目的 建立高良姜提取物指纹图谱，考察其对B16鼠黑素瘤细胞增殖、酪氨酸酶活性及黑素量的影响，并分析其谱效关系。方法 HPLC法建立高良姜醇提物（G1）、G1水沉物（G2）、G2大孔树脂纯化物（G3）、G3聚酰胺精制物（G4）指纹图谱；MTT法检测各提取物对B16细胞增殖的影响；比色法检测其对酪氨酸酶活性和黑素量的影响；灰关联度分析法研究其谱效关系。结果 与对照组相比，G1浓度为1.00～5.00 μmol/L、G2浓度为 0.50～5.00 μmol/L、G3浓度为0.25～5.00 μmol/L，G4浓度为0.25～10.00 μmol/L，能显著促进B16细胞增殖（P＜0.01）；G1、G2浓度为0.50 μmol/L，G3、G4浓度为0.50～1.00 μmol/L，能显著增强酪氨酸酶的活性（P＜0.05、0.01）；G1浓度为0.50、5.00 μmol/L，G2、G4浓度为0.50～5.00 μmol/L，G3浓度为0.25～5.00 μmol/L时，促黑素合成作用显著（P＜0.05）；在相同浓度下，G4促B16细胞增殖和增强酪氨酸酶活性的作用最强。谱效关系分析表明，与药效关联度最高的是高良姜素（7号峰）。G1、G2、G3、G4含高良姜素分别为27.61、88.72、348.53、693.35 mg/g，验证实验结果表明高良姜素在0.25～10.00 μmol/L均有显著的促B16细胞增殖、增强酪氨酸酶活性和促进黑素量升高的作用。结论 随着高良姜素量的增加，高良姜各提取物对B16细胞增殖、酪氨酸酶活性及黑素量的作用逐渐增强，表明高良姜素是高良姜治疗白癜风的有效物质之一。

Objective To establish the fingerprint of the extract from the rhizome of Alpinia officinarum, to investigate the effects on the proliferation, tyrosinase activity, and melanogenesis in melanoma B16 cells, and to analyze the spectrum-effect relasionship. Methods The fingerprint of alcohol extract from the rhizome of A. officinarum (G1), water sediments of G1 (G2), macroporous resin purified extract of G2 (G3), and refined polyamide extract of G3 (G4) were established by HPLC, and the proliferation of each extract on B16 cells was measured by MTT. The tyrosinase activity and melanin content were measured by colorimetry assay. The spectrum-effect relasionship was analyzed by grey relation analysis. Results Compared with the control group, the proliferation of B16 cells was promoted significantly after treatment with 1.00—5.00, 0.50—5.00, 0.25—5.00, and 0.25—10.00 μmol/L of G1, G2, G3, and G4, respectively. The tyrosinase activity was promoted by G1 and G2 with 0.50 μmol/L, and G3 and G4 with 0.50—1.00 μmol/L (P < 0.05, 0.01); The synthesis of melanin was promoted after the treatment with 0.5 and 5.00 μmol/L of G1, 0.50—5.00 μmol/L of G2 and G4, and 0.25—5.00 μmol/L of G3 (P < 0.01); At the same concentration, G4 had the strongest effect to promote the cell proliferation and to increase the tyrosinase activity. The galangin (peak 7) had the highest correlation with pharmacodynamics, and the contents of galangin in G1, G2, G3, and G4 were 27.61, 88.72, 348.53, and 693.35 mg/g, respectively. The proliferation, tyrosinase activity, and galangin content were promoted significantly after the treatment with galangin (0.25—10.00 μmol/L) by the verification experiment. Conclusion The effect of the extract from the rhizome of A. officinarum on the proliferation, tyrosinase activity,and melanogenesis of B16 melanoma cells could be enhanced with the increased content of galangin, which demonstrates that the galangin is one of the efficient substances in the rhizome of A. officinarum for the treatment of vitiligo.

国家自然科学基金资助项目（81160556）