A traditional Chinese medicine composition for preventing and / or treating depression and a preparation method of a fermented wine thereof

Abstract

The application provides a traditional Chinese medicine composition for preventing and / or treating depression and a preparation method of a fermented wine thereof, and belongs to the technical field of traditional Chinese medicine preparations. The traditional Chinese medicine composition is prepared from raw medicinal materials in the following weight ratio: rose 8-55 parts, lily 8-55 parts, dendrobium 5-55 parts, spina date seed 3-25 parts, ginkgo leaf 3-25 parts, jujube 8-55 parts, float wheat 5-55 parts, and licorice 3-25 parts. The traditional Chinese medicine composition has the effects of activating qi, relieving depression, clearing heart and nourishing yin, relieving restlessness and calming the mind, and is used for preventing and treating mild to moderate depression caused by emotional obstruction, heart yin deficiency and liver qi disharmony, and has obvious curative effect. The traditional Chinese medicine composition is fermented to obtain a fermented wine, and the curative effect is further improved; and the fermented wine has bright red color, nose-piercing aroma and excellent taste, and has high public acceptance. In addition, the raw medicinal materials of the traditional Chinese medicine composition are traditional Chinese medicines with the same origin of medicine and food, are safe and reliable to use, and have good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparation technology, specifically relating to a traditional Chinese medicine composition for the prevention and / or treatment of depression and a method for preparing its fermented wine. Background Technology

[0002] Depression is a common mood disorder characterized primarily by low mood, often accompanied by anxiety, sleep disturbances, and loss of appetite. Studies show that there are currently as many as 90 million people with depression in China, meaning one in every 16 people suffers from it. The prevalence of depression is significantly higher in women than in men, approximately two to three times higher. About 200,000 people commit suicide due to depression each year, making it the second leading cause of disease burden on humanity. While the incidence of depression is increasing annually, the cure rate is only 10%. This is not only related to the nature of the disease itself but also closely linked to the large number of people with mild symptoms remaining unaware of their condition. Depression has a high rate of missed diagnosis, high relapse rate, and long duration. If left untreated, symptoms can persist for weeks, months, or even years. In severe cases, it can lead to extreme behaviors such as self-harm and suicide, affecting social stability and endangering human health.

[0003] Currently, the main medications for treating depression are Western medicines, such as imipramine, fluoxetine, and citalopram. However, these often cause significant side effects, including dizziness, headaches, anxiety, insomnia, extrapyramidal reactions, and gastrointestinal reactions. Patients generally believe that antidepressants act directly on the brain, potentially affecting memory, causing anxiety, and leading to insomnia, making them difficult to accept and resulting in poor adherence. Traditional Chinese medicine (TCM) classifies depression into categories such as yuzheng, zangzao, insomnia, globus hystericus, and lily disease. Clinically, commonly used formulas include modified Ganmai Dazao Tang, modified Chaihu Shugan San, and modified Danzhi Xiaoyao San. Commonly used patent medicines include Baishile capsules and Shugan Jieyu capsules, offering limited choices. Functional antidepressant products combining prevention and treatment are even rarer. Therefore, it is essential to develop related health products for the prevention and treatment of depression. Summary of the Invention

[0004] The purpose of this invention is to provide a traditional Chinese medicine composition for the prevention and / or treatment of depression and a method for preparing its fermented wine.

[0005] This invention provides a traditional Chinese medicine composition, which is prepared from raw materials in the following weight ratio: 8-55 parts rose petals, 8-55 parts lily bulbs, 5-55 parts dendrobium, 3-25 parts jujube seed, 3-25 parts bamboo leaf, 8-55 parts jujube, 5-55 parts floating wheat, and 3-25 parts licorice.

[0006] Furthermore, the aforementioned traditional Chinese medicine composition is prepared from the following raw materials in the indicated weight ratios: 10-50 parts rose petals, 10-50 parts lily bulbs, 5-50 parts dendrobium, 5-20 parts jujube seed, 5-20 parts bamboo leaf, 10-50 parts jujube, 5-50 parts floating wheat, and 5-20 parts licorice.

[0007] Furthermore, the aforementioned traditional Chinese medicine composition is prepared from the following raw materials in the indicated weight ratios: 10-50 parts rose petals, 10-30 parts lily bulbs, 5-30 parts dendrobium, 5-15 parts jujube seed, 10-15 parts bamboo leaf, 10-30 parts jujube, 10-30 parts floating wheat, and 5-18 parts licorice.

[0008] Furthermore, the aforementioned traditional Chinese medicine composition is prepared from the following raw materials in the indicated weight ratios: 50 parts rose petals, 30 parts lily bulbs, 30 parts dendrobium, 15 parts jujube seed, 15 parts bamboo leaf, 30 parts jujube, 30 parts floating wheat, and 18 parts licorice. Preferably, The roses mentioned are dark red roses; And / or, the Dendrobium is Dendrobium officinale; And / or, the jujube kernels are roasted jujube kernels.

[0009] The preparation method of the traditional Chinese medicine composition includes the following steps: taking raw materials in various weight proportions and preparing fermented wine through microbial fermentation technology.

[0010] The present invention also provides a method for preparing the aforementioned traditional Chinese medicine composition, which includes the following steps: (1) Weigh each raw material according to the weight ratio; (2) After sterilizing the raw drug, add water, glycogen and fermentation bacteria to ferment. After fermentation, filter coarsely, stop fermentation, age, filter finely, flavor, and bottle to obtain the product; Preferably, In step (2), the weight of the water is 5 to 30 times the total weight of the raw materials; And / or, in step (2), the weight of the glycogen is 10-30% of the total weight of water; And / or, in step (2), the weight of the fermenting bacteria is 0.01% to 0.5% of the total weight of water; And / or, in step (2), the fermenting bacteria are activated fermenting bacteria; And / or, in step (2), the fermentation is carried out at 10-40°C for 5-15 days until the sugar content drops to 4-9%; And / or, in step (2), the fermentation termination is achieved by adding 0.1~0.5% of the coarse filtrate by weight of a terminator; And / or, in step (2), the aging is a sealed aging for 20 to 90 days; And / or, in step (2), the seasoning is 0.01%~0.1% of the weight of the filtrate added as a seasoning agent; More preferably, In step (2), the glycogen is one or more of the following: granulated sugar, maltose, fructose syrup, glucose, or starch hydrolysis products; And / or, in step (2), the fermenting bacteria are yeast. Saccharomycs cerevisiae ; And / or, in step (2), the activation method of the fermentation bacteria is to add the fermentation bacteria to a white sugar aqueous solution with a mass fraction of 20~30% and activate it at 40~50℃ for 20~30 min; And / or, in step (2), the coarse filtration is performed using 10-50 mesh gauze; And / or, in step (2), the terminating agent is one or more of sodium metabisulfite, sodium benzoate, and potassium sorbate; And / or, in step (2), the fine filtration is performed by filtration with a diatomaceous earth filter, and the diatomaceous earth mesh size is selected as 50-200 mesh; And / or, in step (2), the flavoring agent is selected from one or more of sucrose, steviol glycosides, sucralose, and acesulfame potassium.

[0011] The present invention also provides the use of the aforementioned traditional Chinese medicine composition in the preparation of medicaments for the prevention and / or treatment of depression; Preferably, the drug is for the prevention and / or treatment of mild to moderate depression caused by emotional distress, heart yin deficiency, and liver qi disharmony.

[0012] The formula of this invention comprises the following: Rose petals regulate qi and relieve depression, lily bulbs clear the heart and nourish yin, serving as the principal herbs; jujube seed nourishes the heart and liver, calms the mind and soothes the nerves; bamboo leaves clear the heart and relieve irritability; dendrobium nourishes yin, clears heat, benefits the stomach and generates fluids, serving as the assistant herbs; jujube tonifies the middle jiao and replenishes qi, nourishes blood and calms the mind; wheat bran tonifies qi and nourishes yin, relieves irritability and calms the mind, serving as the adjuvant herbs; and licorice root tonifies qi and harmonizes the middle jiao, moistens dryness and relieves spasms, serving as the guiding herb. The combined effects of these herbs are to regulate qi and relieve depression, clear the heart and nourish yin, relieve irritability and calm the mind. This formula is used for symptoms such as depression, insomnia, irritability, anxiety, and forgetfulness caused by heart yin deficiency and liver qi disharmony, as well as menopausal syndrome and neurasthenia.

[0013] Indications: Regulates Qi and relieves depression, clears the heart and nourishes Yin, eliminates irritability and calms the mind. Used for symptoms such as depression, insomnia, irritability, anxiety, and forgetfulness caused by deficiency of heart Yin and disharmony of liver Qi, as well as menopausal syndrome and neurasthenia.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) By examining the process parameters of fermentation, coarse filtration, termination of fermentation, aging, and flavoring, this invention determines the optimal parameters, which can retain the active ingredients to the greatest extent. The resulting wine has a bright red color, a fragrant aroma, and a smooth taste, which fully meets the selection criteria of contemporary women and greatly increases the patient's medication compliance.

[0015] (2) This invention obtains Bai Mei Jie Yu traditional Chinese medicine fermented wine by fermenting the medicinal materials involved in the entire prescription of Bai Mei Jie Yu. Compared with the decoction used in clinical practice, this wine has a 1.4-fold increase in total flavonoid extraction rate, a 1.5-fold increase in the extraction rate of antidepressant active monomer catechin, a 5.7-fold increase in the extraction rate of puerarin, a 4.1-fold increase in the extraction rate of hesperidin, and the addition of shampoosin and naringin active monomers, thereby realizing the effect of increasing the efficacy of traditional Chinese medicine fermentation.

[0016] (3) The Chinese medicine formula of this invention is scientific and reasonable, and has significant effects on mild to moderate depression caused by emotional distress, heart yin deficiency and liver qi disharmony. All the Chinese medicines used are food and medicine of the same origin, with a high safety factor. The prepared products are of excellent quality and have definite curative effects. They are suitable as medicines that combine prevention and treatment of depression and have broad market application prospects.

[0017] In summary, this invention provides a traditional Chinese medicine composition for the prevention and / or treatment of depression. This composition has the effects of regulating qi and relieving depression, clearing the heart and nourishing yin, relieving irritability and calming the mind. It is used to prevent and treat mild to moderate depression caused by emotional distress, heart yin deficiency, and liver qi disharmony, with significant efficacy. Fermenting this composition to obtain fermented wine further enhances its effect in preventing and / or treating depression, making it significantly more effective than decoctions. Furthermore, the raw materials of this composition are all traditional Chinese medicines that are both food and medicine, making it safe and reliable to use and showing good application prospects.

[0018] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0019] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0020] Figure 1 The names and structures of five antidepressant active ingredients in the fermented wine of this invention are as follows.

[0021] Figure 2 The high-performance liquid chromatogram of the mixed reference standard in Experiment Example 1 is shown below; 1: catechin; 2: puerarin; 3: schaferoside; 4: naringin; 5: hesperidin.

[0022] Figure 3 The high-performance liquid chromatogram of the test solution of Bai Mei Jie Yu fermented wine (batch number: 20230901) in Experiment Example 1; 1: catechin; 2: puerarin; 3: schaferoside; 4: naringin; 5: hesperidin.

[0023] Figure 4 The high-performance liquid chromatogram of the test solution of Bai Mei Jie Yu Tang (batch number: J20230901) in Comparative Example 1; 1: catechin; 2: puerarin; 5: hesperidin. Detailed Implementation

[0024] The raw materials and equipment used in the specific embodiments of the present invention are all known products, obtained by purchasing commercially available products.

[0025] Dark red rose ( Rosa chinensis Jacq 'Crimson Glory' HT is an edible double-petaled red rose.

[0026] Floating wheat is a plant belonging to the genus *Triticum* of the family Poaceae. Triticum aestivum The dried, light, and shriveled fruit of L. It is a traditional Chinese medicine that can be used both as food and medicine.

[0027] Lily bulb, Dendrobium officinale, stir-fried jujube seed, Lophatherum gracile, jujube, and licorice are all Chinese medicinal herbs recorded in the 2020 edition of the Chinese Pharmacopoeia. They are all Chinese medicinal herbs that can be used as both food and medicine.

[0028] Lily bulb (batch number: C047230101), Dendrobium officinale (batch number: C509230301), stir-fried jujube seed (batch number: C525230401), Lophatherum gracile (batch number: C156221201), jujube (batch number: C152221201), wheat bran (batch number: C192230201), and licorice root (batch number: C851230201) were purchased from Chengdu Xinfuyuan Chinese Herbal Medicine Pieces Co., Ltd.; dark red rose was purchased from Yunnan Qizi Technology Co., Ltd.

[0029] Example 1: Preparation method of the Hundred Roses Depression-Relieving Fermented Wine of the present invention Ingredient formula (parts by weight): 50 parts dark red rose, 30 parts lily, 30 parts Dendrobium officinale, 15 parts roasted jujube seed, 15 parts light bamboo leaf, 30 parts jujube, 30 parts floating wheat, 18 parts licorice.

[0030] Preparation method: Weigh the raw materials according to the above weight proportions. Except for the dark red rose, sterilize the other medicinal materials at 110℃ for 30 minutes. Add the dark red rose, pour in purified water at 12 times the total weight of the raw materials, add white sugar at 20% of the weight of the purified water, and activated yeast at 0.01% of the weight of the purified water. Saccharomyces cerevisiaeThe yeast activation method involves adding yeast to a 20% (w / w) white sugar aqueous solution and activating it at 40°C for 30 minutes. After stirring and dissolving, the yeast is fermented in a fermentation tank at room temperature for 7-10 days, stirred once a day. When the sugar content drops to 7% as measured by a saccharimeter, the mixture is strained through coarse gauze to remove the residue. Sodium benzoate (0.2% of the weight of the filtrate) is added to the filtrate to terminate fermentation. The mixture is then sealed and aged at room temperature for 30 days. The supernatant is filtered through a diatomaceous earth filter, and steviol glycosides are added for flavoring (0.01% of the weight of the filtrate). The mixture is then bottled to obtain the fermented wine of this invention (Bai Mei Jie Yu Fermented Wine).

[0031] Example 2: Preparation method of the Hundred Roses Depression-Relieving Fermented Wine of the present invention Ingredient formula (parts by weight): 30 parts dark red rose, 20 parts lily, 20 parts fresh Dendrobium officinale, 10 parts stir-fried jujube seed, 10 parts light bamboo leaf, 20 parts jujube, 20 parts floating wheat, 10 parts licorice.

[0032] Preparation method: Weigh the raw materials according to the above weight proportions. Except for the dark red rose and fresh Dendrobium officinale, add 8 times the amount of water and 6 times the amount of water to the remaining raw materials and decoct twice, 1.5 hours each time. Combine the filtrates, cool to room temperature, and then add the dark red rose, fresh Dendrobium officinale, 20% of the weight of white sugar in the filtrate, and 0.01% of the weight of activated yeast in the filtrate. Saccharomyces cerevisiae The yeast is activated by adding yeast to a 20% (w / w) white sugar aqueous solution and activating it at 40°C for 30 minutes. After stirring and dissolving, the yeast is fermented in a fermentation tank at room temperature for 7-10 days, stirred once a day. When the sugar content drops to 7% as measured by a saccharimeter, the mixture is coarsely filtered through gauze to remove the dregs. Sodium metabisulfite (0.5% of the weight of the filtrate) is added to the filtrate to stop fermentation. The mixture is then sealed and aged at room temperature for 30 days. The supernatant is filtered through a diatomaceous earth filter, and sucrose is added for flavoring (1% of the weight of the filtrate). The mixture is then bottled to obtain the fermented wine of this invention (Bai Mei Jie Yu Fermented Wine).

[0033] Example 3: Preparation method of the Hundred Roses Depression-Relieving Fermented Wine of the present invention Ingredient formula (parts by weight): 10 parts dark red rose, 10 parts lily, 5 parts Dendrobium officinale, 5 parts roasted jujube seed, 10 parts light bamboo leaf, 10 parts jujube, 10 parts floating wheat, 5 parts licorice.

[0034] Preparation method: Weigh the raw materials according to the above weight proportions, mix all raw materials, add 8 times and 6 times the total weight of water respectively, and decoct twice, 1.5 hours each time. Combine the filtrates, cool the filtrates to room temperature, and add 20% of the weight of white sugar and 0.01% of the weight of activated yeast (by weight of filtrate). Saccharomyces cerevisiaeThe yeast activation method involves adding yeast to a 20% (w / w) white sugar aqueous solution and activating it at 40℃ for 30 minutes. After stirring to dissolve, ferment in a fermenter at room temperature for 7-10 days, stirring once a day. When the sugar content drops to 7% as measured by a saccharimeter, filter coarsely through gauze to remove the residue. Concentrate the filtrate under reduced pressure to a clear extract with a relative density of 1.2, and store it for later use. The obtained clear extract is then mixed with excipients according to the corresponding dosage form to prepare the desired formulation. For example, an alcoholic beverage can be prepared by adding an appropriate amount of water and grain liquor, with an alcohol content of 5-55°.

[0035] Comparative Example 1: Preparation Method of Bai Mei Jie Yu Decoction The ingredients and formula of Bai Mei Jie Yu Tang are the same as those in Example 1.

[0036] Preparation method: Weigh the appropriate amount of raw materials, decoct twice, adding 6 times the amount of water each time, heat over high heat to boiling, then simmer over low heat for 1 hour. Filter out the dregs, combine the filtrates, and the product is ready.

[0037] The following specific experimental examples demonstrate the beneficial effects of the present invention.

[0038] Experimental Example 1: Study on Antidepressant Active Components in Fermented Wine of the Present Invention 1. Instruments and reagents 1.1 Instruments Ultimate 3000 high-performance liquid chromatograph (Thermo Fisher Scientific, USA); TU-1901 ultraviolet-visible spectrophotometer (Beijing Purkinje General Instrument Co., Ltd.); ES1035B analytical balance, FA2004 analytical balance (Shanghai Lichen Technology Instrument Co., Ltd.); G-100S ultrasonic cleaner (Shenzhen Geneng Cleaning Equipment Co., Ltd.); LCK2000 decoction machine (Tianjin Sanyan Precision Machinery Co., Ltd.).

[0039] 1.2 Test Drugs Bai Mei Jie Yu fermented wine (three batches, batch numbers 20230901, 20230902, and 20230903) was prepared according to the method in Example 1. Bai Mei Jie Yu decoction (three batches, batch numbers J20230901, J20230902, and J20230903) was prepared according to the method in Comparative Example 1. Catechin reference standard (batch number: 110877-202306, purity: 96.6%), puerarin reference standard (batch number: 110752-202217, purity: 96.8%), and shampoosine reference standard (S...) were also prepared. Chaftoside (batch number: 111912-202204, purity: 94.9%), naringin (batch number: 110722-202116, purity: 93.5%), hesperidin (batch number: 110721-202220, purity: 97.2%), and rutoside (batch number: 100080-202012, purity: 92.2%) were all purchased from the China National Institutes for Food and Drug Control. Acetonitrile was of chromatographic grade, water was Yibao purified water, and all other reagents were of analytical grade.

[0040] 2. Methods and Results Three batches of Bai Mei Jie Yu fermented wine test samples with batch numbers 20230901, 20230902, and 20230903 were prepared according to the method described in Example 1. Three batches of Bai Mei Jie Yu decoction test samples with batch numbers J20230901, J20230902, and J20230903 were prepared according to the method described in Comparative Example 1.

[0041] 2.1 Solution Preparation 2.1.1 Preparation of reference solution Accurately weigh catechin, puerarin, shampoosin, naringin, and hesperidin reference standards into separate volumetric flasks. Dissolve and dilute to volume with 50% tetrahydrofuran-acetone for naringin and hesperidin, and dissolve and dilute to volume with methanol for the other components, preparing stock solutions with concentrations of 4480.32, 1261.87, 1597.76, 1589.33, and 199.04 μg / mL, respectively. Accurately pipette 4.0, 0.6, 1.0, 1.5, and 2.5 mL of each stock solution into the same 10 mL volumetric flask, dilute to volume with methanol, and prepare stock solutions containing 1792.13 μg, 75.71 μg, 159.78 μg, 238.40 μg, and 49.76 μg / mL of catechin, puerarin, shampoosin, naringin, and hesperidin, respectively. μg of mixed reference solution was prepared and further diluted with methanol to form a series of gradient mixed reference solutions of varying concentrations for detection by high performance liquid chromatography.

[0042] Accurately weigh rutin reference standard into a volumetric flask, dissolve and dilute to volume with 50% methanol-water to prepare a reference standard solution containing 520.29 μg per mL. Further dilute with methanol to prepare gradient reference standard solutions with concentrations of 20.81, 31.22, 41.62, 52.03, 62.43, 72.84, and 83.25 μg / mL for UV-Vis spectrophotometric detection.

[0043] 2.1.2 Preparation of test solution Take an appropriate amount of the sample to be tested, filter it through a 0.45 μm microporous membrane, and collect the filtrate to obtain the test solution for detection by high performance liquid chromatography.

[0044] Take an appropriate amount of the sample to be tested, dilute it 10 times with water, mix well, filter it through a 0.45 μm microporous membrane, and take the filtrate to obtain the test solution for detection by ultraviolet-visible spectrophotometry.

[0045] 2.2 Detection Method 2.2.1 High Performance Liquid Chromatography (HPLC) for Detection The chromatographic column was an Agilent Eclipse XDB-C18 (4.6 mm × 250 mm, 5 μm); mobile phase A was acetonitrile, and mobile phase B was 0.2% phosphoric acid aqueous solution; the flow rate was 1.0 mL / min; the injection volume was 5 μL; the column temperature was 30℃; the detection wavelength was 280 nm; and the gradient elution program is shown in Table 1.

[0046] Table 1. Gradient elution program for high performance liquid chromatography of fermented wine of the present invention In Table 1, -15-0 min is the solvent equilibrium time. No chromatographic peaks are generated during this time and are not included in the chromatographic peak diagram.

[0047] The determination method involves accurately injecting 5 μL each of the mixed reference solution and the test solution into a liquid chromatograph and determining the concentrations under chromatographic conditions. A standard curve is plotted using the standard curve method, with the concentration of each component in the series of reference solutions as the abscissa and the peak area of ​​each component as the ordinate, to obtain the linear regression equation. The contents of catechin, puerarin, succinate, naringin, and hesperidin in the fermented wine of this invention are calculated based on the linear regression equation.

[0048] 2.2.2 Ultraviolet-Vis Spectrophotometry Detection Accurately pipette 5 mL of the test sample into a 10 mL volumetric flask, add 0.5 mL of 5% NaNO2 solution, shake well and let stand for 6 min; add 0.5 mL of 10% Al(NO3)3 solution, shake well and let stand for 6 min; add 4 mL of 1 mol / L NaOH solution, shake well and let stand for 15 min. Use the aqueous solution without the test sample as a blank control, and measure the absorbance of the test sample at a wavelength of 510 nm using a UV-Vis spectrophotometer.

[0049] A standard curve was plotted using the standard curve method, with the concentration of a series of rutin reference solutions as the abscissa and the absorbance value as the ordinate, to obtain a linear regression equation. The total flavonoid content in the fermented wine of this invention was calculated based on the linear regression equation.

[0050] 2.3 Methodological Examination 2.3.1 Linear Relationship Take the mixed reference solution and dilute it with methanol to prepare 7 concentration gradient solutions. The concentration gradient of each component is shown in Table 2. Take the rutin reference solution, and its concentration gradient is shown in Table 2. Detect them according to the detection method in "2.2". Calculate the peak area integral value ( Y ) on mass concentration ( X Linear regression analysis was performed on the samples (μg / mL), and the results are shown in Table 3.

[0051] Table 2. Concentration gradient of each reference solution Table 3. Experimental results of linear relationships of each component 2.3.2 Precision Test The same mixed reference solution was subjected to six consecutive injections using a high-performance liquid chromatograph (HPLC) according to the method described in section 2.2.1. The peak areas of each reference standard were recorded, and the RSD values ​​of the peak areas of catechin, puerarin, shampoosin, naringin, and hesperidin were calculated. The results showed that the instrument precision was good (see Table 4).

[0052] Six parallel samples were prepared using the same rutin reference solution according to the method described in section 2.2.2, and the absorbance values ​​were recorded. The RSD value of the rutin absorbance was calculated. The results showed that the precision of the UV-Vis spectrophotometer was good, as shown in Table 4.

[0053] 2.3.3 Repeatability Test The same test solution was subjected to six consecutive injections using high-performance liquid chromatography (HPLC) according to the method described in section 2.2.1. The peak areas of each target were recorded, and the RSD values ​​of the contents of catechin, puerarin, shampoosin, naringin, and hesperidin were calculated. The results showed that the detection method had good repeatability, as shown in Table 4.

[0054] Six parallel samples were prepared using the same test solution and prepared according to the method described in section 2.2.2 using a UV-Vis spectrophotometer. The absorbance values ​​were recorded, and the RSD value of the total flavonoid content was calculated. The results showed that the detection method had good repeatability, as shown in Table 4.

[0055] 2.3.4 Stability Test The same test solution was analyzed by high-performance liquid chromatography (HPLC) at 0, 2, 4, 6, 12, and 24 h at room temperature, according to the method described in section "2.2.1". The peak area of ​​each target at each time point was recorded, and the RSD values ​​of the peak areas of catechin, puerarin, shampoosin, naringin, and hesperidin were calculated. The results show that the test solution is stable within 24 h at room temperature (see Table 4).

[0056] Six parallel samples were prepared using the same test solution according to the method described in section 2.2.2. The absorbance values ​​were measured at 0, 5, 10, 15, 30, and 45 min using a UV-Vis spectrophotometer, and the RSD values ​​were calculated. The results show that the test solution prepared from the test solution is stable at room temperature for 45 min (see Table 4).

[0057] 2.3.5 Spiking Recovery Test Accurately pipette 10 mL of the test solution with known concentrations of each active ingredient into a 25 mL volumetric flask, repeating this process six times. Accurately pipette appropriate amounts of each reference stock solution from section "2.1.1" into separate 25 mL volumetric flasks, adding amounts approximately equal to the amounts of each active ingredient in the test solution with known concentrations. Dilute to volume with methanol, filter through a 0.45 μm microporous membrane, and collect the filtrate. Analyze the filtrate using high-performance liquid chromatography (HPLC) according to the method described in section "2.2.1," and calculate the recovery rate. See Table 4.

[0058] Accurately pipette 10 mL of the test solution with a known total flavonoid content into a 25 mL volumetric flask, and repeat in six parallel applications. Accurately pipette an appropriate amount of the rutin reference solution from section 2.1.1 into a 25 mL volumetric flask, the amount added being approximately the same as the total flavonoid content in the test solution. Dilute to volume with methanol, filter through a 0.45 μm microporous membrane, and collect the filtrate. Determine the total flavonoid content using a UV-Vis spectrophotometer according to the method described in section 2.2.2, and calculate the recovery rate. See Table 4.

[0059] Table 4. Results of Precision, Repeatability, Stability, and Accuracy Tests 2.4 Sample content determination Three batches of Bai Mei Jie Yu fermented wine and three batches of Bai Mei Jie Yu decoction were taken respectively. Test solutions were prepared according to the method described in section 2.1.2, and the contents of catechins, puerarin, shampoosin, naringin, hesperidin, and total flavonoids in the Bai Mei Jie Yu fermented wine and Bai Mei Jie Yu decoction were calculated according to the detection method described in section 2.2. The amount of active ingredients extracted per gram of raw herb was then calculated based on the volume of the medicinal solution and the amount of raw herb consumed. The results are shown in Table 5.

[0060] Table 5. Results of assays on formulation samples 3. Conclusion This experimental case specifically selected antidepressant active ingredients (catechins, puerarin, shampooside, naringin, hesperidin, and total flavonoids, among which the structures of catechins, puerarin, shampooside, naringin, and hesperidin are as follows). Figure 1 As shown in the figure, two detection methods (high performance liquid chromatography and ultraviolet-visible spectrophotometry) were established to compare the fermented wine and decoction of Bai Mei Jie Yu obtained by fermentation and traditional decoction, respectively. The results showed that the total flavonoids extracted by fermentation were 1.4 times that extracted by decoction, the catechin content was 1.5 times, the puerarin content was 5.7 times, and the hesperidin content was 4.1 times. Notably, the active ingredients shampoosin and naringin were abundant in the fermented wine of Bai Mei Jie Yu, but were not detected in the decoction. Therefore, the traditional Chinese medicine formula of this invention, extracted through fermentation, can generate new antidepressant active ingredients through the action of microbial flora. Thus, this experiment confirms that the traditional Chinese medicine formula of this invention, extracted through fermentation, can increase the dissolution of antidepressant active ingredients and generate new active ingredients, thereby increasing the therapeutic effect.

[0061] Experimental Example 2: Antidepressant Experiment of Fermented Wine of the Present Invention Depression is a mental disorder characterized by low mood, loss of interest, pessimism, slowed thinking, and lack of initiative. The hypothalamic-pituitary-adrenal axis is closely related to stress; the adrenal hormones released by this axis primarily stimulate the secretion of adrenocorticotropic hormone (ACTH), thereby increasing the secretion of glucocorticoids and regulating emotional and behavioral responses during stress. PC12 cells, derived from rat adrenal medullary tumors, possess typical neuroendocrine cell characteristics. Their surface contains abundant, highly expressed glucocorticoid receptors, making them widely used in antidepressant research. High-concentration corticosterone-induced PC12 cell damage models are widely applied, and different types of antidepressants have protective effects against corticosterone-damaged PC12 cells. This experiment uses a corticosterone-induced PC12 cell damage model to investigate the antidepressant activity of the above-mentioned Examples 1-3 and Comparative Example 1 (Bai Mei Jie Yu Tang).

[0062] 1. Instruments and reagents Multiskan FC microplate reader (Thermo Fisher Scientific (China) Co., Ltd.); LC-18N-50A freeze dryer (Shanghai Lichen Technology Instrument Co., Ltd.); CO2 incubator (Wuxi Woxin Instrument Co., Ltd.); HR40-IIA2 biosafety cabinet (Qingdao Haier Co., Ltd.); ES1035B analytical balance (Shanghai Lichen Technology Instrument Co., Ltd.); RE-1102 rotary evaporator (Shanghai Jiapeng Technology Co., Ltd.); G-100S ultrasonic cleaner (Shenzhen Geneng Cleaning Equipment Co., Ltd.); DGL-75X vertical steam sterilizer (Jiangsu Dengguan Medical Instrument Co., Ltd.); RPMI 1640 (Thermo Fisher Scientific (China) Co., Ltd.); CCK-8 reagent (Nanjing Jiancheng Bioengineering Institute); rat adrenal pheochromocytoma bead PC12 cells were purchased from the Cell Bank of the Committee for the Preservation of Typical Cultures of the Chinese Academy of Sciences; corticosterone (Shanghai Shidander Standard Technology Co., Ltd.).

[0063] 2. Methods 2.1 Preparation of test group solutions The solutions were prepared according to the methods in Examples 1-3. In Example 3, the obtained extract was added to grain liquor and water to adjust the alcohol content to 10%. An appropriate amount was taken, concentrated under reduced pressure until no alcohol odor remained, and then freeze-dried to obtain the powder of each test sample. An appropriate amount was taken, dissolved in water, and prepared into a solution containing 5 mg of the test sample per 1 mL, thus obtaining test groups 1-3.

[0064] 2.2 Preparation of control group solution The filtrate obtained from Comparative Example 1 was concentrated under reduced pressure and then freeze-dried to obtain a powder. 250 mg of the powder was then added to water to prepare a control solution containing 5 mg per mL.

[0065] 2.3 Detection Methods The experiment included a blank group, a model group, a control group, and test groups 1-3 (test group 1 for Example 1, test group 2 for Example 2, and test group 3 for Example 3), with 4 replicates per group and 3 repetitions. PC12 cells were placed in T25 cell culture flasks containing complete culture medium (including DMEM high-glucose medium, 10% fetal bovine serum, and 1% penicillin antibiotics) and incubated at 37°C, 5% CO2, and 90% relative humidity. Cells in the logarithmic growth phase were used for testing. Cell suspension (1×10⁻⁶ cells / mL) was collected. 5Corticosterone (100 μL / mL) was seeded in 96-well plates. After incubation for 24 h, except for the blank group, all other groups were treated with corticosterone at a final concentration of 20 mg / L. The blank group was treated with an equal volume of culture medium. After induced injury for 48 h, the culture medium was discarded. The blank group and the model group were treated with 100 μL of serum-free culture medium. The control group and different test groups were treated with 100 μL of serum-free culture medium containing different test products at a concentration of 5 mg / mL. After 24 h of culture, the absorbance of each well was measured at 450 nm using the CCK-8 assay. The cell viability was calculated as: Cell viability (%) = A value of experimental group / A value of blank group × 100%. Experimental data are presented as mean ± standard deviation. ± s The data were statistically analyzed using SPSS 22.0 software. One-way ANOVA was used to compare differences between groups. P A value <0.05 indicates a statistically significant difference.

[0066] 3 Results The experimental results (Table 6) showed that, compared with the blank group, the survival rate of PC12 cells in the model group was significantly reduced, with a survival rate of 52.12%. Compared with the model group, the control group and test products 1-3 significantly improved the survival rate of PC12 cells induced by corticosterone. Compared with the control group, test groups 1 and 2 significantly improved the survival rate of PC12 cells induced by corticosterone, with test group 1 showing the highest survival rate at 81.78%. All differences were statistically significant. P <0.05).

[0067] Table 6. Effects of each group on corticosterone-induced PC12 cell formation ( n = 3, ± s ) Note: Compared with the blank group, a) P <0.05; compared with the model group, b) P <0.05; compared with the control group, c) P <0.05; compared with test group 2, d) P <0.05.

[0068] Experimental Conclusion: This experiment investigated the antidepressant activity of the control group and test groups 1-3 using a corticosterone-induced PC12 cell damage model. The results showed that both the control group and test groups 1-3 protected PC12 cells, confirming that Bai Mei Jie Yu Tang and the samples prepared using Examples 1-3 of this invention have antidepressant effects. Furthermore, test groups 1 and 2 showed significantly better activity than the control group, indicating that the samples prepared using Examples 1 and 2 of this invention are superior to clinically used decoctions. Additionally, test group 1 exhibited the best activity, demonstrating that Example 1 provided by this invention has the best antidepressant effect. The example used in test group 1 did not involve decoction extraction during preparation; instead, it involved direct fermentation while fully preserving the original medicinal materials, resulting in better and more complete extraction of active ingredients and conversion of inactive ingredients into active ingredients, thereby enhancing its antidepressant activity and better leveraging the synergistic effect of fermentation.

[0069] Experimental Example 3: Clinical efficacy of the fermented wine of the present invention 1. Clinical Data Research subjects The experimental group consisted of 66 patients with mild to moderate depression admitted to the outpatient department of Nanchong Traditional Chinese Medicine Hospital from March 2022 to September 2023. The patients were diagnosed by outpatient physicians and were found to have depression caused by emotional distress, heart yin deficiency, and liver qi disharmony.

[0070] 1.2 Diagnostic criteria The diagnostic criteria for this disease were developed by referring to the diagnostic content of "depression", "grief", "worry" and "insomnia" in the "Guidelines for Clinical Diagnosis and Treatment of Mental Disorders in Traditional Chinese Medicine" (2015 edition) and combining the diagnostic experience of clinicians.

[0071] Main symptoms: depressed mood, anxiety, and unexplained sadness.

[0072] Secondary symptoms include: irritability, fatigue, sluggishness, passivity, loss of interest, social aversion, forgetfulness, insomnia, light sleep, and difficulty falling back asleep.

[0073] (The above-mentioned primary evidence must be present, and any two or more of the secondary symptoms must be met.) 1.3 Case Inclusion Criteria (1) Meets the above-mentioned TCM diagnostic criteria; (2) Age 18-70 years old; (3) The patient is willing to undergo regular follow-up visits.

[0074] 1.4 Case Exclusion Criteria (1) Patients with severe or unstable internal medical conditions; (2) Other types of schizoaffective disorder, schizophrenia patients (3) Suffering from serious illnesses such as organ or cardiovascular diseases.

[0075] 2 Treatment methods Experimental group: The fermented wine prepared in Example 1 of this invention was administered orally, 100 ml twice daily, for 8 consecutive weeks. The efficacy was then statistically analyzed.

[0076] 3. Observation of therapeutic effects 3.1 Evaluation of therapeutic effect Before and after treatment, the severity of depression was comprehensively evaluated using the Traditional Chinese Medicine Symptom Scale and the Self-Rating Depression Scale. The evaluation index used was the Depression Severity Index, which was calculated by dividing the cumulative score of each item by the highest total score. The higher the index, the higher the degree of depression. An index below 0.50 indicates no depression, 0.50-0.59 indicates mild depression, 0.60-0.69 indicates moderate depression, and above 0.70 indicates severe depression.

[0077] 3.2 Efficacy Evaluation Criteria According to the "Standards for Diagnosis and Efficacy Evaluation of Diseases and Syndromes in Traditional Chinese Medicine" (State Administration of Traditional Chinese Medicine, 1994 edition) issued by the Ministry of Health and the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine Drugs" (China Medical Press, 2002 edition), the following standards apply to the evaluation of the efficacy of TCM syndromes: Cure: The score reduction rate after treatment is >75%, and clinical symptoms disappear or basically disappear; Significant effect: The reduction rate after treatment is 51%~75%, and clinical symptoms are greatly reduced or disappear; Effective: The reduction rate after treatment was 25%~50%, and some clinical symptoms were improved; Ineffective: The reduction rate after treatment is <25%, and there is no change in clinical symptoms.

[0078] 3.3 Treatment Efficacy Table 7. Treatment Efficacy Experimental Conclusion: This invention was used to treat 66 patients, of whom 6 were excluded due to non-compliance during treatment or taking other medications. 60 patients completed the full treatment. The therapeutic effect of this invention was evaluated using the Traditional Chinese Medicine Syndrome Scale and the Self-Rating Depression Scale before and after treatment. The results (Table 7) show that after 8 weeks of use, the cure rate reached over 85%, and the total effective rate reached 100%. No adverse reactions were observed. The results indicate that the traditional Chinese medicine formula of this invention is scientifically sound and significantly effective for mild to moderate depression caused by emotional distress, heart yin deficiency, and liver qi disharmony, and is worthy of widespread application.

[0079] In summary, this invention provides a traditional Chinese medicine composition for the prevention and / or treatment of depression. This composition has the effects of regulating qi and relieving depression, clearing the heart and nourishing yin, relieving irritability and calming the mind. It is used to prevent and treat mild to moderate depression caused by emotional distress, heart yin deficiency, and liver qi disharmony, with significant efficacy. Fermenting this composition to obtain fermented wine further enhances its effect in preventing and / or treating depression, making it significantly more effective than decoctions. Furthermore, the raw materials of this composition are all traditional Chinese medicines that are both food and medicine, making it safe and reliable to use and showing good application prospects.

Claims

1. A traditional Chinese medicine composition for the prevention and / or treatment of depression, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: 50 parts rose petals, 30 parts lily bulbs, 30 parts dendrobium, 15 parts jujube seed, 15 parts bamboo leaf, 30 parts jujube, 30 parts floating wheat, and 18 parts licorice. The roses mentioned are dark red roses; The dendrobium mentioned is Dendrobium officinale; The jujube kernels mentioned are stir-fried jujube kernels; The preparation method of the traditional Chinese medicine composition includes the following steps: (1) Weigh each raw material according to the weight ratio; (2) After sterilizing the raw drug, add water, glycogen and fermentation bacteria to ferment. After fermentation, filter coarsely, stop fermentation, age, filter finely, flavor, and bottle to obtain the product; In step (2), the fermenting bacteria is yeast. Saccharomycs cerevisiae .

2. The method for preparing the traditional Chinese medicine composition according to claim 1, characterized in that: It includes the following steps: (1) Weigh each raw material according to the weight ratio; (2) After sterilizing the raw drug, add water, glycogen and fermentation bacteria to ferment. After fermentation, filter coarsely, stop fermentation, age, filter finely, flavor, and bottle to obtain the product; In step (2), the fermenting bacteria is yeast. Saccharomycs cerevisiae .

3. The preparation method according to claim 2, characterized in that: In step (2), the weight of the water is 5 to 30 times the total weight of the raw materials; And / or, in step (2), the weight of the glycogen is 10-30% of the total weight of water; And / or, in step (2), the weight of the fermenting bacteria is 0.01% to 0.5% of the total weight of water; And / or, in step (2), the fermenting bacteria are activated fermenting bacteria; And / or, in step (2), the fermentation is carried out at 10-40°C for 5-15 days until the sugar content drops to 4-9%; And / or, in step (2), the fermentation termination is achieved by adding 0.1~0.5% of the coarse filtrate by weight of a terminator; And / or, in step (2), the aging is a sealed aging for 20 to 90 days; And / or, in step (2), the seasoning is 0.01% to 0.1% of the weight of the filtrate added as a seasoning agent.

4. The preparation method according to claim 3, characterized in that: In step (2), the glycogen is one or more of the following: granulated sugar, maltose, fructose syrup, glucose, or starch hydrolysis products; And / or, in step (2), the activation method of the fermentation bacteria is to add the fermentation bacteria to a white sugar aqueous solution with a mass fraction of 20~30% and activate it at 40~50℃ for 20~30 min; And / or, in step (2), the coarse filtration is performed using 10-50 mesh gauze; And / or, in step (2), the terminating agent is one or more of sodium metabisulfite, sodium benzoate, and potassium sorbate; And / or, in step (2), the fine filtration is performed by filtration with a diatomaceous earth filter, and the diatomaceous earth mesh size is selected as 50-200 mesh; And / or, in step (2), the flavoring agent is selected from one or more of sucrose, steviol glycosides, sucralose, and acesulfame potassium.

5. Use of the traditional Chinese medicine composition according to claim 1 in the preparation of a medicament for the prevention and / or treatment of depression.

6. The use according to claim 5, characterized in that: The medication is for the prevention and / or treatment of mild to moderate depression caused by emotional distress, heart yin deficiency, and liver qi disharmony.

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