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 results 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.

Key words: Plastrum testudinis effective constituent, GC-MS, HPLC, rat marrow-derived mesenchymal stem cell, MTT assay