Application of schizophyllatin in preparation of nerve beauty or emotion skin care product

Abstract

The invention provides an application of schizophyllan in preparation of a nerve beautifying or emotion skin care product, and the schizophyllan can improve the generation of oxytocin and enorphin, so that the schizophyllan can be used in the nerve beautifying or emotion skin care product. The traditional Chinese medicine composition has the effects of regulating brain waves, increasing the pain threshold value of the skin, reducing external pressure, reducing skin sensitivity, relieving the tension state of the skin and improving the skin quality, and can reduce skin pain and pruritus, reduce external stimulation and pressure on the skin and improve the sensitive skin quality.

Description

Technical Field

[0001] The present application relates to the technical field of neurocosmetology or emotional skin care, and in particular to the use of schizophyllan in the preparation of neurocosmetology and emotional skin care products. Background Art

[0002] The skin is the body's largest organ and its most important defense system. Its health is closely linked to the rest of the body, particularly the brain, nerves, and intestines, forming the "brain-skin axis" or "gut-brain-skin axis." As early as 1930, dermatologists Stokes and Pillsbury of the University of Pennsylvania proposed the gut-brain-skin unified theory (Reference: Stokes JH, Pillsbury DH. The effect on the skin of emotional and nervous states: Theoretical and practical consideration of agastrointestinal mechanism. Ars Curandi Em Odontol, 1930, 4:962–993). This theory posits that the gut-brain-skin axis is a dynamic interactive network between the skin and the brain, involving the nervous, endocrine, and immune systems. This network enables bidirectional communication and regulation through mental states, neural signals, hormonal regulation, and immune responses. The bidirectional regulation of the "gut-brain-skin axis" reveals that the skin is not only a "protective organ", but also an integration hub of "neuro-immune-endocrine" and an external manifestation of the body's health (References: Xin Yuchen et al., Principles and Research Progress of Neurocosmetology, Cosmetics and Neuroscience, Issue 1, 2025 (Total Issue 252), 140-155; JIN R, LUO LZHENG J. The trinity of skin: skin homeostasis as aneuro-endocrine-immune organ [J]. Life (Basel), 2022, 12(5): 725.).

[0003] The skin and brain originate from the ectoderm and share a variety of hormones, neurotransmitters, and receptors. They have a close connection and a rich two-way communication mechanism (reference: Carnicu S, Jafferany M. Psychophysiological disorders and the skin [J]. Dermatological Reviews, 2023, 4 (4): 147-151; Xin Yuchen et al. Principles and research progress of neurocosmetology. Cosmetics and Neuroscience. 2025, No. 1 (Total No. 252), 140-155). The feeling of the skin can affect the change of emotions, and the change of emotions will also affect the skin, which has given rise to the discipline of "psychodermatology". In the field of skin care, it is transformed into emotional skin care (reference: Wei Juan et al., A review of the physiological basis of emotional skin care, Daily Chemicals Science, Vol. 47, No. 6, 2024, 47-52). At the same time, the new EU regulations in 2024 also allow the claim of "relieving stress-related skin problems", which is also a claim and embodiment of emotional skin care. The two-way communication between emotions and skin is achieved through psychological stress, circadian rhythm, neurotransmitter network, and endocrine system. Among them, the levels of hormones such as cortisol, oxytocin, melatonin, dopamine, and endorphins in the epidermis or dermis are important markers for judging whether emotional skin care plays a role (References: KIM JE, CHO BK, CHO DH, et al. Expression of hypothalamic–pituitary–adrenal axis in common skin diseases: Evidence of its association with stress-related disease activity [J]. Acta Dermato-Venereologica, 2013, 93(4): 387-393; Wei Juan et al., A review of the physiological basis of emotional skin care, Science of Daily Chemicals, Vol. 47, No. 6, 2024, 47-52).

[0004] Furthermore, the "neuro-immune-endocrine" network also reveals that the human skin is closely related to the nervous system (reference: JIN R, LUO L, ZHENG J. The trinity of skin: skin homeostasis as a neuro-endocrine-immune organ [J]. Life (Basel), 2022, 12 (5): 725.). The nervous system is also distributed in the skin, with nerve endings reaching the upper layer of the epidermis, and nerve fibers in direct contact with skin cells. Neurotransmitters are the medium for information exchange between the nervous system and the immune and endocrine systems. Regulating the skin condition through neurotransmitters is the realization of neurobeauty. Neurotransmitters can be divided into small molecule neurotransmitters and neuropeptides, such as nerve growth factor, adrenocorticotropic hormone, endorphins, acetylcholine, oxytocin, etc.

[0005] It can be seen that oxytocin and endorphins are the biochemical bridge connecting neurobeauty (skin science) and emotional skin care (psychological experience).

[0006] Oxytocin (OXT) is an endogenous neuropeptide hormone that is mainly synthesized by the paraventricular nucleus and supraoptic nucleus of the hypothalamus and acts on the entire central nervous system and periphery. In addition, studies have found that OXT and its receptor can be expressed in primary human dermal fibroblasts (FIB) and keratinocytes (KER) (References: Kanta Fujimoto, Kosuke Inada, Kotaro Oka, Etsuro Ito. Revisiting oxytocin generation inkeratinocytes. Biophysics and Physicobiology. 2023, Vol. 20; Verena Deing, Dennis Roggenkamp, ​​Jochen Kühnl, Alisa Gruschka, Franz Horst Wenck, Alexander Bürkle, and Gitta Neufang. Oxytocin modulates proliferation and stress responses of human skin cells: implications for atopic dermatitis. Experimental Dermatology, 2013, 22, 399-405. OXT can directly or indirectly improve skin condition and promote skin repair by regulating emotional stress, resisting and alleviating skin sensitivity, anti-aging, regulating inflammation, and providing neuroprotection. Long-term stress on the skin can lead to elevated cortisol levels. OXT can reduce HPA axis activity, thereby reducing ACTH and cortisol levels and enhancing relaxation (References: Markus Heinrichs, Thomas Baumgartner, Clemens Kirschbaum, and Ulrike Ehlert. Social Support and Oxytocin Interact to Suppress Cortisol and Subjective Responses to Psychosocial Stress. BIOL PSYCHIATRY. 2003, 54, 1389-1398; Kerstin Moberg, Linda Handlin, Kathleen Kendall-Tackett, Maria Petersson. Oxytocin is a principal hormone that exerts part of its effects through active fragments. Medical Hypotheses, 2019, 133, 109394. Nrf2 is a key regulator of intracellular redox homeostasis. OXT can increase intracellular Nrf2 levels to maintain intracellular redox homeostasis. OXT reduces and inhibits TRPV1 activity through vasopressin receptor 1a. In turn, thermal stimulation of TRPV1 can release oxytocin. OXT is a TRPV1 agonist that can desensitize the receptor and effectively relieve pain (References: Biswa M. Padhy and Vijay L. Kumar. Inhibition of Calotropis procera Latex-Induced Inflammatory Hyperalgesia by Oxytocin and Melatonin. Mediators of Inflammation, 2005, 6, 360-36; Yelena Nersesyan, Lusine Demirkhanyan, Deny Cabezas-Bratesco, et al. Oxytocin Modulates Nociception as an Agonist of Pain-Sensing TRPV1. Cell Reports, 2017, 21, 1681-1691; Phuoc-Tan Diep, TRPV1, Nrf2, and COVID-19: Could Oxytocin Have a Beneficial Role to Play? Int Arch Allergy Immunol 2, 2021, DOI: 10.1159 / 000521246; Abimael González-Hernández, Antonio Espinosa De Los Guadalupe Martínez-Lorenzana, Miguel Condés-Lara. Recurrent antinociception induced by intrathecal or peripheral oxytocin in an aneuropathic pain rat model. Experimental Brain Research, 2019, 12). OXT can inhibit SASP-induced senescence in normal human skin fibroblasts through the OXTR-mediated extracellular signal-regulated kinase / Nrf2 pathway, thereby preventing skin aging (reference: S.-Y. Cho, AY Kim, J. Kim, D.-H. Choi, ED Son and D.W. Shin. Oxytocin alleviates cellular senescence through oxytocin receptor-mediated extracellular signal-regulated kinase / Nrf2 signaling. British Journal of Dermatology, 2019, 181, 1122-1123). OXT can inhibit the loss of OXTR in keratinocytes, which leads to increased release of proinflammatory cytokines IL6, CCL5, and CXCL10. Reduced OXT levels lead to increased ROS levels and decreased GSH levels, reducing the proliferation of dermal fibroblasts and keratinocytes. This suggests that the OXT system can regulate inflammation, oxidative stress, and proliferative responses. The OXT system plays an important role in human skin homeostasis (References: Verena Deing, Dennis Roggenkamp, ​​Jochen Kühnl, Alisa Gruschka, Franz Horst Wenck, Alexander Bürkle and Gitta Neufang. Oxytocin modulates proliferation and stress responses of human skin cells: implications for atopic dermatitis. Experimental Dermatology, 2013, 22, 399-405.). The neuroprotective effect of OXT is mainly mediated by its anti-inflammatory properties. Due to its antioxidant, anti-inflammatory and anti-apoptotic properties, it has been proposed as a potential neuroprotective and neurobehavioral therapeutic agent, including regulating mood disorders and cognitive barriers (reference: Roya Kamrani-Sharif, A. Wallace Hayes,, Mina Gholami, et al. Oxytocin as neuro-hormone and neuro-regulator exert neuroprotective properties: A mechanistic graphical review. Neuropeptides, 2023, 10, 102352).

[0007] β-Endorphin (β-EP) is also an endogenous neuropeptide hormone and one of the "happy hormones," with analgesic, stress-relieving, and euphoric effects. β-EP and its receptors are synthesized not only by nerve cells but also by cells of the skin and immune system (Reference: Xin Yuchen et al., Principles and Research Progress of Neurocosmetology. Cosmetics and Neuroscience, 2025, Vol. 1 (No. 252), pp. 140-155). β-EP's effects on skin health are primarily reflected in promoting tissue repair, combating inflammation, resisting damage, protecting the skin barrier, accelerating wound healing, and relieving pain. Studies have shown that β-EP, as a μ-opioid receptor activator, promotes the migration of human keratinocytes by binding to the μ-opioid receptor, accelerating re-epithelialization, tissue repair, and wound healing (Reference: Paul L. Bigliardi, Stanislaus Büchner, Theo Rufli, and Mei Bigliardi-Qi. Specific Stimulation of Migration of Human Kerationcytes by μ-Opiate Receptor Agonists. Journal of Receptors and Signal Transduction, 2002, Vol. 22, Nos. 1-4, pp. 191-199). β-EP alleviates UVB-induced inflammatory responses in human keratinocytes by blocking NF-κB signaling, reducing the secretion of inflammatory factors such as IL-1β, IL-6, IL-8, and TNF-α, indicating that β-EP can protect against UVB-induced epidermal inflammatory responses. In addition, β-EP reduces UVB-induced epidermal hyperplasia and protects against barrier damage (Reference: Hyung-Su Kim, Hyoung-June Kim, Yong-Deog Hong, et al. β-Endorphin Suppresses Ultraviolet Birradiation-induced Epidermal Barrier Damage by Regulating Inflammation-dependent mTORC1 Signaling., Nature Portfolio, 2023, 13, 22357). β-EP has been used to treat wounds in animal models. It can promote the expression of cytokeratin CK16 and transforming growth factor β type II receptor in human skin, which are associated with epidermal proliferation and differentiation, thereby inducing fibroblast proliferation and capillary growth, and accelerating the maturation of granulation tissue and epithelialization of defects. In addition, β-EP can fine-tune pain and inflammatory responses during the healing process.This suggests that β-endorphin plays a crucial role not only in pain control but also in wound healing (P.L. Bigliardi, L.T. Sumanovski, S. Büchner, et al. Different Expression of μ-Opiate Receptor in Chronic and Acute Wounds and the Eiect of β-Endorphin on Transforming Growth Factor b Type II Receptor and Cytokeratin Expression. The Journal of Investigative Dermatology, 2002, 120, 145-152). Reports have confirmed that after tissue injury, the transport of β-EP and μ-opioid receptors to the injured tissue is enhanced. This unique simultaneous upregulation of receptors and ligands helps to cope with or prevent skin inflammatory pain (Reference: Shaaban A. Mousa, Qin Zhang, Nicolle Sitte, et al. β-Endorphin-containing memory-cells and μ-opioid receptors undergo transport to peripheral inflamed tissue. Journal of Neuroimmunology, 2001, 115, 71-78).

[0008] In summary, by targeted regulation of the synthesis of neurotransmitters and hormones such as OXT and β-EP, the product can not only improve the pathological state of the skin, but also reshape the user's emotional experience, and promote the beauty industry to evolve into the 3.0 era of "neuro-emotion-skin" three-in-one integration. Summary of the Invention

[0009] The inventors of this application discovered in experiments that schizophyllan can upregulate the synthesis of oxytocin and endorphins, and can be used in neurobeauty or emotional skin care products to reduce skin pain and itching, reduce external stimulation and pressure on the skin, relieve skin tension, and improve sensitive skin quality and skin emotions, thereby completing this application.

[0010] The specific technical solutions of this application are as follows:

[0011] 1. Use of schizophyllan in the preparation of a product for increasing the production of endorphins and / or oxytocin.

[0012] 2. Use of schizophyllan as a promoter for increasing the production of endorphins and / or oxytocin in cells.

[0013] 3. Use of schizophyllan in the preparation of neurobeauty or emotional skin care products.

[0014] 4. Use of schizophyllan in the preparation of products for regulating brain waves and / or increasing the pain threshold of the skin and / or reducing external stimulation and pressure.

[0015] 5. Use of schizophyllan in the preparation of products for relieving skin tension and / or repairing or improving sensitive skin and / or reducing skin sensitivity and itching and / or improving skin function.

[0016] 6. The use according to any one of items 3 to 5, wherein the schizophyllan is used to increase the production of endorphins and / or oxytocin.

[0017] 7. The use according to any one of items 3 to 5, wherein the concentration of the schizophyllan is 0.0005-5.0 wt.%, preferably 0.0025-0.1 wt.%.

[0018] 8. The use according to any one of items 1 to 7, wherein the dosage form of the product comprises a cream, lotion, lotion, powder, spray, gel, mask or freeze-dried powder;

[0019] Optionally, the product comprises cosmetics, food, device products or daily chemicals.

[0020] 9. The use according to items 1 to 8, wherein the endorphin comprises β-endorphin.

[0021] 10. A method for promoting the formation of endorphins and / or oxytocin for non-therapeutic purposes, comprising administering an effective amount of schizophyllan, preferably administering the effective amount of schizophyllan to the skin.

[0022] Effects of the Invention

[0023] The schizophyllan described in the present application can increase the production of oxytocin and endorphins, and thus can be used in neurobeauty or emotional skin care products to regulate brain waves, increase the skin's pain threshold, reduce external pressure, reduce skin sensitivity, relieve skin tension, and improve skin texture. It can also reduce skin pain and itching, reduce external stimulation and pressure on the skin, and improve sensitive skin texture. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Schematic diagram of β-endorphin fluorescence intensity in different groups of cells

[0025] Figure 2 Schematic diagram of oxytocin fluorescence intensity in different groups of cells. DETAILED DESCRIPTION

[0026] The present application is described in detail below in conjunction with the described embodiments. Although specific embodiments of the present application are shown, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0027] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the difference in the functions of the components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of this application shall be as defined by the attached claims.

[0028] The present application provides a method for using schizophyllan in increasing the production of endorphins and / or oxytocin.

[0029] Schizophyllan (SPG) has a β-1,3-pyranose backbone, with one of every three glycosyl groups linked to a β-1,6-pyranose. When dissolved in water, SPG forms a trimer composed of three sugar chains of similar length, forming a rigid, rod-shaped triple helix (a unique triple helix structure).

[0030] In the present application, the schizophyllan can be commercially available or prepared by methods known in the art, such as conventional microbial fermentation methods or Schizophyllan fruiting body extraction methods in the art.

[0031] In some embodiments, the weight average molecular weight of the schizophyllan is 600,000-3000,000 Da, preferably 1200,000-1800,000 Da.

[0032] For example, the weight average molecular weight of the schizophyllan can be 600,000 Da, 650,000 Da, 700,000 Da, 750,000 Da, 800,000 Da, 850,000 Da, 900,000 Da, 950,000 Da, 1000,000 Da, 1050,000 Da, 1100,000 Da, 1150,000 Da, 0Da, 1200,000Da, 1250,000Da, 1300,000Da, 1350,000Da, 1400,000Da, 1450,000D a. 1500,000Da, 1550,000Da, 1600,000Da, 1650,000Da, 1700,000Da, 1750,000Da, 1 800,000Da, 1850,000Da, 1900,000Da, 1950,000Da, 2000,000Da, 2050,000Da, 210 0,000Da, 2150,000Da, 2200,000Da, 2250,000Da, 2300,000Da, 2350,000Da, 2400,0 00Da, 2450,000Da, 2500,000Da, 2550,000Da, 2600,000Da, 2650,000Da, 2700,000D a, 2750,000Da, 2800,000Da, 2850,000Da, 2900,000Da, 2950,000Da, 3000,000Da, etc.

[0033] In the present application, there is no limitation on the method for determining the weight-average molecular weight of schizophyllin. It can be determined by conventional methods in the art, for example, high performance liquid chromatography (HPLC), multi-angle laser scattering (GPC-MALLS), viscometry, and the like.

[0034] In one embodiment, the weight average molecular weight of schizophyllan is determined using multi-angle laser light scattering (GPC-MALLS).

[0035] Endorphins, also known as endorphins or brain endorphins, are a group of neuromodulator chemicals secreted by the pituitary gland. They are "painkillers" spontaneously produced by the body after people persist in doing something for a long time, making people feel euphoric, satisfied and calm. Endorphins play an important role in regulating emotional behaviors (including anxiety, fear, tension and pleasure) and pain control. They are closely related to drug addiction, excitement after exercise, appetite control, sexual response, and mental illnesses such as depression and schizophrenia. β-endorphins and their receptors can not only be synthesized by nerve cells, but can also be produced by cells of the skin and immune system. They play a role in repairing the skin, fighting inflammation, resisting damage, protecting the barrier, accelerating wound healing, reducing external pressure, increasing the skin's pain threshold, reducing sensitivity and itching, and relieving skin tension.

[0036] Oxytocin (OXT) is an endogenous neuropeptide hormone primarily synthesized in the paraventricular and supraoptic nuclei of the hypothalamus, acting throughout the central nervous system and periphery. Studies have also found that OXT and its receptors are expressed in primary human dermal fibroblasts (FIBs) and keratinocytes (KERs). In the skin, OXT can regulate inflammation, oxidative stress, neuroprotection, and skin mood. It also plays a role in products that increase the skin's pain threshold, reduce external stimuli and stress, reduce skin sensitivity and itching, relieve skin tension, and repair or improve sensitive skin and mood.

[0037] In some embodiments, the endorphin is beta-endorphin.

[0038] During the experiment, the inventors of the present application creatively discovered that schizophyllan can increase the production of endorphins and / or oxytocin, and thus can be used in neurobeauty or emotional skin care.

[0039] The present application provides the use of schizophyllan as a promoter in increasing the production of endorphins and / or oxytocin in cells. In some embodiments, the cells include skin cells, nerve cells, or tissue cells, preferably skin cells, more preferably keratinocytes or dermal fibroblasts.

[0040] The schizophyllan described in the present application can act as a promoter to increase the production of endorphins and / or oxytocin in cells.

[0041] The present application provides the use of schizophyllan in the preparation of neurobeauty or emotional skin care products.

[0042] The schizophyllan described in the present application can increase the production of endorphins and / or oxytocin, and endorphins and / or oxytocin are closely related to neurobeauty and emotional skin care. Therefore, schizophyllan can be used to prepare neurobeauty or emotional skin care products.

[0043] The present application provides the use of schizophyllan in the preparation of products for regulating brain waves, increasing the pain threshold of the skin, reducing external stimulation and pressure, reducing skin sensitivity and itching, improving skin function, relieving skin tension, and repairing or improving sensitive skin.

[0044] As described above, the mycophenolate mofetil described in the present application can increase the production of endorphins and / or oxytocin, and endorphins and / or oxytocin are related to regulating brain waves, increasing the pain threshold of the skin, reducing external stimuli and pressure, reducing skin sensitivity and itching, improving skin function, relieving skin tension, and repairing or improving sensitive skin. Therefore, it can be used to prepare products that regulate brain waves, increase the pain threshold of the skin, reduce external stimuli and pressure, reduce skin sensitivity and itching, improve skin function, relieve skin tension, and repair or improve sensitive skin.

[0045] In some embodiments, the schizophyllan is used to increase the production of endorphins and / or oxytocin.

[0046] In some embodiments, the concentration of the schizophyllan is 0.0005-5.0 wt.%, preferably 0.0025-0.1 wt.%.

[0047] For example, the concentration of the mycophenolate mofetil can be 0.0005wt.%, 0.001wt.%, 0.005wt.%, 0.05wt.%, 0.1wt.%, 0.2wt.%, 0.3wt.%, 0.4wt.%, 0.5wt.%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, etc.

[0048] In some embodiments, the dosage form of the product comprises a cream, lotion, solution, powder, spray, gel, mask or freeze-dried powder;

[0049] Optionally, the product comprises cosmetics, food, device products or daily chemicals. Optionally, the cosmetics can be skin care products, makeup, scalp care products, sunscreen products, etc.

[0050] The present application provides a composition for neurobeauty or emotional skin care, which contains schizophyllan.

[0051] The present application also provides a composition for regulating brain waves, increasing the pain threshold of the skin, reducing external pressure and stimulation, reducing skin sensitivity and itching, and repairing or improving sensitive skin, which contains schizophyllan.

[0052] In some embodiments, the mass percentage of the schizophyllan is 0.0005-5.0%, preferably 0.0025-0.1% in the composition.

[0053] For example, in terms of the mass percentage in the composition, the mycophenolate mofetil can be 0.0005%, 0.001%, 0.005%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, etc.

[0054] The present application provides a method for promoting the formation of endorphins and / or oxytocin for non-therapeutic purposes, which comprises administering an effective dose of schizophyllan, preferably, administering an effective dose of schizophyllan on the skin.

[0055] In the present application, there is no limitation on the effective dose, and those skilled in the art can select an effective dose that can be applied to act on the relevant site.

[0056] Example

[0057] This application provides general and / or specific descriptions of the materials and experimental methods used in the experiments. In the following examples, unless otherwise specified, % represents wt%, i.e., percentage by weight. Reagents or instruments used without manufacturer indication are commercially available conventional reagents. The raw materials used are shown in Table 1.

[0058] Table 1

[0059] Experimental Materials supplier Remark DMEM medium Gibco β-endorphin antibody Invitrogen Oxytocin Antibody Invitrogen schizophyllin Bloomage Biotech Co., Ltd. Molecular weight 1461K Da

[0060] Example 1: Schizophyllan as a promoter to upregulate the expression of endorphins

[0061] 1) Detection method: Immunofluorescence was used to evaluate the effect of the sample on the expression of β-endorphin in fibroblasts (FB cells) after UVA irradiation. 2 After UVA irradiation, the cells were exposed to the samples and cultured for 72 h to evaluate the effects of the samples on β-endorphin expression in FB cells.

[0062] 2) Detection index: The relative mean fluorescence intensity (%) of the target protein was used as the evaluation index. The relative mean fluorescence intensity of the target protein after sample treatment was calculated by comparing with the blank control (BC).

[0063] 3) Group settings

[0064] Table 2

[0065]

[0066] 4) Detection steps:

[0067] Cell seeding: Fibroblasts in the logarithmic growth phase were cultured in DMEM medium containing 10% FBS. When the cell density reached approximately 80%, cells were digested with 0.05% trypsin-EDTA solution and collected by centrifugation at 1000 rpm for 5 minutes. After centrifugation, cells were resuspended in DMEM medium containing 10% FBS and counted using a hemocytometer. Cells were seeded at an appropriate density in a 24-well cell culture plate equipped with a slide and cultured in a 37°C, 5% CO2 incubator to ensure normal cell attachment and growth.

[0068] UVA irradiation: After 24 hours of cell inoculation, the culture medium was removed and 1× DPBS solution was added to each well. The BC group was placed in a dark environment, and the cells of the MC and sample groups were placed in a solar simulator at 2.8 J / cm 2 UVA irradiation was performed. After irradiation, the 1× DPBS solution was removed. BC and MC were added to DMEM medium containing 10% FBS. Sample groups were added with different concentrations of sample solution and incubated in a constant temperature incubator at 37°C and 5% CO2 for 72 hours.

[0069] Detection: After 72 hours of sample treatment, the culture medium was aspirated and ice-cold methanol was added to each well to fix the cells for 15 minutes. After fixation, the cells were gently washed once with 1× DPBS solution in each well. The 1× DPBS solution was aspirated and the cells were blocked with blocking solution in each well for 1 hour. After blocking, β-endorphin antibody was added to each well and incubated overnight. After the antibody incubation, the cells were gently washed three times with 1× DPBS solution in each well. The 1× DPBS solution was aspirated and Alexa Fluor 488-labeled anti-rabbit secondary antibody was added to each well and incubated for 1.5 hours. After the secondary antibody incubation, the cells were gently washed three times with 1× DPBS solution in each well. The slides were transferred to a slide with DAPI mounting medium and the fluorescence channel corresponding to Alexa Fluor 488 was photographed using a fluorescence microscope. Ten observation areas were selected for each group, and the β-endorphin fluorescence intensity was recorded.

[0070] 5) Data analysis: The relative mean fluorescence intensity (%) was calculated using the following formula:

[0071]

[0072] Images were processed using Graphpad Prism software, and the results are expressed as mean ± SD. Student's t-test was used. For comparisons with BC, *P < 0.05 indicated statistical significance, **P < 0.01 indicated significant statistical significance, and ***P < 0.001 indicated extremely significant statistical significance. For comparisons with MC, #P < 0.05 indicated statistical significance, ##P < 0.01 indicated significant statistical significance, and ###P < 0.001 indicated extremely significant statistical significance.

[0073] 6) Result determination: If the relative mean fluorescence intensity (%) is greater than MC and is statistically significant compared with MC (P<0.05), it is determined that the sample at this test concentration can increase the expression of β-endorphin in FB cells after UVA irradiation.

[0074] 7) Results and analysis, the results are shown in Table 3 and Figure 1 As shown in the figure, Merge is a merged image of the β-endorphin group and the DAPI group. Both cell and endorphin staining can be displayed in the same image. Green fluorescence indicates the expression of the target protein. The stronger the green fluorescence, the more the target protein is expressed.

[0075] Table 3

[0076]

[0077] From Table 3 and Figure 1 It can be seen that the fluorescence intensity of β-endorphin in FB cells in the MC group was significantly reduced, indicating that the model was successful. Compared with MC, schizophyllan at test concentrations of 0.005%, 0.01%, and 0.025% increased the fluorescence intensity of β-endorphin in FB cells under UVA irradiation model, with the relative average fluorescence intensity increase rates being 14%, 20%, and 18%, respectively. The results were statistically significant (P < 0.001), indicating that schizophyllan can enhance and upregulate the expression of β-endorphin in FB cells.

[0078] Example 2: Schizophyllan as a promoter to upregulate the expression of oxytocin

[0079] 1) Detection method: Immunofluorescence assay was used to evaluate the effect of the sample on the expression of oxytocin in keratinocytes HaCaT cells after UVB irradiation. 2 After UVB irradiation, the cells were exposed to the samples and cultured for 72 hours to evaluate the effects of the samples on oxytocin expression in HaCaT cells.

[0080] 2) Detection index: The relative average fluorescence intensity (%) of the target protein is used as the evaluation index.

[0081] 3) Group settings

[0082] Table 4

[0083]

[0084] 4) Detection steps

[0085] Cell seeding: Keratinocytes in the logarithmic growth phase were cultured in DMEM medium containing 10% FBS. When the cell density reached approximately 80%, cells were digested with 0.05% trypsin-EDTA solution and collected by centrifugation at 1000 rpm for 5 minutes. After centrifugation, the cells were resuspended in DMEM medium containing 10% FBS and counted using a hemocytometer. The cells were seeded at an appropriate density in a 24-well cell culture plate equipped with a slide and cultured in a 37°C, 5% CO2 incubator to ensure normal cell attachment and growth.

[0086] UVB irradiation: After 24 hours of cell inoculation, the culture medium was removed and 1× DPBS solution was added to each well. The BC group was placed in a dark environment, and the cells of the MC and sample groups were placed in a solar simulator at 60 mJ / cm 2 UVB irradiation was performed. After irradiation, the 1× DPBS solution was removed. BC and MC were added to DMEM medium containing 10% FBS. Sample groups were added with different concentrations of sample solution and cultured in a constant temperature incubator at 37°C and 5% CO2 for 72 hours.

[0087] Detection: After 72 hours of sample treatment, the culture medium was aspirated and ice-cold methanol was added to each well to fix the cells for 15 minutes. After fixation, 1× DPBS solution was added to each well to gently wash the cells once. The 1× DPBS solution was aspirated and the cells were blocked with blocking solution for 1 hour in each well. After blocking, oxytocin antibody was added to each well and incubated overnight. After the antibody incubation, 1× DPBS solution was added to each well to gently wash the cells 3 times. The 1× DPBS solution was aspirated and Alexa Fluor 488-labeled anti-rabbit secondary antibody was added to each well to incubate for 1.5 hours. After the secondary antibody incubation, 1× DPBS solution was added to each well to gently wash the cells 3 times. The slides were transferred to a slide with DAPI mounting medium and the fluorescence channel corresponding to Alexa Fluor 488 was photographed using a fluorescence microscope. Ten observation areas were selected for each group, and the oxytocin fluorescence intensity was recorded.

[0088] 5) Data analysis: The relative mean fluorescence intensity (%) was calculated using the following formula:

[0089]

[0090] Images were processed using Graphpad Prism software, and the results are expressed as mean ± SD. Student's t-test was used for comparisons with MC. *P < 0.05 indicates statistical significance, **P < 0.01 indicates significant statistical significance, and ***P < 0.001 indicates extremely significant statistical significance. For comparisons with MC, #P < 0.05 indicates statistical significance, ##P < 0.01 indicates significant statistical significance, and ###P < 0.001 indicates extremely significant statistical significance.

[0091] 6) Result determination: If the relative mean fluorescence intensity (%) is greater than MC and is statistically significant compared with MC (P<0.05), the sample is determined to be able to increase oxytocin expression in HaCaT cells after UVB irradiation at this test concentration.

[0092] 7) Results and analysis, the results are shown in Table 5 and Figure 2 As shown in the figure, Merge is a merged image of the oxytocin group and the DAPI group. Both cell and endorphin staining can be displayed in the same image. Green fluorescence indicates the expression of the target protein. The stronger the green fluorescence, the more the target protein is expressed.

[0093] Table 5

[0094]

[0095] From Table 3 and Figure 1 As can be seen, the fluorescence intensity of oxytocin in HaCaT cells in the MC group decreased significantly, indicating that the model was successfully established. Compared with MC, schizophyllan at test concentrations of 0.005%, 0.01%, and 0.025% increased the fluorescence intensity of oxytocin in HaCaT cells under UVB irradiation, with relative mean fluorescence intensity increases of 16%, 24%, and 21%, respectively. These results were statistically significant (P < 0.01), indicating that schizophyllan can enhance the expression of oxytocin in HaCaT cells.

[0096] In summary, schizophyllan as a promoter can upregulate the expression of β-endorphin and oxytocin, which, as key mediators in the "gut-brain-skin axis", are directly involved in the regulation of the "neuro-immune-endocrine" system and play a role in neurobeauty and emotional skin care products.

[0097] Example 3: Clinical testing experiment

[0098] Experimental study on the effect of schizophyllin on reducing external irritation and sensitivity, repairing and improving skin texture

[0099] Note: This trial was approved by the Xiamen University Medical Ethics Committee under the experimental plan "Study on the Cosmetic Efficacy of Schizophyllin Raw Materials" with the approval number XDYX202410K62.

[0100] 1. Sample information is shown in Table 6:

[0101] Table 6

[0102] Serial number Sample name Sample properties batch number Shelf life Storage conditions 1 Gel containing 0.01% schizophyllin Transparent colorless gel 20240801 2 years normal temperature 2 Placebo Transparent colorless gel 20240802 2 years normal temperature

[0103] Placebo formula: Carbomer 0.2%, 1,2-hexanediol 2%, ethylhexylglycerin 0.05%, triethanolamine 0.2%, balance water

[0104] Schizophyllin gel: Carbomer 0.2%, 1,2-hexanediol 2%, ethylhexylglycerin 0.05%, triethanolamine 0.2%, schizophyllin 0.01%, balance water

[0105] 2. Usage:

[0106] When using, take an appropriate amount of product on the palm of your hand, apply evenly to the face, and massage gently until completely absorbed. The sample usage period is 28 days, twice a day, that is, once in the morning and once in the evening.

[0107] 3. Volunteer inclusion criteria (sensitive skin population, 29 volunteers)

[0108] 1) Have signed the informed consent form and voluntarily participated in this test.

[0109] 2) Inclusion criteria

[0110] A. Subjects were Chinese females aged between 18 and 40 years (based on the test date);

[0111] B. Self-reported facial skin type as sensitive (lactic acid sting test score ≥ 3 points), expert-assessed wrinkle grade of 2 or higher, and facial redness, dryness, and sagging;

[0112] C. The subject has no history of allergy to cosmetics or drugs;

[0113] D. The subjects promise not to change their existing skin care product usage habits during the testing period;

[0114] E. The subjects guarantee that they will not use any facial skin care products with the same functions as the test samples during the trial period;

[0115] F. The subject did not participate in prolonged outdoor exercise during the testing period, nor did he or she plan to go to the seaside area with strong sunlight;

[0116] G. The subjects have a certain awareness of makeup;

[0117] H. The subject has not undergone laser or chemical peels.

[0118] 4. Test method:

[0119] Reference: T / CAB 0152-2022. Test Methods for Seven Cosmetic Efficacy Functions: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing[S]. Beijing: China Association for Promoting Industry-University-Research Cooperation, 2022. Transepidermal water loss testing (TEWAMETER™ HEX) was performed at the intersection of the midline of the eye and the tip of the nose.

[0120] And fill in the consumer experience evaluation questionnaire, as follows:

[0121] Evaluation Method: A questionnaire survey was conducted via mobile phone. Participants self-assessed their experience and satisfaction with the samples 14 and 28 days after using them. The scoring was based on a 5-point scale. See Table 7 for specific scoring criteria:

[0122] Table 7

[0123] score Evaluation content 1 point Strongly disagree / strongly dislike / strongly unwilling 2 points Disagree / dislike / unwilling 3 points Neither agree nor disagree 4 points Agree / Like / Willing 5 points Very satisfied / very like / very willing

[0124] Results Evaluation: Quantify the respondent's satisfaction with the product using satisfaction ratings. Count the proportion of respondents who responded to each question at each time point and calculate satisfaction (see the formula below).

[0125] The higher the value, the higher the satisfaction.

[0126]

[0127] Adverse symptom monitoring:

[0128] During the use of the samples, any skin discomfort symptoms (such as skin redness, stinging, itching, burning) that occur are monitored and recorded.

[0129] 5. Test results:

[0130] 5.1 Transepidermal water loss and transepidermal water loss rate are shown in Table 8

[0131] Table 8

[0132]

[0133]

[0134] Compared with the placebo, the transepidermal water loss rate of 0.01% mycophenolate mofetil gel was significantly reduced after 30 minutes, 2 hours and 28 days of use (P < 0.010), indicating that it has immediate and long-term repair effects, can reduce skin sensitivity and enhance the skin's ability to resist stress and irritation.

[0135] 5.2 Consumers’ usage experience evaluation is shown in Table 9.

[0136] Table 9

[0137]

[0138]

[0139] As can be seen from the table above, the product was found to have no adverse effects on the skin and physical health of the subjects, and there was no significant difference in skin feel, texture, color, and smell between the experimental group and the placebo.

[0140] Compared with the placebo, consumers were more satisfied with the skin's hydration, desquamation, elasticity, wrinkle reduction and improved skin texture; there was a significant improvement in resistance to external pressure, resistance to cold water stimulation, improvement of itching and red blood streaks, satisfaction, comfort and skin function.

[0141] According to clinical tests, schizophyllin can increase the skin's pain threshold, reduce external stimulation and pressure, relieve skin tension, improve skin sensitivity and itching, and play a role in repairing and enhancing skin function, and improving sensitive skin quality and skin mood.

[0142] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application in any other manner. Any person skilled in the art may utilize the above disclosed technical content to modify or modify the present application into equivalent embodiments with equivalent variations. However, any simple modifications, equivalent variations, and modifications to the above embodiments that do not depart from the technical content of the present application and are based on the technical essence of the present application shall still fall within the scope of protection of the present application.

Claims

1. Use of schizophyllan in the preparation of a product for increasing the production of endorphins and / or oxytocin.

2. Use of schizophyllan as a promoter for increasing the production of endorphins and / or oxytocin in cells.

3. Use of schizophyllan in the preparation of neurobeauty or emotional skin care products.

4. Use of schizophyllan in the preparation of products for regulating brain waves and / or increasing the pain threshold of the skin and / or reducing external stimulation and pressure.

5. Use of schizophyllan in the preparation of products for relieving skin tension and / or repairing or improving sensitive skin and / or reducing skin sensitivity and itching and / or improving skin function.

6. The use according to any one of claims 3 to 5, wherein the schizophyllan is used to enhance the production of endorphins and / or oxytocin.

7. The use according to any one of claims 3 to 5, wherein The concentration of the schizophyllan is 0.0005-5.0 wt.%, preferably 0.0025-0.1 wt.%.

8. The use according to any one of claims 1 to 7, wherein The dosage form of the product includes cream, lotion, solution, powder, spray, gel, mask or freeze-dried powder; Optionally, the product comprises cosmetics, food, device products or daily chemicals.

9. The use according to claims 1-8, wherein the endorphin comprises beta-endorphin.

10. A method for promoting the formation of endorphins and / or oxytocin for non-therapeutic purposes, wherein: This comprises administering an effective amount of schizophyllin, preferably, to the skin.

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