New use of yujie tablet in treating anxiety

Abstract

This invention belongs to the field of biomedical technology, specifically relating to a novel use of Yueju tablets in anxiolytic activity. Specifically, the Yueju tablets provided by this invention exhibited significant anxiolytic activity in a mouse acute anxiety model experiment, showing superior efficacy compared to the positive control drug fluoxetine in some models, and can be used to develop anxiolytic drugs. Preliminary mechanistic studies have shown that it exerts a significant anxiolytic effect by reducing neuroinflammation and protecting neuronal integrity.

Description

[0001] Priority application This application claims priority to Chinese invention patent application No. 2026104581020, filed on April 8, 2026, entitled "A Novel Use of Yueju Tablets in Anti-Anxiety," which is incorporated herein by reference in its entirety. Technical Field

[0002] This invention belongs to the field of biomedical technology, specifically relating to a new use of Yueju tablets in anti-anxiety. Background Technology

[0003] Anxiety neurosis, or anxiety disorder for short, is a group of mental disorders characterized by persistent worry, fear, tension, and autonomic nervous system dysfunction. The core features of anxiety disorder are the long duration and high severity of symptoms, which significantly interfere with the patient's daily life.

[0004] It is generally believed that anxiety disorders are measurable changes in the physical structure and function of the brain (biological basis), and also the result of a vicious cycle of intertwined specific thought and behavioral patterns (psychological processes). The core issues are altered neuroplasticity and neurotransmitter imbalances in the brain. Examples include insufficient function of the gamma-aminobutyric acid (GABA) system, dysregulation of the serotonin system, and overactivity of the norepinephrine system. Current mainstream drug treatments primarily focus on correcting neurological function. Fluoxetine is a classic selective serotonin reuptake inhibitor (SSRI) and is a first-line drug for treating anxiety. However, this class of drugs has many drawbacks, including excessive sedation, headaches, dizziness, gastrointestinal discomfort, and liver damage. Furthermore, approximately 30%–40% of patients develop treatment-resistant depression due to insensitivity to fluoxetine.

[0005] Acute anxiety is a symptom fundamentally different from chronic, diffuse anxiety disorders such as generalized anxiety disorder. Its core characteristic is an intense, explosive anxiety response that occurs after an individual experiences a sudden, significant, or traumatic event. This particular type of anxiety is often accompanied by neuroinflammation, such as the abnormal activation of microglia and astrocytes in the central nervous system, releasing large amounts of pro-inflammatory cytokines (such as IL-1β, IL-6, and TNF-α). This, in turn, disrupts the blood-brain barrier, interferes with neurotransmitter balance, inhibits hippocampal neurogenesis, and impairs the function of emotional regulation brain regions such as the prefrontal cortex and amygdala.

[0006] Currently, there are no approved drugs specifically for alleviating anxiety-induced behaviors caused by neuroinflammation. Therefore, a new approach is needed to improve this specific type of anxiety disorder. Summary of the Invention

[0007] The purpose of this invention is to provide a new use of Yueju tablets in anti-anxiety, partially solving or alleviating the above-mentioned deficiencies in the prior art. The invention specifically adopts the following technical solution.

[0008] One aspect of this invention is to provide the application of Yueju tablets in the treatment of a specific type of anxiety disorder.

[0009] The application of Yueju tablets in the preparation of drugs for treating anxiety, wherein the anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation.

[0010] Furthermore, the ingredients of the Yueju tablets include vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu (medicated leaven).

[0011] Furthermore, the Yueju tablets also include pharmaceutical excipients or additives used in the preparation of the drug.

[0012] Another aspect of the present invention aims to provide the application of Yueju tablets in the prevention of a specific anxiety disorder.

[0013] The application of Yueju tablets in the preparation of drugs for preventing anxiety, wherein the anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation.

[0014] Furthermore, the ingredients of the Yueju tablets include vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu (medicated leaven).

[0015] Furthermore, the Yueju tablets also include pharmaceutical excipients or additives used in the preparation of the drug.

[0016] Another aspect of the present invention can provide the application of a traditional Chinese medicine composition in the prevention or treatment of anxiety.

[0017] The application of a traditional Chinese medicine composition in the preparation of a drug for the prevention or treatment of anxiety, wherein the anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation; the traditional Chinese medicine composition includes vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu (medicated leaven).

[0018] Furthermore, the traditional Chinese medicine composition includes other pharmaceutically acceptable excipients.

[0019] Furthermore, the dosage form of the traditional Chinese medicine composition includes tablets, powders, pills, or aqueous solutions.

[0020] Furthermore, the mass ratio of the vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu is 1:1:1:1:1.

[0021] Beneficial technical effects: This invention proposes a novel use for Yueju tablets in the prevention or treatment of acute anxiety; specifically, this invention addresses the accompanying neuroinflammation associated with this acute anxiety. Traditionally, Yueju tablets are used to regulate Qi and relieve depression, alleviate abdominal distension, and treat symptoms such as chest tightness, abdominal distension, food stagnation, belching, and acid reflux. This invention discovers that Yueju tablets can exert a significant anti-acute anxiety effect by reducing neuroinflammation and protecting neuronal integrity. Specific experiments show that the Yueju tablets provided by this invention exhibit significant anti-anxiety activity in a mouse acute anxiety model, and in some models, their efficacy is superior to the positive control drug fluoxetine, suggesting their potential for developing anti-acute anxiety drugs. Preliminary mechanistic studies indicate that they exert a significant anti-anxiety effect by reducing neuroinflammation and protecting neuronal integrity. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0023] Figure 1 The effect of Yueju tablets on anxiety-like behavior in mice (Yue Ju Low represents the low-dose group of Yueju tablets, and Yue Ju High represents the high-dose group of Yueju tablets). Figure 2 Morphological results of anti-anxiety effects of Yueju tablets; Figure 3 Iba-1 and NLRP3 staining of brain tissue from anxious mice treated with Yueju tablets. Detailed Implementation

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0025] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0026] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0027] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, and even more typically + / -0.5% of the value.

[0028] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0029] Detailed description of the attached images: Figure 1 (A) Feeding latency in the novelty inhibition feeding experiment; (B) Central region time in the open field experiment; (C) Bright field dwell time in the light-dark box experiment; n = 10; compared to the model group, * P <0.05, ** P <0.01, *** P <0.001; compared with the control group, ### P <0.001.

[0030] Figure 2 (A) HE staining of brain tissue from anxious rats; (B) Nissl staining of brain tissue from anxious rats; Figure 3 (A) Iba-1 staining of brain tissue from anxious mice; (B) NLRP3 staining of brain tissue from anxious mice.

[0031] Definition of noun: The acute anxiety described in this invention refers to a situational stimulus or an acute event that occurs in a special situation, and this special type of anxiety is accompanied by the occurrence of neuroinflammation.

[0032] Example 1 1. Test drug Yueju tablets: Commercially available standard tablets are used. The dosage is determined by referring to the drug instructions and combining the animal body surface area conversion formula. It is divided into two dosage groups: low dose group 1 g / kg and high dose group 4 g / kg.

[0033] Positive control drug: Fluoxetine, 10 mg / kg 2. Acute lipopolysaccharide-induced anxiety animal model Fifty male C57BL-6J mice were randomly divided into four groups (n=10 per group) in a serpentine pattern, ordered by the number of spontaneous activities during screening: a control group (same volume of distilled water), a model group, a positive control group (fluoxetine), a low-dose group (Yueju tablets), and a high-dose group (Yueju tablets). All drugs were prepared as suspensions in distilled water and administered to each group once daily by gavage at a volume of 0.2 mL / 10 g body weight for 7 consecutive days. From day 6 to day 7, all groups except the control group received an intraperitoneal injection of 100 μL of lipopolysaccharide (LPS, 1 mg / kg), while the control group received an equal volume of physiological saline. Open field and novelty-induced feeding inhibition tests were performed on day 6, and open field and light-dark chamber tests were performed on day 7.

[0034] 3. Behavioral assessment (1) Open Field Test (OFT) OFT is primarily used to assess spontaneous activity levels and exploratory behavior in animals, and can also reflect anxiety-like behaviors (central area avoidance) and whether drugs cause nonspecific motor inhibition or excitation. In this study, it was used to rule out whether the anxiolytic effect was caused by changes in motor ability. Mice were placed in the center of a 40×40×30 cm white square open box, and the total movement distance, time spent in the central area, and number of entries were recorded over 5 minutes.

[0035] (2) Light-Dark Box Test (LDB) LDB utilizes mice's instinct to prefer dark environments, reflecting their anxiety levels through their willingness to explore in bright areas. Anti-anxiety drugs significantly prolong the time spent in the bright area and increase the number of times they move between areas. The device is divided into a bright area (20×20×20 cm, bright light illumination) and a dark area (20×20×20 cm, dark), connected by a small door at the bottom. Mice are placed in the bright area, and the latency to enter the dark box, the number of times they enter the bright area, and the time spent in the bright area are recorded within 5 minutes.

[0036] (3) Novelty-Suppressed Feeding Test (NSFT) NSFT is a classic behavioral method for assessing anxiety-like behavior in animals and has high sensitivity for screening anxiety-related drugs. Mice that have been fasted for 24 hours are placed in the center of an unfamiliar open field, with a food trough containing food placed in the center of the field. The latency period from when the mice are placed in the open field to when they first eat is recorded.

[0037] NSFT is a classic behavioral method for assessing anxiety-like behavior in animals and has high sensitivity for screening anxiety-related drugs. Mice that have been fasted for 24 hours are placed in the center of an unfamiliar open field, with a food trough containing food placed in the center of the field. The latency period from when the mice are placed in the open field to when they first eat is recorded.

[0038] 4. Statistical Analysis All data are expressed as mean ± standard error (Mean ± SEM). Statistical analysis was performed using Graphpad Prism 10.1.2 software. One-way ANOVA was used for comparisons among multiple groups. Dunnett's method was used when variances were homogeneous, and Dunnett's T3 method was used when variances were unequal. P A value <0.05 is considered statistically significant.

[0039] 5. Morphological analysis (1) HE staining (Hematoxylin-Eosin Staining, HE) HE staining is the most commonly used routine staining method in histological studies, mainly used to observe the morphology, cell arrangement, and pathological changes of tissues. It can clearly distinguish between cell nuclei and cytoplasm, clarify the histological integrity of brain tissues such as the prefrontal cortex, and determine whether there are abnormalities such as cell damage, edema, or inflammatory infiltration. In this study, it was used to evaluate the effect of Yueju tablets on the morphology of prefrontal cortex tissue in anxiety model mice. Prefrontal cortex tissue from mice was processed through dewaxing, hydration, hematoxylin staining, eosin staining, dehydration and clearing, and mounting. The tissues were then observed and photographed under an optical microscope to analyze morphological changes.

[0040] (2) Nissl staining Nissl staining is primarily used to visualize the cell bodies and nucleoli of neurons, clearly showing the number, morphology, and distribution of neurons. It is a classic method for assessing neuronal damage and survival. In this study, it was used to detect the integrity of neurons in the prefrontal cortex of anxiety model mice and to determine whether Yueju slices have a neuroprotective effect. Processed paraffin sections of the prefrontal cortex were dewaxed, hydrated, stained with Nissl stain, differentiated, dehydrated, cleared, and mounted. The sections were then observed under a microscope, and the number of neurons and the proportion of morphological abnormalities were statistically analyzed.

[0041] (3) Immunohistochemistry (IHC) Immunohistochemistry is a technique that utilizes the principle of specific antigen-antibody binding to locate the expression of specific proteins in tissues. It can accurately detect the expression site and intensity of target proteins, and is characterized by high specificity and high sensitivity. Paraffin sections are dewaxed, hydrated, antigen-retrievaled, blocked, incubated with primary antibody, incubated with secondary antibody, DAB staining, hematoxylin counterstaining, dehydration and clearing, and then mounted. The expression intensity of BDNF-positive cells is then observed and quantitatively analyzed under a microscope.

[0042] Experimental results: see Figure 1 And Table 1-3. Animal experimental results showed that in the novelty inhibition feeding experiment, both Yueju tablets and the positive control drug fluoxetine significantly shortened the feeding latency period. Yueju tablets showed a dose-dependent effect and at high doses, they were superior to the positive control drug fluoxetine. Figure 1 A, Table 1); In the open field test, Yueju tablets significantly increased the time mice spent exploring the decentral area at low doses, and showed a trend of increasing the exploration time at high doses, but there was no statistically significant difference ( Figure 1 B, Table 2); In the light-dark box experiment, both low and high doses of Yueju tablets significantly increased the time mice spent exploring in the light box (B, Table 2). Figure 1 (C, Table 3). The above results indicate that Yueju tablets have a significant anti-anxiety effect.

[0043] Table 1. Feeding latency in novelty inhibition feeding experiments (mean ± standard error) (Compared with the blank group, ### P <0.001, compared with the model group, ** P <0.01, *** P <0.001) Table 2. Central region time in open field experiments (mean ± standard error) (Compared with the blank group, ## P <0.01, compared with the model group, * P <0.05, ** P <0.01) Table 3. Dwell time in the bright field during the dark-field experiment (mean ± standard error) (Compared with the blank group, ### P <0.001, compared with the model group, * P <0.05) Figure 2The morphological results of the mouse brain after treatment with Yueju tablets are shown. HE staining results showed that neurons in the brain tissue sections of the blank control group were morphologically intact, with neatly arranged cells, clear nuclei, and no obvious inflammatory infiltration or cellular edema. Neurons in the model group showed obvious pathological damage, manifested as disordered cell arrangement, cell body swelling, nucleus condensation or dissolution, and inflammatory cell infiltration and interstitial edema were also observed, indicating that the model successfully induced brain injury and inflammatory response. Neuronal morphology in the fluoxetine group was significantly improved compared to the model group, with more regular cell arrangement, improved nucleus clarity, and significantly reduced inflammatory infiltration and cellular edema, leaving only a few scattered abnormal cells. Neurons in the Yueju tablet treatment group had neatly arranged cells, clear nuclei, and almost complete disappearance of inflammatory infiltration and edema, with only a very few abnormal cells observed, suggesting that Yueju tablets have a significant repair effect on pathological damage.

[0044] Nissl staining results showed that neurons in the blank control group had abundant Nissl bodies, which appeared as dark blue granules, were evenly distributed, and had plump cell morphology. In the model group, Nissl bodies were significantly reduced, dissolved, or even disappeared; cell bodies were shrunken, and cell outlines were blurred, indicating severe neuronal damage and dysfunction. In the fluoxetine group, the number of Nissl bodies was significantly restored compared to the model group; the granules were clear, and the cell morphology was plump, with only a small number of neurons showing reduced Nissl bodies remaining. In the Yueju tablet-treated group, Nissl bodies were abundant and evenly distributed, and neuronal morphology was intact, indicating that Yueju tablets can effectively protect neuronal integrity.

[0045] Figure 3 The results show the changes in signaling proteins in mouse brain tissue after treatment with Yueju tablets. Iba-1 immunohistochemical staining results showed that brain tissue sections from the control group were light brownish-yellow, with only a few scattered Iba-1 positive cells visible under high magnification, indicating that microglia were in a resting state. Compared with the control group, brain tissue sections from the model group showed significantly deeper staining, appearing dark brownish-yellow; under high magnification, a large number of densely distributed Iba-1 positive cells were visible, with enlarged cell bodies and thickened processes, exhibiting typical activated morphology, indicating that microglia were widely activated and the neuroinflammatory response was significantly enhanced in the model group. The staining intensity in the fluoxetine group and the Yueju tablet treatment group was significantly weaker than that in the model group, with fewer positive cells visible under high magnification, indicating that microglia activation was significantly inhibited and neuroinflammatory response was reduced.

[0046] Immunohistochemical staining of NLRP3 showed that brain tissue sections from the control group mice were light brownish-yellow, with only a few scattered positive staining areas under high magnification, indicating a low basal expression level of NLRP3 inflammasomes in the brain. Brain tissue sections from the model group mice showed significantly deeper staining, appearing dark brownish-yellow; extensive and dense positive staining areas were visible under high magnification, indicating that NLRP3 inflammasomes were significantly activated and their expression level was greatly increased, resulting in a significantly enhanced neuroinflammatory response. Sections from both the fluoxetine group and the Yueju tablet-treated group showed light staining, with scattered positive staining areas, indicating that Yueju tablets can significantly reduce the expression level of NLRP3 inflammasomes and alleviate neuroinflammatory symptoms, with efficacy comparable to fluoxetine.

[0047] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0048] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. The application of Yueju tablets in the preparation of drugs for treating anxiety, characterized in that, The anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation.

2. The application as described in claim 1, characterized in that, The ingredients of Yueju tablets include vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu (medicated leaven).

3. The application as described in claim 1, characterized in that, The Yueju tablets include pharmaceutical excipients or additives used in the preparation of the drug.

4. The application of Yueju tablets in the preparation of drugs for preventing anxiety, characterized in that, The anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation.

5. The application as described in claim 4, characterized in that, The ingredients of Yueju tablets include vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu (medicated leaven).

6. The application as described in claim 4, characterized in that, The Yueju tablets include pharmaceutical excipients or additives used in the preparation of the drug.

7. The application of a traditional Chinese medicine composition in the preparation of a drug for the prevention or treatment of anxiety, characterized in that, The anxiety is acute anxiety; the acute anxiety is accompanied by the occurrence of neuroinflammation; the traditional Chinese medicine composition includes vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu.

8. The application as described in claim 7, characterized in that, The traditional Chinese medicine composition includes other pharmaceutically acceptable excipients.

9. The application as described in claim 7, characterized in that, The dosage forms of the traditional Chinese medicine composition include tablets, powders, pills, or liquids.

10. The application as described in claim 7, characterized in that, The mass ratio of the vinegar-processed Cyperus rotundus, Ligusticum chuanxiong, stir-fried Gardenia jasminoides, stir-fried Atractylodes lancea, and stir-fried Shenqu is 1:1:1:1:1.

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