滇山茶、银线草、车桑子、重楼对黑素细胞株增殖活性和酪氨酸酶影响的研究

Effects of Camellia retic ulata, Chloranthus japonicas, Dodonaea viscosa and Paris polyphylla on the proliferation of and tyrosinase activity in a melanocyte cell line

目的 探讨滇山茶、银线草、车桑子、重楼提取物的美白作用.方法 7种植物提取物（滇山茶枝叶提取物、滇山茶枝叶提取物70％乙醇洗脱物、滇山茶花蕾提取物、滇山茶花蕾提取物70％乙醇洗脱物、银线草提取物、车桑子提取物、重楼提取物）分别设立10、25、50、100、200、400、800 mg/L 7个浓度作用于体外培养人表皮黑素细胞株,同时设立阴性对照组、空白对照组和100 μg/ml熊果苷阳性对照组.采用噻唑蓝比色法测定植物提取物对黑素细胞增殖影响,体外氧化多巴反应法测定提取物对酪氨酸酶活性的影响.结果 对黑素细胞增殖活性的影响：400 mg/L滇山茶枝叶提取物,10 ～ 100 mg/L滇山茶枝叶提取物70％乙醇洗脱物,10 ～ 25 mg/L滇山茶花蕾提取物,10 mg/L滇山茶花蕾提取物70％乙醇洗脱物和10 ～ 25 mg/L车桑子提取物对黑素细胞增殖活性的抑制作用与100 mg/L熊果苷相当（P＞0.05）.800 mg/L滇山茶枝叶提取物对HEM-m细胞增殖活性的抑制作用低于熊果苷（P＜0.05）.余不同浓度提取物对HEM-m细胞增殖的抑制作用均强于熊果苷（P＜0.05或P＜0.01）.对黑素细胞酪氨酸酶活性的影响：10 mg/L滇山茶枝叶提取物,10 ～ 400 mg/L滇山茶枝叶提取物70％乙醇洗脱物,10～ 25 mg/L滇山茶花蕾提取物,50～100 mg/L滇山茶花蕾提取物70％乙醇洗脱物,10～50和400 mg/L银线草提取物,10 ～ 800 mg/L车桑子提取物,10和100 ～ 200 mg/L重楼提取物对酪氨酸酶活性的抑制作用与100 mg/L熊果苷相当（P＞0.05）.10～ 25 μg/ml滇山茶花蕾提取物70％乙醇洗脱物对酪氨酸酶的抑制作用明显低于熊果苷（P＜0.05）.其余不同浓度的提取物对酪氨酸酶的抑制活性均优于熊果苷组（P＜0.05或P＜0.01）.结论 滇山茶、车桑子提取物于特定浓度时,能抑制酪氨酸酶活性.

Objective To estimate the whitening effect of extracts of Camellia reticulata,Chloranthus japonicas,Dodonaea viscosa and Paris polyphylla.Methods A human epidermal melanocyte cell line HEM-m was cultured in vitro,and classified into several groups to be treated with 7 concentrations （10,25,50,100,200,400,800 mg/L） of different plant extracts,including Camellia reticulata leaf extract,70% ethanol-eluted fraction of Camellia reticulata leaf extract,extract of Camellia reticulata alabastrum,70% ethanol-eluted fraction of extract of Camellia reticulata alabastrum,Chloranthus japonicas extract,Dodonaea viscosa extract,and Paris polyphylla extract.At the same time,the negative control group,blank control group and positive control group （treated with 100 mg/L arbutin） were set up.After 72 hours of treatment,MTT assay was performed to detect cell proliferation,and dopa-oxidation assay to estimate tyrosinase activity in melanocytes.Results As far as the inhibitory effect on the proliferation of HEM-m cells was concerned,400 mg/L Camellia reticulata leaf extract,70% ethanol-eluted fraction of Camellia reticulata leaf extract at 10-100 mg/L,extract of Camellia reticulata alabastrum at 10-25 mg/L,70% ethanol-eluted fraction of extract of Camellia reticulata alabastrum at 10 mg/L and Dodonaea viscosa extract at 10-25 mg/L were equivalent to （all P 〉 0.05）,800 mg/L Camellia reticulata leaf extract was significantly weaker than （P 〈 0.05）,while the other concentrations of these plant extracts were significantly stronger than （P 〈 0.05 or 0.01）,100 mg/L arbutin.In the case of inhibitory effect on tyrosinase activity,100 mg/L arbutin was similar to 10 mg/L Camellia reticulata leaf extract,70% ethanol-eluted fraction of Camellia reticulata leaf extract at 10-400 mg/L,extract of Camellia reticulata alabastrum at 10-25 mg/L,70% ethanol-eluted fraction of extract of Camellia reticulata alabastrum at 50-100 mg/L,Chloranthus japonicas extract at 10-50 mg/L and 400 mg/L,Dodonaea viscosa extract at 10-800 mg/L and Pads polyphylla extract at 10 and 100-200 mg/L （all P 〉 0.05）,stronger than 70% ethanol-eluted fraction of extract of Camellia reticulata alabastrum at 10-25 mg/L （P 〈 0.05）,but weaker than the other concentrations of these plant extracts （P 〈 0.05 or 0.01）.Conclusion Special concentrations of Camellia reticulate and Dodonaea viscose can inhibit tyrosinase activity.