Application of ganoderic acid A in preparation of repairing agent for skin barrier function impairment

Abstract

The application relates to the field of biological medicine, and particularly relates to application of ganoderic acid A in preparation of a repairing agent for skin barrier function impairment. The ganoderic acid A can repair the impaired barrier from the source by up-regulating the expression of barrier function proteins and improving the cell resistance value, has a significant improvement effect on the barrier impairment caused by UVB irradiation, age growth and high-fat and high-sugar diet, is suitable for a wide range of scenes, has high biological safety and no obvious toxic side effects, can be used for preparing various product forms, and provides a safe and reliable solution for preventing and improving skin barrier function recession.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically, to the application of ganoderic acid A in the preparation of repair agents for impaired skin barrier function. Background Technology

[0002] The skin barrier mainly refers to the brick-and-mortar structure formed by keratinocytes during keratinization, composed of intercellular lipids such as ceramides, barrier-related proteins, and keratinocytes. They work together to effectively prevent moisture loss and the invasion of harmful external substances into the skin.

[0003] Impaired skin barrier function is a common clinical skin abnormality and a core cause of various skin diseases. When the barrier structure is damaged, skin permeability becomes abnormal, allowing external antigens to easily penetrate the epidermis, activating the skin's immune inflammatory response, triggering the release of inflammatory factors, and thus inducing or aggravating various skin diseases.

[0004] Among them, barrier dysfunction is an important pathological feature of skin diseases such as atopic dermatitis, sensitive skin, seborrheic dermatitis, acne vulgaris, and psoriasis: acne patients have abnormal expression of barrier-related proteins such as tight junction proteins and filaggrin in their skin, and the stratum corneum barrier structure is disordered; psoriasis and eczema patients have abnormal proliferation and differentiation of keratinocytes, and damage to the epidermal brick wall structure, forming a vicious cycle of barrier damage and inflammatory response.

[0005] Experiments have confirmed that the skin barrier function declines and becomes damaged with age. Simultaneously, external environmental stimuli, improper skincare practices, and metabolic abnormalities can all disrupt the skin barrier's integrity, further inducing the accumulation of reactive oxygen species, activating inflammatory signaling pathways such as NF-κB, disrupting the skin's immune balance, and exacerbating discomfort symptoms such as dryness, redness, and stinging, severely impacting skin health and quality of life.

[0006] Therefore, targeted repair and improvement of skin barrier function are key to treating barrier-related skin diseases and maintaining healthy skin homeostasis.

[0007] Current skincare and pharmaceutical products primarily focus on antioxidant, anti-inflammatory, and exogenous moisturizing ingredients, primarily addressing direct symptoms such as dryness and redness on the skin's surface. However, research and development of intervention products targeting the repair of the skin barrier structure and improvement of its function is limited. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide the application of ganoderic acid A in the preparation of a repair agent for impaired skin barrier function.

[0009] The technical solution of the present invention to solve the above-mentioned technical problems is as follows: This invention provides the application of ganoderic acid A in the preparation of a repair agent for impaired skin barrier function.

[0010] Based on the above technical solution, the present invention can be further improved as follows.

[0011] Furthermore, impaired skin barrier function includes decreased resistivity of human immortalized epidermal keratinocytes and / or reduced expression of skin barrier-related proteins.

[0012] Furthermore, the skin barrier-related proteins include at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein.

[0013] Furthermore, the factors that can cause impaired skin barrier function include UVB radiation, aging, or a high-fat, high-sugar diet.

[0014] The present invention also provides a method for regulating the expression level of skin barrier-related proteins, wherein ganoderic acid A is applied to skin cells to increase the expression level of skin barrier-related proteins; the skin barrier-related proteins include at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein.

[0015] The present invention also provides a topical formulation for repairing UVB irradiation-induced skin barrier damage, wherein the topical formulation contains ganoderic acid A at a concentration of 8-12 μM; the topical formulation further includes pharmaceutically or cosmetically acceptable excipients.

[0016] Furthermore, the topical preparation is one of the following: cream, lotion, serum, gel, spray, or mask.

[0017] The present invention also provides an oral formulation for repairing age-induced skin barrier damage, wherein the oral formulation contains ganoderic acid A, wherein the mass percentage of ganoderic acid A is 0.01%-0.02%; the oral formulation also includes pharmaceutically or food-grade excipients. Furthermore, the oral preparation is a functional food.

[0018] The present invention also provides an injectable formulation for repairing skin barrier damage induced by a high-fat, high-sugar diet. The injectable formulation comprises a solvent system and ganoderic acid A, wherein the mass percentage of ganoderic acid A is 0.4%-0.5% of the solvent system; the solvent system comprises DMSO, polyethylene glycol, and PBS.

[0019] The beneficial effects of this invention are as follows: This invention utilizes ganoderic acid A to prepare a repair agent for impaired skin barrier function. As a naturally derived active ingredient, ganoderic acid A has the advantage of targeted repair, acting directly on the core of the skin barrier. It repairs the damaged barrier from the source by upregulating the expression of barrier function proteins and increasing cell resistance. It also has significant improvement effects on barrier damage caused by ultraviolet radiation, aging, and high-fat, high-sugar diets. It has a wide range of applications and high biosafety with no obvious toxic side effects. It can be used to prepare various product forms, providing a safe and reliable solution for preventing and improving the decline of skin barrier function. Attached Figure Description

[0020] Figure 1 The experimental results are for Example 1 of the present invention. Figure 1 Figure A shows a comparison of the changes in HaCaT cell resistance values ​​in each group; Figure 1 Figure B shows the immunofluorescence detection results of DSG2 protein in each group; Figure 1 The middle C figure shows the Western blot (WB) results of barrier-related proteins in each group; Figure 1 The middle section (D) shows a comparison of the expression levels of barrier-related proteins in each group; Figure 2 This is a distribution diagram of the content of ganoderic acid A in the serum and major tissues of mice in the ganoderic acid A intervention group in Example 2 of the present invention; Figure 3 This is a comparative diagram of the qualitative analysis of the expression levels of skin barrier-related proteins in the skin tissues of mice in Example 2 of the present invention, obtained by immunohistochemical detection. Figure 4 This is a comparative diagram showing the quantitative analysis of immunohistochemical detection of skin barrier-related protein expression levels in mouse skin tissues of different groups in Example 2 of the present invention. Figure 4 In this context, A stands for LOR. Figure 4 B in the middle is DSG2. Figure 4 C in the middle is AQP3. Figure 4 D is CK10. Figure 4 E in the middle is CK14; Figure 5 This is an image showing the immunohistochemical detection results of Example 3 of the present invention. Figure 5 Figure A shows a comparison of the qualitative detection results of LOR protein, DSG2 protein, and AQP3 protein in mice from different groups. Figure 5 Figure B shows a quantitative comparison of LOR protein expression levels in different groups of mice. Figure 5 The figure in C is a quantitative comparison of DSG2 protein expression levels; Figure 5 The middle section (D) shows a quantitative comparison of AQP3 protein expression levels. Detailed Implementation

[0021] The principles and features of the present invention are described below. The examples given are only for explaining the present invention and are not intended to limit the scope of the present invention.

[0022] This invention provides the application of ganoderic acid A in the preparation of a repair agent for impaired skin barrier function.

[0023] Ganoderic acid A (GAA) is a triterpenoid lanostane derivative, primarily derived from Ganoderma fungi. It is one of the important active components of Ganoderma lucidum, belonging to the tetracyclic triterpenoid family. Its molecular structure contains multiple functional groups, including carboxyl, hydroxyl, and carbonyl groups. It is soluble in organic solvents such as methanol, ethanol, and chloroform, but has low solubility in water. Ganoderic acid A exhibits broad biological activity and good biocompatibility.

[0024] At the cellular level, ganoderic acid A can reverse UVB-induced barrier damage in human immortalized epidermal keratinocytes (HaCaT), significantly increasing the cell resistivity that decreased after UVB irradiation, while upregulating the expression of barrier-related proteins such as DSG2, AQP3, and FLG, thereby effectively repairing the barrier structure and function of keratinocytes.

[0025] At the animal level, ganoderic acid A can effectively accumulate in skin tissue, enabling it to target the skin barrier and reduce the risk of exposure to non-target organs. For skin barrier damage induced by natural aging, intervention with ganoderic acid A can effectively restore the expression levels of skin barrier-related proteins and promote keratinocyte proliferation and differentiation. For barrier damage induced by metabolic abnormalities, intervention with ganoderic acid A can improve skin barrier function in obese mice induced by a high-fat, high-sugar diet and restore the expression levels of skin barrier-related proteins.

[0026] In summary, ganoderic acid A, as a naturally derived active ingredient, has the advantage of targeted repair. It can directly act on the core of the skin barrier, repairing the damaged barrier from the source by upregulating the expression of barrier function proteins and increasing cell resistance. It also has significant improvement effects on barrier damage caused by ultraviolet radiation, aging, and high-fat, high-sugar diets. It has a wide range of applications and high biosafety with no obvious toxic side effects. It can be used to prepare various product forms, providing a safe and reliable solution for preventing and improving the decline of skin barrier function.

[0027] Preferably, the skin barrier-related proteins include at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein.

[0028] Preferred factors contributing to impaired skin barrier function include UVB radiation, aging, or a high-fat, high-sugar diet.

[0029] The present invention discloses a method for regulating the expression level of skin barrier-related proteins by acting ganoderic acid A on skin cells to increase the expression level of skin barrier-related proteins, including at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein. This regulation method is effective and has the advantages of being safe and non-toxic.

[0030] The topical formulation of the present invention for repairing UVB irradiation-induced skin barrier damage contains ganoderic acid A at a concentration of 8-12 μM.

[0031] Preferably, the topical formulation also includes pharmaceutically or cosmetically acceptable excipients.

[0032] Preferably, the topical preparation is one of the following: cream, lotion, serum, gel, spray, or mask.

[0033] The oral formulation of the present invention for repairing age-induced skin barrier damage contains ganoderic acid A, wherein the mass percentage of ganoderic acid A is 0.01%-0.02%, preferably 0.015%.

[0034] Preferably, the oral formulation also includes pharmaceutically or food-grade excipients.

[0035] Preferably, the oral formulation is a functional food.

[0036] For functional foods, the ingredients can also include synergistic components. For example, polysaccharides and other Ganoderma lucidum acids, which belong to the Ganoderma lucidum family, can work synergistically to resist oxidation, inflammation, and aging when combined. They can also contain natural antioxidants, such as vitamin C, vitamin E, polyphenols (tea polyphenols, grape seed extract), astaxanthin, etc., which can enhance the effect of scavenging free radicals. In addition, they can include small molecule active peptides, such as fish collagen and elastin peptides, which can directly help improve skin condition.

[0037] In addition, the following ingredients should not be added to functional foods: First, strongly alkaline substances, as ganoderic acid A is an acidic terpene compound that is prone to structural damage, decomposition, and inactivation in a strongly alkaline environment; second, high-concentration metal ion chelating agents and chemical additives with strong oxidizing properties, which will oxidize and destroy the functional groups of ganoderic acid A, accelerating its degradation; and third, substances high in tannins and tannins, such as strong tea and persimmon extract, which will combine with ganoderic acid A in the gastrointestinal tract to form insoluble complexes, reducing the solubility of the ingredients and the intestinal absorption rate.

[0038] The present invention relates to an injectable formulation for repairing skin barrier damage induced by a high-fat, high-sugar diet, comprising a solvent system and ganoderic acid A, wherein the mass percentage of ganoderic acid A is 0.4%-0.5% of the solvent system; the solvent system comprises DMSO, polyethylene glycol, and PBS.

[0039] Preferably, the mass percentages of each component in the solvent system are: 10% DMSO, 40% polyethylene glycol, and 50% PBS, with the mass percentage of ganoderic acid A being approximately 0.45% of the solvent system.

[0040] The effects of the present invention will be illustrated by specific embodiments below.

[0041] Example 1: Repair of UVB-induced skin barrier damage by ganoderic acid A This embodiment uses human immortalized epidermal keratinocytes (HaCaT) as the experimental subject to investigate the effect of ganoderic acid A on UVB irradiation-induced skin barrier damage.

[0042] In this embodiment, the experiment specifically included a ganoderic acid A intervention group (UVB+GAA), an untreated control group (CON), and a group treated with only 60 mJ / cm². 2 A model group of HaCaT-induced damage caused by UVB irradiation.

[0043] The intervention group (UVB+GAA) was established using the following method: 60 mJ / cm 2 Human immortalized epidermal keratinocytes (HaCaT) were irradiated with UVB and then pretreated with 10 μM GAA for 24 h.

[0044] The skin barrier function of HaCaT cells in the above groups was detected. Specifically, the cell resistance level was detected using a RE1600 transmembrane cell resistance meter, and the expression levels of proteins related to skin barrier structure and function were detected using Western blot and immunofluorescence.

[0045] like Figure 1 As shown in Figure A, compared to the control group CON, the UVB-induced HaCaT cell resistivity in the model group decreased to 56.76 Ω·cm. 2 The expression of barrier-related proteins was reduced; while in the intervention group, the resistance value of HaCaT cells increased by 23.5% after treatment with ganoderic acid A (P<0.001).

[0046] Regarding the expression of barrier-related proteins DSG2, AQP3, and FLG, such as Figure 1 As shown in the B-D diagram, the expression levels of all the above proteins were significantly increased (all P<0.05), indicating that ganoderic acid A can effectively repair UVB-induced skin barrier damage.

[0047] Example 2: Repair of Age-Induced Skin Barrier Damage by Ganoderic A This study investigated the effect of ganoderic acid A on age-induced skin barrier damage in mice of different ages.

[0048] The mice used in this embodiment were male SPF-grade C57BL / 6J mice, which were divided into a young group, an old group, and an intervention group (old+GAA). Each group was fed a normal maintenance diet (21.5% protein, 11.1% fat, and 67.4% carbohydrates; 120 mg / kg·diet). Mice fed to 2 months of age were in the young group, and mice fed to 23 months of age were in the old group. After being fed the normal maintenance diet to 16 months of age, the mice were then fed a maintenance diet containing ganoderic acid A (120 mg / kg·diet) until they reached 23 months of age.

[0049] The levels of ganoderic acid A in the serum and major tissues of mice in the intervention group were detected by UPLC / MS. The results are as follows: Figure 2 As shown in the figure, the accumulation of ganoderic acid A in each gram of skin tissue of the intervention group mice reached 15.76 ng.

[0050] The expression levels of skin barrier-related proteins DSG2, AQP3, LOR, CK10, and CK14 were detected using immunohistochemistry. The results are as follows: Figure 3 and Figure 4 As shown.

[0051] Figure 3 The red arrow in the image indicates a positive precipitate, which, when combined with... Figure 4 The results from the AC study showed that intervention with ganoderic acid A promoted the proliferation and differentiation of keratinocytes and enhanced skin barrier function. Compared with 2-month-old mice, 23-month-old mice showed decreased expression levels of skin barrier-related proteins (all P < 0.01), while the intervention group treated with ganoderic acid A showed significantly increased expression levels of these skin barrier-related proteins.

[0052] pass Figure 4 As shown in Figures D and E, compared with the young 2-month-old mice, the protein expression levels of cytokeratin 14 and cytokeratin 10 in the skin tissue of the old 23-month-old mice were reduced by 31.9% and 20.0%, respectively (all P<0.01). However, in the intervention group treated with ganoderic acid A, the protein expression levels of cytokeratin 14 and cytokeratin 10 were significantly increased compared with the old group, indicating that the intervention of ganoderic acid A can repair age-induced skin barrier damage.

[0053] Example 3: Repair of skin damage in high-fat, high-glucose induced obese mice by ganoderic acid A This embodiment uses a high-fat, high-glucose induced obese mouse model to investigate the repair effect of ganoderic acid A on skin barrier damage.

[0054] The mouse model used in this embodiment was a male SPF-grade C57BL / 6J mouse. The model was established by randomly dividing 6-month-old mice according to body weight into a normal diet group (21.5% protein, 11.1% fat, and 67.4% carbohydrates), a high-fat, high-sugar diet group (HFHS-Diet), and an intervention group (HFHS-Diet+GAA). The housing environment, lighting, water intake, and circadian rhythm were completely identical for all groups of mice.

[0055] The diet formulation for the high-fat, high-sugar diet group (HFHS-Diet) and the intervention group (HFHS-Diet+GAA) was 14% protein, 42% fat, 44% carbohydrates, 0.2% cholesterol (energy ratio), and high-sugar water (23.1 g / L D-fructose and 18.9 g / L L-glucose). The feeding period lasted for 4 months to complete the modeling.

[0056] The intervention group (HFHS-Diet+GAA) received intraperitoneal injections of 15 mg / kg·bw of ganoderic acid A every three days for three months after modeling.

[0057] Immunohistochemistry was used to detect the expression levels of skin barrier-related proteins DSG2, AQP3, and LOR in mice of each group. The results are as follows: Figure 5 As shown.

[0058] Figure 5 The red arrow in A indicates a positive precipitate, which, when combined with... Figure 5 The results showed that, compared with the normal diet group, the protein expression levels of LOR and AQP3 in the skin tissue of obese mice induced by the high-fat and high-sugar diet decreased by 29.0% and 8.6%, respectively (all P<0.01), while the intervention group effectively restored the expression levels of the above proteins.

[0059] Therefore, intervention with ganoderic acid A can effectively restore skin barrier damage induced by high-fat and high-sugar diets in obese mice.

[0060] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. Application of ganoderic acid A in the preparation of repair agents for impaired skin barrier function.

2. The application of ganoderic acid A according to claim 1 in the preparation of a repair agent for impaired skin barrier function, characterized in that, The impaired skin barrier function includes decreased resistivity of human immortalized epidermal keratinocytes and / or reduced expression of skin barrier-related proteins.

3. The application of ganoderic acid A according to claim 2 in the preparation of a repair agent for impaired skin barrier function, characterized in that, The proteins associated with impaired skin barrier function include at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein.

4. The application of ganoderic acid A according to claim 1 in the preparation of a repair agent for impaired skin barrier function, characterized in that, The factors that can impair skin barrier function include UVB radiation, aging, or a high-fat, high-sugar diet.

5. A method for regulating the expression level of skin barrier-related proteins, characterized in that, Ganoderma lucidum acid A was applied to skin cells to increase the expression of skin barrier-related proteins; the skin barrier-related proteins include at least one of DSG2 protein, AQP3 protein, FLG protein, LOR protein, CK10 protein, and CK14 protein.

6. A topical formulation for repairing UVB irradiation-induced skin barrier damage, characterized in that, The topical formulation contains ganoderic acid A at a concentration of 8-12 μM; the topical formulation also includes pharmaceutically or cosmetically acceptable excipients.

7. A topical formulation for repairing UVB irradiation-induced skin barrier damage according to claim 5, characterized in that, The topical preparation is one of the following: cream, lotion, serum, gel, spray, or mask.

8. An oral formulation for repairing age-induced skin barrier damage, characterized in that, The oral preparation contains ganoderic acid A, with a mass percentage of 0.01%-0.02%; the oral preparation also includes pharmaceutically or food-grade excipients.

9. An oral formulation for repairing age-induced skin barrier damage according to claim 8, characterized in that, The oral preparation is a functional food.

10. An injectable formulation for repairing skin barrier damage induced by a high-fat, high-sugar diet, characterized in that, The injectable formulation comprises a solvent system and ganoderic acid A, wherein the mass percentage of ganoderic acid A is 0.4%-0.5% of the solvent system; the solvent system comprises DMSO, polyethylene glycol, and PBS.

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