Next Article in Journal
Sequestration Effect on the Open-Cyclic Switchable Property of Warfarin Induced by Cyclodextrin: Time-Resolved Fluorescence Study
Next Article in Special Issue
Identification of a Quality Marker (Q-Marker) of Danhong Injection by the Zebrafish Thrombosis Model
Previous Article in Journal
Chemometric Analysis of Lavender Essential Oils Using Targeted and Untargeted GC-MS Acquired Data for the Rapid Identification and Characterization of Oil Quality
Previous Article in Special Issue
Time- and NADPH-Dependent Inhibition on CYP3A by Gomisin A and the Pharmacokinetic Interactions between Gomisin A and Cyclophosphamide in Rats
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Molecules
Volume 22
Issue 8
10.3390/molecules22081331
molecules-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Anti-Anxiety Effect of (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside from Albizzia julibrissin Durazz (Leguminosae)

by
Jie Liu
1,
Yue-Wei Lv
1,
Jin-Li Shi
1,*,
Xiao-Jie Ma
1,
Yi Chen
1,
Zhi-Quan Zheng
1,
Sheng-Nan Wang
1 and
Jian-You Guo
2,*
1
School of Chinese Materia Medica, Beijing University of Chinese Medicine, 11ANorth Third Ring East Road, Chaoyang District, Beijing 100029, China
2
Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, 4ADatun Road, Chaoyang District, Beijing 100101, China
*
Authors to whom correspondence should be addressed.
Molecules 2017, 22(8), 1331; https://doi.org/10.3390/molecules22081331
Submission received: 4 July 2017 / Revised: 4 August 2017 / Accepted: 10 August 2017 / Published: 11 August 2017
(This article belongs to the Collection Herbal Medicine Research)
Browse Figures
Versions Notes

Abstract

:
Albizzia julibrissin Durazz, a Chinese Medicine, is commonly used for its anti-anxiety effects. (−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG) is the main ingredient of Albizzia julibrissin Durazz. The present study investigated the anxiolytic effect and potential mechanisms on the HPA axis and monoaminergic systems of SAG on acute restraint-stressed rats. The anxiolytic effect of SAG was examined through an open field test and an elevated plus maze test. The concentration of CRF, ACTH, and CORT in plasma was examined by an enzyme-linked immune sorbent assay (ELISA) kit while neurotransmitters in the cerebral cortex and hippocampus of the brain were examined by High Performance Liquid Chromatography (HPLC). We show that repeated treatment with SAG (3.6 mg/kg, p.o.) significantly increased the number and time spent on the central entries in the open-field test when compared to the vehicle/stressed group. In the elevated plus maze test, 3.6 mg/kg SAG could increase the percentage of entries into and time spent on the open arms of the elevated plus maze. In addition, the concentration of CRF, ACTH, and CORT in plasma and neurotransmitters (NE, 5-HT, DA and their metabolites 5-HIAA, DOPAC, and HVA) in the cerebral cortex and hippocampus of the brain were decreased after SAG treatment, as compared to the repeated acute restraint-stressed rats. These results suggest that SAG is a potential anti-anxiety drug candidate.

1. Introduction

Neuropsychiatric disorders are expected to rise sharply [1]. Anxiety disorders are the most common mental disorders, affecting nearly one in five adults in the U.S. alone [2]. The development of anxiolytic drugs is important for the treatment of anxiety disorders. Benzodiazepines have been used for the treatment of several forms of anxiety, but these compounds have well-known side effects, including sedation, muscle relaxation, amnesia, and dependence potential [3]. Albizzia julibrissin Durazz (Leguminosae, the cortex of Albizia julibrissin Durazz. officially recognized in the Chinese Pharmacopoeia), commonly named mimosa or silk trees, are widely distributed in Asia. Asians administered A. julibrissin soup to patients as a folk medicine to treat insomnia, diuresis, sthenia, and confusion [4]. The extract of A. julibrissin has an anxiolytic-like effect due to 5-HT1A receptor activation, and because it has no unwanted adverse effects [5]. (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG) is the main ingredient of the cortex of A. julibrissin, and we speculate that it may be the effective component in treating anxiety disorders [6].
Monoamine neurotransmitters including 5-HT, norepinephrine (NE), and dopamine (DA) are believed to be involved in pathogenes is of emotional disorders, and play an important role in mediating behavioral effects [7]. Several preclinical and clinical reports provided evidence to support that a dysfunction of the monoaminergic system may be implicated as a promising mechanism in the pathophysiology of anxiety disorders [8,9,10].Furthermore, anxiety disorders were closely related to the hippocampus and cerebral cortex of the brain. Meanwhile, a growing body of evidence suggests connections between dysregulation of the hypothalamic-pituitary-adrenocortical (HPA) axis and affective behaviors [11]. For example, hyperactivity of the HPA axis [12], enlarged adrenal glands, altered daily rhythm of corticoids secretion, and impairments in the HPA negative feedback mechanism have been implicated in emotional disorders. It is well known that HPA axis activation is a key component of the physiological response to stress and anxiety [13], while the monoamine neurotransmitters in the cerebral cortex, such as NE, DA and 5-HT, and plasma CORT are considered to be involved in generating symptoms of anxiety [14].
Stress, including chronic and acute restraint stress, is the most important and widely used animal model of anxiety [15]. Stress can induce behavioral, physiological, cognitive, and neural changes, potentially altering homeostasis and promoting vulnerability to illness [16]. Acute stress is an adaptive response to a multitude of adverse stimuli that are perceived to be noxious or threatening [17]. Acute restraint stress (ARS) can induce both psychological and physical effects. This results in a broad range of behavioral and physiological changes, including anxiogenic-like effects [18], endocrine, and autonomic alterations [19]. Continued stressful situations can be responsible for activation of the hypothalamic-pituitary-adrenal (HPA) axis, which subsequently results in the development of anxiety and depression [20].
In the present study, we examined the anxiolytic-like effect of SAG on repeated acute restraint-stressed rats using the elevated plus maze and open field test. Furthermore, in order to explore the potential mechanisms on monoaminergic systems and the HPA axis, we also examined the influences on the levels of the plasma corticotropin releasing factor (CRF), adrenocortico tropic hormone (ACTH), and corticosterone (CORT). We also studied the level of monoamines noradrenaline (NE), dopamine (DA), serotonin (5-HT) and their metabolites 5-HIAA, DOPAC, and HVA in the whole of the cerebral cortex and hippocampus of rats.

2. Results

2.1. Effect of SAG in the Open Field Test

The results of the OFT are shown in Figure 1 (Supplementary Materials). There were significant differences in the number of central entries (F(5,53) = 3.743, p < 0.01), and time spent in central areas (F(5,53) = 2.637, p < 0.05). Following the ARS, the number of central entries (p < 0.01) and the time spent in central areas (p < 0.01) of rats in the V/S group significantly decreased when compared with the V/US group. Compared with the V/S group, diazepam, and SAG (3.6 mg/kg) could significantly increase the number of central entries (p < 0.05) and the time spent in central areas (p < 0.05). However, there was no difference with SAG at the doses of 0.9 mg/kg and 1.8 mg/kg.

2.2. Effect of SAG in the Elevated Plus Maze

As shown in Figure 2, there were significant differences among groups in the percentage of open arm entries (F(5,53) = 2.751, p < 0.05) and the time spent on the open arms (F(5,53) = 2.602, p < 0.05). The V/S group significantly decreased the percentage of open arm entries (p < 0.01) and the time spent on the open arms (p < 0.01) of rats compared with the V/US group. Compared to the V/S group, diazepam and SAG (3.6 mg/kg) could significantly increase the percentage of open arm entries (both p < 0.01) and the time spent on the open arms (p < 0.01, p < 0.05) (Supplementary materials are available online).

2.3. Effect of SAG on Serum CRF, ACTH, and CORT Levels

As shown in Figure 3, the one-way ANOVA revealed significant differences in the concentration of CRF (F(5,54) = 3.377, p < 0.05), ACTH (F(5,54) = 5.092, p < 0.01), and CORT (F(5,54) = 6.625, p < 0.01). With the acute restraint stress, the V/S group significantly increased the content of plasma CRF, ACTH, and CORT compared with the V/US group (all p < 0.01). Diazepam treated rats could significantly decrease the plasma CRF, ACTH, and CORT level compared with the V/S group (all p < 0.01). SAG at the doses of 1.8 and 3.6 mg/kg could significantly decrease the plasma CRF level (p < 0.05). The level of ACTH could be significantly decreased by SAG at the dose of 3.6 mg/kg, and the level of CORT could be significantly decreased by SAG at the doses of 0.9, 1.8, and 3.6 mg/kg. (Supplementary materials are available online).

2.4. Effect of SAG on Monoamine Neurotransmitters and Their Metabolites

In Table 1, the one-way ANOVA indicated significant differences in the levels of monoamine neurotransmitters and their metabolites in the cerebral cortex of rats (F(5, 53) = 12.561, p < 0.01, NE; F(5,53) = 3.065, p < 0.05, DA; F(5, 53) = 2.562, p < 0.05, HVA; F(5, 53) = 8.324, p < 0.01, 5-HT; F(5,53) = 11.435, p < 0.01, 5-HIAA. No differences were found in the level of DOPAC (F(5,53) = 1.258, p > 0.05). The V/S group could significantly increase the levels of NE (p < 0.01), DA (p < 0.05), HVA (p < 0.05), 5-HT (p < 0.05) and 5-HIAA(p < 0.05) when compared with the V/US group. SAG at the doses of 1.8 and 3.6 mg/kg could significantly decrease the levels of NE (p < 0.05, p < 0.01) and 5-HT (p < 0.05, p < 0.01). At the dose of 3.6 mg/kg, SAG could significantly decrease the levels of DA (p < 0.05), HVA (p < 0.05) and 5-HIAA (p < 0.01). (Supplementary materials are available online).
In Table 2, the one-way ANOVA indicated significant differences in the levels of monoamine neurotransmitters and their metabolites in the hippocampus of rats (F(5,53) = 3.716, p < 0.01, NE; F(5,53) = 4.879, p < 0.05, 5-HT; F(5,53) = 2.656, p < 0.05, 5-HIAA). No differences were found in the levels of DA (F(5,53) = 1.421, p > 0.05), DOPAC (F(5,53) = 1.624, p > 0.05) and HVA (F(5,53) = 1.245, p > 0.05). The V/S group could significantly increase the levels of NE (p < 0.01), 5-HT (p < 0.05) and 5-HIAA (p < 0.05) compared with the V/US group. SAG at the doses of 1.8 and 3.6 mg/kg could significantly decrease the levels of NE (both p < 0.01) and 5-HT (both p < 0.01). At the dose of 3.6 mg/kg, SAG could significantly decrease the level of 5-HIAA (p < 0.01). (Supplementary materials are available online).

3. Discussion

The results of this behavioral investigation revealed the anxiolytic-like effect of SAG in animal models of anxiety. Moreover, the results support the designed hypothesis that the HPA axis and monoaminergic systems play an important role in pathogenesis of anxiety disorders. Repeated ARS-subjected rats exhibited anxiety-like behavior. This is evidenced by the significant decline of central entries and time spent in central areas of OFT, the open arm entries, and time spent in open arm in EPM. Treatment with SAG at the dose of 3.6 mg/kg prevented the decrease resulting from the ARS by regulating the levels of CRF, ACTH, and CORT in serum and monoamine neurotransmitters and their metabolites.
Stress refers to any given condition affecting the integrity of biological systems. Since acute stress is more related with the expression of adaptive responses, it is characterized by early compensatory responses oriented to restore homeostatic conditions, and it provides relevant information on the origin and nature of ongoing harmful events in the nervous system. Stress is often associated with psychiatric (depression, anxiety, and panic) and neurodegenerative disorders (Alzheimer’s disease, Parkinson’s disease, etc.). Earlier reports have suggested that animals submitted to acute RS for 3 h induced long lasting anxiety-like behavior in the EPM [21] and acute RS for 60 min could activate the AT1-angiotensin receptors in the PVN [22], which is related to anxiety-like behavior in the brain. Of particular interest to this study is the aim to investigate the anti-anxiety effect of SAG in repeated acute restraint-stressed rats. In the present study, the procedure of repeated ARS is improved on Ortiz (1996) [23], and is verified by Lv (2015) [24]. Our findings show that repeated acute restraint stress could lead to anxiety and interfere with the HPA axis.
The dried stem bark of A. julibrissin has been used in China, Japan and Korea as a tonic, to ease the state of mind and calm the nerves [25]. Previous work has reported that the water extract [5] and n-butyl alcohol extract of A. julibrissin have obvious anti-anxiety effects [6]. Flavonol glycoside isolated from A. julibrissin showed sedative activity in mice [26]. Kinjo et al. reported that syringesinol glycoside isolated from the A. julibrissin cortex had a tonic effect. It is inferred that the lignan might be the valid anti-anxiety effective components [6]. Meanwhile, the SAG is the main lignan of A. julibrissin. The present study has investigated the anxiolytic effect of SAG and determined which neuronal mechanism is primarily involved. Our results defined the anti-anxiety effect and interpret the mechanism of SAG.
The open field test is widely used to examine the behavioral effects of drugs and anxiety [27]. The test is based on rodents’ natural tendency to remain near the periphery of a novel environment, and their aversion to open and illuminated spaces. Activity in the central area of the open field is thought to be correlated with the degree of fear, whereas activity in the peripheral zone and along the walls of the apparatus is thought to reflect general activity [28]. To demonstrate anxiogenic or anxiolytic effects of specific treatments, the control animals should display a reasonably high number of entries into the aversive parts of the apparatus, and be active in the peripheral areas of the open field. Anxiety-like behavior is reflected by diminished exploration of the aversive space with relatively unaffected ambulatory activity in the safe areas [29]. Rats that were treated with 3.6 mg/kg SAG could significantly increase the number of central entries and the time spent in central areas. Therefore, 3.6 mg/kg of SAG appeared to exert significant anxiolytic-like effects in this paradigm.
The elevated plus maze is considered an ethologically valid animal model of anxiety because it uses natural stimuli (e.g., fear of novel open spaces and fear of balancing on a relatively narrow, raised platform) that can induce anxiety in humans [30]. The elevated plus maze is based on the natural aversion of rodents to height and open spaces. In the present study, oral administration of SAG could induce anxiolytic-like effects on repeated acute restraint-stressed rats by increasing the number of entries into, and time spent on the open arms of the elevated plus maze.
A prominent mechanism by which the brain reacts to acute and chronic stress is activation of the hypothalamic-pituitary-adrenal (HPA) axis. The activity of the HPA axis is controlled by several brain pathways, including the hippocampus (which exerts an inhibitory influence on hypothalamic CRF-containing neurons via a polysynaptic circuit) and the amygdala (which exerts a direct excitatory influence) [31]. CRF is a critical mediator of fear conditioning and other forms of emotional memory to both aversive and rewarding stimuli. The well documented hyperactivity of the HPA axis in depression seems to be related to hypothalamic secretion of CRF [32]. Acute and chronic stress can increase CRF levels in the locus coeruleus and anxiolytic drugs can reduce them [33]. The current study has shown that the repeated ARS rats have a high level of CRF, and SAG can reduce them. Furthermore, they also show an increase in basal ACTH and CORT in the repeated ARS rats. By comparison, varying alterations in ACTH and/or CORT have been reported in studies employing the widely-used Chronic Variable Stress (CVS)/Chronic Mild Stress (CMS) model. Most have shown either no change or an increase in basal ACTH [34] and no change or an increase in basal CORT [35]. Changes in HPA responses to acute stress challenge after CVS/CMS are variable. One study reported an increased ACTH response to acute restraint stress, but no change in CORT response. From the above, the present study revealed that ARS could significantly increase the CRF, ACTH and CORT levels, and this effect could be retrained by DZP and SAG (3.6 mg/kg).Furthermore, SAG at the dose of 1.8 mg/kg could also reverse the increase of CRF and CORT caused by ARS. SAG at the dose of 0.9 mg/kg only could decrease the level of CORT. Our findings are the first to evaluate the mechanism of SAG from the aspect of the HPA axis.
Most of the researches have concentrated on the circuits that engage in rapid signaling. However, anxiogenic responses have long been known to engage complex neuromodulatory systems, and many current therapies for anxiety disorders are based on this activity. The central systems responsible for the response to stress appear to be the NE system and the corticotropin-releasing factor (CRF) system [36]. The central monoaminergic system (including serotonin (5-HT), NE and DA) has been widely implicated in the pathophysiology and therapeutic strategies for emotional disorders [37,38]. The present study is consistent with these reports. In addition, the synthesis and release of neurotransmitters in the hippocampus and cerebral cortex are closed related to neuropsychiatric disorders [20,39]. Furthermore, the brain removed and dissected after 7 days administration is according to Lv, Li and Liu [24,40,41]. Our data shows that repeated acute restraint-stressed rats could significantly increase the concentration of NE, DA, HVA, 5-HT, and 5-HIAA in the whole cerebral cortex while NE, 5-HT, and 5-HIAA act in the hippocampus of the brain. DZP and SAG (3.6 mg/kg) could reverse this effect of repeated ARS by decreasing the concentration of NE, DA, HVA, 5-HT, and 5-HIAA in the whole cerebral cortex and NE, 5-HT, and 5-HIAA in the hippocampus of the brain.
Notably, a previous study found that the extract of Albizzia julibrissin (at 100 or 200 mg/kg) significantly increased time-spent and arm entries into the open arms of the EPM in rats [5], and the content of SAG in the bark of Albizzia julibrissin was 0.04% [6]. According to the preliminary experiments, the 3.6 mg/kg dose was chosen as the highest dose in the present study. However, our data showed that SAG exerted an anxiolytic effect only at this high dose. Testing higher doses is warranted to further elucidate the anxiolytic effect of SAG. Meanwhile, the repeated treatment of SAG for 7 days in the present study had exerted the anxiolytic effect, the effect of acute SAG will be investigated in the future. Furthermore, in order to reduce the number of animals used, and to compare the effect between the SAG and the stressed group, the SAG with the unstressed group was not designed in the present experiment, which was consistent with the previous studies [24,42]. We detected the content of neurotransmitters only in the whole cerebral cortex and hippocampus, and many researches of anxiety were also only detected the two important central regions of the brain [20,39]. Meanwhile, the amygdala plays an important role in altering monoamines such as norepinephrine and serotonin in several animal models of anxiety [43].The synthesis and release of neurotransmitters in the amygdala should also be investigated in the next step and to estimate whether the content of neurotransmitters in the amygdala is consistent with the present results. In addition, the present study proves that the mechanism of SAG involves monoaminergic systems, including NE ergic, DAergic and 5-HTergic systems, and the further progress will be made by using an antagonist.

4. Materials and Methods

4.1. Animals

A total of 60 8-week-old male Sprague-Dawley rats from Laboratory Animal Center of the Academy of Military Medical Sciences (Beijing, China) were used. Each animal was housed in individual cages in a temperature-controlled environment (22 ± 1 °C) with unlimited access to food and water. They were maintained on a 12-h light/dark cycle (light phase: 07:00–19:00). All rats were allowed to acclimatize to the standard conditions for 7 days before experimental procedures were initiated. Behavior experiments were performed between 09:00 and 14:00. The experimental procedures were approved by the Institutional Animal Care and Use Committee of the Institute of Psychology of the Chinese Academy of Sciences and in accordance with the National Institutes of Health Guide for Care and Use of Laboratory Animals.

4.2. Drugs and Reagents

SAG (Figure 4) (PubChem CID: 91973808) was purchased from Weikeqi Biological Technology (Sichuan, China), the purity of SAG (Figure 5) was 98%, which was quantificationally analysed employing HPLC-UV method carried on a Waters 2695 HPLC system equipped with UV-vis detector.
Diazepam was obtained from Yimin Pharmaceutical Factory (Beijing, China).Norepinephrine (NE), 5-HT, 5-hydroxy-3-indoleacetic acid (5-HIAA), dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC)and homovanillic acid (HVA) were purchased from Sigma (St. Louis, MO, USA). The Elisa kits of CRF, ACTH and CORT were obtained from R&D (Minneapolis, MN, USA). All of the other reagents were of analytical grade.

4.3. Treatment

All drugs were prepared immediately before use and were given orally in a volume of 1 mL/100 g body weight. The vehicle/unstressed (V/US) group was orally administered physiological saline. Diazepam at the dose of 1 mg/kg was chosen as a positive control drug. DZP and SAG were dissolved in physiological saline. To evaluate the anxiolytic effect of SAG, the rats were orally administered SAG (0.9, 1.8, and 3.6 mg/kg) [5,6] 60 min prior to behavioral testing or diazepam 30 min before behavioral testing. The treatment protocol of doses and the administration route used for SAG and diazepam were adopted according to previous studies [6,44]. All animals were given daily administration for 7 days [5,40,41] (Figure 6).

4.4. AcuteRestraint Stress(ARS)

Animals were submitted to repeated restraint for 3 days by placing each rat into a plastic cylindrical restraint tube (diameter 6.5 cm, length 15 cm), ventilated by holes (1-cm diameter) that comprised approximately 20% of the tube surface. Restraint lasted for 30 min. After the last restraint, the rats were transported to the closed experiment room that was under low light, noise, and other external disturbances for 30 min.

4.5. Behavioral Tests

4.5.1. Open Field Test (OFT)

The OFT apparatus was a 180 cm diameter cylinder with 60 cm high walls. The center of the bottom of the apparatus had a 52 cm diameter section. The rats were placed into the field at the same point against the wall and allowed to freely explore the apparatus for 5 min. The number of central entries, and the time spent in the center were recorded by an automatic video tracking system. The apparatus was thoroughly cleaned with 70% methanol after each trial.

4.5.2. Elevated Plus Maze (EPM)

Immediately after the OFT, the rats were transported to the EPM room. After the adaptation of 5 min, anxiolytic activity was measured using the EPM, which consisted of two open arms (50.8 cm × 10.2 cm × 1.3 cm) and two closed arms (50.8 cm × 10.2 cm × 40.6 cm) that extended from a central platform (10.2 cm × 10.2 cm). The maze was elevated to a height of 72.4 cm above the floor. The entire maze was constructed of clear Plexiglas. Each rat was placed on the central square facing an open arm and was allowed to freely explore the maze for 5 min. Arm entries were defined as the entry of all four paws into an arm. A computer recorded the time spent on, and number of entries into the open and closed arms by means of infrared photocells. The apparatus was wiped clean with a 70% ethanol solution and dried after each subject.

4.6. Estimation of Serum CRF, ACTH and CORT

Immediately after the completion of the two behavioral tests, the rats were sacrificed by decapitation and blood samples were collected in EDTA coated tubes kept in ice and centrifuged at 3000 r·min−1 for 15 min at 4 °C. Plasma was separated and supernatants were stored at −80 °C for estimation. The contents of CRF, ACTH, and CORT were determined by a commercially available, enzyme-linked immunesorbent assay (ELISA) kit according to the manufacturer’s instructions. The absorbance of each sample was measured at a wavelength of 450 nm. The results of CRF and ACTH are presented as ng/L, and the results of CORT are presented as ng/mL.

4.7. Estimation of Monoamines and Metabolites

Following the collection of blood, the rats’ brains were rapidly removed and dissected, the cerebral cortex and hippocampus were isolated on an ice plate. The tissue samples were weighed and stored at −80 °C until homogenization. Levels of monoamines and metabolites (NE, DA, DOPAC, HVA, 5-HT and 5-HIAA) were estimated using High Performance Liquid Chromatography (HPLC) with an Electrochemical detector as described previously [45]. In brief, the brain tissue samples were homogenized in 0.10 Mperchloric acid by Polytron (Swedesboro, NJ, USA) homogenizer. Homogenates were then centrifuged at 1200 r·min−1 for 10 min at 4 °C. Twenty microliters of supernatant was injected via a HPLC pump (Model 1525, Binary Gradient Pump, Waters, Milford, MA, USA) into a column (2.1 mm ×150 mm at 30 °C, 3 μm, Waters Atlantis) connected to a Electrochemical detector (Model 2465, Waters, Milford, MA, USA) at a potential of +0.75 V with a glassy carbon working electrode Vs Ag/AgCl reference electrode. The mobile phase consists of 50 mM citric acid, 0.3 mM Na2-EDTA, 1.8 mM dibutylamine, and 4% methanol (pH 3.5) at a flow rate of 0.35 mL/min.

4.8. Statistical Analysis

The data is expressed as mean ± standard error of the mean (SEM) individual value of the rats from each group. The statistical analysis was performed using one-way analysis of variance (ANOVA), followed by the Student-Newman-Keuls post hoc test and Graph Pad Prism 5.0 software (Graphpad Sofware Inc, La Jolla, CA, USA). In cases of significant variation, the individual values were compared using Dunnett’s test. Values of p < 0.05 were considered statistically significant.

5. Conclusions

To summarize, the present data indicates that SAG induced anxiolytic-like effects on repeated acute restraint-stressed rats in the elevated plus maze and open field test. Additionally, its mechanism of action appears to be related with the HPA axis and monoaminergic systems.

Supplementary Materials

Supplementary materials are available online.

Acknowledgments

This work was supported by the Key New Drugs Innovation project from Ministry of Science and Technology (2012ZX09102201-018).

Author Contributions

Jie Liu and Yue-Wei Lv were responsible for the pharmacological activity, and Jie Liu was responsible for the manuscript writing. Xiao-Jie Ma and Yi Chen were responsible for the mechanism research; Zhi-Quan Zheng and Sheng-Nan Wang conducted the analyzed and interpreted the data; Jin-Li Shi suggested the research idea; and, Jin-Li Shi and Jian-You Guo conceived and designed the experiments, contributed to the discussion of results and reviewed the entire manuscript. All authors read and approved the final manuscript.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Naismith, S.L.; Norrie, L.M.; Mowszowski, L.; Hicki, I. The neurobiology of depression in later-life: Clinical, neuropsychological, neuroimaging and pathophysiological feateres. Prog. Neurobiol. 2012, 98, 99–143. [Google Scholar] [CrossRef] [PubMed]
  2. McLean, C.P.; Asnaani, A.; Litz, B.T.; Hofmann, S.G. Gender differences in anxiety disorders: Prevalence, course of illness, comorbidity and burden of illness. J. Psychiatr. Res. 2011, 45, 1027–1035. [Google Scholar] [CrossRef] [PubMed]
  3. Youssef, N.A.; Rich, C.L. Does acute treatment with sedatives/hypnotics for anxiety in depressed patients affect suicide risk? A literature review. Ann. Clin. Psychiatry 2008, 20, 157–169. [Google Scholar] [CrossRef] [PubMed]
  4. Zhu, Y.P. Chinese Materiamedica Chemistry, Pharmacology and Applications; Harwood Academic Publishers: Boca Raton, The Netherlands, 1999; pp. 519–520. [Google Scholar]
  5. Kim, W.K.; Jung, J.W.; Ahn, N.Y.; Oh, H.R.; Lee, B.K.; Oh, J.K.; Cheong, J.H.; Chun, H.S.; Ryu, J.H. Anxiolytic-like effects of extracts from Albizziajulibrissin bark in the elevated Plus-maze in rats. Life Sci. 2004, 75, 2787–2795. [Google Scholar] [CrossRef] [PubMed]
  6. Bo, F. Study on the Antianxiety Effects of Albiziajulibrissin on Mice. Master’s Thesis, Henan College of Traditional Chinese Medicine, Zhengzhou, China, 2014. [Google Scholar]
  7. Millan, M.J. The role of monoamines in the actions of established and “novel” antidepressant agents: A critical review. Eur. J. Pharmacol. 2004, 500, 371–384. [Google Scholar] [CrossRef] [PubMed]
  8. Aghajanian, G.K.; Sprouse, J.S.; Sheldon, P.; Rasmussen, K. Electrophysiology of the central serotonin system: Receptor subtypes and transducer mechanisms. Ann. N. Y. Acad. Sci. 1990, 600, 93–103. [Google Scholar] [CrossRef] [PubMed]
  9. Revelli, A.; Tesarik, J.; Massobrio, M. Nongenomic effects of neurosteroids. Gynecol. Endocrinol. 1998, 12, 61–67. [Google Scholar] [CrossRef] [PubMed]
  10. Zhang, Y.; Wang, W.; Zhang, J. Effects of novel anxiolytic 4-butyl-alpha-agarofuran on levels of monoamine neurotransmitters in rats. Eur. J. Pharmacol. 2004, 504, 39–44. [Google Scholar] [CrossRef] [PubMed]
  11. Gold, P.W. The organization of the stress system and its dysregulation in depressive illness. Mol. Psychiatry 2015, 20, 32–47. [Google Scholar] [CrossRef] [PubMed]
  12. Vreeburg, S.A.; Hoogendijk, W.J.G.; Van Pelt, J.; DeRijk, R.H.; Verhagen, J.C.M.; Van Dyck, R.; Smit, J.H.; Zitman, F.G.; Penninx, B.W. Major depressive disorder and hypothalamic-pituitary-adrenal axis activity: Results from a large cohort study. Arch. Gen. Psychiatry 2009, 66, 617–626. [Google Scholar] [CrossRef] [PubMed]
  13. Adinoff, B.; Junghanns, K.; Keifer, F.; Krishnan-Sarin, S. Suppression of the HPA axis stress-response: Implications for relapse. Alcohol. Clin. Exp. Res. 2010, 29, 1351–1355. [Google Scholar] [CrossRef]
  14. Kent, J.M.; Mathew, S.J.; Gorman, J.M. Molecular targets in the treatment of anxiety. Biol. Psychiatry 2002, 52, 1008–1030. [Google Scholar] [CrossRef]
  15. Luo, Y.W.; Xu, Y.; Cao, W.Y.; Zhong, X.L.; Duan, J.; Wang, X.Q.; Hu, Z.L.; Li, F.; Zhang, J.Y.; Zhou, M.; et al. Insulin-like growth factor 2 mitigates depressive behavior in a rat model of chronic stress. Neuropharmacology 2015, 89, 318–324. [Google Scholar] [CrossRef] [PubMed]
  16. McEwen, B.S. Stress, adaptation, and disease. Allostasis and allostatic load. Ann. N. Y. Acad. Sci. 1998, 840, 33–44. [Google Scholar] [CrossRef] [PubMed]
  17. Jereme, G.S.; Hsiao-Jou, C.C.; James, S.M.C.; Conrad, S.; Nickolas, A.L. Acute restraint stress induces rapid changes in central redox status and protective antioxidant genes in rats. Psychoneuroendocrinology 2016, 67, 104–112. [Google Scholar]
  18. Padovan, C.M.; Del Bel, E.A.; Guimaraes, F.S. Behavioral effects in the elevated plus maze of an NMDA antagonist injected into the dorsal hippocampus: Influence of restraint stress. PharmacolBiochem. Behav. 2000, 67, 325–330. [Google Scholar] [CrossRef]
  19. Busnardo, C.; Alves, F.H.; Crestani, C.C.; Scopinho, A.A.; Resstel, L.B.; Correa, F.M. Paraventricular nucleus of the hypothalamus glutamate neurotransmission modulates autonomic, neuroendocrine and behavioral responses to acute restraint stress in rats. Eur. Neuropsychopharmacol. 2013, 23, 1611–1622. [Google Scholar] [CrossRef] [PubMed]
  20. Vishnu, N.T.; Valmik, D.D.; Bhoomika, M.P. Potential antidepressant-like activity of silymarin in the acute restraint stress in mice: Modulation of corticosterone and oxidative stress response in cerebral cortex and hippocampus. Pharmacol. Rep. 2016, 68, 1020–1027. [Google Scholar]
  21. Vila-verde, C.; Marinho, A.L.Z.; Lisboa, S.F.; Guimaraes, F.S. Nitric oxide in the prelimbic medial prefrontal cortex is involved in the anxiogenic-like effect induced by acute restraint stress in rats. Neuroscience 2016, 320, 30–42. [Google Scholar] [CrossRef] [PubMed]
  22. Busnardo, C.; Tavares, R.F.; Correa, F.M. Angiotensinergic neurotransmission in the paraventricular nucleus of the hypothalamus modulates the pressor response to acute restraint stress in rats. Neuroscience 2014, 270, 12–19. [Google Scholar] [CrossRef] [PubMed]
  23. Ortiz, J.; Fitzgerald, L.W.; Lane, S.; Terwilliger, R.; Eric, J.N. Biochemical Adaptations in the Mesolimbic Dopamine System in Response to Repeated Stress. Neuropsychopharmacology 1996, 14, 443–452. [Google Scholar] [CrossRef]
  24. Lv, Y.W.; Liu, J.; Shi, S.N.; Guo, J.Y.; Liu, Y.; Shi, J.L. Anxiolytic effect of antianxietic compound prescription capsule on acute stress in rats and influence upon expression of ERK/CREB signaling pathway and BDNF in the brain of rats. Chin. Pharmacol. Bull. 2015, 31, 1614–1619. [Google Scholar]
  25. Kinjo, J.; Fukui, K.; Higuchi, H.; Nohara, T. The first isolation of lignan tri- and tetra- glycosides. Chem. Pharm. Bull. 1991, 39, 1623–1625. [Google Scholar] [CrossRef]
  26. Kang, T.H.; Jeong, S.J.; Kim, N.Y.; Higuchi, R.; Kim, Y.C. Sedative activity of two flavonol glycosides isolated from the flowers of Albizziajulibrissin Durazz. J. Ethnopharmacol. 2000, 71, 321–323. [Google Scholar] [CrossRef]
  27. Choleris, E.; Thomas, A.W.; Kavaliers, M.; Prato, F.S. A detailed ethological analysis of the mouse open field test: Effects of diazepam, chlordiazepoxide and an extremely low frequency pulsed magnetic field. Neurosci. Biobehav. Rev. 2001, 25, 235–260. [Google Scholar] [CrossRef]
  28. Belzung, C.; Griebel, G. Measuring normal and pathological anxiety-like behaviour in mice: A review. Behav. Brain Res. 2001, 125, 141–149. [Google Scholar] [CrossRef]
  29. Artur, H.S.; Adrian, J.D. Effects of interleukin-1β and lipopolysaccharide on behavior of mice in the elevated plus-maze and open field tests. Pharmacol. Biochem. Behav. 2007, 86, 651–659. [Google Scholar]
  30. Dawson, G.R.; Tricklebank, M.D. Use of the elevated plus maze in the search for novel anxiolytic agents. Trends Pharmacol. Sci. 1995, 16, 33–36. [Google Scholar] [CrossRef]
  31. Nestler, E.J.; Barrot, M.; DiLeone, R.J.; Eisch, A.J.; Gold, S.J.; Monteggia, L.M. Neurobiology of depression. Neuron 2002, 34, 13–25. [Google Scholar] [CrossRef]
  32. Pitts, A.F.; Samuelson, S.D.; Meller, W.H.; Bissette, G.; Nemeroff, C.B.; Kathol, R.G. Cerebrospinal fluid corticotropin-releasing hormone, vasopressin, and oxytocin concentrations in treated patients with major depression and controls. Biol. Psychiatry 1995, 38, 330–335. [Google Scholar] [CrossRef]
  33. Van Bockstaele, E.J.; Colago, E.E.; Valentino, R.J. Corticotropin-releasing factorcontaining axon terminals synapse onto catecholamine dendrites and may presynaptically modulate other afferents in the rostral pole of the nucleus locus coeruleus in the rat brain. J. Comp. Neurol. 1996, 364, 523–534. [Google Scholar] [CrossRef]
  34. Kioukia-Fougia, N.; Antoniou, K.; Bekris, S.; Liapi, C.; Christofidis, I.; PapadopoulouDaiafoti, Z. The effects of stress exposure on the hypothalamic-pituitary -adrenal axis, thymus, thyroid hormones, and glucose levels. Prog. Neuropsychopharmacol. Biol. Psychiatry 2002, 26, 823–830. [Google Scholar] [CrossRef]
  35. Christiansen, S.; Bouzinova, E.V.; Palme, R.; Wiborg, O. Circadian activity of the hypothalamic-pituitary-adrenal axis is differentially affected in the rat chronic mild stress model of depression. Stress 2012, 15, 647–657. [Google Scholar] [CrossRef] [PubMed]
  36. Sullivan, G.M.; Coplan, J.D.; Kent, J.M.; Gorman, J.M. The noradrenergic system in pathological anxiety: A focus on panic with relevance to generalized anxiety and phobias. Biol. Psychiatry 1999, 46, 1205–1218. [Google Scholar] [CrossRef]
  37. Bondi, C.O.; Jett, J.D.; Morilak, D.A. Beneficial effects of desipramine on cognitive function of chronically stressed rats are mediated by alpha1-adrenergic receptors in medial prefrontal cortex. Prog. Neuropsychopharmacol. Biol. Psychiatry 2010, 34, 913–923. [Google Scholar] [CrossRef] [PubMed]
  38. Arnsten, A.F.; Li, B.M. Neurobiology of executive functions: Catecholamine influences on prefrontal cortical functions. Biol. Psychiatry 2005, 57, 1377–1384. [Google Scholar] [CrossRef] [PubMed]
  39. Wang, Y.L. The Study on Pharmacodynamics and Mechanism of Compound Ma Ti Xiang (CMTX). Master’s Thesis, Beijing University of Chinese Medicine, Beijing, China, 2011. [Google Scholar]
  40. Li, J.; Liu, Q.T.; Chen, Y.; Liu, J.; Shi, J.L.; Liu, Y.; Guo, J.Y. Involvement of 5-HT1A Receptors in the Anxiolytic-Like Effects of Quercitrin and Evidence of the Involvement of the Monoaminergic System. Evid.-Based Compl. Alt. Med. 2016. [Google Scholar] [CrossRef]
  41. Liu, J.; Zhai, W.M.; Yang, Y.X.; Shi, J.L.; Liu, Q.T.; Liu, G.L.; Fang, N.; Li, J.; Guo, J.Y. GABA and 5-HT systems are implicated in the anxiolytic-like effect of spinosin in mice. Pharmacol. Biochem. Behav. 2015, 128, 41–49. [Google Scholar] [CrossRef] [PubMed]
  42. Chen, W.W.; He, R.R.; Li, Y.F.; Li, S.B.; Tsoi, B.; Kurihara, H. Pharmacological studies on the anxiolytic effect of standardized Schisandralignans extract on restraint-stressed mice. Phytomedicine 2011, 18, 1144–1147. [Google Scholar] [CrossRef] [PubMed]
  43. Roche, M.; Kerr, D.M.; Hunt, S.P.; Kelly, J.P. Neurokinin-1 receptor deletion modulatesbehavioural and neurochemical alterations in an animal model of depression. Behav. Brain Res. 2012, 228, 91–98. [Google Scholar] [CrossRef] [PubMed]
  44. Wang, Y.L.; Shi, J.L.; Yong, L.; Zhao, R.; Zhai, Y.J.; Guo, J.Y. Anxiolytic-like effects of compound Zhi Zhu Xiang in rats. Evid.-Based Compl. Alt. Med. 2012, 3, 1–7. [Google Scholar] [CrossRef] [PubMed]
  45. Kim, C.; Speisky, M.B.; Kharouba, S.N. Rapid and sensitive method for measuring norepinephrin, dopamine, 5-hydroxytryptamine and their major metabolites in rat brain by high performance liquid chromatography. Differential effect of probenecid, haloperidol and yohimbine on the concentrations of biogenic amines and metabolites in various regions of rat brain. J. Chromatogr. A 1987, 386, 25–35. [Google Scholar]
Sample Availability: Samples of the SAG ((−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside)are available from the authors.
Molecules 22 01331 g001 550
Figure 1. Effect of (−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG) on the number of central entries and the time spent in central areas in the open field test in rats. (A) Number of central entries; (B) Time spent in central areas. Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Figure 1. Effect of (−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG) on the number of central entries and the time spent in central areas in the open field test in rats. (A) Number of central entries; (B) Time spent in central areas. Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Molecules 22 01331 g001
Molecules 22 01331 g002 550
Figure 2. Effect of SAG on the open arm entries and the percentage of time spent in open arms in the elevated plus maze in rats. (A) Open arm entries; (B) Percentage of time spent in open arms. Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Figure 2. Effect of SAG on the open arm entries and the percentage of time spent in open arms in the elevated plus maze in rats. (A) Open arm entries; (B) Percentage of time spent in open arms. Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Molecules 22 01331 g002
Molecules 22 01331 g003a 550Molecules 22 01331 g003b 550
Figure 3. Effect of SAG (0.9, 1.8, 3.6 mg/kg) on serum CRF, ACTH and CORT levels after the behavior tests. (A) Concentration of CRF (ng/L); (B) Concentration of ACTH (ng/L); (C) Concentration of CORT (ng/mL). Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Figure 3. Effect of SAG (0.9, 1.8, 3.6 mg/kg) on serum CRF, ACTH and CORT levels after the behavior tests. (A) Concentration of CRF (ng/L); (B) Concentration of ACTH (ng/L); (C) Concentration of CORT (ng/mL). Bars represent mean ± SEM. ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Molecules 22 01331 g003aMolecules 22 01331 g003b
Molecules 22 01331 g004 550
Figure 4. Structure of (−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG).
Figure 4. Structure of (−)-syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside (SAG).
Molecules 22 01331 g004
Molecules 22 01331 g005 550
Figure 5. The purity of SAG detected by HPLC. SAG was analyzed by HPLC with UV detection at 204 nm. The analysis was performed with a C-18 column (4.6 mm × 250 mm) at 35 °C. It was eluted (eluent A: acetonitrile; eluent B: 0.2% phosphate solution) at a flow rate of 1.0 mL/min.
Figure 5. The purity of SAG detected by HPLC. SAG was analyzed by HPLC with UV detection at 204 nm. The analysis was performed with a C-18 column (4.6 mm × 250 mm) at 35 °C. It was eluted (eluent A: acetonitrile; eluent B: 0.2% phosphate solution) at a flow rate of 1.0 mL/min.
Molecules 22 01331 g005
Molecules 22 01331 g006 550
Figure 6. Experimental groups and procedure.
Figure 6. Experimental groups and procedure.
Molecules 22 01331 g006
Table
Table 1. Effect of SAG on the monoamines neurotransmitters and their metabolites in the whole cerebral cortex of rats.
Table 1. Effect of SAG on the monoamines neurotransmitters and their metabolites in the whole cerebral cortex of rats.
GroupsNE (μg/g)DA (μg/g)DOPAC (μg/g)HVA (μg/g)5-HT (μg/g)5-HIAA (μg/g)
V/US121.35 ± 8.3239.33 ± 5.3835.99 ± 4.6134.03 ± 3.5820.31 ± 1.8948.11 ± 3.27
V/S188.87 ± 9.45 ##65.24 ± 6.69 #49.51 ± 5.0847.76 ± 5.15 #28.03 ± 2.43 #58.15 ± 3.92 #
DZP/S129.78 ± 4.60 **44.88 ± 6.00 *38.40 ± 4.2631.32 ± 4.23 *15.57 ± 0.97 **31.32 ± 4.23 **
SAG/S
(0.9 mg/kg)		167.45 ± 13.1755.44 ± 6.5745.87 ± 3.2237.89 ± 3.4121.14 ± 1.01 *55.21 ± 1.84
SAG/S
(1.8 mg/kg)		164.30 ± 6.12 *52.93 ± 4.6041.59 ± 4.0136.47 ± 3.3417.89 ± 1.01 **50.92 ± 2.91
SAG/S
(3.6 mg/kg)		136.10 ± 2.96 **46.76 ± 4.44 *40.64 ± 4.5234.00 ± 2.97 *17.69 ± 1.13 **34.00 ± 2.97 **
Values are expressed as the mean ± S.E.M. # p < 0.05 or ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.
Table
Table 2. Effect of SAG on the monoamine neurotransmitters and their metabolites in the hippocampus of rats.
Table 2. Effect of SAG on the monoamine neurotransmitters and their metabolites in the hippocampus of rats.
GroupsNE (μg/g)DA (μg/g)DOPAC (μg/g)HVA (μg/g)5-HT (μg/g)5-HIAA (μg/g)
V/US148.71 ± 7.9428.36 ± 2.5818.71 ± 1.3019.35 ± 2.8823.44 ± 3.2474.29 ± 2.69
V/S188.87 ± 9.45 ##65.24 ± 6.69 #49.51 ± 5.0847.76 ± 5.15 #28.03 ± 2.43 #58.15 ± 3.92 #
DZP/S124.13 ± 4.47 **23.77 ± 1.2014.78 ± 1.0911.95±1.2419.57 ± 1.76 **72.54 ± 2.53 **
SAG/S
(0.9 mg/kg)		153.39 ± 9.5328.06 ± 1.9619.93 ± 2.3917.37 ± 1.4425.76 ± 4.9586.17 ± 3.88
SAG/S
(1.8 mg/kg)		139.09 ± 4.89 **28.49 ± 1.6721.48 ± 4.6613.28 ± 2.1220.18 ± 1.40 *81.13 ± 10.86
SAG/S
(3.6 mg/kg)		135.37 ± 14.78 **26.45 ± 1.2415.93 ± 1.4314.71 ± 1.5919.50 ± 1.56 **76.99 ± 4.56 *
Values are expressed as the mean ± S.E.M. # p < 0.05 or ## p < 0.01 vs. vehicle/unstressed group; * p < 0.05 or ** p < 0.01 vs. vehicle/stressed group. One-way ANOVA with Student-Newman-Keuls post hoc test.

Share and Cite

MDPI and ACS Style

Liu, J.; Lv, Y.-W.; Shi, J.-L.; Ma, X.-J.; Chen, Y.; Zheng, Z.-Q.; Wang, S.-N.; Guo, J.-Y. Anti-Anxiety Effect of (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside from Albizzia julibrissin Durazz (Leguminosae). Molecules 2017, 22, 1331. https://doi.org/10.3390/molecules22081331

AMA Style

Liu J, Lv Y-W, Shi J-L, Ma X-J, Chen Y, Zheng Z-Q, Wang S-N, Guo J-Y. Anti-Anxiety Effect of (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside from Albizzia julibrissin Durazz (Leguminosae). Molecules. 2017; 22(8):1331. https://doi.org/10.3390/molecules22081331

Chicago/Turabian Style

Liu, Jie, Yue-Wei Lv, Jin-Li Shi, Xiao-Jie Ma, Yi Chen, Zhi-Quan Zheng, Sheng-Nan Wang, and Jian-You Guo. 2017. "Anti-Anxiety Effect of (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside from Albizzia julibrissin Durazz (Leguminosae)" Molecules 22, no. 8: 1331. https://doi.org/10.3390/molecules22081331

APA Style

Liu, J., Lv, Y.-W., Shi, J.-L., Ma, X.-J., Chen, Y., Zheng, Z.-Q., Wang, S.-N., & Guo, J.-Y. (2017). Anti-Anxiety Effect of (−)-Syringaresnol-4-O-β-d-apiofuranosyl-(1→2)-β-d-glucopyranoside from Albizzia julibrissin Durazz (Leguminosae). Molecules, 22(8), 1331. https://doi.org/10.3390/molecules22081331

Article Metrics

Back to TopTop