睾酮对梅花鹿鹿茸间充质细胞增殖与分化的影响

Effects of testosterone on proliferation and differentiation of sika deer antler mesenchymal cells

通过原位杂交检测雄激素受体(androgen receptor,AR)mRNA在茸角中的表达情况;添加适当质量浓度的睾酮(testosterone,T)作用于体外分离培养的鹿茸间充质细胞后,应用MTS法检测睾酮对细胞增殖能力的影响,并通过荧光定量PCR方法检测软骨细胞标志物Ⅱ型胶原(typeⅡcollagen,ColⅡ)、聚集蛋白聚糖(aggrecan,AGC)和软骨寡聚基质蛋白(cartilage oligomeric matrix protein,COMP)的表达变化。结果显示,AR在茸角顶端的表皮层、真皮层和间充质层中均有表达;用质量浓度为10^(-6)g/L的睾酮处理鹿茸间充质细胞48 h后,MTS结果显示睾酮并未明显改变间充质细胞的增殖能力;相同质量浓度的睾酮处理间充质细胞24,48,72 h后,荧光定量PCR结果显示ColⅡ、AGG及COMP的表达水平均随时间推移逐渐增强,并在72 h时达到最大值。结果表明,睾酮对梅花鹿鹿茸间充质细胞的增殖能力无明显影响,但能显著促进其向软骨细胞的分化。

The aim of this study was to explore the role of testosterone(T)on the regeneration of sika deer antlers.In situ hybridization was used to detect the expression of androgen receptor(AR)mRNA in sika deer antlers.After the appropriate concentration of testosterone acted on the antler mesenchymal cells in vitro,the effect of testosterone on the proliferation of antler mesenchymal cells was estimated by MTS method,and the expression of chondrocyte markers type Ⅱ collagen(ColⅡ),aggrecan(AGC)and cartilage oligomeric matrix protein(COMP)were examined by fluorescence quantitative PCR.The results showed that AR was expressed in the epidermal layer,dermis and mesenchymal layer at the top of deer antler.After antler mesenchymal cells were treated with 10^(-6)g/L testosterone for 48 hours,MTS results showed that testosterone could not significantly change the proliferation activity of mesenchymal cells.After treatment with the same concentration of testosterone for 24,48 and 72 hours,the expression levels of ColⅡ,AGC and COMP were increased in a time-dependent manner,and reached the maximum at 72 hours.These results indicated that testosterone had no significant effect on the proliferation of antler mesenchymal cells,but could obviously promote the differentiation from mesenchymal cells to chondrocytes.