中药龟板提取物浸膏经硅胶柱层析,用石油醚-乙酸乙酯作洗脱剂,梯度洗脱,得到16个组分;采用MTT法和流式细胞技术研究了它们对鼠骨髓间充质干细胞(rMSCs)的增殖作用,实验结果显示组分Ts-12具有促进rMSCs增殖的作用(p〈0.05),而组分Ts-4则表现出抑制rMSCs增殖的作用(p〈0.05),其它样品对rMSCs的增殖作用不显著(p〉0.05),不具统计学意义.采用了GC-MS分析各组分化学成分,并用HPLC和9个标准物质鉴定确证了组分Ts-12中含有十六酸甲酯(s-1)、十六酸乙酯(s-3)、十八酸甲酯(s-4)和甾醇(s-7)以及十四酸甾醇酯(s-8),组分Ts-4主要含有十八酸(s-5).对标准品也采用了MTT法和流式细胞仪研究了它们对rMSCs的增殖作用,结果表明十四酸甾醇酯和十六酸甲酯具有促进rMSCs增殖的作用,而十八酸具有抑制作用.
Ointment extracted from Plastrum Testudinis was separated by silica gel column chromatography, gradiently eluted by petroleum ether-ethyl acetate, and 16 (Ts-1-Ts-16) components were obtained, which were biologically evaluated by MTT assay and flow cytometry on the proliferation of rat marrow-derived mesenchymal stem cells (rMSCs). Results showed that Ts-12 induced proliferation compared with the control group (p〈0.05), while Ts-4 showed inhibitive effect. Other groups showed no significance (p〉0.05). GC-MS was used to analyze the chemical components in different groups. The experimental resuits of the analysis suggest that palmitic acid methyl ester (S-1), palmitic acid ethyl ester (S-3), stearic acid methyl ester (S-4), stearic acid ethyl ester (S-6), cholesterol (S-7) and cholesterol myristate (S-8) are identified from Ts-12, while stearic acid (S-5) from Ts-4 by HPLC method and compared to 9 standard materials. The proliferation effects of standard materials were investigated by MTT method and flow cytometry. Cholesterol myristate and palmitic acid methyl ester showed proliferation bioactivity while stearic acid showed inhibition effect.