The effect of the elevated plus maze test-keptin on the anxiety behavior of rats

Effect of scorpion scorpion on anxiety behavior in rats

Abstract: Objective To further clarify whether anthraquinone has an anxiolytic effect. Methods The rat model of elevated plus maze was used as an animal model to evaluate the anxiolytic activity and characteristics of single subcutaneous injection and continuous subcutaneous injection. Results The scorpion scorpion could significantly increase the number of open arms and the percentage of staying time in the open arm of the elevated cross maze (P < 0.05, P < 0.01). The autonomous movement of the scorpion scorpion at the experimental dose Activity has no effect (P > 0.05). Conclusion The scorpion venom may have an anxiolytic effect.
Key words: scorpion; anxiety; elevated cross maze

Anxiety disorders, also known as anxiety disorders, or anxiety mental disorders, are a type of mental illness characterized by episodic or persistent emotional anxiety and stress. The incidence of anxiety disorders has increased year by year, seriously affecting people's quality of life. Drug therapy is currently the most widely used and most effective treatment. However, classic first-line anti-anxiety drugs often have drug-dependent, sedative, sexual dysfunction and other adverse reactions while treating anxiety disorders, which limits their clinical application. Therefore, it is urgent to develop effective and safe new anti-anxiety drugs. The hook kiss is a genus of the genus Macaque, and the Chinese crocodile is abundant in the carp, cassia, and other places. Its main component is steroidal alkaloids, which contain dozens of alkaloid monomers, of which koumine is An active ingredient with the highest content of domestic alkaloids. In the early stage of the laboratory, the mouse elevated cross maze test and the light and dark shuttle box test method were used. It was observed for the first time that the scorpion venom has a significant anti-anxiety effect. However, the anti-anxiety effect of scrotonin needs to be confirmed in more animal models of anxiety. Based on this, this paper intends to investigate the anxiolytic effect and characteristics of arbutin in rat elevated plus maze model by single administration and continuous administration.

1 Instruments and materials

1.1 Instruments XR-XG201 rat elevated cross maze (provided by Shanghai Xinsoft Information Technology Co., Ltd.)

1 . 2 The drug chelating prostaglandin powder was separated and purified by our research group and identified by high performance liquid chromatography with a mass fraction of 99.2%. When used, it was formulated with physiological saline to prepare the solution of the required mass concentration; diazepam injection, Tianjin Jinyao Amino Acid Co., Ltd., batch number 1010181, was prepared by using physiological saline to prepare a solution of the required concentration.

1 . 3 animals clean male Wistar rats weighing 180 ~ 200 g, license number: SCXK (Shanghai) 2012--0002, Shanghai SLAC Laboratory Animal Co. provided. Feed in a light-dark cycle 12/12 h (lighting time 8:00 to 20:00), constant humidity (50% ± 5%), constant temperature (24 ± 2 °C ), free to eat and drink, change 1 every 2 days The litter is kept in the animal room. Rats were strobed for 1 to 2 minutes per day 7 days before the experiment to reduce the effect of irrelevant stimulation on the experiment. Rats were transferred to the laboratory 1 h before dosing to familiarize them with the environment and eliminate the effects of unfamiliar environment on rats. All experimental procedures for rats are in compliance with regulations promulgated by the International and Local Laboratory Animal Use and Protection Committee.

2 methods

2 . 1 Animal Grouping and administration The rats were randomly divided into control group (saline), diazepam (diazepam, DZP) positive control group (0.5 mg · kg -1) and 0.75, 1.5 koumine And 3 mg·kg -1 dose group, 7-19 rats in each group, administered with a single injection of 0.005 mL·g -1 subcutaneously for 1 hour , and then from 9:00 to 13:00. Behavioral test; rats were randomly divided into control group ( salt saline), DZP group (0.5 mg·kg -1 ) and scrotonin 0.1875, 0.75, 1.5 and 3 mg·kg. -1 dose group, 7 to 15 cases per group. 0.005 mL · g -1, subcutaneous injection administered once a day for 7 d (wherein the DZP group of 1 ~ 6 d administered an equal volume of saline, administered first 7 d DZP 0.5 mg · kg -1), each group was subjected to a behavioral test at 9:00 to 13:00 after the last administration of d 7 for 1 hour .
2 . 2 Behavioral test The experimental animals were quickly placed in the central open part of the maze, with their heads facing one of the open arms, and the activity of the animals within 5 minutes was photographed by the upper camera . The following indicators were recorded: 1 Number of closed arms entering ( closearmentry , CE ); 2 open arm entry times ( openarmentry , OE ); 3 open arm time ( openarmtime , OT ). The following indicators were calculated from the above behavioral indicators: the total number of animals entering the open and closed arms ( to ‐ talarmentries ); the proportion of animals entering the open arms ( OE %), ie OE /( CE + OE ) × 100%; animals The ratio of residence time in the open arm ( OT %: OT / total time × 100%).
2 . 3 Statistical processing The experimental data was calculated using SPSS 13.0 for Windows software. Data were expressed as mean ± standard error ( mean ± S . E . M ). The mean comparison between the two groups was performed by two-sided t test. One -way ANOVA method was used for statistical processing among multiple groups of data . When the difference was significant, it was reused. LSD ‐ posthoc for statistical analysis. P <0.05 was considered statistically significant.

4 discussion
At present, the animal model of anxiety includes more than 30 kinds of various improvements, which mainly include two categories according to the behavior pattern: a model based on animal non-conditioned reflection and a model based on conditioned reflex (traditional learning mode). Elevated plus maze (elevatedplus - mazetest, EPM) is an animal principle conflict state of fear hanging open arms formed based on the novel to explore the behavior of animals and the environment, non-conditioned reflex model, is the most widely used One of the classic models for detecting animal anxiety behavior is widely used in psychopharmacological research. With the development of psychopharmacology, EPM can be used to evaluate anxiety behaviors of other animals belonging to rodents, such as guinea pigs , in addition to anxiety behavior in rats and mice. Evaluating whether a drug has an anxiolytic effect requires verification from multiple perspectives on multiple models. Therefore, this paper uses the rat elevated cross maze model to further evaluate the anxiolytic effect of the scorpion. In the EPM , the number of times the animal enters the arm and the time of stay in the arm fully reflects the anxiety state of the animal. In contrast, the total number of arms reflects the animal's motor activity. At the same time, we also found that EPM models based on animal non-conditioned reflex behavior are easily affected by experimental conditions throughout the experiment, such as: selection of experimental animals (species, age, gender, health status), animal-to-environment The adaptation and experimental conditions are basically the same (experimental operator, lighting, smell), feeding conditions, testing time, and so on. Diazepam is a classic benzodiazepine anti-anxiety drug with fast onset and significant effect. It is one of the most commonly used positive control drugs in animal models of anxiety. In single-dose and continuous-dose EPM models, we It can be observed that diazepam can significantly increase the number of entry of the animal's open arms and the residence time of the open arms, consistent with the literature reports, indicating that the model is stable.

Studies have shown that the total alkaloids of North American hook and kiss at a dose of 150 mg·kg -1 can significantly increase the number and time of mice entering the open arms in the elevated plus maze. In the mouse-dark boxing experiment, it was found that the total alkaloids of North American hook and kiss can significantly increase the residence time of the mice in the bright box and the number of shuttles in the light and dark box, showing significant anti-anxiety effects. The hook-knot alkaloids, scutellarin and scorpion, also showed significant anti-anxiety effects in the mouse-dark boxing model. In addition, the scorpion venom is relatively less toxic than other chelating alkaloid monomers (mouse LD 50 is 99.0 mg·kg -1 , sc ), suggesting that scrotonin may be highly efficient and low toxic. Anxiolytic effect. On this basis, this paper uses the rat elevated cross maze as an animal model to further explore the hook alkaloids from different angles.
Anxiolytic effects and characteristics.

The results suggest that the scorpion venom exhibits an anxiolytic effect consistent with diazepam in a rat model of elevated plus maze, in a dose-dependent manner. Interestingly, we found that the anxiolytic effects of scorpion scorpion were not a classic linear relationship in the single and continuous administration of the rat elevated plus maze. The reason may be that the scorpion scorpion has reached the maximum effect of anti-anxiety effect within a certain range, and its anti-anxiety effect no longer increases with the increase of the drug dose, so the brain area associated with anxiety appears to be high dose of oxin. Not sensitive. It can be seen that the dose is strictly controlled when applying the anesthesia. In addition, we also observed that the scorpion venom has no significant effect on the animal's motor activity, indicating that it exerts an anxiolytic effect and has no sedative and hypnotic effects.
In summary, the scorpion venom may have an anti-anxiety effect, and has the characteristics of high efficiency and low toxicity, and is expected to become a novel drug for treating anxiety. In view of the diversity of animal models of anxiety, its anti-anxiety effect has yet to be confirmed on more models, and the mechanism of its anti-anxiety effect remains to be explored.

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