柴胡疏肝散体外抗氧化活性成分与调控抑郁症相关酶的关联性分析

Correlation Between Antioxidant Components of Chaihu Shugansan and Regulation on Depression-related Enzymes

目的：研究柴胡疏肝散（CSGS）体外抗氧化活性能力,识别抗氧化活性成分并评价其对CSGS治疗抑郁症的调控能力。方法：将CSGS进行拆方研究,采用1,1-diphenyl-2-picryl-hydrazyl（DPPH）法测定各配伍方自由基清除能力,以半数抑制浓度（IC50）（清除率为50%时的质量浓度）为评价指标;高效液相色谱法（HPLC）建立特征图谱,利用正交投影偏最小二乘法（orthogonal partial least squares,OPLS）研究特征峰与药效的相关性,根据变异权重系数（variable importance of projection,VIP）来辨识显著活性成分。运用SYBYL 2.0软件对CSGS中的抗氧化活性成分与CSGS治疗抑郁症所调控的相关酶进行分子对接研究,根据Total score值判断小分子与大分子的结合能力。结果：CSGS组方药材对抗氧化药效贡献的大小依次为枳壳〉陈皮〉川芎〉柴胡〉香附〉白芍〉甘草,鉴定出3个具有显著抗氧化活性的成分,分别为橙皮苷、新橙皮苷和芸香柚皮苷。3个抗氧化活性成分与CSGS治疗抑郁症所调控的相关酶的分子对接打分结果均显示了良好的结合能力,其中橙皮苷与色氨酸酶（TPH）,新橙皮苷与超氧化物歧化酶（SOD）,芸香柚皮苷与色氨酸2,3-加二氧酶（TPHD）的结合能分别是9.088 7,8.734 0,9.271 8。结论：橙皮苷,新橙皮苷和芸香柚皮苷是CSGS体外抗氧化活性的主要有效成分且在CSGS治疗抑郁症中可能发挥了一定的调控作用。

Objective： To research the antioxidant activity of Chaihu Shugansan（ CSGS） from different compatibility rules in vitro,further identify its antioxidant components and evaluate its regulation ability in the treatment of CSGS in depression. Method： After CSGS was divided into eight compatibility groups,their free radical scavenging activity was determined respectively by 1,1-diphenyl-2-picryl-hydrazyl（ DPPH） method,with the mass concentration value at a clearance rate of 50%（ IC50） as an evaluation index. Orthogonal partial least squares（ OPLS） method was used to investigate the correlation between the characteristic peaks and the efficacy of CSGS,and subsequently the significant active ingredients were identified by the variable importance of projection（ VIP） based on the high performance liquid chromatography（ HPLC） characteristic chromatograms. SYBYL 2. 0 software was used for molecular docking studies of the antioxidant components in CSGS and regulation of CSGS treatment on depression-related enzymes. Finally,the binding capacity of small molecules and macromolecules was determined on the basis of total score value. Result： The results demonstrated that the contribution order of eight herbs for antioxidant activity was as follows： Aurantii Fructus Citri Reticulatae Pericarpium Chuanxiong Rhizoma Bupleuri Radix Cyperi Rhizoma Paeoniae Radix Alba Glycyrrhizae Radix et Rhizoma. Besides,three ingredients including hesperidin,neohesperidin and narirutin were identified as the antioxidant activity components. Docking results of both antioxidant components in CSGS and the enzymes regulated by CSGS in the treatment of depression showed a good binding ability. Among them,the binding energy of hesperidin and tryptophanase（ TPH）, neohesperidin and superoxide dismutase（ SOD）, narirutin and tryptophan 2, 3-dioxygenase（ TPHD） was 9. 088 7,8. 734 0,9. 271 8,respectively. Conclusion： Hesperidin,neohesperidin and narirutin might be the main effective components of CSGS for the antioxidant activity in vitro and they play a regulatory role in the CSGS treatment of depression.