页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
广东省自然科学基金资助项目(S2012010010616)
广东省科技计划资助项目(2012A030400036)
广州市科技计划资助项目(20121A021009)
广州市医药卫生科技重点资助项目(2012J4100027)
