Osmanthus fragrans fragrance type anti-depression fragrance and preparation method thereof

Abstract

The invention discloses sweet-scented osmanthus fragrance type anti-depression fragrance and a preparation method thereof, belongs to the technical field of aromatherapy, and aims to solve the problems that traditional anti-depression therapy takes effect slowly and has large side effects and an existing aromatic product mechanism is not clear. The fragrance comprises the following components in percentage by mass: 10-15% of borneol essential oil, 5-10% of sandalwood oil, 1-5% of ink safflower essential oil, 1-3% of tree orchid extract, 2-6% of michelia alba leaf oil, 2-5% of sweet-scented osmanthus extract, 2-5% of jasminum sambac essential oil, 1-3% of Leyburg extract, 1-3% of lavender oil, 1-3% of dendranthema indicum essential oil and the balance of sweet-scented osmanthus essential oil. The preparation method comprises the following steps: extracting the osmanthus essential oil, mixing the components, sealing and standing at room temperature for 7 days for stabilization, and carrying out quality inspection. Olfactory administration avoids the liver first-pass effect, takes effect quickly, has few side effects, takes sweet-scented osmanthus as a leading fragrance, is pleasant and lasting, is suitable for long-term adjuvant therapy of depression, and provides a new scheme for non-drug intervention.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of aromatherapy and its application in the adjuvant treatment of depression, in particular to an osmanthus fragrance type anti-depression fragrance and its preparation method. BACKGROUND

[0002] Depression, as a high-incidence psychological disease worldwide, has affected the physical and mental health and quality of life of hundreds of millions of people. Current mainstream treatment methods focus on drug therapy and psychological therapy, among which drug therapy mainly uses selective serotonin reuptake inhibitors (SSRIs) and the like, but has significant limitations: first, the onset is delayed, patients need to wait for several weeks or even months to show the therapeutic effect, and during the waiting period, they still face the risk of continuous emotional depression and suicide; second, the side effects are obvious, which can easily cause sexual dysfunction, abnormal body weight, gastrointestinal discomfort and other problems, leading to reduced patient compliance, and about 30%-50% of patients discontinue treatment due to intolerance of side effects; third, there are large individual differences, and some patients do not respond to the drug, making it difficult to guarantee the treatment effect.

[0003] Although psychological therapy can improve the cognitive and behavioral patterns of patients, it relies on the guidance of professional physicians, has a long treatment cycle, high cost, and is limited by regional medical resources, making it difficult to meet the needs of a large number of people. Under this background, aromatherapy, as a non-drug intervention method, has gradually attracted attention due to its non-invasive, easy-to-operate, and high safety advantages. Existing research has confirmed that essential oils such as lavender and mint can relieve depression symptoms by regulating emotion-related neural pathways, but related research has mostly stopped at behavioral verification of single essential oils, lacking systematic exploration of the synergistic effects of compound essential oils, and the anti-depression mechanism of osmanthus fragrance type essential oils is particularly scarce, and a complete theory system of "component-target-disease" has not yet been formed.

[0004] The existing aromatherapy products on the market are diverse in types, but most of them lack scientific efficacy verification and mechanism analysis, and only use vague concepts such as "relieving emotions" for promotion, which cannot provide precise selection basis for patients with depression. At the same time, existing research ignores the synergistic effects of different essential oil components, and fails to fully tap the potential advantages of compound essential oils in multi-target regulation and reduction of side effects. In addition, the mechanism of aroma inhalation on depression is still at the level of behavioral observation, lacking in-depth research at the molecular level (such as neurotransmitters, inflammatory factors, and neural plasticity-related proteins), making it difficult to determine the target and signaling pathway, which restricts the standardized application of aromatherapy in the adjuvant treatment of depression.

[0005] Therefore, the development of an osmanthus fragrance type compound fragrance with clear anti-depression efficacy, clear mechanism of action, and no side effects not only fills the research gap of osmanthus fragrance type essential oils in the field of anti-depression, but also provides a new adjuvant treatment scheme with fast onset and high compliance for patients with depression, which has important clinical value and market prospect. Summary of the Invention

[0006] The core objective of this invention is to provide an osmanthus-scented antidepressant fragrance and its preparation method. Through the synergistic effect of compound essential oil components, combined with network pharmacology mechanism analysis and animal experiment verification, it aims to rapidly relieve depressive symptoms, reduce side effects, and improve treatment safety, thus providing a novel non-drug intervention solution for patients with depression.

[0007] An osmanthus-scented antidepressant fragrance, comprising, by weight percentage:

[0008] Borneol essential oil 10%-15%, sandalwood oil 5%-10%, safflower essential oil 1%-5%, tree flower extract 1%-5%, magnolia leaf oil 2%-6%, osmanthus extract 2%-5%, jasmine essential oil 2%-5%, lysimachia extract 1%-3%, lavender oil 1%-3%, chrysanthemum essential oil 1%-3%, with the remainder being osmanthus essential oil;

[0009] The osmanthus essential oil contains dihydro-β-ionone, β-ionone, and linalool as active ingredients.

[0010] Furthermore, by weight percentage, it includes:

[0011] Borneol essential oil 10%-15%, sandalwood oil 5%-10%, safflower essential oil 1%-5%, tree flower extract 1%-5%, magnolia leaf oil 2%-6%, osmanthus extract 2%-5%, jasmine essential oil 2%-5%, lysimachia extract 1%-3%, lavender oil 1%-3%, chrysanthemum essential oil 1%-3%, with the remainder being osmanthus essential oil;

[0012] The osmanthus essential oil contains dihydro-β-ionone, β-ionone, and linalool as active ingredients.

[0013] Furthermore, the aforementioned osmanthus-scented antidepressant fragrance can be further applied to various scented products, such as scented candles and flameless diffusers.

[0014] Furthermore, the fragrance is administered via olfactory delivery. The active ingredients enter the bloodstream through the nasal mucosa and act on the limbic system of the brain, achieving an antidepressant effect by regulating neurotransmitters, inhibiting inflammatory factors, and enhancing neuroplasticity. The neurotransmitters are serotonin and norepinephrine, the inflammatory factors are IL-6 and TNF-α, and the neuroplasticity-related protein is BDNF.

[0015] Furthermore, the fragrance can be used via a diffuser or spray, 1-2 times daily, for 20-40 minutes each time, suitable for long-term use by patients with depression.

[0016] Further, the core target points of the fragrance effect include STAT3, AKT1 and HSP90AA1, and the key signal pathways of the effect include the serotonin synaptic pathway, the TNF signal pathway and the MAPK signal pathway.

[0017] The beneficial effects of the present application are:

[0018] 1. Multi-target synergistic anti-depression, clear efficacy

[0019] The fragrance of the present application regulates three core anti-depression pathways through the synergistic effect of various natural ingredients such as osmanthus absolute, borneol essential oil and sandalwood oil.

[0020] Neurotransmitter regulation: significantly increases the levels of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) in the brain, improves monoamine neurotransmitter imbalance, and quickly relieves emotional depression.

[0021] Inflammation suppression: precisely down-regulates pro-inflammatory factors such as IL-6 and TNF-alpha, breaks the vicious cycle of chronic inflammation accompanying depression, and creates a healthy microenvironment for nerve cells.

[0022] Enhanced neural plasticity: up-regulates BDNF expression, promotes neuron growth, synapse formation and repair, reverses chronic stress-induced neural damage (such as decreased Nissl bodies and nuclear pyknosis), and fundamentally improves brain function.

[0023] Animal experiments have confirmed that the food intake, open field movement ability and elevated plus maze open arm exploration behavior of the mice in the fragrance intervention group are significantly restored, close to the level of the blank control group, and the anti-depression effect is clear.

[0024] 2. Rapid onset, no first-pass effect

[0025] Using olfactory administration, the active ingredients of the fragrance directly enter the blood circulation through the nasal mucosa and quickly act on the brain edge system (emotional regulation center), avoiding the gastrointestinal absorption and liver first-pass effect of oral drugs, shortening the onset time to several hours to 1 day, which is much faster than traditional anti-depression drugs (weeks of onset), and can timely relieve acute depression and anxiety symptoms of patients and reduce the risk of suicide.

[0026] 3. Few side effects, high safety

[0027] The fragrance of the present application is based on 100% natural plant ingredients (osmanthus absolute, borneol essential oil, etc.), and animal experiments have verified that:

[0028] There are no common side effects of traditional drugs such as gastrointestinal discomfort and sexual dysfunction;

[0029] Long-term inhalation (for 30 consecutive days) did not observe abnormal liver and kidney function or tissue damage in mice;

[0030] The fragrance has a pH value close to that of human skin, has no risk of skin irritation, and is suitable for long-term auxiliary use by patients of different ages.

[0031] 4. The fragrance is pleasant and has a long-lasting scent, with high compliance

[0032] The fragrance takes osmanthus as the core fragrance type, combines with the delicate fragrance of jasmine, lavender, etc., realizes the balance of "freshness and richness", and has a pleasant olfactory experience, which can improve the decrease in olfactory sensitivity and emotional resistance of patients caused by depression; at the same time, through the stable effect of isododecane and acetone glycerol, the scent lasts for 8-12 hours, without the need for frequent use; the product form is a liquid fragrance, which can be used through an aromatherapy machine, a spray, etc., and the operation is simple, with a significantly higher patient compliance than traditional drugs and psychological treatment.

[0033] 5. The preparation process is mature and can be industrialized

[0034] The preparation process adopts a patented essential oil extraction process, and parameters such as distillation temperature and standing time are clear and controllable, which is suitable for large-scale production; the quality inspection standard is clear, which can guarantee the batch consistency of the product; the raw material sources are extensive, the cost is controllable, and the product has the potential for large-scale industrialization. BRIEF DESCRIPTION OF DRAWINGS

[0035] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present application, and therefore should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0036] Figure 1 The VENN diagram of the osmanthus fragrance component of the present application and the depression target.

[0037] Figure 2 The protein interaction network diagram of the osmanthus fragrance component of the present application and depression.

[0038] Figure 3 The visualization diagram of the protein interaction network of the osmanthus fragrance component of the present application and depression.

[0039] Figure 4 GO enrichment analysis of the intersection target of the osmanthus fragrance of the present application and depression.

[0040] Figure 5 KEGG enrichment analysis of the intersection target of the osmanthus fragrance of the present application and depression.

[0041] Figure 6Effects of the osmanthus fragrance type anti-depression aroma inhalation of the present application on the novel environment food intake inhibition experiment behavior of mice.

[0042] Figure 7 Effects of the osmanthus fragrance type anti-depression aroma inhalation of the present application on the open field experiment behavior of mice.

[0043] Figure 8 Effects of the osmanthus fragrance type anti-depression aroma inhalation of the present application on the elevated plus maze experiment behavior of mice.

[0044] Figure 9 Effects of the osmanthus fragrance type anti-depression aroma of the present application on the inflammatory factors of mice.

[0045] Figure 10 Effects of the osmanthus fragrance type anti-depression aroma of the present application on the brain nerve regulatory substances of mice.

[0046] Figure 11 Nissl staining diagram of the brain tissue of mice after the osmanthus fragrance type anti-depression aroma inhalation of the present application.

[0047] Figure 12 HE staining diagram of the brain tissue of mice after the osmanthus fragrance type anti-depression aroma inhalation of the present application. DETAILED DESCRIPTION

[0048] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application. The components of the embodiments of the present application described and shown in the drawings herein can be arranged and designed in various different configurations.

[0049] An osmanthus fragrance type anti-depression aroma, the mass percentage of each component and the main effects are shown in the following table, wherein osmanthus essential oil is used as the basic matrix to ensure the dominant aroma characteristics of the osmanthus fragrance type, and the anti-depression effect is realized through the synergy of multiple components.

[0050] (I) Formula composition of the osmanthus fragrance type anti-depression aroma

[0051] Table 1 Formula and effects

[0052]

[0053]

[0054] (II) Preparation method of the osmanthus fragrance type anti-depression aroma

[0055] Osmanthus fragrans essential oil, Osmanthus fragrans extract, jasmine essential oil, borneol essential oil, lavender essential oil, Michelia alba leaf oil, tree orchid extract, labdanum extract, Osmanthus fragrans fragrance base from Shanghai Zixin Biotechnology Co., Ltd., and other fragrance raw materials from Beida Zhengyuan Fragrance Co., Ltd., stored at 4℃ in the dark.

[0056] 1. Essential oil extraction step

[0057] Fresh Osmanthus fragrans petals were selected, and the essential oil was extracted using a patented process (refer to the patent "Method for extracting and separating fragrance and medicinal ingredients from the whole plant of plant Shennong Chrysanthemum"). The distillation temperature was controlled at 100-120℃ to avoid high-temperature damage to the aroma components (such as dihydroethyl ionone and linalool), ensuring the activity and aroma purity of the essential oil. After extraction, the essential oil was stored at 4℃ in the dark to prevent oxidation and deterioration.

[0058] 2. Mixing step

[0059] According to the above formula ratio, the extracted Osmanthus fragrans essential oil was first mixed evenly with Shennong Chrysanthemum essential oil, and then borneol essential oil, sandalwood oil, labdanum essential oil, tree orchid extract, Michelia alba leaf oil, Osmanthus fragrans extract, small flower jasmine essential oil, labdanum extract, and lavender oil were added in sequence. The mixture was stirred at room temperature (10-27℃) for 30-60 minutes to ensure that all components were fully dissolved and mixed.

[0060] 3. Stabilization treatment step

[0061] The mixed fragrance solution was transferred to a sealed container and stored at room temperature (10-27℃) in the dark for 7 days to allow the aroma of each component to stabilize and reduce solvent residue, improving product safety.

[0062] 4. Quality inspection step

[0063] After standing, the fragrance was subjected to two core tests:

[0064] Aroma detection: sensory evaluation by a professional fragrance blender confirmed that the aroma was pure Osmanthus fragrans type without odor, stratification, or abnormal aroma volatilization.

[0065] Stability detection: The fragrance was stored at 4℃ and 40℃ for 30 days, and no stratification, discoloration, or abnormal aroma volatilization was observed, ensuring the stability of the product at different temperatures.

[0066] (Three) Examples of fragrance application

[0067] 1. Antidepressant aromatherapy candle production process

[0068] The solid wax (mass ratio 80%~95%) is added to the electric stirring pot, heated and stirred, the temperature is raised to about 90°C, the auxiliary volatile agent is added after the raw and auxiliary materials are completely melted, and then the natural cooling is continued. When the temperature is reduced to 55~60°C, the anti-depression fragrance (mass ratio 5%~20%) is added and stirred. A small amount of dried osmanthus is added during the stirring process, and the dried osmanthus is dispersed by continuous stirring. Then, the product is filled. After the surface solidifies, a small amount of dried osmanthus is sprinkled on the surface. After the whole solidifies, the lid is covered and the product is plastic-wrapped.

[0069] 2. Anti-depression incense production process

[0070] The anti-depression fragrance 5-20%, acetone glycerol 70-95%, water 0-10%, and ethanol 0-5% are mixed uniformly, aged and stabilized for 24 hours, and then filled and packaged.

[0071] The anti-depression fragrance 5-20%, MMB (3-methoxy-3-methyl-1-butanol) 70-95%, water 0-10%, and ethanol 0-5% are mixed uniformly, aged and stabilized for 24 hours, and then filled and packaged.

[0072] The anti-depression fragrance 5-20%, isomeric alkanes 80-95%, and water 0-10% are mixed uniformly, aged and stabilized for 24 hours, and then filled and packaged.

[0073] (IV) Verification method of anti-depression mechanism:

[0074] 1. Prediction of the effect of osmanthus fragrance anti-depression fragrance on depression targets based on network pharmacology

[0075] The volatile active ingredients in the fragrance are identified by GC-MS, and the SMILES number of each component is obtained through the PubChem database;

[0076] The potential targets of active ingredients are predicted by using SwissTargetPrediction database.

[0077] 1.1 Screening of potential pharmacologically active ingredient targets in osmanthus fragrance anti-depression fragrance

[0078] Based on the GC-MS analysis data, target prediction analysis is carried out through SwissTargetPrediction database, and 495 potential action targets are obtained after deduplication.

[0079] 1.2 Acquisition of depression-related targets

[0080] The depression-related search results of TTD, GeneCards and OMIM databases are combined, and 2670 targets are obtained after deleting duplicate values. The Venn diagram of osmanthus fragrance anti-depression fragrance and depression targets is taken, as shown in Figure 1As shown, a total of 220 overlapping targets of osmanthus-scented antidepressant fragrance and depression were obtained. A "component-target-disease" network was constructed, and it was found that many active ingredients of osmanthus fragrance are closely related to key targets of depression.

[0081] 1.3 Construction and Visualization of Protein-Protein Interaction (PPI) Networks

[0082] By introducing osmanthus-scented antidepressant fragrances and identifying 220 drug interaction targets related to diseases into STRING, a protein-protein interaction network was established, such as... Figure 2 As shown. Protein interactions on the STRING platform were visualized using Cytoscape software. Figure 3 Each circle represents a protein; the size and color of the circle indicate the protein's importance, with darker colors and larger volumes indicating higher importance. Results showed that proteins such as STAT3, AKT1, and HSP90AA1 had high degree values, forming the core hubs of the PPI network. STAT3 mediates IL-6 inflammatory signal transduction and participates in neuroimmune regulation; AKT1 regulates neuronal survival and glucose metabolism, affecting neuronal plasticity; and HSP90AA1 maintains stress protein homeostasis, protecting nerve cells from stress damage. These core proteins may play a key regulatory role in the antidepressant effects of osmanthus-scented antidepressant fragrances.

[0083] 1.4 Pathway enrichment analysis

[0084] The 220 intersection targets of osmanthus-scented antidepressant fragrance and depression were imported into Metascape for gene function enrichment analysis (GO). Figure 4 and pathway enrichment analysis (KEGG) such as Figure 5From the GO plot, the significantly enriched functions in molecular function (MF) include ion channel activity, transmembrane signaling receptor activity, etc. The dysfunction of ion channels (sodium, potassium, calcium channels) may lead to abnormal neuronal excitability, which is related to the electrophysiological changes of manic / depressive episodes. The significantly enriched processes in biological process (BP) include chemical synaptic transmission, behavior, response to steroid hormone, etc. Abnormal synaptic transmission and hormone imbalance (such as estrogen, thyroid hormone) are related to the neural plasticity and mood regulation of bipolar disorder. The significantly enriched components in cellular component (CC) include postsynaptic membrane, membrane, and synaptic receptor complex (such as NMDA receptor, AMPA receptor), which affect neural signal transmission.

[0085] The significantly enriched pathways in KEGG plot include cancer pathway, TNF signaling pathway, MAPK signaling pathway, etc. Among them, the serotonergic synapse (hsa04726: Serotonergic synapse) is the pathway with the highest significance. Serotonin (5-HT) is an important neurotransmitter that plays a key role in mood regulation. Many antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), work by regulating the level of serotonin in the synaptic cleft. The dysfunction of serotonergic synapse may be closely related to the development of depression. For example, polymorphism of serotonin transporter gene and dysregulation of presynaptic membrane serotonin release and reuptake may lead to changes in serotonin signaling in the brain, affecting mood state.

[0086] 2. The effect of osmanthus fragrance anti-depression fragrance on the behavior of CUMS mice

[0087] In this study, a chronic unpredictable mild stress (CUMS) method was used to establish a mouse model of depression. This method can simulate various chronic and unpredictable stressors in human daily life, thereby inducing mice to exhibit behavior and physiological changes similar to human depression, with high observability and predictability.

[0088] 2.1 The effect of osmanthus fragrance anti-depression fragrance on the behavior of mice in the novel environment feeding experiment

[0089] From the GO plot, the significantly enriched functions in molecular function (MF) include ion channel activity, transmembrane signaling receptor activity, etc. The dysfunction of ion channels (sodium, potassium, calcium channels) may lead to abnormal neuronal excitability, which is related to the electrophysiological changes of manic / depressive episodes. The significantly enriched processes in biological process (BP) include chemical synaptic transmission, behavior, response to steroid hormone, etc. Abnormal synaptic transmission and hormone imbalance (such as estrogen, thyroid hormone) are related to the neural plasticity and mood regulation of bipolar disorder. The significantly enriched components in cellular component (CC) include postsynaptic membrane, membrane, and synaptic receptor complex (such as NMDA receptor, AMPA receptor), which affect neural signal transmission. Figure 6It can be seen that the mice in the CUMS group showed food intake inhibition and fecal volume reduction, which is consistent with the characteristics of depressive-like behavior in known CUMS models. The food intake and fecal volume of the osmanthus fragrance anti-depression fragrance treatment group were restored compared with the CUMS group, and their food intake and fecal volume were close to the blank group level, indicating that the osmanthus fragrance anti-depression fragrance may have the effect of alleviating CUMS-induced depressive-like behavior.

[0090] 2.2 Effect of osmanthus fragrance anti-depression fragrance on the behavior of mice in the open field test

[0091] Figure 7 The data showed that compared with the blank group, the total movement distance, central movement distance and central activity time of the mice in the CUMS group in the open field test were reduced, while the osmanthus fragrance anti-depression fragrance treatment group showed different degrees of recovery, indicating that the depressive-like behavior induced by CUMS could be alleviated by the osmanthus fragrance anti-depression fragrance.

[0092] 2.3 Effect of osmanthus fragrance anti-depression fragrance on the behavior of mice in the elevated plus maze test

[0093] As Figure 8 shown, in the elevated plus maze behavior test of the CUMS model group of mice, the open arm entry frequency ratio (OE%) and residence time ratio (OT%) were reduced by 61.08% and 65.23% respectively compared with the normal control group, and the data change showed that the chronic stress state successfully induced the depressive-like behavior characteristics of the experimental animals. After the osmanthus fragrance anti-depression fragrance olfactory intervention, the above behavior indexes showed a recovery improvement trend: the OE% and OT% of the osmanthus fragrance anti-depression fragrance group were restored to 113.96% and 153.82% of the blank group respectively. It can be seen that the osmanthus fragrance anti-depression fragrance group significantly reduced the anxiety behavior of mice.

[0094] 2.4 Effect of osmanthus fragrance anti-depression fragrance on inflammatory factors in mice

[0095] The content of cytokines in the serum of mice was detected by ELISA, Figure 9The results showed that the levels of IL-6 and TNF-a in the brain tissue homogenate of the model group mice were significantly increased (P < 0.01), indicating that chronic stress triggered neuroinflammatory response. The levels of IL-6 and TNF-a in the group of mice treated with osmanthus fragrance antidepressant ambience were significantly decreased (P < 0.05), close to the levels of the blank group (P > 0.05), indicating that the osmanthus fragrance antidepressant ambience olfaction accurately down-regulated the levels of pro-inflammatory cytokines, created a relatively mild inflammatory environment for nerve cells, and broke the vicious cycle of chronic inflammation accompanied by depression. This study revealed the mechanism and pathway of the osmanthus fragrance antidepressant ambience olfaction in playing an antidepressant role. The osmanthus fragrance antidepressant ambience olfaction accurately down-regulated the levels of pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-a), created a relatively mild inflammatory environment for nerve cells, and broke the vicious cycle of chronic inflammation accompanied by depression.

[0096] 2.5 Influence of osmanthus fragrance antidepressant ambience on nerve regulatory substances in the brain of mice

[0097] As Figure 10 The results showed that in the dimension of neural plasticity, the expression level of BDNF in the hippocampus tissue of the model group mice was significantly decreased (P < 0.01), indicating impaired neural plasticity. The expression level of BDNF in the group of mice treated with osmanthus fragrance antidepressant ambience was significantly increased (P < 0.05), and there was no significant difference with the blank group (P > 0.05), indicating that the osmanthus fragrance antidepressant ambience olfaction significantly up-regulated the expression of BDNF, promoted the growth, development and synaptic plasticity of neurons, and provided favorable conditions for the recovery of brain function. In terms of neurotransmitter transmission, the levels of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) in the brain of the model group mice were significantly decreased (P < 0.01), indicating dysfunction of monoamine neurotransmitters. The levels of 5-HT and NE in the group of mice treated with osmanthus fragrance antidepressant ambience were significantly increased (P < 0.05), close to the levels of the blank group (P > 0.05), indicating that the osmanthus fragrance antidepressant ambience olfaction optimized the neural signal transmission network and regulated the emotional state of the brain.

[0098] 2.6 Influence of osmanthus fragrance antidepressant ambience on brain tissue of mice

[0099] Nissl staining, as a classic technique of neural histology, highlights Nissl bodies (mainly composed of rough endoplasmic reticulum and ribosomes) in neuron cell bodies, directly showing the morphological characteristics and functional state of neuron cell bodies and dendrites. Figure 11The mouse brain section staining results shown in the middle clearly reveal the health status of nerve cells in different groups. In the blank control group, the neurons have full morphology, clear outline, and clear cytoplasm and dendritic structure; the staining shows a high degree of uniform deep staining, indicating that the Nissl bodies in the cytoplasm are rich and uniformly distributed, which reflects that the neuron structure is intact, the metabolic activity is vigorous, and it is in a healthy and stable physiological state. In sharp contrast to this is the CUMS (chronic unpredictable mild stress) model group, which shows significant pathological changes: the staining is uneven, some neuron cell bodies appear obvious shrinkage, and more critically, the number of Nissl bodies is significantly reduced, and even completely disappears in some areas, and vacuolar degeneration can be seen in the cytoplasm of some neurons. These characteristic changes strongly suggest that chronic stress has caused substantial damage to neurons, leading to decreased protein synthesis capacity and cell metabolic disorder, which is a typical morphological marker of neuron degenerative disease or stress-induced injury. After intervention treatment with osmanthus flower fragrance type antidepressant fragrance, the Nissl body density of neurons in the osmanthus flower fragrance type antidepressant fragrance group is significantly restored, close to the normal level, and only scattered mild staining unevenness is observed, indicating that the osmanthus flower fragrance type antidepressant fragrance intervention effectively alleviates or reverses the chronic stress-induced neuronal damage, and maintains the structural and functional integrity of Nissl bodies.

[0100] Hematoxylin-eosin (HE) staining, as the cornerstone of histopathological diagnosis, fully and clearly presents the overall architecture and cellular details of the tissue through the deep staining of cell nucleus nucleic acid by hematoxylin and the light staining of cytoplasmic protein by eosin. Figure 12The HE staining results of representative mouse brain sections provide detailed information on the microstructure of brain tissues in each group. The blank control group shows an ideal healthy brain tissue morphology: nerve cells are arranged closely and orderly, with clear hierarchy, no accumulation or sparseness; cell morphology is regular, size is consistent, no swelling or atrophy; cell nucleus structure is particularly clear, nuclear membrane is smooth and complete, nucleolus is obvious, and chromatin is distributed in fine and uniform granules; the entire neural tissue structure maintains continuity and integrity, the intercellular space width is normal, and there is no sign of edema, hemorrhage, necrotic focus or inflammatory cell (such as lymphocytes, neutrophils) infiltration. These observations fully meet the morphological standards of healthy brain tissue, providing a reliable baseline reference for the experiment. The HE staining images of the CUMS model group reveal significant pathological damage: the uniformity of staining is destroyed, and some areas are over-stained or under-stained; more critically, obvious karyopyknosis (cell nucleus volume reduction, staining deepening, irregular morphology) is observed, which is an early indication of apoptosis or severe damage; at the same time, the tissue structure becomes loose, and the intercellular space widens, suggesting tissue edema or cell loss. These changes collectively indicate that chronic stress has caused damage to the brain parenchyma, possibly involving neuronal apoptosis, glial cell reaction, and even mild inflammatory processes. The staining results of the osmanthus fragrance type antidepressant fragrance intervention group show a positive improvement trend: compared with the CUMS group, staining unevenness and karyopyknosis are significantly reduced, the compactness of the tissue structure is restored, and the intercellular space tends to be normal, indicating that the osmanthus fragrance type antidepressant fragrance has a certain protective or repair effect on the brain tissue damage induced by chronic stress, reversing some of the pathological morphological changes.

[0101] The present application clearly identifies the core action targets (STAT3, AKT1, etc.) and pathways (5-hydroxytryptaminergic synapse, etc.) through network pharmacology, and verifies through CUMS animal experiments that it can regulate serotonin and norepinephrine levels, inhibit IL-6 and TNF-α inflammatory factors, enhance BDNF-mediated neural plasticity, and significantly alleviate depressive behaviors; olfactory administration avoids the liver first-pass effect, has a fast onset and fewer side effects, the fragrance is mainly osmanthus-based and pleasant, and is suitable for long-term adjuvant treatment of depression, providing a new solution for non-drug intervention.

[0102] The above merely describes preferred embodiments of the present application and is not intended to limit the present application. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A fragrance with an osmanthus-scented antidepressant effect, characterized in that, By weight percentage, including: Borneol essential oil 10%-15%, sandalwood oil 5%-10%, safflower essential oil 1%-5%, tree flower extract 1%-5%, magnolia leaf oil 2%-6%, osmanthus extract 2%-5%, jasmine essential oil 2%-5%, lysimachia extract 1%-3%, lavender oil 1%-3%, chrysanthemum essential oil 1%-3%, with the remainder being osmanthus essential oil; The osmanthus essential oil contains dihydro-β-ionone, β-ionone, and linalool as active ingredients.

2. The osmanthus-scented antidepressant fragrance according to claim 1, characterized in that: The preparation method includes the following steps: S1. Osmanthus essential oil extraction: Fresh golden osmanthus petals are selected, and osmanthus essential oil is extracted using the process of "a method for extracting and separating fragrance and medicinal components from the whole herb of a plant called Shennong Xiangju". The distillation temperature is controlled at 100℃-120℃ during the extraction process. S2. Compounding and mixing: According to the formula ratio, first mix the osmanthus essential oil and Shennong chrysanthemum essential oil evenly, then add the borneol essential oil, sandalwood oil, safflower essential oil, tree orchid extract, magnolia leaf oil, osmanthus extract, jasmine essential oil, lysimachia extract, and lavender oil in sequence, and stir at 10℃-27℃ for 30-60 minutes to fully integrate the ingredients; S3. Stabilization treatment: The mixture obtained in step S2 is placed in a sealed container and left to stand for 7 days at 10℃-27℃ in the dark. S4. Quality Inspection: The mixture after standing in step S3 is subjected to aroma and stability testing. After passing the tests, it is bottled to obtain the finished osmanthus-scented antidepressant fragrance. The aroma test is confirmed by the sensory evaluation of a professional perfumer to have a rich and fresh osmanthus fragrance with no off-odor. The stability test is conducted by storing the mixture at 4℃, 25℃, and 40℃ for 30 days and observing no layering or discoloration.

3. The osmanthus-scented antidepressant fragrance according to claim 1, characterized in that: This fragrance can be further applied to a variety of scented products, including scented candles and flameless diffusers.

4. The application of the osmanthus-scented antidepressant fragrance as described in claim 1 in the adjunctive treatment of depression, characterized in that, The fragrance is administered via olfactory delivery. The active ingredients enter the bloodstream through the nasal mucosa and act on the limbic system of the brain, achieving an antidepressant effect by regulating neurotransmitters, inhibiting inflammatory factors, and enhancing neuroplasticity. The neurotransmitters are serotonin and norepinephrine, the inflammatory factors are IL-6 and TNF-α, and the neuroplasticity-related protein is BDNF.

5. The application according to claim 4, characterized in that, The fragrance can be used via diffuser or spray, 1-2 times daily, for 20-40 minutes each time, and is suitable for long-term use by patients with depression.

6. The application according to claim 5, characterized in that, The core targets of the fragrance's effects include STAT3, AKT1, and HSP90AA1, and the key signaling pathways involved include the 5-hydroxytryptamine synaptic pathway, the TNF signaling pathway, and the MAPK signaling pathway.

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