Study on Anti-anxiety Effect and Mechanism of Schisandra chinensis
OBJECTIVE: To study the anti-anxiety effect and mechanism of Schisandra Chinensis Baill Lignans (SCL). Methods The low-, medium-, and high-dose lignans of Schisandra Chinensis were observed by light and dark box test and elevated plus maze test (35, 70, 140). The effect of mg/kg on the behavior of mice after 7 days of intragastric administration; a blank control group (administered with normal saline) and diazepam (2 mg / kg) positive control group. Enzyme-linked immunosorbent assay (ELISA) The serum γ-aminobutyric acid (GABA) content was determined. Results The results of the black box test and the elevated plus maze test showed that the middle and high dose groups of Schisandra chinensis had significant anti-anxiety effects (P<0.05). ), and in a dose-dependent manner, this anxiolytic effect can be antagonized by flumazenil. Enzyme-linked immunosorbent assay showed that the serum levels of γ-aminobutyric acid (GABA) were significantly increased in the middle and high dose groups. Conclusion Schisandra lignan has significant anti-anxiety effect, and its mechanism may be through γ-aminobutyric acid (GABA-R).
Key words: Schisandra chinensis lignans; anxiolytic; light and dark box; elevated plus maze; γ-aminobutyric acid
Schisandra chinensis (Turcz Baill) is a traditional herbal medicine in China. Its main pharmacological activity is Schisandra Chinensis Baill Lignans (SCL). At present, seven pharmacological activities have been isolated from Schisandra chinensis. The ingredients are Schisandra A, B, A, A, B, E, E. B. schisandra has a variety of pharmacological effects, which can eliminate free radicals, protect the liver and improve immunity. Force and anti-tumor, etc. In recent years, some studies have found that: Schisandra lignan has significant central inhibition and soothing effect. According to our previous research: Schisandra lignan has a good sedative and hypnotic effect, Schisandra chinensis It has a strong sedative and hypnotic effect on experimental mice. Based on its sedative and hypnotic effects, this study used a variety of anxiety models to observe the effect of Schisandra chinensis on anxiety in mice, and to explore its mechanism of action. To develop a theoretical basis for the development of functional foods and medicines for the protection of anxiolytics in Schisandra chinensis, and to promote the comprehensive research, development and utilization of Schisandra chinensis.
Experimental material
1. 1 experimental animals
ICR mice, body weight 18 ~ 22 g (Changchun Yisi Experimental Animal Center, animal license number SCXK (Ji) 2011-0004). 20 ~ 25 °C caged for 5 days, free diet to adapt the mice lab environment.
1. 2 Drugs and reagents Schisandra lignan (SCL) (Laboratory of Food Science and Engineering Research, College of Forestry, Beihua University); Sodium Chloride Injection (Shandong Jinyang Pharmaceutical Co., Ltd.); Diazepa Tablet (0806007) , Beijing Yimin Pharmaceutical Co., Ltd.); flumazenil (054k37041, Sigma, USA); mouse gamma aminobutyric acid (GABA) ELISA kit (US RD).
1. 3 experimental instrument mouse with black and white box (XR-XB120 type, provided by Shanghai Xinsoft Information Technology Co., Ltd.); mouse elevated plus maze (XR-XG201 type, provided by Shanghai Xinsoft Information Technology Co., Ltd.); enzyme-linked immunosorbent assay Detector (Biotek, USA).
2. Method
2. 1 Grouping and administration of experimental animals ICR mice were randomly divided into low, medium and high doses of SCL (35, 70, 140 mg / kg), blank control group (physiological saline solution) and positive control group (diazepam, 2 Mg / kg), a total of 5 groups, SCL group administered intraperitoneal morning and evening, a total of 2 times, the blank control group was given the same volume of normal saline, the positive control group was administered 2 mg / (kg · d-1) The above groups were continuously administered (or physiological saline) for 7 days. On the 7th day, the SCL group and the blank control group were administered for 1 h, and the positive control group was administered for 0.5 h after the test. All tests were at 8: 00 ~ 14:00 in a quiet environment.
2. 2 Mouse black and white box test Before the light and dark box experiment, in order to reduce the influence of some external non-essential stimuli on the experimental results, the mice were touched every day for 1 ~ 2 min, and the test was started after 5-7 days. The experimental mice were placed in the dark box to the center of the bright box, and then the time of the mouse in the bright box and the number of shuttles between the bright box and the black box were recorded within 5 min.
2. 3 mouse elevated plus maze test Before the elevated cross maze test, in order to ensure the reliability and accuracy of the test results, the mice were first allowed to move freely in a separate cage for 5 min, and then tested. The mouse is placed in the center of the maze, facing the open arm, and then recording begins. The main recorded data includes the number of times the mouse entered the open arm and the time spent in the open arm within 5 min, the number of times the arm was closed and the time spent in the closed arm. During the whole test process, the mouse was used to exit the representative to complete an activity in both arms. In order to ensure the accuracy of the test, the maze should be cleaned after each completion before the next test can be performed. Processing, the main indicators include: the percentage of the number of open arms and the total number of times to enter the two arms (OE); the percentage of activity time in the open arm and the total activity time into the two arms (OT).
2. 4 Half an hour before the intraperitoneal injection of flumazenil, animals in the low, medium and high doses of SCL and the control group of diazepam were intraperitoneally injected with flumazenil (3 mg / kg).
2. 5 Determination of serum gamma aminobutyric acid concentration After the end of the experiment, the mice were decapitated and taken at 4 °C, 3 000 r / min to obtain the supernatant, and the mouse γ aminobutyric acid ELISA detection reagent was used. The gamma aminobutyric acid content of the serum was detected by the cartridge, and the absorbance (OD) value at 450 nm was measured by a microplate reader. The final gamma aminobutyric acid content was expressed in μg / mL.
2. 6 Statistical analysis Experimental data were statistically analyzed using SPSS 13. 0 software, and one-way ANOVA was used to perform a comparison test between multiple groups. All data were in mean ± standard deviation ( x ■± s ) indicates that P < 0.05 is statistically significant.
4 Discussion
Based on anxiety factors can be induced by a variety of factors, such as psychological factors, social stress, mental stimulation and other factors, this study uses a common anxiety model light box and elevated plus maze can be a good examination of the anxiolytic effect of SCL. In this study The results of the black and white box test showed that when the dose of schisandra lignans was 70,140 mg / kg, it had obvious anti-anxiety effect, and its anxiolytic effect increased with the increase of SCL dose. In the elevated plus maze test, it is usually used. Mouse OT and OE to reflect the anxiety state of the mice. In the elevated plus maze test of this study, when the dose of schisandra lignans reached 70 mg / kg, OT and OE were > 26% and 28%, respectively, with the blank control group. The 19% and 23% of the drug were significantly increased. In this experiment, the benzodiazepine drug diazepam was used as a positive control to initially explore the anti-anxiety mechanism of SCL. The anti-anxiety mechanism of diazepam was through the increase of γ-amino group. The content of butyric acid (GABA) receptor enhances the post-synaptic inhibition effect, thereby achieving anti-anxiety. In recent years, studies have found that γ-aminobutyric acid plays an important role in the γ-aminobutyric acid neural pathway. Type A Body (GABA-A receptor). GABA is the main neurotransmitter in the brain, which acts by inhibiting GABA-A receptors. Conversely, flumazenil is an antagonist of this receptor and can block benzene. The combination of diazoxides (such as diazepam) with the receptor weakens or even reverses the effects of anti-anxiety drugs. The results of the light and dark box test and the elevated plus maze test in this study show that: Schisandra chinensis The anxiolytic effects of din and diazepam can be inhibited by flumazenil. Therefore, the mechanism by which SCL exerts an anxiolytic effect may be achieved by gamma-aminobutyric acid receptor. In addition to gamma-aminobutyric acid neurotransmission In addition to neurotransmitters, dopamine (DA), norepinephrine (NE), and serotonin (5-HT) are associated with anxiety disorders. Therefore, the exact mechanism of action against olfactorin in Schisandra chinensis is still Need further research.
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