Composition for preventing, ameliorating or treating post-traumatic stress disorder, depression, and cognitive function disorder comprising composite herbal medicine extract as effective component

Abstract

The present invention relates to a composition for improving or treating post-traumatic stress disorder, depression, and cognitive impairment, comprising a complex herbal extract as an active ingredient. When the hot water extracts of ginseng, astragalus, atractylodes, or licorice, respectively, and the complex herbal extract according to the present invention were administered to animal models induced with post-traumatic stress disorder, and changes in behavior in an open space were observed, it was confirmed that compared to the single extracts of ginseng, astragalus, atractylodes, or licorice, the total distance traveled and movement paths (number of times passing through the center, time spent in the center) significantly increased when the complex herbal extract composed of ginseng, astragalus, atractylodes, and licorice was administered. Furthermore, as a result of the Y-maze experiment, the behavioral score reduced by post-traumatic stress disorder was improved by the administration of the complex herbal extract of the present invention, and the expression level of proteins related to brain (hippocampus) function was enhanced; thus, it can be usefully utilized as a health functional food or pharmaceutical for the improvement or treatment of post-traumatic stress disorder, depression, and cognitive impairment.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention, improvement, or treatment of post-traumatic stress disorder, depression, and cognitive impairment, comprising a complex herbal medicine extract as an active ingredient. Background Technology

[0002] Post-traumatic stress disorder (PTSD) is a mental illness that develops following psychological trauma resulting from an event involving physical injury or anxiety about life. It is also known as traumatic stress disorder, post-traumatic stress disorder, post-traumatic syndrome, trauma, or post-traumatic stress. It primarily occurs after experiencing events that deviate from everyday occurrences, such as natural disasters, fires, war, physical assault, torture, rape, sexual assault, hostage situations, child abuse, accidents involving cars, airplanes, trains, or ships, or other major disasters. The timing of symptom onset varies from person to person; symptoms may begin immediately after the shock or may appear after several days, weeks, months, or years. A diagnosis of PTSD is made only if symptoms persist for at least one month; if symptoms appear within one month and last for less than three months, it is classified as acute stress disorder. Symptoms manifest as hyperalertness or hyperarousal, re-experience or intrusion of the trauma, and avoidance or emotional numbness. Hyperalertness includes being easily startled, feeling anxious, unable to sleep, and having difficulty concentrating. For example, Vietnam War veterans often keep their backs against a wall, while victims of rape remain on high alert as if the perpetrator were present. Symptoms of re-experience of the trauma involve the replay of memories, dreams, or hallucinations of the event, which are perceived as real, accompanied by physical symptoms. Even in cases where there is almost no memory, physical symptoms may appear if faced with a similar situation. Symptoms of emotional avoidance or numbness refer to the absence of normal emotions; because only unrealistic feelings arise, anger, victim mentality, and shame develop.Furthermore, symptoms such as headaches, indigestion, vomiting, stomach pain, tremors, and allergic reactions caused by hormonal changes may occur. These manifest in various ways, including difficulty with bowel movements, separation anxiety (a reluctance to be apart), school phobia, and fear of strangers. Unrealistic emotions can lead to reliance on alcohol and drugs, resulting in abuse and addiction, as well as autonomic nervous system disorders. Sometimes, hallucinations occur, accompanied by dissociative disorders or panic attacks. In young children, dreams about ghosts may occur rather than dreams about actual experiences.

[0003] Meanwhile, ginseng ( Panax Ginseng Ginseng is a medicinal herb belonging to the genus Panax of the family Araliaceae, which has been widely known and used in both traditional Korean medicine and folk remedies. It is classified into fresh ginseng, white ginseng, and red ginseng depending on its state. Fresh ginseng refers to the roots of ginseng that have not been dried after harvesting, while white ginseng refers to the roots of fresh ginseng that have had their fine roots removed, the skin peeled off, and then dried in the sun. Red ginseng is produced by steaming and drying the roots of ginseng. Ginseng saponins possess antioxidant activity by promoting the secretion of catecholamines in the adrenal glands and inhibiting the lipid peroxidation process. Additionally, protective effects against radiation, protection against chromosomal abnormalities and micronuclei induced by alkylating agents, and reductions in mutations and cell deformation in cultured cells have been reported.

[0004] Astragalus is a perennial herbaceous plant belonging to the Leguminosae family. Astragalus membranceusIt is the root of *Bunge* with the outer skin almost completely removed, and its components include sucrose, glucuronic acid, various amino acids, bitter substances, mucilage, choline, betaine, and folic acid. In traditional Korean medicine, it is known to be effective as a tonic for physical weakness, spontaneous sweating, boils, thirst, abdominal pain, kidney debilitation, deafness, shortness of breath, draining pus and relieving pain, loss of taste in food, loss of appetite, hemorrhoids, conjunctivitis, malaria, prolapse of the anus, uterine bleeding, leukorrhea, menstrual irregularities, prenatal and postnatal diseases, expectorant, headache, heart fire in the lungs, dysentery, and various childhood diseases.

[0005] Atractylodes Atractylodes japonica Koidzumi refers to the dried, trimmed roots of the Atractylodes plant, also known as Sangae or Cheongae. It is very bitter but has a slightly sweet taste. It is best when the inside is white and the bulb is full, and it is known that those harvested in the autumn are particularly effective.

[0006] licorice( Glycyrrhiza uralensis Glycyrrhiza glabra is a perennial plant belonging to the legume family, and its roots and the lower part of its stems are used as a sweetener and a treatment for gastric and duodenal ulcers. Because it softens the effects of other drugs, it is used for various purposes in traditional Korean medicine. In particular, the glablidin component in licorice has skin whitening activity and is used as an ingredient in cosmetics; it has also been reported to have antioxidant and lipid metabolism physiological activities.

[0007] Meanwhile, regarding technologies related to post-traumatic stress disorder, Korean Registered Patent No. 1887578 discloses a composition for the prevention or treatment of depression or post-traumatic stress disorder containing a melanin aggregation hormone as an active ingredient, and Korean Registered Patent No. 2424765 discloses a method for treating post-traumatic stress disorder; however, there has not yet been any disclosure of a composition for the prevention, improvement, or treatment of post-traumatic stress disorder, depression, and cognitive impairment containing the complex herbal medicine extract of the present invention as an active ingredient. The problem to be solved

[0008] The present invention was derived from the above-mentioned needs. The present invention provides a composition for improving or treating post-traumatic stress disorder, depression, and cognitive impairment, comprising a complex herbal medicine extract as an active ingredient. The invention was completed by administering a hot water extract of ginseng, astragalus, atractylodes, or licorice, respectively, and a hot water extract of the complex herbal medicine according to the present invention to an animal model induced with post-traumatic stress disorder, and observing changes in behavior in an open space. It was confirmed that, compared to the single extracts of ginseng, astragalus, atractylodes, or licorice, the total distance traveled and movement paths (number of times passing through the center, time spent in the center) significantly increased when the complex herbal medicine hot water extract consisting of ginseng, astragalus, atractylodes, and licorice was administered. Furthermore, the invention was completed by confirming, through Y-maze experiments, that the behavioral score reduced by post-traumatic stress disorder was enhanced by the administration of the complex herbal medicine extract of the present invention, and that the expression level of proteins related to brain (hippocampus) function was increased. means of solving the problem

[0009] To achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0010] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of anxiety disorders, depression, or cognitive decline caused by post-traumatic stress disorder, comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0011] In addition, the present invention provides a health functional food composition for the prevention or improvement of post-traumatic stress disorder comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0012] In addition, the present invention provides a health functional food composition for the prevention or improvement of anxiety, depression, or cognitive decline caused by post-traumatic stress disorder, comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0013] In addition, the present invention provides a veterinary composition for the prevention or treatment of post-traumatic stress disorder comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0014] In addition, the present invention provides a feed additive for the prevention or improvement of post-traumatic stress disorder comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0015] In addition, the present invention provides a method for preventing or treating post-traumatic stress disorder, comprising the step of administering a pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder to an individual other than a human who has developed or is likely to develop post-traumatic stress disorder.

[0016] In addition, the present invention provides a method for preventing or treating anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder, comprising the step of administering a pharmaceutical composition for the prevention or treatment of anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder to an individual other than a human who has developed or is likely to develop anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder. Effects of the invention

[0017] The present invention relates to a composition for improving or treating post-traumatic stress disorder, depression, and cognitive impairment, comprising a complex herbal extract as an active ingredient. When the hot water extracts of ginseng, astragalus, atractylodes, or licorice, respectively, and the complex herbal extract according to the present invention were administered to an animal model induced with post-traumatic stress disorder, and changes in behavior in an open space were observed, it was confirmed that compared to the single extracts of ginseng, astragalus, atractylodes, or licorice, the total distance traveled and movement paths (number of times passing through the center, time spent in the center) significantly increased when the complex herbal extract composed of ginseng, astragalus, atractylodes, and licorice was administered. Furthermore, as a result of the Y-maze experiment, the behavioral score reduced by post-traumatic stress disorder was improved by the administration of the complex herbal extract of the present invention, and there was an effect of enhancing the expression level of proteins related to brain (hippocampus) function. Brief explanation of the drawing

[0018] Figure 1 shows the results of confirming behavioral changes in an open space after administering a complex herbal extract (hot water extract of ginseng, Astragalus membranaceus, Atractylodes macrocephala, and licorice) to an animal model induced with post-traumatic stress disorder. Fluoxetine is the positive control, and extracts of ginseng, Astragalus membranaceus, Atractylodes macrocephala, and licorice alone are the comparison groups. The figures show the total movement path (A), total distance traveled (B), movement speed (C), time spent in the center (D), and the number of times crossing the center (E). ### and #### indicate that the total distance traveled, time spent in the center, and number of times crossing the center were statistically significantly reduced in the control group compared to the normal group, with ### being p<0.001 and #### being p<0.0001. *, **, and **** indicate that the total distance traveled, time spent in the center, and number of times crossing the center increased significantly for the group administered the complex herbal medicine extract of the present invention or fluoxetine (positive control) compared to the control group, with * being p<0.05, ** being p<0.01, and **** being p<0.0001. Figure 2 shows the results of a Y-maze experiment in which a complex herbal extract (hot water extract of ginseng, Astragalus membranaceus, Atractylodes macrocephala, and licorice) was administered to an animal model with induced post-traumatic stress disorder, and the results are represented as behavioral scores. It shows the total number of entries (A) and the change behavioral score (B) for each branch. # and ### indicate that the total number of entries or the change behavioral score of the control group was statistically significantly reduced compared to the normal group, with # being p<0.05 and ### being p<0.001. * and **** indicate that the change behavioral score of the group administered the complex herbal extract of the present invention or fluoxetine (positive control) was statistically significantly increased compared to the control group, with * being p<0.05 and **** being p<0.0001. Figure 3 shows the results of confirming changes in the expression levels of brain (hippocampus) function-related proteins using immunohistochemistry after administering a complex herbal medicine extract (hot water extract of ginseng, Astragalus membranaceus, Atractylodes macrocephala, and licorice) to an animal model induced with post-traumatic stress disorder. It shows the changes in the expression levels of Prox1 and DCX (doublecortin), markers of dentate granule cells and immature neurons in the hippocampus (A, B), and the expression levels of c-fos and p-CREB (C, D). Specific details for implementing the invention

[0019] The present invention relates to a pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0020] In addition, the present invention relates to a pharmaceutical composition for the prevention or treatment of anxiety disorders, depression, or cognitive decline caused by post-traumatic stress disorder, comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0021] The complex herbal extract composed of ginseng, astragalus, atractylodes, and licorice can be prepared by a method including the following steps, but is not limited thereto:

[0022] (1) A step of extracting by adding an extraction solvent to a complex herbal medicine composed of dried ginseng, Astragalus membranaceus, Atractylodes macrocephala and licorice;

[0023] (2) A step of filtering the extract of step (1); and

[0024] (3) A step of preparing an extract by drying the filtered extract of step (2).

[0025] In step (1) above, the extraction solvent is preferably selected from water, C1-C4 lower alcohols, or a mixture thereof, more preferably water or ethanol, and even more preferably water, but is not limited thereto. In the above manufacturing method, the extraction method may utilize any conventional method known in the art, such as hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. It is preferable to extract by adding the extraction solvent in an amount of 1 to 20 times the weight of the complex herbal medicine. The extraction temperature is preferably 20 to 110°C, but is not limited thereto. In addition, the extraction time is preferably 1 to 5 hours, and more preferably 2 to 4 hours, but is not limited thereto. In step (3) above, drying is preferably performed by reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze-drying, and more preferably by reduced pressure drying, but is not limited thereto.

[0026] The above-mentioned complex herbal medicine composed of ginseng, astragalus, atractylodes, and licorice is preferably mixed in a weight ratio of 1 to 10:1 to 10:1 to 10:1 to 10:1, more preferably in a weight ratio of 1 to 5:1 to 5:1 to 5:1 to 5:1, and even more preferably in a weight ratio of 1:1:1:1, but is not limited thereto.

[0027] A pharmaceutical composition comprising a complex herbal extract according to the present invention as an active ingredient is preferably in any one of the formulations selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols, but is not limited thereto. In addition to the complex herbal extract, it may further include a pharmaceutically acceptable carrier, excipient, or diluent. Examples of carriers, excipients, or diluents that may be included in a pharmaceutical composition comprising a complex herbal extract include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulating, it may be prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, humectants, disintegrants, and surfactants. The preferred dosage of the complex herbal extract of the present invention varies depending on the patient's condition and body weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by those skilled in the art. However, for a desirable effect, it is preferable to administer the pharmaceutical composition containing the complex herbal extract of the present invention at a dose of 0.0001 to 500 mg / kg per day, preferably 0.001 to 250 mg / kg. Administration may be performed once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0028] In addition, the present invention relates to a health functional food composition for the prevention or improvement of post-traumatic stress disorder comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0029] In addition, the present invention relates to a health functional food composition for the prevention or improvement of anxiety, depression, or cognitive decline caused by post-traumatic stress disorder, comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0030] The above-mentioned health functional food composition is preferably prepared in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto. The health functional food composition of the present invention may be prepared by adding the complex herbal medicine extract as is or by mixing it with other foods or food ingredients, and may be appropriately prepared according to conventional methods. Examples of foods to which the complex herbal medicine extract may be added include any one form selected from caramel, meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense. That is, there are no special restrictions on the types of foods. The above health functional food composition may contain various nutritional supplements, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, coloring agents and promoters (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, it may contain fruit pulp for the production of natural fruit juices and vegetable beverages. The above ingredients may be used independently or in combination.

[0031] The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, and the natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Although the proportion of the natural carbohydrates is not of great importance, it is preferable to include 0.01 to 0.04 g per 100 g of the composition of the present invention, and more preferably 0.02 to 0.03 g, but is not limited thereto. As sweeteners, natural sweeteners such as taumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used.

[0032] In addition, the present invention relates to a veterinary composition for the prevention or treatment of post-traumatic stress disorder comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0033] The veterinary composition of the present invention may further include suitable excipients and diluents according to conventional methods. Examples of excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, ethanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil. The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, humectants, spices, emulsifiers, preservatives, etc. The veterinary composition according to the present invention may be formulated using methods well known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, etc. The effective amount of the veterinary composition according to the present invention may be appropriately selected according to the individual animal. It may be determined according to the severity of the disease or condition, the sensitivity to the active ingredient of the present invention according to the age, weight, health status or sex of the individual, the route of administration, the duration of administration, factors including other compositions combined with or used simultaneously with said composition, and other factors well known in the physiological or veterinary field.

[0034] In addition, the present invention relates to a feed additive for the prevention or improvement of post-traumatic stress disorder comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient.

[0035] The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act. In the present invention, the term "feed" may refer to any natural or artificial prescribed food, single meal, etc., or the components of said single meal, intended for or suitable for animals to eat, consume, and digest. The types of said feed are not particularly limited, and feeds commonly used in the relevant technical field may be used. Non-limiting examples of said feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, single-cell proteins, zooplankton, or food waste. These may be used individually or in a mixture of two or more types.

[0036] In addition, the present invention relates to a method for preventing or treating post-traumatic stress disorder, comprising the step of administering a pharmaceutical composition for the prevention or treatment of said post-traumatic stress disorder to an individual other than a human who has developed or is likely to develop post-traumatic stress disorder.

[0037] In addition, the present invention relates to a method for preventing or treating anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder, comprising the step of administering a pharmaceutical composition for the prevention or treatment of anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder to an individual other than a human who has developed or is likely to develop anxiety disorder, depression, or cognitive decline caused by post-traumatic stress disorder.

[0038] The present invention will be described in more detail below using examples. These examples are solely for the purpose of more specifically explaining the present invention, and it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

[0039] Example 1. Preparation of a complex herbal medicine extract

[0040] A complex herbal medicine mixture of ginseng (250g), astragalus (250g), atractylodes (250g), and licorice (250g) was reflux-extracted in 10 times the amount of distilled water at 101±1℃ for 3 hours. Afterward, the complex herbal medicine extract was filtered, concentrated, freeze-dried to powder, and stored frozen until administration to animals.

[0041] In addition, single extracts of ginseng, astragalus, atractylodes, and licorice used as comparative examples were also prepared under the same conditions as the above-mentioned complex herbal medicine extract.

[0042] Example 2. Establishment of an animal model for inducing post-traumatic stress disorder.

[0043] (1) Classification of animal models and experimental groups

[0044] For animal experiments aimed at improving and treating posttraumatic stress disorder (PTSD), 20–22g C57BL / 6J mice (7 weeks old, Daehan BioLink Co., Eumseong-gun, Chungcheongbuk-do, Korea) were used. The mouse rearing environment maintained a 12-hour light / dark cycle (lights turned on at 8:00 am and off at 8:00 pm), a temperature of 21±2℃, and a relative humidity of 55±10%. All of the above methods and procedures were approved by the Animal Care Committee of the Korea Institute of Oriental Medicine, and all procedures were performed in accordance with the guidelines for the care of experimental animals.

[0045] In the above animal model, after being exposed to SPS+FS by applying a single prolonged stress (SPS) stimulus and a foot shock (FS) to the soles of the feet, a hot water extract of a complex herbal medicine dissolved in 0.9% saline (250 mg / kg) and a positive control fluoxetine hydrochloride (20 mg / kg) were orally administered once a day for 14 days, and the experimental group not exposed to SPS+FS (normal group) and the SPS+FS-induced mouse group were orally administered the same dose of saline as above to serve as the control group.

[0046] In addition, 8 mice per group were randomly divided into a normal group, a PTSD-induced group (control group), a PTSD+ginseng group, a PTSD+Astragalus group, a PTSD+Atractylodes group, and a PTSD+licorice group. The subjects were classified into a PTSD+complex herbal extract administration group and a PTSD+fluoxetine administration group (positive control group).

[0047] (2) Single prolonged stress and foot shock

[0048] Among the groups classified in (1) above, mice excluding the normal group were placed in a holder for 4 hours, and then immediately forced to swim for 20 minutes. Afterwards, the mice were allowed to recover for 15 minutes, and then a single prolonged stress (SPS) stimulus was applied by exposing them to ether until they lost consciousness. After they fully regained consciousness, an electric shock (Foot shock; FS, 1 mA, 5 sec, twice) was applied to the soles of their feet. Afterwards, the FS was applied to the mice every two days.

[0049] Example 3. Verification of behavioral changes in an open space (Open field test; OFT)

[0050] An open field test (OFT) was conducted to confirm behavioral changes following SPS+FS stimulation and the administration of the complex herbal extract.

[0051] A black plastic square box (100cm × 100cm × 40cm) and a white acrylic plate were prepared and divided into a central area and a periphery. The movement of the mice was measured using a video-tracking system with the EthoVision program (Noldus, Wageningen, Netherlands), and activity was measured by the total distance traveled in the box, speed, the number of times crossing the central area over 20 minutes, and the time spent there.

[0052] As a result, the total distance traveled by the control group was significantly reduced compared to the normal group, and the total distance traveled by the group administered the complex herbal extract was increased compared to the control group (Figs. 1A and 1B), while there was no difference in the speed of movement among all experimental groups (Fig. 1C).

[0053] Meanwhile, it was confirmed that the time spent in the center and the number of times crossing the center were significantly reduced in the control group compared to the normal group, and that the time spent in the center and the number of times crossing the center were significantly increased in the group administered the complex herbal extract of the present invention compared to the control group. In addition, it was confirmed that there was almost no difference in behavioral changes with the single extracts of ginseng, astragalus, atractylodes, or licorice compared to the control group (Figs. 1D and 1E).

[0054] Example 4. Evaluation of cognitive function using the Y-maze test

[0055] A Y-maze experiment was conducted to evaluate cognitive function induced by SPS+FS stimulation and herbal medicine extracts.

[0056] The apparatus for the Y-maze experiment consists of three arms (designated as A, B, and C). The length of each arm is 40 cm, the width is 12 cm, and the height is 30 cm. The angle at which three arms meet adjacent arms is 120 degrees, and the entire experimental apparatus is composed of black plastic and white acrylic plates. A mouse was carefully placed in the center of the maze and allowed to move freely for 8 minutes. The arm the mouse entered was then recorded, but only if the mouse had entered completely up to its tail; repeated entry into an arm was also recorded. The number of times and the order of entry into each arm were measured to evaluate the mouse's total arm entry and spontaneous alteration score (% correction). The spontaneous alteration score was defined as entering the three arms sequentially, i.e., ABC, BCA, CAB, etc., and was calculated using the following Equation 1.

[0057] [Mathematical Formula 1]

[0058] Change Action Points (% correction) = [Total number of alterations] / [Total number of times entered in each arm - 2] × 100

[0059] As a result, as shown in Figure 2, the cognitive function of the control group was impaired compared to the normal group, resulting in a low change behavior score, whereas the group administered the complex herbal medicine extract showed improved memory and recovered to the level of the normal group.

[0060] Example 5. Confirmation of changes in hippocampal function using immunostaining

[0061] The anxiety, depression, and memory impairment associated with PTSD are closely related to the temporal lobe of the brain, particularly involving the hippocampus. The dentate gyrus of the hippocampus is the site where hippocampal neurogenesis—the generation of new neurons throughout life—occurs, as well as learning and memory.

[0062] Brains excised from mice were sectioned in a cryostat in a 1-in-6 series. To inactivate intracellular peroxidase, the sections were treated with 0.6% hydrogen peroxide, washed, and blocked. They were then incubated overnight with a primary antibody at 4°C, followed by incubation with a biotinylated secondary antibody for 2 hours at room temperature. After washing, the sections were incubated with ABC solution for 1 hour at room temperature, stained with a DAB kit, and observed under a light microscope. Sections 6 through 10 of the rostrocaudal sections of the entire hippocampus were analyzed.

[0063] Changes in the expression levels of Prox1 and DCX (doublecortin), markers of dentate granule cells and immature neurons in the hippocampus, were confirmed using immunohistochemistry.

[0064] As a result, the number of Prox-positive cells expressing Prox1 and DCX-positive cells expressing DCX in the hippocampal hilus of the control group increased compared to the normal group (arrow). A pattern of significant decrease was observed in the group administered the complex herbal extract and fluoxetine (Figures 3A and 3B).

[0065] In addition, changes in the expression levels of c-fos and p-CREB, markers that can confirm the activity of dental granule cells and neural progenitor cells present in the hippocampus, were confirmed by immunohistochemistry.

[0066] As a result, the expression of c-fos in the hippocampus of the control group was significantly increased compared to the normal group and decreased by the administration of the complex herbal extract and fluoxetine (Fig. 3C). In the case of p-CREB, expression decreased in the control group compared to the normal group and was confirmed to recover in the group administered the complex herbal extract and fluoxetine (Fig. 3D).

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, comprising as an active ingredient a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice. Claim 2 A pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, characterized in that, in claim 1, the extraction solvent of the complex herbal medicine extract is water, any one lower alcohol selected from C1 to C4, or a mixture thereof. Claim 3 delete Claim 4 delete Claim 5 A health functional food composition for the prevention or improvement of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, comprising a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice as an active ingredient. Claim 6 delete Claim 7 A veterinary composition for the prevention or treatment of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, comprising as an active ingredient a complex herbal medicine extract consisting of ginseng, astragalus, atractylodes, and licorice. Claim 8 A feed additive for the prevention or improvement of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, comprising a complex herbal medicine extract composed of ginseng, astragalus, atractylodes, and licorice as an active ingredient. Claim 9 A method for the prevention or treatment of post-traumatic stress disorder accompanied by anxiety, depression, or cognitive decline, comprising the step of administering the pharmaceutical composition of claim 1 or 2 to an individual other than a human who has developed or is likely to develop post-traumatic stress disorder. Claim 10 delete

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