22味酸味药性中药提取物影响黑素合成的实验研究

Melanin Synthesis Was Affected by Extracts of 22 Kinds Chinese Herbs of Acid Taste: an Experimental Study

目的证实酸味药性中药对黑素的合成具有抑制作用。方法以碱提酸沉、醇提、水提3种方法,提取22味酸味中药有效成分,分为25、12.5、6.25 g/L 3个浓度,用蘑菇酪氨酸酶多巴速率氧化法检测各提取物对酪氨酸酶活性的影响,比较3种不同提取物不同浓度酪氨酸酶活性抑制率分别与熊果苷水溶液1.0、0.5、0.1 mmol/L的差异。结果 22味酸味中药(山茱萸、山楂、乌梅、五味子、木瓜、虎杖、牛膝、地榆、酸枣仁、墨旱莲、覆盆子、枳实、枳壳、绿萼梅、枳椇子、桑椹、石榴皮、白芍、金樱子、马齿苋、诃子、五倍子)碱提物除墨旱莲外,余21味对酪氨酸酶均有不同程度的抑制作用,其中乌梅在25 g/L时抑制率最高(88.49±9.98)%,枳实在6.25 g/L时抑制率最低(11.22±3.36)%;醇提物除墨旱莲无抑制作用外,其余21味中药均可抑制酪氨酸酶活性,以枳椇子在25 g/L时抑制率最高(75.92±5.57)%,枳壳在6.25 g/L时抑制率最低(9.60±1.15)%;水提物均有抑制酪氨酸酶活性的作用。其中桑椹在25 g/L时抑制率最高(54.23±3.56)%,酸枣仁在6.25 g/L时抑制率最低(10.25±1.83)%。与1 mmol/L熊果苷水溶液比较,乌梅、五味子、五倍子、金樱子、覆盆子、木瓜、马齿苋、桑椹、牛膝、石榴皮碱提物、乌梅、五倍子、石榴皮、枳椇子、山楂、牛膝醇提物及木瓜、覆盆子、桑葚水提物在25 g/L浓度时酪氨酸酶抑制率明显升高,差异有统计学意义(P<0.05,P<0.01);与0.5 mmol/L熊果苷水溶液比较,木瓜碱提物及乌梅醇提物在12.5 g/L时酪氨酸酶抑制率明显升高,差异有统计学意义(P<0.05,P<0.01)。结论具有酸味药性的一类中药均具有抑制黑素合成的作用,其机制与抑制酪氨酸酶活性有关。

Objective To confirm the inhibitory effect of Chinese herbs of acid taste on melanin synthesis. Methods Active ingredients of 22 kinds Chinese herbs of acid tastes were extracted by alkali extraction and acid precipitation, alcohol extraction, and water extraction, respectively, which was then dispensed into 25.00, 12.50, and 6.25 g/L suspension. Their effects on activities of tyrosinase were de- tected using mushroom-tyrosinase-DOPA speed oxidation. Their inhibition rates on activities of tyrosinase were respectively compared with inhibition rates of 1.0, 0.5, and 0.1 mmol/L arbutin. Results The 22 kinds Chinese herbs of acid taste included Comus Officinalis, Crataegus pinnatifida, dark plum fruit, Schisandra Chinensis , Chaenomeles sinensis Koehne, Reynoutria japonica Houtt, Achyranthes Bidenta- ta, Sanguisorba officinalis L., Semen Ziziphi Spinosae, Herba Ecliptae, blueberry, immature bitter or- ange, submature bitter orange, Prunus mume Var, Hovenia acerba Lindl., Fructus Mori, Pomegranate Rind, white paeony root, Rosa laevigata Michx., Portulaca oleracea L, Terminalia chebula Retz, Rhus chinensis Mill. Their alkaline extractions showed inhibition to activities of tyrosinase to different degrees except Herba Ecliptae. Of them, the highest inhibition rate （88.49%±9.98%.） was got by dark plum fruit at25 g/L, while the lowest inhibition rate （11.22%±3.36%） was got by immature bitter orange at 6.25 g/L. Their alcohol extractions showed inhibition to activities of tyrosinase to different degrees except Herba Ecliptae. Of them, the highest inhibition rate （75.92%±5.57%） was got by Hovenia acerba Lindl. at 25 g/L, while the lowest inhibition rate （9.60%±1.15% ） was got by submature bitter orange at 6.25 g/L. Their water extractions all had inhibition on activities of tyrosinase. Of them, the highest inhibition rate （54.23%±3.56%） was got by Fructus Mori at 25 g/L, while the lowest inhibition rate （10.25%±1.83% ） was got by Semen Ziziphi Spinosae at 6.25 g/L. Compared with 1 mmol/L arbutin water solution, alkaline extractions of dark plum fruit, Schisandra Chinensis, Rhus chinensis Mill., Rosa laevigata Michx., blueberry, Chaenomeles sinensis Koehne, Portulaca oleracea L, Fructus Mori, Achyranthes Bidentata, Pomegranate Rind; alcohol extractions of dark plum fruit, Rhus chinensis Mill., Pomegranate Rind, Hovenia acerba Lindl., Crataegus pinnatifida, Achyranthes Bidentata; water extractions of Chaenomeles sinensis Koehne, blueberry, and Fructus Mori at 25 g/L got obviously higher inhibition rates （P 〈0.05, P 〈0.01 ）. Compared with 0.5 mmol/L arbutin water solution, alcohol extraction of Chaenomeles sinensis Koehne and alcohol extraction of dark plum fruit at 12.5 g/L got obviously higher inhibition rates （P 〈 0.05, P 〈0.01 ）. Conclusion Chinese herbs of acid taste could inhibit melanin synthesis, and its mechanism was related to inhibiting activities of tyrosinase.