补骨脂有效成分的提取及治疗白癜风药效评价

Extraction of effective components of Psoralea corylifolia and efficacy evaluation in the treatment of vitiligo

目的提取补骨脂的有效成分并评价其治疗白癜风的药效。方法利用超高效液相色谱法测定补骨脂提取液中补骨脂素、异补骨脂素、新补骨脂异黄酮、补骨脂异黄酮、补骨脂定、补骨脂乙素和补骨脂酚的浓度。在层次分析法和Plackett-Burman实验设计基础上,以上述7种成分浓度为评价指标,药材粉碎度、乙醇体积分数、浸渍时间为考察因素,采用Box-Behnken响应面法优化补骨脂的提取工艺并验证。将斑马鱼分为空白对照组、阳性对照组(8-甲氧基补骨脂素,10.8μg/mL)和补骨脂提取液低、中、高浓度组(500、1000、2000μg/mL),每组6条,通过密度测定研究补骨脂提取液对斑马鱼黑色素生成的影响。结果补骨脂提取的最优工艺为药物粉碎度60目,用80%乙醇浸渍72 h;3次验证实验综合评分的平均值为98.27,RSD为1.36%,与拟合方程预测值(97.28)的相对误差为1.02%。与空白对照组比较,补骨脂提取液低、中、高浓度组斑马鱼的黑色素沉着均显著增加(P<0.01)。结论优化所得的补骨脂提取工艺合理可行,所得补骨脂提取液能显著促进斑马鱼黑色素的生成。

OBJECTIVE To extract the effective components of Psoralea corylifolia and evaluate its efficacy in the treatment of vitiligo.METHODS The concentrations of psoralen,isopsoralen,neobavaisoflavone,corylin,psoralidin,corylifolinin,and bakuchiol in P.corylifolia extract were determined by ultra-performance liquid chromatography.Based on the analytic hierarchy process(AHP)and Plackett-Burman design,with the concentrations of the 7 components as evaluation indexes and the crushing degree,ethanol concentration,and soaking time as factors,the extraction process of P.corylifolia was optimized by Box-Behnken response surface methodology and the validation test was conducted.Zebrafish were divided into blank control group,positive control group(8-methoxypsoralen,10.8μg/mL),and low-,medium-,and high-concentration groups of P.corylifolia extract(500,1000,2000μg/mL),with 6 fish in each group.The effects of P.corylifolia extract on the melanin production of zebrafish were studied by density analysis.RESULTS The best extraction process was P.corylifolia powder over 60 meshes and soaked in 80%ethanol for 72 hours.The average comprehensive score of three validation experiments was 98.27,with an RSD of 1.36%,and the relative error was 1.02%compared with the predicted value of the fitting equation(97.28).Compared with the blank control group,the melanin pigmentation of zebrafish in the low-,medium-,and high-concentration groups of P.corylifolia extract was significantly increased(P<0.01).CONCLUSIONS The optimized extraction process of P.corylifolia is reasonable and feasible,and the obtained P.corylifolia extract can significantly promote the production of melanin in zebrafish.