Target for treating and diagnosing anxiety disorder and application thereof

Abstract

The invention belongs to the technical field of biological medicine, and particularly provides a target spot for treating and diagnosing anxiety disorder, and the target spot comprises IL-1beta. The expression level of the target is closely related to the anxiety state and can be used as one of standards for screening the anxiety emotional state, and a brand-new drug target and a new treatment means and thought are provided for research and development of anxiety drugs. The invention further provides a method for rapidly constructing the anxiety disorder animal model based on the target, the problems that an existing anxiety disorder mouse model is long in construction period and unstable in effect, resistant mice can appear, and much manpower and material resources need to be consumed are solved, and great convenience is provided for follow-up research on the drug target for treating anxiety disorder.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to a target for anxiety treatment and diagnosis and application thereof. BACKGROUND

[0002] Anxiety, also known as anxiety neurosis, is the most common and highest incidence neurosis in mental illness, and is mainly characterized by anxiety emotion, with worry as the core symptom. It mainly includes generalized anxiety, acute anxiety attack, phobia, post-traumatic stress disorder, acute stress disorder, and obsessive-compulsive disorder. It is mainly manifested as nervous worry without clear objective object, restlessness, and autonomic nervous system symptoms (palpitation, hand tremor, sweating, frequent urination, etc.).

[0003] Since the pathogenesis of anxiety is very complex, there is a lack of safe and effective treatment methods, and therefore, in-depth exploration of the pathogenesis is an important basis for treating anxiety. A mouse disease model plays a very important role in studying the pathogenesis of human diseases and drug screening. In exploring human cognitive function, neurodegenerative diseases, neuropsychiatric diseases, and the like, constructing an anxiety mouse model is conducive to the research. A classic anxiety mouse model is currently achieved by long-term chronic stress modeling, and the construction method has a long cycle, unstable effect, and resistant mice may appear, which requires more human and material resources to be spent for screening. SUMMARY

[0004] The purpose of the present application is to provide a new drug target for treating anxiety disorders, and to overcome the problems of the existing anxiety mouse model, such as a long construction cycle, unstable effect, resistant mice, and the need for more human and material resources.

[0005] To this end, the present application provides a target for anxiety treatment and diagnosis, which comprises IL-1β.

[0006] Specifically, the above-mentioned early warning and diagnostic reagent for screening anxiety.

[0007] Specifically, the above-mentioned diagnostic reagent is used for detecting the expression level of IL-1β.

[0008] Specifically, the above-mentioned target can be used for preparing an anti-anxiety drug, and the drug comprises an effective dose of an IL-1β antagonist and a pharmaceutically acceptable carrier.

[0009] The present application also provides a method for screening an anti-anxiety drug, comprising the step of screening IL-1β as a therapeutic target.

[0010] Specifically, the above-mentioned method takes the decrease or absence of the expression level of IL-1β as a screening index.

[0011] The application further provides a method for rapidly constructing an anxiety animal model, comprising the following steps: continuously injecting IL-1β into the basolateral amygdaloid brain area of an animal, preferably embedding a cannula in the basolateral amygdaloid brain area of the animal bilaterally, and delivering IL-1β to the basolateral amygdaloid brain area through the cannula.

[0012] Specifically, IL-1β is injected into the basolateral amygdaloid brain area for 3 consecutive days, and 300 nL of IL-1β is injected every day.

[0013] Specifically, the anxiety animal model constructed by the method can be used for screening drugs for treating anxiety.

[0014] Compared with the prior art, the application has the following advantages and beneficial effects:

[0015] The target expression level provided by the application for anxiety treatment and diagnosis is closely related to the anxiety state, can be used as one of the standards for screening the anxiety emotional state, and provides a brand-new drug target and a new treatment method and idea for the research on drug targets for treating anxiety disorders.

[0016] The construction method provided by the application can rapidly prepare a mouse model of anxiety, overcomes the problems of long construction period, unstable effect, occurrence of resistant mice and consumption of a large amount of manpower and material resources of the existing mouse model of anxiety, and provides great convenience for subsequent research on drug targets for treating anxiety disorders.

[0017] The application will be further described in detail below with reference to the drawings. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is the detection result of a mouse induced by chronic stress in the embodiment of the application; wherein, A, a flowchart of chronic stress induction; B, total distance in an open field test; C, number of times of entering a central area in the open field test; D, total time of staying in the central area in the open field test; E, open arm staying time in an elevated plus maze test; F, number of times of entering the open arm in the elevated plus maze test; G, light box staying time in a black and white shuttle box test; H, crawling distance in the light box in the black and white shuttle box test; and I, number of times of conversion.

[0019] Figure 2 is the IL-1β expression concentration of the basolateral amygdaloid brain area of an anxiety mouse in the embodiment of the application.

[0020] Figure 3 is the result of improving anxiety of an anxiety mouse by neutralizing IL-1β in Anti-IL-1β-IgG in the embodiment of the application; wherein, A, an experimental flowchart; B, number of times of entering the open arm in the elevated plus maze test; and C, open arm staying time in the elevated plus maze test.

[0021] Figure 4 These are the results of the IL-1β anxiety mouse model constructed in the examples of the present invention; wherein, A, experimental process; B, speed, number of entries and total time spent in the central area in the open field test; C, total distance traveled, number of entries into the open arm and time spent in the open arm in the cross elevated maze test. DETAILED DESCRIPTION

[0022] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Although the representative embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited to the embodiments, but should be defined by the appended claims and their equivalents.

[0023] The target for the treatment and diagnosis of anxiety disorders and the effects of their application are studied below through specific examples.

[0024] Example 1:

[0025] Chronic stress can disrupt body homeostasis and cause abnormal changes in brain state and function, leading to anxiety. This example used a chronic stress-induced anxiety mouse model to detect IL-1β expression in the basolateral amygdala. The specific steps are as follows.

[0026] Six-week-old male mice were selected and subjected to a 28-day chronic stress model.

[0027] There are five types of chronic stressors: one is wet bedding overnight, sprinkle water on the bedding to ensure it is moist; the second is restraint, drill a few small holes in a 50mL centrifuge tube, and then stuff the mouse into the centrifuge tube to ensure its breathing is unobstructed, for 2 hours; the third is crowding, squeeze multiple mice into a box, also for 2 hours; the fourth is tilting the cage 45°; the fifth is reversing the mouse's circadian rhythm, that is, turning on the lights at night.

[0028] After 28 days, anxiety-like behavior tests were performed using open field, cross elevated maze, and black and white shuttle box. Figure 1 As shown, the model was confirmed to be successful.

[0029] After the basolateral amygdala of normal mice and anxious mice was dissected out, the expression level of IL-1β was detected using ELISA kit. The results are as follows: Figure 2 As shown in Figure 3, IL-1β expression was significantly increased in the basolateral amygdala of anxious mice.

[0030] Example 2:

[0031] In this example, specific compounds were used to regulate the expression level of IL-1β in astrocytes in the basolateral amygdala region of a classic anxiety mouse model, and their effect on improving the anxiety behavior of mice was studied, as follows.

[0032] The classic anxiety mouse model constructed in Example 1 was randomly divided into an experimental group and a control group. Cannulas were implanted bilaterally in the basolateral amygdala of the anxious mice, and the Anti-IL-1β-IgG antagonist (experimental group) and control IgG (control group) were delivered to the basolateral amygdala through the cannulas. At the same time, cannulas were implanted bilaterally in the basolateral amygdala of the normal mice, and control IgG was delivered to the basolateral amygdala through the cannulas. Each group was injected with 300 nL per day for 3 days. A cross elevated maze test was performed 30 minutes after the last injection. The results are as follows: Figure 3 As shown, S-anti IL-1β represents the experimental group injected with anti-IL-1β-IgG, S-ctrl represents anxious mice injected with control IgG, and N-ctrl represents normal mice injected with control IgG. The study found that neutralizing elevated IL-1β levels in the basolateral amygdala of anxious mice using an anti-IL-1β-IgG antagonist improved anxiety in these mice. This confirms that IL-1β is an important target for anxiety disorders and plays a crucial role in the development and treatment of anxiety drugs.

[0033] Example 3:

[0034] In this example, IL-1β was used to construct an anxiety mouse model, and the specific steps are as follows.

[0035] Cannulas were implanted bilaterally in the basolateral amygdala of normal mice. IL-1β (experimental group) and saline (control group) were delivered to the basolateral amygdala through the cannulas, with an injection volume of 300 nL for 3 days. The open field and cross elevated maze tests were performed 30 minutes after the last injection. Figure 4 The study found that IL-1β can indeed be used to construct an anxiety mouse model. This method can complete the construction of an anxiety mouse model in just three days, overcoming the problems of existing anxiety mouse models, such as long construction cycles, unstable results, the emergence of resistant mice, and the need for a large amount of manpower and material resources. This provides great convenience for subsequent research on drug targets for the treatment of anxiety disorders.

[0036] The above examples are merely illustrative of the present invention and do not limit the scope of protection of the present invention. Any design that is identical or similar to the present invention falls within the scope of protection of the present invention.

Claims

1. A target for the treatment and diagnosis of anxiety disorders, characterized by: The targets include IL-1β.

2. Use of the target as claimed in claim 1 in screening early warning and diagnostic reagents for anxiety disorders.

3. Use of the target site according to claim 2 in screening for early warning and diagnostic reagents for anxiety disorders, characterized in that: The diagnostic reagent is used to detect the expression level of IL-1β.

4. The use of the target site according to claim 1 in the preparation of antianxiety drugs, characterized in that: The medicine comprises an effective dose of IL-1β antagonist and a pharmaceutically acceptable carrier.

5. A method for screening antianxiety drugs, characterized in that: The method includes the step of screening IL-1β as a therapeutic target.

6. The method for screening antianxiety drugs according to claim 5, wherein: The decrease or absence of IL-1β expression level was used as the screening indicator.

7. A method for rapidly constructing an anxiety disorder animal model, characterized in that: The method comprises the following steps: continuously injecting IL-1β into the basolateral amygdala brain region of the animal.

8. The method for rapidly constructing an anxiety disorder animal model according to claim 7, wherein: IL-1β was injected into the basolateral amygdala for 3 consecutive days, with 300 nL injected each day.

9. An animal model of anxiety disorder, characterized by: The animal model is constructed using the method described in any one of claims 7 or 8.

10. Use of the anxiety disorder animal model according to claim 9 in screening drugs for treating anxiety disorders.

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