Cong Meng-Yu, Liang Xiao-Xia, Chen Feng-Lian, Cao Cheng, Wang Shu-Ling
Guangzhou University of Chinese Medicine Guangzhou 510000,China Jimo District Qingdao Hospital of Traditional Chinese Medicine Qingdao 266000,China.
Guangzhou University of Chinese Medicine Guangzhou 510000,China.
Zhongguo Zhong Yao Za Zhi. 2021 Jul;46(14):3633-3642. doi: 10.19540/j.cnki.cjcmm.20210120.402.
This article is to explore the antidepressant mechanism of Shugan Lipi recipe in regulating tryptophan metabolism,and to find out their common pharmacodynamic substances. UPLC-Q-TOF-MS technology was used to establish fingerprints of Shugan Lipi recipe,and 124 components were identified. The depressed mouse model was replicated by triple-one multiple stress method. Chaihu Shugan Powder,Sini Powder and Xiaoyao Powder were administered in groups to observe the changes in body weight and behavior of the mice. The results showed that compared with the model group,the body weight,sucrose preference percentage and autonomous activity behavior of each administration group were improved. Among them,the effect of Chaihu Shugan Powder was better than that of Sini Powder and Xiaoyao Powder. LC-MS/MS method was used to determine the contents of 5-hydroxytryptamine( 5-HT),kynurenine( KYN) and tryptophan( TPP) in blood,liver,brain,colon and other tissues,as well as TDO enzyme activity in liver. Western blot and RT-PCR were used to detect the protein and gene expression of TDO enzyme,respectively. It was found that the three prescriptions increased the ratio of 5-HT/KYN in different degrees,decreased the ratio of KYN/TRP in liver,colon and brain,and decreased the expression level and activity of TDO enzyme in liver. The order of their ability to regulate tryptophan metabolism was Chaihu Shugan Powder>Sini Powder>Xiaoyao Powder. In addition,the correlation between the chromatographic peaks in the fingerprints of Shugan Lipi recipes and the pharmacodynamic indexes of tryptophan metabolism was analyzed by the grey relation analysis. The grey relation analysis found that the chemical components with the highest correlation with tryptophan metabolism were mainly from Paeoniae Radix Alba,Citri Reticulatae Pericarpium,Aurantii Fructus Immaturus and Aurantii Fructus. UPLC-Q-TOF-MS was used to analyze the migration components in the plasma of mice after administration of Shugan Lipi recipe,and to verify the common pharmacodynamic substances of Shugan Lipi recipe. The migration of these detected components in plasma was studied,and a total of 18 prototype components and 36 metabolites were identified. Therefore,it was believed that Chaihu Shugan Powder,Sini Powder and Xiaoyao Powder could play an antidepressant role by reducing the expression of TDO enzyme in the liver and regulating the metabolism of tryptophan.The components contained in Paeoniae Radix Alba,Citri Reticulatae Pericarpium,Aurantii Fructus Immaturus and Aurantii Fructus were the common pharmacodynamic substances of Shugan Lipi recipe,which played an important role in regulating tryptophan metabolism.
本文旨在探讨疏肝理气方调节色氨酸代谢的抗抑郁机制，并找出其共同的药效物质。采用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF-MS）技术建立疏肝理气方指纹图谱，共鉴定出124个成分。采用三联应激法复制抑郁小鼠模型，分组给予柴胡疏肝散、四逆散和逍遥散，观察小鼠体重及行为变化。结果显示，与模型组相比，各给药组小鼠体重、蔗糖偏好百分比及自主活动行为均有所改善。其中，柴胡疏肝散的效果优于四逆散和逍遥散。采用液相色谱-串联质谱（LC-MS/MS）法测定血液、肝脏、脑、结肠等组织中5-羟色胺（5-HT）、犬尿氨酸（KYN）和色氨酸（TPP）的含量，以及肝脏中色氨酸2,3-双加氧酶（TDO）的活性。采用蛋白质免疫印迹法（Western blot）和逆转录-聚合酶链反应（RT-PCR）分别检测TDO酶的蛋白和基因表达。发现三方均不同程度提高了5-HT/KYN比值，降低了肝脏、结肠和脑中KYN/TRP比值，降低了肝脏中TDO酶的表达水平和活性。三方调节色氨酸代谢的能力顺序为柴胡疏肝散>四逆散>逍遥散。此外，通过灰色关联分析疏肝理气方指纹图谱中的色谱峰与色氨酸代谢药效学指标之间的相关性。灰色关联分析发现，与色氨酸代谢相关性最高的化学成分主要来自白芍、陈皮、枳实和枳壳。采用UPLC-Q-TOF-MS分析疏肝理气方给药后小鼠血浆中的移行成分，验证疏肝理气方的共同药效物质。研究了这些检测成分在血浆中的移行情况，共鉴定出18个原型成分和36个代谢产物。因此，认为柴胡疏肝散、四逆散和逍遥散可通过降低肝脏中TDO酶的表达，调节色氨酸代谢发挥抗抑郁作用。白芍、陈皮、枳实和枳壳所含成分是疏肝理气方的共同药效物质，在调节色氨酸代谢中起重要作用。
