Preparation for relieving depressive mood as well as preparation method and application thereof

Abstract

The invention discloses a preparation for relieving depressive mood, which is prepared from the following three medicinal materials: black sesame, arillus longan and folium ginkgo, and the weight ratio of extracts of the black sesame, the arillus longan and the folium ginkgo is (90-110): (90-110): 1. The invention further relates to application and a preparation method of the preparation for relieving the depression. The preparation for relieving the depression mood has the effect of remarkably enhancing the depression mood relieving effect. The preparation method of the preparation for relieving the depression mood has the advantages of being high in extraction efficiency, easy to prepare and low in cost, and the prepared preparation has the beneficial effect of remarkably relieving the depression mood.

Description

Technical Field

[0001] This application relates to the technical field of biomedicine, and particularly relates to a preparation for relieving depressive mood, a preparation method thereof, and an application thereof. Background Art

[0002] Nerve growth factor (NGF) is the earliest discovered and currently the most thoroughly studied among neurotrophic factors. It is a nerve cell growth regulatory factor with dual biological functions of neuronal nutrition and promoting neurite growth. It plays an important regulatory role in the development, differentiation, growth, regeneration, and expression of functional characteristics of central and peripheral neurons. NGF contains three subunits, α, β, and γ. The active region is the β subunit, which is a dimer formed by non-covalent binding of two single chains composed of 118 amino acids each, and has a high homology with the structure of human NGF, and there is no obvious interspecies specificity in biological effects. NGF is essential for the survival of effector neurons. In vitro experiments have confirmed that if NGF is not added to the culture medium, nerve cells cannot grow axons and cannot survive. NGF and its receptors are also widely distributed in the central nervous system. NGF produced by the hippocampus and cerebral cortex can be retrogradely transported to the basal forebrain nucleus through cholinergic nerves to maintain the survival and function of cholinergic neurons. In the early stage of embryonic development, the content of central NGF determines the density of cholinergic nerves. In the cerebellar region and hypothalamus without cholinergic innervation, the content of NGF is also relatively high, indicating that in addition to cholinergic nerves, NGF also has a trophic effect on other types of neurons. We have discovered the relationship between the axonal growth response of cells to NGF and depression from a large number of recent reports (Zhu et al., 2010; Zhu et al., 2011; Zhu et al., 2012; Zhu et al., 2013).

[0003] Black sesame (scientific name: Sesami Semen Nigrum) is the seed of the sesame plant in the Pedaliaceae family. The seeds are oblate and flat, black on the surface, sweet in taste and fragrant with oil. It is mainly produced in Shandong, Henan, Hubei, Sichuan, Anhui, Jiangxi, Hebei and other places. It is harvested when the fruits are ripe in autumn, dried, the impurities are removed, and then dried again. Black sesame is flat in nature and sweet in taste, and is mainly used to tonify the liver and kidney, benefit essence and blood, and moisten the intestines and relieve constipation. It is applicable to reducing blood pressure, promoting the adrenal gland, anti-inflammatory, inducing diarrhea, reducing cholesterol, etc.

[0004] Longan Arillus (scientific name: Longan Arillus) is the aril of the longan, a plant of the Sapindaceae family. It is also known as Yizhi, Mipi, and Guiyuan. It is mainly produced in Guangdong, Fujian, Taiwan, Guangxi, Yunnan, Guizhou, Sichuan and other places. It is picked when the fruit is ripe from July to October, dried or sun-dried, and the flesh is taken and the pit is removed and sun-dried until it is dry and non-sticky. Longan Arillus is an official drug in the Pharmacopoeia of the People's Republic of China and an ideal tonic. Traditional Chinese medicine believes that longan is sweet in taste and warm in nature. It enters the heart and spleen meridians. It has the effects of tonifying the heart and spleen, nourishing blood and calming the nerves, strengthening the spleen and stopping diarrhea, and diuretic and detumescence. It is applicable to symptoms such as weakness after illness, sallow complexion due to blood deficiency, qi and blood deficiency, neurasthenia, palpitations, forgetfulness and insomnia.

[0005] Ginkgo Leaf (scientific name: Follium Ginkgo) is the leaf of the Ginkgo biloba L. of the Ginkgoaceae family. The leaf blade is fan-shaped, with a shallow wavy upper edge and a long petiole. It is cultivated throughout the country. It is distributed in Guangxi, Sichuan, Henan, Shandong, Hubei, Liaoning and other places. The leaves are harvested from June to September. Traditional Chinese medicine believes that its leaves can be used as medicine to astringe the lungs, relieve asthma, activate blood circulation and remove stasis, and relieve pain. It is applicable to treating lung deficiency cough, coronary heart disease, angina pectoris, hyperlipidemia, etc.

[0006] There is no information or report in the prior art on preparing a preparation from black sesame, longan arillus and ginkgo leaf as raw materials and using it to soothe depressive mood.

[0007] Therefore, there is a need for a preparation for soothing depressive mood, a preparation method and its application to at least partially solve the above problems. Summary of the Invention

[0008] A series of simplified concepts are introduced in the content part of this application, which will be further described in detail in the specific implementation part. The content part of this application does not mean to attempt to define the key features and essential technical features of the claimed technical solution, nor does it mean to attempt to determine the protection scope of the claimed technical solution.

[0009] The technical problem to be solved by the present invention is: to provide a preparation for soothing depressive mood, a preparation method and its application using black sesame, longan arillus and ginkgo leaf as raw materials.

[0010] To solve the above technical problems, the technical solution adopted by the present invention is:

[0011] A preparation for soothing depressive mood is prepared from three medicinal materials: black sesame, longan arillus and ginkgo leaf. Among them, the weight ratio of the extracts of black sesame, longan arillus and ginkgo leaf is 90 - 110:90 - 110:1.

[0012] Further, the weight ratio of the extracts of black sesame, longan arillus and ginkgo leaf is 100:100:1.

[0013] Further, the black sesame seeds, longan pulp, and ginkgo leaves are respectively extracts that have been successively subjected to alcohol dissolution, concentration, freeze-drying, and re-dissolution treatments.

[0014] The present invention also discloses a preparation method of the above-mentioned preparation for relieving depressive mood. The black sesame seeds, longan pulp, and ginkgo leaves are respectively placed in an aqueous ethanol solution with a concentration of 45 - 55% (v%) for soaking to obtain a soaking solution. The volume ratios of the black sesame seeds, longan pulp, and ginkgo leaves to ethanol are respectively 1:15 - 25, and the soaking time is 65 - 80 hours. The soaking solution is concentrated to the original weight of the medicinal materials under vacuum conditions and then freeze-dried to a powder to obtain a powder. The powder is re-dissolved with pure water. The powders obtained from the black sesame seeds, longan pulp, and ginkgo leaves are re-dissolved to the same mass concentration to obtain a re-dissolution solution. The re-dissolution solutions obtained from the black sesame seeds, longan pulp, and ginkgo leaves are mixed in a weight ratio of 90 - 110:90 - 110:1 to obtain the preparation.

[0015] Further, the temperature of the concentration is 50 - 70°C.

[0016] Further, the concentration of the ethanol is 48 - 52% (v%), the volume ratios of the black sesame seeds, longan pulp, and ginkgo leaves to ethanol are 1:18 - 22, and the soaking time is 70 - 74 hours.

[0017] The present invention also discloses the application of the above-mentioned preparation for relieving depressive mood in the preparation of a product capable of relieving depressive mood.

[0018] Further, the product capable of relieving depressive mood is a medicine, health product, food, food additive, or beauty product.

[0019] The beneficial effects of the preparation for relieving depressive mood of the present invention are as follows:

[0020] (1) Using black sesame seeds, longan pulp, and ginkgo leaves as the original medicinal materials to prepare a formulated preparation, thereby obtaining the beneficial effect of relieving depressive mood;

[0021] (2) The effect of relieving depressive mood of the formulated combination of black sesame seeds, longan pulp, and ginkgo leaves is significantly enhanced compared to that of individual black sesame seeds, longan pulp, and ginkgo leaves.

[0022] The beneficial effects of the preparation method of the preparation for relieving depressive mood of the present invention are as follows:

[0023] (1) By using the technical means of an aqueous ethanol solution with a concentration of 45 - 55% (v%), the volume ratios of the black sesame seeds, longan pulp, and ginkgo leaves to ethanol being respectively 1:15 - 25, and the soaking time being 65 - 80 hours, it has the advantage of high extraction efficiency;

[0024] (2) The preparation method of the present invention has the advantages of simple preparation and low cost;

[0025] (3) The prepared preparation has the beneficial effect of significantly relieving depressive mood. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The following drawings of the present application are hereby incorporated as part of the present application for understanding the present application. The embodiments of the present application shown in the drawings and their descriptions are used to explain the principles of the present application.

[0027] In the drawings:

[0028] Figure 1 is a scanned image of the experimental reaction tank of black sesame seeds;

[0029] Figure 2 is a scanned image of the experimental reaction tank of longan pulp;

[0030] Figure 3 is a scanned image of the experimental reaction tank of ginkgo leaves;

[0031] Figure 4 is the percentage increase in the average axon length measured under a microscope at a magnification of 100 times after co-culturing PC12 cells with black sesame seeds, longan pulp, and ginkgo leaves;

[0032] Figure 5 is the percentage increase in the average axon length measured under a microscope at a magnification of 100 times after co-culturing PC12 cells with two medicinal materials;

[0033] Figure 6 is the percentage increase in the average axon length measured under a microscope at a magnification of 100 times after co-culturing PC12 cells with black sesame seeds + longan pulp + ginkgo leaves;

[0034] Figure 7 is the result of the sugar water preference and forced swimming experiments in animal experiments;

[0035] Figure 8 is the result of the effect of black sesame seeds + longan pulp + ginkgo leaves on the quantity of neurotransmitters;

[0036] Figure 9 is the result of the effect of black sesame seeds + longan pulp + ginkgo leaves on the expression of neurotrophic factors;

[0037] Figure 10 is the result of the effect of black sesame seeds + longan pulp + ginkgo leaves on the expression of neurotrophic factor receptors;

[0038] Figure 11 is the result of the effect of black sesame seeds + longan pulp + ginkgo leaves on the expression of neuron factor markers;

[0039] Figure 12Results of the effects of black sesame + longan pulp + ginkgo leaf on neurotransmitter regulatory enzyme expression;

[0040] Figure 13 Results of the effects of black sesame + longan pulp + ginkgo leaf on contactin expression;

[0041] Figure 14 Results of the effects of black sesame + longan pulp + ginkgo leaf on neurotransmitter expression. Detailed implementation manners

[0042] In the following description, numerous specific details are given to provide a more thorough understanding of the present application. However, it is obvious to those skilled in the art that the present application can be implemented without one or more of these details. In other instances, some technical features well known to those skilled in the art are not described to avoid confusion with the present application.

[0043] To thoroughly understand the present application, a detailed description will be presented in the following. Obviously, the implementation of the embodiments of the present application is not limited to the specific details familiar to those skilled in the art. The preferred embodiments of the present application are described in detail below. However, in addition to these detailed descriptions, the present application may have other embodiments.

[0044] It should be noted that the terms used herein are only for describing specific implementation manners and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular forms are also intended to include the plural forms. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they specify the presence of the described features, wholes, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, wholes, steps, operations, elements, components, and / or their combinations.

[0045] The ordinal numbers such as "first" and "second" cited in the present application are only identifiers and do not have any other meanings, such as a specific order, etc. Moreover, for example, the term "first component" itself does not imply the existence of a "second component", and the term "second component" itself does not imply the existence of a "first component".

[0046] To illustrate the technical content, the achieved objectives, and the effects of the present invention in detail, the following is described in conjunction with the embodiments and with reference to the accompanying drawings.

[0047] The most crucial concept of the present invention lies in: by combining black sesame, longan pulp, and ginkgo leaf, a beneficial effect of significantly relieving depressive mood can be achieved.

[0048] A preparation for relieving depressive mood of the present invention is prepared from three medicinal materials: black sesame seeds, longan pulp, and ginkgo leaves. The weight ratio of the extracts of black sesame seeds, longan pulp, and ginkgo leaves is 90 - 110:90 - 110:1.

[0049] Further, the weight ratio of the extracts of black sesame seeds, longan pulp, and ginkgo leaves is 100:100:1.

[0050] Further, the black sesame seeds, longan pulp, and ginkgo leaves are respectively extraction liquids that have been successively subjected to alcohol dissolution, concentration, freeze - drying, and re - dissolution treatments.

[0051] The beneficial effects of the preparation for relieving depressive mood of the present invention are as follows:

[0052] (1) Black sesame seeds, longan pulp, and ginkgo leaves are used as raw medicinal materials to prepare a formulated preparation, thereby obtaining the beneficial effect of relieving depressive mood;

[0053] (2) The effect of relieving depressive mood of the formulation combining black sesame seeds, longan pulp, and ginkgo leaves is significantly enhanced compared to that of black sesame seeds, longan pulp, and ginkgo leaves alone.

[0054] The present invention also discloses a preparation method of the above - mentioned preparation for relieving depressive mood. The black sesame seeds, longan pulp, and ginkgo leaves are respectively placed in an ethanol aqueous solution with a concentration of 45 - 55% (v%) for soaking to obtain soaking solutions. The volume ratios of black sesame seeds, longan pulp, and ginkgo leaves to ethanol are respectively 1:15 - 25, and the soaking time is 65 - 80 hours. The soaking solutions are concentrated to the original weight of the medicinal materials under vacuum conditions and then vacuum freeze - dried into powders to obtain powder substances. The powder substances are re - dissolved with pure water, and the powder substances obtained from black sesame seeds, longan pulp, and ginkgo leaves are re - dissolved to the same mass concentration to obtain re - dissolved solutions. The re - dissolved solutions obtained from black sesame seeds, longan pulp, and ginkgo leaves are mixed in a weight ratio of 90 - 110:90 - 110:1 to obtain the preparation.

[0055] The beneficial effects of the preparation method of the preparation for relieving depressive mood of the present invention are as follows:

[0056] (1) By using the technical means of an ethanol aqueous solution with a concentration of 45 - 55% (v%), the volume ratios of black sesame seeds, longan pulp, and ginkgo leaves to ethanol are respectively 1:15 - 25, and the soaking time is 65 - 80 hours, it has the advantage of high extraction efficiency;

[0057] (2) The preparation method of the present invention has the advantages of simple preparation and low cost;

[0058] (3) The prepared preparation has the beneficial effect of significantly relieving depressive mood.

[0059] Further, the concentration temperature is 50-70°C.

[0060] Further, the concentration of the ethanol is 48-52% (v%), the volume ratio of black sesame seeds, longan pulp and ginkgo leaves to ethanol is 1:18-22, and the soaking time is 70-74 hours.

[0061] The present invention also discloses an application of the above-mentioned preparation for relieving depressive mood in the preparation of a product capable of relieving depressive mood.

[0062] Further, the product capable of relieving depressive mood is a medicine, a health product, a food, a food additive or a beauty product.

[0063] Example 1 of the present invention is as follows:

[0064] A preparation for relieving depressive mood in this example is prepared from three medicinal materials: black sesame seeds, longan pulp and ginkgo leaves. The weight ratio of black sesame seeds, longan pulp and ginkgo leaves is 90:90:1.

[0065] The preparation method of the preparation for relieving depressive mood in this example is as follows: the black sesame seeds, longan pulp and ginkgo leaves are respectively placed in an ethanol aqueous solution with a concentration of 45% (v%) for soaking to obtain a soaking solution. The volume ratio of black sesame seeds, longan pulp and ginkgo leaves to ethanol is 1:15 respectively, and the soaking time is 65 hours. The soaking solution is concentrated to the original weight of the medicinal materials under vacuum conditions and then freeze-dried to powder to obtain a powder. The powder is redissolved with pure water, and the powders obtained from the black sesame seeds, longan pulp and ginkgo leaves are redissolved to the same mass concentration to obtain a redissolved solution. The redissolved solutions obtained from the black sesame seeds, longan pulp and ginkgo leaves are mixed in a weight ratio of 90:90:1 to obtain the preparation. The concentration temperature is 50°C.

[0066] Example 2 of the present invention is as follows:

[0067] Only the following are different: "the concentration of the ethanol is 50%, the volume ratio of black sesame seeds, longan pulp and ginkgo leaves to ethanol is 1:20 respectively, the soaking time is 72 hours, and the concentration temperature is 60°C", and the rest is the same as Example 1.

[0068] Example 3 of the present invention is as follows:

[0069] Only the following are different: "the concentration of the ethanol is 55%, the volume ratio of black sesame seeds, longan pulp and ginkgo leaves to ethanol is 1:25 respectively, the soaking time is 80 hours, and the concentration temperature is 70°C", and the rest is the same as Example 1.

[0070] Example 4 of the present invention is as follows:

[0071] It is different only in that "the weight ratio of the black sesame seeds, longan pulp and ginkgo leaves is 100:100:1" and "the back solutions separately prepared from the black sesame seeds, longan pulp and ginkgo leaves are mixed in a weight ratio of 100:100:1", and the rest is the same as in Example 1.

[0072] Example 5 of the present invention is as follows:

[0073] It is different only in that "the weight ratio of the black sesame seeds, longan pulp and ginkgo leaves is 110:110:1" and "the back solutions separately prepared from the black sesame seeds, longan pulp and ginkgo leaves are mixed in a weight ratio of 110:110:1", and the rest is the same as in Example 1.

[0074] Example 6 of the present invention is as follows:

[0075] A preparation for relieving depressive mood in this example is prepared from three medicinal materials: black sesame seeds, longan pulp and ginkgo leaves. The weight ratio of the black sesame seeds, longan pulp and ginkgo leaves is 100:100:1. The black sesame seeds, longan pulp and ginkgo leaves are respectively extracts that have been successively subjected to alcohol dissolution, concentration, freeze-drying and back-dissolution treatments.

[0076] The preparation method of the preparation for relieving depressive mood in this example is to place the black sesame seeds, longan pulp and ginkgo leaves respectively in an ethanol aqueous solution with a concentration of 50% (v%) for soaking to obtain soaking solutions. The volume ratios of the black sesame seeds, longan pulp and ginkgo leaves to ethanol are 1:20 respectively, and the soaking time is 72 hours. The soaking solutions are concentrated to the original weight of the medicinal materials under vacuum conditions and then freeze-dried to powder to obtain powder substances. The powder substances are back-dissolved with pure water, and the powder substances separately prepared from the black sesame seeds, longan pulp and ginkgo leaves are back-dissolved to the same mass concentration to obtain back solutions. The back solutions separately prepared from the black sesame seeds, longan pulp and ginkgo leaves are mixed in a weight ratio of 100:100:1 to obtain the preparation. The temperature of the concentration is 60°C. Effect analysis

[0077] The medicinal materials described in Examples 1 to 6 all use herbal chips containing the medicinal materials as the raw material sources, and all the herbal chips are from Yunnan Baiyao Group Co., Ltd.

[0078] 4 ng of biotinylated nerve growth factor (NGF) at a concentration of 200 ng / mL was added to the experimental reaction wells of the herbal chips of black sesame, longan pulp, and ginkgo leaves (10 pieces of each) for hybridization. After 3 hours of hybridization, the chips were washed 4 times with phosphate-buffered saline for 15 minutes each time. Then, 20 ng of STREPTAVIDIN Cy5 CONJUGATE (Life Technologies) at a concentration of 1 μg / mL was added for secondary hybridization. After 3 hours of hybridization, the chips were washed 4 times with phosphate-buffered saline for 15 minutes each time. Finally, the chips were scanned using a chip scanner (GenePix 4100A) under the conditions of an excitation wavelength of 600 nm and an emission wavelength of 670 nm.

[0079] Figures 1 - 3 Scanning diagrams of the black sesame, longan pulp, and ginkgo leaf chips with NGF binding signals obtained by the chip scanner. Figures 1 - 3 The chemical components of black sesame, longan pulp, and ginkgo leaves can bind to NGF.

[0080] According to 5×10 per well 4 PC12 cells were seeded in a 6-well cell culture plate. The next day, the cell culture medium was replaced with a low-serum cell culture medium, which included 1% fetal bovine serum reagent (FBS) and 1% HSDMEM. The extraction solutions of ginkgo leaves, longan pulp, black sesame, ginkgo leaves + longan pulp, ginkgo leaves + black sesame, longan pulp + black sesame, and black sesame + longan pulp + ginkgo leaves were added to each well, and 50 ng / mL NGF was used as a positive control. Among them, the concentration of the longan pulp extraction solution in each group was 1 mg / mL, the concentration of the black sesame extraction solution was 1 mg / mL, and the concentration of the ginkgo leaf extraction solution was 10 μg / mL. The extraction solutions of various herbs in the two- or three-herb mixture groups were mixed in equal volumes. The black sesame + longan pulp + ginkgo leaf group was prepared into a preparation according to Example 6. After 72 hours, the cell culture medium was removed and the cells were rinsed with PBS, and then the cells were fixed with 4% paraformaldehyde (PFA). The cell images were recorded using a microscope equipped with a camera at a magnification of 100 times, and the axon length was measured using the same software and the percentage increase in the average axon length was calculated to obtain Figures 4 - 6 。

[0081] Figure 4 The percentage increase in the average axon length measured under a microscope at a magnification of 100 times for each group of experiments, Figure 4 The abscissa in corresponds to the blank control experiment, NGF, black sesame, longan pulp, and ginkgo leaves in sequence.

[0082] Figure 5 The percentage increase in the average axon length measured under a microscope at a magnification of 100 times for each group of experiments, Figure 5Among them, the abscissa corresponds to the blank control experiment, NGF, longan pulp + ginkgo leaf, black sesame + ginkgo leaf, and black sesame + longan pulp in sequence.

[0083] Figure 6 It is the percentage increase in the average axon length measured under a 100-fold magnification microscope for each group of experiments. Figure 6 Among them, the abscissa corresponds to the blank control experiment, NGF, and black sesame + longan pulp + ginkgo leaf in sequence.

[0084] From Figures 4 - 6 it can be seen that for the PC12 cells added with the extract of black sesame, longan pulp and ginkgo leaf, the percentage increase in the average axon length is 8% higher than that of the black sesame extract; 12% higher than that of the longan pulp extract; 23% higher than that of the ginkgo leaf extract; 24% higher than that of the black sesame + longan pulp extract group; 56% higher than that of the black sesame + ginkgo leaf extract group; and 72% higher than that of the longan pulp + ginkgo leaf extract group.

[0085] Through a series of animal behavior experiments, it can be seen that the preparation of the present invention has the effect of relieving depressive mood. The mice are divided into 4 major groups, namely the non-stressed group, the black sesame + longan pulp + ginkgo leaf group (prepared according to Example 6), the fluoxetine group (positive control), and the non-treatment group (negative control). Within six weeks, each group (except the non-stressed group) is subjected to chronic irregular stress every week. The environmental stress includes 24-hour food deprivation, 24-hour water deprivation, 1-hour empty water bottle without water, 7-hour cage tilt at 45°, 24-hour sunlight exposure, 24-hour exposure to a humid environment, forced swimming in water at 8°C for 6 minutes, 2-hour restraint of the mouse's movement, and allowing the mouse to contact unknown items for 24 hours. After six weeks, the black sesame + longan pulp + ginkgo leaf group is treated with high-concentration (6 g / kg / day) or low-concentration (0.6 g / kg / day) black sesame + longan pulp + ginkgo leaf, and the fluoxetine group is treated with fluoxetine (4 mg / kg). One week later, a sugar water preference test and a forced swimming test are carried out.

[0086] 72 hours before the sugar water preference test, the mice are trained to adapt to 1% sugar water. Two bottles of 1% sugar water are placed in the cage, and one bottle of sugar water is replaced with clean water after 24 hours. After adaptation, the mice are placed in a state of 24-hour food and water deprivation, and then one bottle of 100 ml of 1% sugar water and 100 ml of clean water are presented to the mice. After three hours, the volumes of the sugar water and the clean water are recorded to calculate the sugar water preference.

[0087] Each group of mice is placed in a cylindrical glass container (50 cm high and 20 cm in diameter) containing 30 cm deep water at 22°C. The forced swimming test is divided into two parts, 15 minutes of pre-experiment swimming, and 5 minutes of observation of the mice's dynamics 24 hours later, recording the time the mice float on the water surface and the struggling time.

[0088] Non-depressed mice prefer sugar water and tend to give up swimming for survival in water in a shorter time. Figure 7 Based on the non-stressed group (blank), the sugar water preference and the floating time on the water surface of the four groups were compared. The sugar water preference of the high-concentration black sesame + longan pulp + ginkgo leaf group was similar to that of the fluoxetine group. The floating time on the water surface was longer than that of the fluoxetine group but shorter than that of the non-treatment group (negative control). This indicates that the effect of high-concentration black sesame + longan pulp + ginkgo leaf in treating depressive mood is slightly worse than that of fluoxetine.

[0089] Dissect the heads of the mice in each group, extract brain tissues with Trizol reagent, and then analyze by qPCR.

[0090] Figure 8 It shows that high-concentration black sesame + longan pulp + ginkgo leaf has a similar effect to fluoxetine in effectively increasing dopamine, norepinephrine, and serotonin; compared with the non-treatment group, it can effectively increase dopamine, norepinephrine, and serotonin.

[0091] Among them, dopamine (DA) belongs to catecholamine neurotransmitters and is widely present in the central nervous system. Research shows that dopamine plays an important role in the occurrence and development of depression. The extracellular DA level in patients with depression decreases. The normal physiological function of DA in the body depends on dopamine receptor (DR) and dopamine transporter (DAT). Genetic variations of these two types of molecules are closely related to depression.

[0092] Norepinephrine (NA) belongs to catecholamine. It is a neurotransmitter mainly synthesized and secreted by sympathetic postganglionic neurons and noradrenergic neurons in the brain. Its reduction will lead to insufficient functions of adrenergic α1 receptor and α2 receptor. These two receptors are responsible for wakefulness and cognition respectively. Therefore, the insufficiency will lead to reduced energy, that is, psychomotor retardation and slow thinking; it also leads to insufficient function of β receptor, which is mainly antidepressant. So it will lead to low mood. The things caused by the reduction of the functions of the above three receptors can be summarized as hypokinesia, slow thinking, and low mood. Together, they constitute retarded depression.

[0093] Serotonin changes can reflect the central activity state and are related to functions such as sleep, memory, cognition, emotional response, and hypothalamic endocrine regulation. A large number of clinical studies have shown that reduced serotonin functional activity can cause anorexia, insomnia, circadian rhythm disorders, sexual dysfunction, and various neuropsychiatric diseases, which are closely related to the onset of depression.

[0094] Therefore, dopamine (DA), norepinephrine (NA), and serotonin are negatively correlated with depressive mood, that is, the lower the expression levels of dopamine (DA), norepinephrine (NA), and serotonin, the greater the probability of suffering from depression.

[0095] Figure 9 It is shown that high-concentration black sesame + longan pulp + ginkgo leaf is more effective than fluoxetine in increasing the expression of neurotrophic factors: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), neurotrophin-3 (NT-3), and neurotrophin-4 / 5 (NT4 / 5).

[0096] Among them, nerve growth factor (NGF) is an important member of the neurotrophic factor family, mainly produced in the cortex, hippocampus, and hypothalamus, and also present in the peripheral nervous system and immune system. NGF plays an important role in neuron survival, neural plasticity, and learning and memory, is crucial for the stress response, participates in the regulation of the neuro-endocrine-immune (NEI) system, and changes in neural plasticity and NEI system homeostasis play important roles in the pathophysiology of depression. Therefore, NGF is involved in the pathogenesis of depression.

[0097] Brain-derived neurotrophic factor (BDNF) is one of the members of the neurotrophic factor family. It is mainly synthesized by neurons and is widely distributed in the nervous system, with the highest content in the hippocampal dentate gyrus and cerebral cortex. BDNF is a very important neurotrophic factor that can promote the growth and differentiation of nerve cells. It has a maintaining and promoting effect on the development, differentiation, growth and regeneration of various neurons, especially 5-hydroxytryptamine (5-HT) and dopamine (DA) neurons, and also has the effect of regulating synaptic plasticity. Research shows that a significant decrease in BDNF mRNA can be detected in the serum of patients with depression, the level of BDNF in the serum decreases, and the degree of decrease is proportional to the degree of depression. It has also been found in the nervous system that BDNF is closely related to the occurrence of depression. Under stress, BDNF in the peripheral circulation decreases, and at the same time, the level of BDNF in the limbic system of the brain also decreases significantly. Autopsies of patients with severe suicidal depression show that the BDNF in brain regions such as the hippocampal prefrontal cortex is significantly lower than that in the normal population.

[0098] Glial cell derived neurotrophic factor (GDNF) is a neurotrophic factor of the transforming growth factor-β family. It is widely expressed throughout the brain, regulates the noradrenergic and γ-aminobutyric acid (GABA)ergic systems, and is one of the most effective neurotrophic factors affecting the development, survival and activity of midbrain dopaminergic neurons. It also protects neurons and glial cells from oxidative stress damage and plays an important role in the development and functional integrity of hippocampal cells.

[0099] Neurotrophin-3 (NT-3) belongs to the nerve growth factor (NGF) family and has regulatory activity on specific neurons in the central nervous system and peripheral nervous system. In addition, it can support the survival and differentiation of existing surviving nerve cells and promote the growth and differentiation of newly generated neurons.

[0100] Neurotrophin-4 / 5 (NT4 / 5) belongs to the nerve growth factor (NGF) family and has regulatory activity on specific neurons in the central nervous system and peripheral nervous system.

[0101] Therefore, NGF, BDNF, GDNF, NT3, and NT4 / 5 are negatively correlated with depressive mood, that is, the lower the expression levels of NGF, BDNF, GDNF, NT3, and NT4 / 5, the greater the probability of suffering from depression.

[0102] Figure 10Show that high-concentration black sesame + longan pulp + ginkgo leaves are more effective than fluoxetine in increasing the expression of neurotrophic factor receptors: tyrosine kinase (TrkA), tyrosine kinase receptor (TrkB), and tyrosine kinase receptor C (TrkC).

[0103] Among them, tyrosine kinase (TrkA) is the functional receptor of NGF. In the absence of the TrkA receptor, NGF cannot bind to specific receptors, cannot conduct intracellular signal transduction, and the developing basal forebrain cholinergic neurons cannot fully develop and mature, resulting in changes in the homeostasis of the EI system and affecting the incidence of depression.

[0104] Tyrosine kinase receptor (TrkB) specifically binds to BDNF and can also bind to NT4 / 5. After BDNF binds to the TrkB receptor, it can activate three intracellular signaling pathways: extracellular regulated protein kinases (MEK-ERK), phosphatidylinositol kinase (PI3K), and phospholipase (CPLC-γ). These three signaling pathways play important roles in maintaining nerve growth, nerve cell differentiation, and neuron survival, and also play roles in maintaining synaptic plasticity, brain neuron structure, and function in adults. Lack of the TrkB receptor will affect the binding of BDNF to the TrkB receptor, reduce the expression level of TrkB on the cell membrane surface, and lead to a decrease in the BDNF signaling pathway.

[0105] Tyrosine kinase receptor C (TrkC) is the functional receptor of NT3. Studies have found that NT3 and its receptor TrkC play important roles in the development and maturation of spinal cord neurons during development and play a key role in the development and function of the nervous system, including neuron survival, growth, and differentiation.

[0106] Therefore, TrK A, TrK B, and TrK C are negatively correlated with depressive mood, that is, the lower the expression levels of TrK A, TrK B, and TrK C, the greater the probability of suffering from depression.

[0107] Figure 11 Show that high-concentration black sesame + longan pulp + ginkgo leaves are more effective than fluoxetine in increasing neuron factor markers: GAP43, RhoA, cell division cycle protein 42 (Cdc42), Rac1, NF200, NF160, NF68.

[0108] Among them, GAP43 is a specific phosphoprotein on the nerve cell membrane and is closely related to nerve regeneration, development, and plasticity, etc.

[0109] RhoA affects axon regeneration in opposite ways in neurons and astrocytes. RhoA in neurons initiates the prevention of axon regeneration, while RhoA in astrocytes is beneficial to axon regeneration. Extracellular inhibitory signals inhibit the size and kinetics of growth cones through RhoA, thereby hindering neuron axon growth.

[0110] Cell division cycle protein 42 (Cdc42), a small GTPase, plays an important role in cell growth, adhesion, proliferation and migration, and is associated with the pathogenesis of neurological diseases such as stroke and epilepsy. Research has shown that by activating bradykinin B2 receptor and its downstream small G protein Cdc42 that regulates synaptogenesis, synaptogenic effects and rapid antidepressant effects can be produced.

[0111] NF200, NF160, and NF68 are differentiation markers of the differentiation ability of nerve cells in the body and are downregulated in the brains of patients with depression.

[0112] Figure 12 It is shown that high-concentration black sesame + longan pulp + ginkgo biloba leaves are more effective than the non-treatment group in enhancing the expression of neurotransmitter-regulating enzymes: tyrosine hydroxylase, aromatic acid decarboxylase, dopamine-β-hydroxylase, tryptophan hydroxylase, and monoamine oxidase A. High-concentration black sesame + longan pulp + ginkgo biloba leaves are more effective than fluoxetine and the non-treatment group in inhibiting the expression of monoamine oxidase B and catechol-O-methyltransferase.

[0113] Among them, tyrosine hydroxylase can synthesize the precursor L-dopa (L-3,4-dihydroxyphenylalanine), and inhibiting the synthesis of the precursor L-dopa leads to the consumption of dopamine and norepinephrine in the brain, thereby affecting depressive mood.

[0114] Aromatic acid decarboxylase is involved in the synthesis of neurotransmitters dopamine and serotonin, and these two factors are closely related to the occurrence of depression. Therefore, aromatic acid decarboxylase also has corresponding effects on depression.

[0115] In the brain, norepinephrine is converted from dopamine via dopamine-β-hydroxylase, and these two factors are closely related to the occurrence of depression. Therefore, dopamine-β-hydroxylase also has corresponding effects on depression.

[0116] Tryptophan hydroxylase is an enzyme that plays a role in serotonin biosynthesis and is the catalytic or initiating step in the chemical reaction that converts tryptophan into serotonin, and serotonin is negatively correlated with the onset of depression. Therefore, tryptophan hydroxylase also has a certain impact on depression.

[0117] Monoamine oxidase A is mainly distributed in catecholaminergic neurons and inactivates monoamine neurotransmitters. In the "monoamine hypothesis of depression", this enzyme can lead to the onset of depression.

[0118] Monoamine oxidase B is mainly distributed in serotonergic neurons, histaminergic neurons and glial cells and can inactivate monoamine neurotransmitters. In the "monoamine hypothesis of depression", this enzyme can lead to the onset of depression.

[0119] Monoamine oxidase is an enzyme that catalyzes the deamination reaction of monoamines and converts them into other substances. After monoamines are deaminated, they lose their role in regulating mood, emotion and stress. When the monoamine oxidase in the body inactivates dopamine, a person's mood will become depressed.

[0120] Catechol-O-methyltransferase is an important protease for inactivating catecholamine neurotransmitters and can inactivate catecholamine hormones or neurotransmitters through methylation. COMT is present in multiple parts such as glial cells in the basal ganglia of the forebrain and the limbic system and has a relatively high activity in the synaptic cleft, especially on the postsynaptic membrane. Catechol-O-methyltransferase can break down catecholamines, especially dopamine, thereby reducing the bioavailability of extracellular dopamine and blunting its neurotransmitter effect.

[0121] Therefore, tyrosine hydroxylase, aromatic acid decarboxylase, dopamine-β-hydroxylase, and tryptophan hydroxylase are negatively correlated with depressive mood. That is, the lower the expression levels of tyrosine hydroxylase, aromatic acid decarboxylase, dopamine-β-hydroxylase, and tryptophan hydroxylase, the higher the probability of suffering from depression. Monoamine oxidase A, monoamine oxidase B, and catechol-O-methyltransferase are positively correlated with depressive mood. That is, the higher the expression levels of monoamine oxidase A, monoamine oxidase B, and catechol-O-methyltransferase, the higher the probability of suffering from depression.

[0122] Figure 13 It is shown that high concentrations of black sesame + longan pulp + ginkgo leaf are more effective than fluoxetine in increasing the expression of syntaphilin: synapsin 1, synaptophysin (SYP), synaptotagmin, synaptosomal-associated protein 25 (SNAP-25), and postsynaptic density protein-95 (PSD-95).

[0123] Among them, synapsin 1, this gene encodes an integral membrane protein associated with presynaptic vesicles in neuronal cells, and it plays a role in synaptic plasticity, thus having an impact on neurotrophic factors and being correspondingly associated with depressive mood.

[0124] Synaptophysin (SYP), it is widely present in all nerve terminals of the body and is specifically distributed on the membrane of presynaptic vesicles. Because it is closely related to nerve growth, repair, regeneration, and synaptic remodeling, it is correspondingly associated with depressive mood.

[0125] Synaptotagmin is located on the synaptic vesicles of brain neurons and plays a key role in the process of neurotransmitter release and membrane transport. There is a significant correlation between research findings and synaptic activities. Therefore, it has an impact on neurotrophic factors and is correspondingly associated with depressive mood.

[0126] Synaptosomal-associated protein 25 (SNAP-25) is a presynaptic plasma membrane protein involved in regulating neurotransmitter release. There is a correlation in its participation in synaptic activities. Therefore, it has an impact on neurotrophic factors and is correspondingly associated with depressive mood.

[0127] Postsynaptic density protein-95 (PSD-95) is widely present in the central nervous system, mainly in mature excitatory glutamatergic synapses, and is involved in the formation of synaptic connections. Therefore, it has an impact on neurotrophic factors and is correspondingly associated with depressive mood.

[0128] Therefore, synapsin 1, synaptophysin, synaptotagmin, SNAP-25, and PSD-95 are negatively correlated with depressive mood, that is, the lower the expression levels of synapsin 1, synaptophysin, synaptotagmin, SNAP-25, and PSD-95, the higher the probability of suffering from depression.

[0129] Figure 14 It shows that high concentrations of black sesame + longan pulp + ginkgo leaf have a similar effect to fluoxetine in enhancing the expression of dopamine receptors; high concentrations of black sesame + longan pulp + ginkgo leaf and fluoxetine inhibit the expression of norepinephrine receptors compared to the untreated group; high concentrations of black sesame + longan pulp + ginkgo leaf and fluoxetine are more effective in enhancing the expression of serotonin receptors than the untreated group.

[0130] Among them, the dopamine receptor is a site on the membrane for the neurotransmitter dopamine to recognize, and dopamine exerts its function with it.

[0131] The norepinephrine receptor is a receptor that reacts with norepinephrine in various organs innervated by the postganglionic fibers of the sympathetic nerve and is associated with norepinephrine. Therefore, it plays a corresponding role in depressive mood.

[0132] Serotonin receptors (5-HT receptors) are a group of G protein-coupled receptors (GPCRs) and ligand-gated ion channels (LGICs) found in the central and peripheral nervous systems. The 5-HT receptors are activated by the neurotransmitter serotonin, which is its natural ligand. At the same time, 5-HT regulates the release of many neurotransmitters and many hormones and affects various neural processes such as aggression, anxiety, appetite, cognition, learning, memory, mood, nausea, sleep, etc.

[0133] Therefore, dopamine receptors, norepinephrine receptors, and serotonin receptors are negatively correlated with depressive mood, that is, the lower the expression levels of dopamine receptors, norepinephrine receptors, and serotonin receptors, the higher the probability of suffering from depression.

[0134] In summary, high-concentration black sesame + longan pulp + ginkgo leaf is similar to the fluoxetine group in terms of effect; compared with the untreated group, it can effectively increase dopamine, norepinephrine, and serotonin. It greatly increases the mRNA levels of growth factors, namely NGF, BDNF, GDNF, NT3, as well as high-affinity neurotrophic factor receptors, namely TrkA and TrkB. In addition, the high-dose extract also significantly increases the mRNA expression levels of differentiation markers NF200, NF160, GAP43, and synaptogenesis markers (i.e., synapsin 1, synaptophysin, synaptotagmin, synaptosome-associated protein 25, and postsynaptic density protein-95). The results support the potential of black sesame + longan pulp + ginkgo leaf in promoting neural differentiation in vivo. In addition, the transcriptional levels of Rho GTPases (i.e., RhoA, Cdc42, and Rac1) also increase, while the changes in dopamine receptors, norepinephrine receptors, and serotonin receptors are not significant. Therefore, it can be concluded that black sesame + longan pulp + ginkgo leaf has a better effect on treating depressive mood.

[0135] Acute toxicity tests and chronic toxicity tests were carried out using 3 g / kg / day (low concentration), 10 g / kg / day (medium concentration), and 30 g / kg / day (high concentration) of black sesame + longan pulp + ginkgo leaf. Black sesame + longan pulp + ginkgo leaf has no toxicity.

[0136] In summary, the preparation for relieving depressive mood provided by the present invention has a significant effect of enhancing the relief of depressive mood. The preparation method of the preparation for relieving depressive mood of the present invention has the beneficial effects of high extraction efficiency, simple preparation, low cost, and the prepared preparation has a significant effect of relieving depressive mood.

[0137] The above are only embodiments of the present invention and do not limit the patent scope of the present invention. Any equivalent transformation made using the content of the specification and drawings of the present invention, or directly or indirectly applied in related technical fields, is equally included in the patent protection scope of the present invention.

[0138] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field of this application. The terms used herein are only for the purpose of describing specific embodiments and are not intended to limit this application. The features described in one embodiment herein can be applied alone or in combination with other features in another embodiment, unless the feature is not applicable or otherwise stated in that other embodiment.

Claims

1. A preparation for relieving depressive mood, characterized in that, Prepared from three medicinal materials, namely black sesame seeds, longan pulp and ginkgo leaves, wherein the weight ratio of the extracts of the black sesame seeds, longan pulp and ginkgo leaves is 90 - 110:90 - 110:

1.

2. The preparation for relieving depressive mood according to claim 1, characterized in that, The weight ratio of the extracts of the black sesame seeds, longan pulp and ginkgo leaves is 100:100:

1.

3. The preparation for relieving depressive mood according to claim 1 or 2, characterized in that, The black sesame seeds, longan pulp and ginkgo leaves are respectively the extraction liquids that have been successively subjected to alcohol dissolution, concentration, freeze-drying and re-dissolution treatments.

4. A method for preparing the preparation for relieving depressive mood according to any one of claims 1 - 3, characterized in that, The black sesame seeds, longan pulp and ginkgo leaves are respectively placed in an ethanol aqueous solution with a concentration of 45 - 55% (v%) for soaking to obtain soaking liquids. The volume ratios of the black sesame seeds, longan pulp and ginkgo leaves to ethanol are respectively 1:15 - 25, and the soaking time is 65 - 80 hours. The soaking liquids are concentrated to the original weight of the medicinal materials under vacuum conditions and then vacuum freeze-dried into powders to obtain powder substances. The powder substances are re-dissolved with pure water, and the powder substances respectively prepared from the black sesame seeds, longan pulp and ginkgo leaves are re-dissolved to the same mass concentration to obtain re-dissolution liquids. The re-dissolution liquids respectively prepared from the black sesame seeds, longan pulp and ginkgo leaves are mixed in a weight ratio of 90 - 110:90 - 110:1 to obtain the preparation.

5. The method for preparing the preparation for relieving depressive mood according to claim 4, characterized in that, The temperature of the concentration is 50 - 70°C.

6. The method for preparing the preparation for relieving depressive mood according to claim 4, characterized in that, The concentration of the ethanol is 48 - 52%, the volume ratios of the black sesame seeds, longan pulp and ginkgo leaves to ethanol are 1:18 - 22, and the soaking time is 70 - 74 hours.

7. Use of the preparation for relieving depressive mood according to any one of claims 1 - 3 in the preparation of a product capable of relieving depressive mood.

8. The use of the preparation for relieving depressive mood according to claim 7 in the preparation of a product capable of relieving depressive mood, characterized in that, The product capable of soothing depressive emotions is a medicine, health product, food, food additive or beauty product.

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