Chaihu-Shugan-San Research Articles

In Vitro and in Vivo Antioxidant Effects and the Possible Relationship between the Antidepression Efficacy of Traditional Chinese Medicine Formulation Chaihu Shugan San

Aim To investigate the in vitro and in vivo antioxidant activities of the traditional Chinese prescription Chaihu Shugan San (CSGS), which may help to understand the relationship between the antioxidant properties and the antidepressive effect of CSGS, to further clarify the possible mechanism of its antidepression efficacy. Methods The antioxidant activity in vitro was evaluated by employing five antioxidant assays including ferric reducing antioxidant power (FRAP) assay, DPPH (1, 1-diphenyl-2-picryl-hydrazyl) radicals, ABTS [2, 2’-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid) diammonium salt] cation radicals, superoxide anion (O 2 ·-) and hydroxyl free radicals (OH ·) scavenging activity assays. Vitamin C (Vc) was used as a positive control. The in vivo study was carried out to investigate the effects of CSGS on mice in restraint stress model by determining glutathione (GSH) contents in blood and malondialdehyde (MDA) values, superoxide dismutase (SOD) and catalase (CAT) activities in liver. Results CSGS showed a ferric reducing antioxidant power (FRAP assay) that was equivalent to 0.24 mmol FeSO 4 per gram CSGS aqueous extract and certain scavenging activities on free radicals in vitro. The IC 50 values of CSGS in scavenging DPPH radicals, ABTS cation radicals, superoxide anion and hydroxyl free radicals were 0.83, 1.03, 10.31 and 7.79 mg(mL −1, respectively. The results of in vivo study showed that the restrained mice with orally administered CSGS had higher SOD and catalase CAT activities, lower malondialdehyde MDA values and higher glutathione GSH levels compared with those of the mice in model group. Conclusion CSGS could inhibit lipid peroxidation, relieve oxidative injury and regulate antioxidant capacity of organisms. The findings suggest that antioxidant activity of CSGS should make contributions to its antidepression effect.

Online identification of the antioxidant constituents of traditional Chinese medicine formula Chaihu-Shu-Gan-San by LC–LTQ-Orbitrap mass spectrometry and microplate spectrophotometer

Chaihu-Shu-Gan-San (CSGS), a traditional Chinese medicine (TCM) formula containing seven herbal medicines, has been used in treatment of gastritis, peptic ulcer, irritable bowel syndrome and depression clinically. However, the chemical constituents in CSGS had not been studied so far. To quickly identify the chemical constituents of CSGS and to understand the chemical profiles related to antioxidant activity of CSGS, liquid chromatography coupled with electrospray ionization hybrid linear trap quadrupole orbitrap (LC–LTQ-Orbitrap) mass spectrometry has been applied for online identification of chemical constituents in complex system, meanwhile, antioxidant profile of CSGS was investigated by the fraction collecting and microplate reading system. As a result, 33 chemical constituents in CSGS were identified. Among them, 13 components could be detected both in positive and in negative ion modes, 20 constituents were determined only in positive ion mode and 2 components were only detected in negative ion mode. Meanwhile, the potential antioxidant profile of CSGS was also characterized by combination of 96-well plate collection of elutes from HPLC analysis and microplate spectrophotometer, in which the scavenging activities of free radical produced by DPPH of each fraction could be directly investigated by the analysis of microplate reader. This study quickly screened the contribution of CSGS fractions to the antioxidant activity and online identified the corresponding active constituents. The results indicated that the combination of LC–MS n and 96-well plate assay system established in this paper would be a useful strategy for correlating the chemical profile of TCMs with their bioactivities without isolation and purification.

Antidepressant effect of Chaihu-Shugan-San extract and its constituents in rat models of depression

Abstract Herbal preparations may be effective alternatives in the treatment of depression, which remains difficult to manage. Chaihu-Shugan-San (CSS), an oriental traditional medicine, has been used as a remedy for Hwa-Byung, a Korean culture-bound syndrome resembling depression. We examined whether aqueous extracts of CSS and its constituent herbs exert antidepressant-like effects in two experimental animal models: the forced swimming test (FST) and the chronic mild stress (CMS) model. The herbal extracts were administered orally for 7 days in the FST and for 21 days during the CMS model; imipramine at 20 mg/kg/day was injected intraperitoneally as a positive control. CSS, Radix Bupleuri (one of the most important constituent plants in CSS), and imipramine had significant anti-immobility effects in the FST and reversed the CMS-induced reduction in sucrose consumption. Rhizoma Cyperi, another constituent of CSS, had antidepressant activity in the FST, while it failed in the CMS model. In conclusion, our results suggest that CSS and its constituent herbs exert antidepressant-like effects comparable to those of imipramine in experimental animal models.