Low-alcohol fruit wine with effects of maintaining beauty, keeping young and improving sleep and preparation method of low-alcohol fruit wine

Abstract

The invention belongs to the technical field of functional fruit wine, and discloses low-alcohol fruit wine with the effects of maintaining beauty and keeping young and improving sleep and a preparation method of the low-alcohol fruit wine. The fruit wine is prepared from the following components in parts by weight: 1-7 parts of collagen tripeptide, 0.1-1 part of walnut peptide, 1-3 parts of golden flower, 0.1-0.5 part of sodium hyaluronate and 500 parts of low-alcohol fruit wine of 10-15 degrees. The fruit wine prepared by the invention is reasonable in formula, and a plurality of experiments prove that the fruit wine can effectively improve sleep and delay senescence, has a relatively remarkable curative effect, is small in toxic and side effects, and is free of toxic and side effects. The health-care effect is good. The fruit wine prepared by the method is suitable for the technical fields of foods and medicines, and has a relatively great development prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of functional fruit wine, in particular to a low-alcohol fruit wine with the effects of beautifying the face and improving sleep, and a preparation method thereof. Background Art

[0002] With the rapid development of society, sleep disorders caused by the fast-paced life have developed into a global public problem. The insomnia population in my country is becoming younger and more professional. This chronic stress state continuously activates the hypothalamic-pituitary-adrenal axis, causing abnormal cortisol levels, and then forming a vicious cycle of "stress-insomnia-skin aging".

[0003] From the perspective of dermatology, dermal fibroblasts are the core effector cells for skin rejuvenation. The type I / III collagen and elastin network they synthesize constitute the biological basis for maintaining the mechanical properties of the skin. However, this delicate balance is easily disrupted by endogenous and exogenous aging factors: chronological aging causes a 50%-70% decrease in the proliferation capacity of fibroblasts, and ultraviolet-induced increased activity of matrix metalloproteinases, resulting in an annual loss rate of extracellular matrix of 1.5%-2.1%. Chronic sleep disorders further inhibit the collagen synthesis rate by 40% by regulating TGF-β / Smad signaling, accelerating the degradation of skin barrier function.

[0004] Faced with this multi-dimensional health challenge, existing solutions face significant technological gaps. While traditional sleep aids can improve sleep structure in the short term, long-term use can lead to downregulation of GABA receptors, triggering drug tolerance and withdrawal symptoms. While oral formulations such as collagen peptides are used in the beauty industry, their bioavailability is less than 15% and they lack the ability to target the dermal stem cell niche. These challenges highlight the urgent need for the health industry to develop a multi-dimensional "sleep-skin-metabolism" intervention system. By modulating circadian gene expression, optimizing extracellular matrix metabolomics networks, and innovating bioactive substance delivery systems, this approach can shift from symptom relief to root cause repair.

[0005] Currently, there are few functional foods on the market that can both improve sleep and beautify the skin. Some functional foods often have limited nutritional value and are often added with chemicals that may be harmful to the human body. Therefore, it is very necessary to develop a low-alcohol fruit wine that can improve beauty and sleep. Summary of the Invention

[0006] The present invention aims to provide a low-alcohol fruit wine with the effects of beautifying the face and improving sleep, and a preparation method thereof. The fruit wine has a reasonable formula and significant therapeutic effects.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The invention discloses a low-alcohol fruit wine with the effects of beautifying the face and improving sleep. The fruit wine is composed of the following components in parts by weight: 1-7 parts of collagen tripeptide, 0.1-1 part of walnut peptide, 1-3 parts of golden flower, 0.1-0.5 part of sodium hyaluronate, and 500 parts of low-alcohol fruit wine with a degree of 10-15.

[0009] Furthermore, the fruit wine is composed of the following components in parts by weight: 1-5 parts of collagen tripeptide, 0.5-1 parts of walnut peptide, 1-2 parts of golden flower, 0.1-0.4 parts of sodium hyaluronate, and 500 parts of 10-15 degree low-alcohol fruit wine.

[0010] Furthermore, the fruit wine is composed of the following components in parts by weight: 3 parts of collagen tripeptide, 0.7 parts of walnut peptide, 1.1 parts of golden flower, 0.2 parts of sodium hyaluronate, and 500 parts of 10-15 degree low-alcohol fruit wine.

[0011] A method for preparing low-alcohol fruit wine having the effects of beautifying the face and improving sleep comprises the following steps:

[0012] S1. Mix 150 parts of 40-45 vol% liquor, 320 parts of water, and 30 parts of concentrated fruit juice to prepare a low-alcohol fruit wine;

[0013] S2, collagen tripeptide is prepared by enzymatic hydrolysis, filtration, concentration and drying;

[0014] S3, walnut peptide is prepared by enzymatic hydrolysis, filtration, concentration, drying, ethanol polarization treatment, filtration, and drying;

[0015] S4, golden flower is prepared by crushing, enzymatic hydrolysis, ethanol polarization treatment, filtration, concentration and drying;

[0016] S5. Add sodium hyaluronate and the collagen tripeptide, walnut peptide and golden flower in steps S2-S4 into the low-alcohol fruit wine in proportion.

[0017] Furthermore, a method for preparing low-alcohol fruit wine with the effects of beautifying the skin and improving sleep is provided, wherein the alcohol content of the liquor in step S1 is 42 vol%.

[0018] The invention discloses a low-alcohol fruit wine having the effects of beautifying the face and improving sleep, and the fruit wine is used in the fields of food and medicine.

[0019] The principles and beneficial effects of this technical solution are as follows: 1. The peptide components in this invention are hydrolyzed with specific proteases and are highly active and stable. Golden flower extracts are enzymatically extracted, resulting in a high concentration of active ingredients. The resulting low-alcohol fruit wine has significant benefits for improving sleep and beautifying the complexion.

[0020] 2. The fruit wine ingredients of the present invention can consume DPPH free radicals, hydroxyl free radicals, and superoxide anion free radicals, thereby improving the body's antioxidant capacity.

[0021] 3. The fruit wine component of the present invention can inhibit the activity of elastase and play a role in delaying aging.

[0022] 4. The fruit wine ingredients in the present invention can inhibit and prevent UVB-induced oxidative stress response in HSF cells and improve the antioxidant capacity of cells.

[0023] 5. The fruit wine ingredients in the present invention can inhibit the increase in IL-6 and MMP-1 levels during UVB-induced cell senescence.

[0024] 6. The use of high concentrations of fruit wine ingredients can improve the UVB-induced decrease in COL-1 content and delay the aging process of cells.

[0025] 7. Using high doses of fruit wine ingredients can effectively inhibit tyrosinase activity and melanin synthesis in B16 cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the effect of fruit wine on the hypnotic effect of a suprathreshold dose of chloral hydrate in Example 1 of the present invention;

[0027] Figure 2 This is the effect of fruit wine on the hypnotic effect of a subthreshold dose of chloral hydrate in Example 1 of the present invention;

[0028] Figure 3 This is the effect of the fruit wine ingredients on free radicals in Example 2 of the present invention;

[0029] Figure 4 The inhibitory effect of the fruit wine component on elastase in Example 2 of the present invention;

[0030] Figure 5 This is the effect of the fruit wine ingredients on HSF cell viability in Example 2 of the present invention;

[0031] Figure 6 This is the effect of the fruit wine ingredients in Example 2 of the present invention on β-galactosidase staining of HSF cells after UVB induction;

[0032] Figure 7 This is the effect of the fruit wine ingredients in Example 2 of the present invention on the MDA activity in HSF cells;

[0033] Figure 8 This is the effect of the fruit wine ingredients in Example 2 of the present invention on the SASP content in UVB-induced HSF cells;

[0034] Figure 9 This is the effect of the fruit wine ingredients in Example 2 of the present invention on the content of type I collagen in UVB-induced HSF cells;

[0035] Figure 10This is the determination of the effect of fruit wine ingredients on B16 cell viability in Example 2 of the present invention;

[0036] Figure 11 This is the effect of the fruit wine ingredients in Example 2 of the present invention on melanin synthesis and tyrosinase activity in B16 cells. DETAILED DESCRIPTION

[0037] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0038] In the following examples, unless otherwise specified, the raw materials and equipment used can be purchased from the market or are commonly used in the art. The methods in the following examples, unless otherwise specified, are all conventional methods in the art.

[0039] A method for preparing low-alcohol fruit wine having the effects of beautifying the face and improving sleep comprises the following steps:

[0040] S1. Mix 150 parts of 42 vol% liquor, 320 parts of water, and 30 parts of concentrated fruit juice to prepare a low-alcohol fruit wine;

[0041] S2, collagen tripeptide is prepared by enzymatic hydrolysis, filtration, concentration and drying;

[0042] S3, walnut peptide is prepared by enzymatic hydrolysis, filtration, concentration, drying, ethanol polarization treatment, filtration, and drying;

[0043] S4, golden flower is prepared by crushing, enzymatic hydrolysis, ethanol polarization treatment, filtration, concentration and drying;

[0044] S5. Add sodium hyaluronate and the collagen tripeptide, walnut peptide and golden flower in steps S2-S4 to 500 parts of low-alcohol fruit wine in a ratio of: 3 parts of collagen tripeptide, 0.7 parts of walnut peptide, 1.1 parts of golden flower and 0.2 parts of sodium hyaluronate.

[0045] The fruit wine obtained by the preparation was subjected to experimental research. The following is the implementation process and the results obtained by the implementation:

[0046] Improved sleep research

[0047] All experimental data in this study are presented as mean ± standard deviation (mean ± SD). Data analysis was performed using SPSS 22.0 statistical software. The experimental procedures strictly adhered to the following statistical procedures: Before intergroup comparisons, data normality was verified using the Shapiro-Wilk test, and homogeneity of variance was assessed using the Levene test. For data that were normally distributed and had homogeneous variances, one-way analysis of variance (ANOVA) was used to compare differences between multiple groups. Statistical significance was determined as follows: *P < 0.05 indicated a statistically significant difference, and **P < 0.01 indicated a highly significant difference.

[0048] Example 1

[0049] 1.1 Materials

[0050] Analytical balance; electronic timer; chloral hydrate: provided by Wuhan Kerui Biotechnology Co., Ltd.

[0051] 1.2 Animals

[0052] Sixty specific pathogen-free (SPF) male Kunming mice, 4-5 weeks old and weighing (22 ± 2) g, were used. The animals were housed in an environment with a temperature of (23 ± 2)°C, a humidity of 55% ± 5%, and a 12-h light-dark cycle, with free access to food and water.

[0053] 1.3 Animal grouping and model establishment

[0054] Seventy-two male Kunming mice were randomly divided into three groups (n=24) after a 7-day acclimation period: a blank group, a control group, and a fruit wine group. Each group was gavaged with the corresponding test substance at 0.2 mL / 20 g body weight (determined according to the pharmacological dose conversion relationship) for 10 consecutive days.

[0055] 1.4 Judgment Indicators

[0056] Loss of righting reflex occurs when a mouse does not turn over within 1 minute while lying on its back with its limbs facing upward. The time from chloral hydrate injection to loss of righting reflex is the sleep onset latency, and the time from loss of righting reflex to recovery of righting reflex is the sleep duration.

[0057] 1.5 Direct sleep experiment

[0058] The three groups of mice were gavaged with the corresponding test substance at 0.2 mL / 20 g body weight, and the sleepiness of the mice in each group was observed within 30 minutes.

[0059] 1.6 Effects of fruit wine on sleep latency and sleep duration in mice exposed to suprathreshold doses of chloral hydrate

[0060] The suprathreshold sleep dose is the minimum dose that results in loss of righting reflex in 100% of mice. The suprathreshold dose of chloral hydrate is known to be 400 mg / kg. Thirty minutes after gavage with fruit wine, 12 mice randomly selected from each group were intraperitoneally injected with a subthreshold dose of chloral hydrate (0.2 mL / 20 g). The sleep latency and sleep duration of each group were observed and recorded.

[0061] 1.7 Effect of fruit wine on the sleep rate of mice after subthreshold dose of chloral hydrate

[0062] The subthreshold sleep dose is the maximum dose at which 80-90% of mice maintain their righting reflex. A subthreshold dose of chloral hydrate is 260 mg / kg. Thirty minutes after oral gavage with fruit wine, 12 randomly selected mice from each group were intraperitoneally injected with a suprathreshold dose at a volume of 0.2 mL / 20 g. The sleep onset of mice in each group within 30 minutes after administration was observed and recorded, along with the number of mice that fell asleep in each group. The sleep onset rate (%) for each group was calculated as follows: number of animals that fell asleep / total number of animals × 100%.

[0063] 2. Results

[0064] 2.1. Observation results of direct sleep effects on mice

[0065] The experimental results are shown in Table 1. After oral administration of the test substance, mice in the blank group showed no unusual behavior, while mice in the control and fruit wine groups showed decreased activity. When placed in a dorsal recumbent position, the mice were able to right themselves, exhibiting no loss of righting reflex. All mice were awake, with a zero sleep rate (Table 1), indicating that fruit wine has no direct hypnotic effect on mice.

[0066] Table 1 Observation results of direct sleep effects on mice (n=12, )

[0067]

[0068] 2.2 Effects of suprathreshold doses of chloral hydrate on the hypnotic effect

[0069] The experimental results are as follows Figure 1 As shown in Table 2, there was no significant difference in sleep latency between the control and blank groups (p>0.05), while the sleep latency in the fruit wine group was significantly reduced compared to the blank group (p<0.05). The sleep time of mice in the control and fruit wine groups was significantly prolonged compared to the blank control group (p<0.001). These results indicate that fruit wine can effectively prolong the sleep time of mice injected with chloral hydrate and significantly shorten their sleep latency.

[0070] Table 2 Sleep latency and sleep time of mice in each group at suprathreshold dose of chloral hydrate (n=12, x±s)

[0071]

[0072] 2.3 Effects of subthreshold doses of chloral hydrate on the hypnotic effect

[0073] The statistical experimental results are as follows Figure 2 As shown in Table 3, a subthreshold sleep agent is defined as the maximum dose at which 80-90% of mice maintain their righting reflex. None of the mice in the blank group fell asleep after receiving chloral hydrate. The number of mice in the experimental group that fell asleep was significantly higher than that in the blank group, with the fruit wine group showing the highest number of sleep episodes. This suggests that fruit wine can effectively increase sleep rates and aid in falling asleep.

[0074] Table 3 Number of mice falling asleep and sleeping rate in each group of mice with subthreshold dose of chloral hydrate

[0075]

[0076] Example 2

[0077] 1.1 Materials

[0078] HSF cells were purchased from Wuhan Yuansheng Primary Biopharmaceutical Technology Co., Ltd. (Batch No. HUM-CELL-0132); B16 cells were purchased from Wuhan Punosai Life Science Technology Co., Ltd.; 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH·), sivelestat sodium, and N-succinyl-L-alanyl-L-alanyl-L-alanine (NAAAPN) were all purchased from Shanghai MacLean Company; ferrous sulfate (FeSO4) and sivelestat sodium were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; ascorbic acid (VC), salicylic acid, hydrogen peroxide, and pyrogallol were all purchased from Sinopharm Chemical Reagent Co., Ltd.; cell proliferation and activity detection-8 (CCK-8) kit was purchased from Dojindo Chemical Research Institute, Japan; β-galactosidase staining kit, reactive oxygen species detection kit (ROS), and malondialdehyde (MDA) content detection kit were all purchased from Beijing Solaibao Technology Co., Ltd.; human type I collagen (Col I) Enzyme-linked immunosorbent assay (ELISA) kits were purchased from Shanghai ELISA Biotechnology Co., Ltd.; human matrix metalloproteinase 1 (MMP-1) ELISA kit was purchased from Wuhan Huamei Bioengineering Co., Ltd.; and human interleukin (IL-6) quantitative detection kit was purchased from Ruixin Biotechnology Co., Ltd.

[0079] 1.2. DPPH free radical scavenging experiment

[0080] 100 μL of sample solutions of varying concentrations were mixed with a 0.2 mmol / L DPPH ethanol solution in a 96-well plate, designated A1. 100 μL of deionized water was added to replace the wine component in A1 and mixed thoroughly, designated A2. An equal amount of ethanol solution was added to replace the DPPH ethanol solution in A1 and mixed thoroughly, designated A0. The plates were then incubated at 25°C in the dark for 30 minutes. The absorbance was measured at 517 nm. VC was used as a positive control at the same concentration as the wine component. The DPPH scavenging rate was calculated for three samples per group.

[0081]

[0082] Where: A1 is the absorbance of the test group; A2 is the absorbance of the blank control group; A0 is the absorbance of the sample control group.

[0083] 1.3 Hydroxyl radical scavenging experiment

[0084] 50 μL of fruit wine components of varying concentrations were placed in a 96-well plate. 9 mmol / L FeSO₄, salicylic acid ethanol solution, 4.4 mmol / L H₂O₂ solution, and 100 μL distilled water were added, followed by mixing. This was designated A1. 50 μL of deionized water was added to replace the healthy wine components in A1, mixed thoroughly, and designated A2. An equal amount of deionized water was added to replace the H₂O₂ solution in A1, mixed thoroughly, and designated A0. The plates were then incubated at 37°C for 60 minutes. The absorbance was measured at 510 nm. Vc was used as a positive control at the same concentration as the fruit wine components. Three samples were used in each group, and the OH scavenging rate was calculated.

[0085]

[0086] Where: A1 is the absorbance of the test group; A2 is the absorbance of the blank control group; A0 is the absorbance of the sample control group.

[0087] 1.4. Superoxide anion free radical scavenging experiment

[0088] Take 0.05 mol / L tris-hydroxymethylaminomethane hydrochloride (Tris-HCl, pH 8.2) buffer solution preheated in a 25℃ water bath for 20 minutes and place it in a 96-well plate. Add 25 μL of different concentrations of fruit wine components and 25 mmol / L pyrogallol solution in sequence, mix well, react in a 25℃ water bath for 3 minutes, add 25 μL of 10 mol / L HCL to terminate the reaction, and detect the absorbance at 325 nm. For the blank group, use the same volume of blank solvent instead of the sample solution and repeat the above steps. Use Vc as a positive control with the same concentration as the fruit wine component. Calculate O for each group of 3 samples. 2- Clearance rate.

[0089]

[0090] Where: A1 is the absorbance of the test group; A2 is the absorbance of the blank control group; A0 is the absorbance of the sample control group.

[0091] 1.5. Elastase activity inhibition experiment

[0092] In a 96-well plate, 25 μL of elastase solution was added to 50 μL of fruit wine components at varying concentrations. Then, 100 μL of Tris-HCl buffer (pH 8.2) was added and incubated at 25°C for 20 minutes. Afterward, 25 μL of substrate (NAAAPN) was added, the mixture was immediately shaken and mixed. The absorbance (A) of each group was measured at 405 nm. The absorbance (A') was then measured again at 25°C for 10 minutes. ΔA (A'-A) was calculated for each group, and the enzyme inhibition rate was calculated. Sivelestat sodium (15 μg / mL) was used as a positive control. Three samples were included in each group, and the elastase inhibition rate was calculated.

[0093]

[0094] Where: ΔA1 is the absorbance of the test group; ΔA2 is the absorbance of the blank control group; ΔA0 is the absorbance of the sample control group.

[0095] 1.6 Cell Culture

[0096] Select 6-10 generations of vigorously growing HSF cells and culture and subculture them in a medium containing 10% FBS, 1% double antibody, and 1% additives, and place them in an incubator at 37°C with 5% CO2. Change the culture medium regularly, and when the cell confluence reaches more than 80%, add 1×10 4 The density of cells / well was inoculated in 96-well plates and 1×10 cells / well in 6-well plates. 5 The cells were seeded at a density of 100 cells / mL.

[0097] 1.7. Determination of Antioxidant Enzyme Activity and Peroxide Content in Cells

[0098] The cells were divided into 1×10 5 Cells were seeded at a density of 100 cells / well in a 6-well plate, with triplicate wells per group. Twenty-four hours after illumination, cells in each group in the 6-well plate were trypsinized and transferred to a 1.5 mL centrifuge tube. The cells were centrifuged at 9000 rpm for 5 minutes at room temperature to collect the cell pellet. The corresponding reagents were added using the MDA and SOD kits, and the absorbance was measured to calculate the enzyme activity and superoxide content.

[0099] 1.8. Determination of Senescence-Associated Secretory Phenotype (SASP) and Type I Collagen Content

[0100] The cells were divided into 1×10 4The cells were seeded at a density of 1000 / well in a 96-well plate, with 3 replicates per group for treatment. 24 h after illumination, the contents of IL-6, MMP-1, and COL-1 in the supernatant of HSF cells in the blank group, model group, and experimental group were determined according to the instructions of the enzyme-linked immunosorbent assay (ELISA) kit, and the absorbance was measured at 450 nm.

[0101] 1.9 Effects on melanin synthesis in B16 cells

[0102] B16 cells in good logarithmic growth phase were counted and inoculated into T25 culture flasks, and cultured overnight in an incubator at 37°C and 5% CO2. The culture medium was replaced and samples of different concentrations were added. The negative control group was not treated. After 48 h of culture, the culture medium was discarded, the cells were washed with PBS, and 250 μL of trypsin was added for 4 min. The digestion was terminated by adding 1 mL of culture medium and the cells were rotated at 1500 r·min. -1 Centrifuge for 3 minutes, discard the supernatant, add 1 mL of 10% (volume fraction) DMSO 1 mol·L -1 Sodium hydroxide solution, ultrasonically disperse for 3 minutes, take 20 μL of the above solution to determine the BCA kit, and the remaining solution is water bathed at 80℃ for 30 minutes; shake and mix, 10000r·min -1 Centrifuge for 2 minutes, pipette 200 μL of the supernatant into a 96-well plate, and read the absorbance at 405 nm. Repeat five times for each component. The change in melanin concentration requires correction by BCA concentration.

[0103] Melanin mass fraction change (%) = (OD value of the assay well - OD value of the blank control) / (OD value of the cell control group - OD value of the blank control) × 100%.

[0104] 1.10 Effects on tyrosinase activity in B16 cells

[0105] B16 cells in good logarithmic growth phase were counted and inoculated into T25 culture flasks, and cultured overnight in an incubator at 37°C and 5% CO2. The culture medium was replaced and samples of different concentrations were added. The negative control group was not treated. After 48 h of culture, the culture medium was discarded, the cells were washed with PBS, and 250 μL of trypsin was added for 4 min. The digestion was terminated by adding 1 mL of culture medium and the cells were rotated at 1500 r·min. -1Centrifuge for 3 minutes, discard the supernatant, add 300 μL of 1% Triton X-100, and sonicate for 5 minutes. Transfer 20 μL of this solution to the BCA assay kit. Transfer 100 μL of the remaining solution to a 96-well plate and prewarm at 37°C for 5 minutes. Add 100 μL of 0.25% L-DOPA solution, incubate at 37°C for 1 hour, and read the absorbance at 490 nm. Perform five replicates for each component. Variations in tyrosinase activity require correction with BCA mass fraction.

[0106] Change in tyrosinase activity (%) = (OD value of the assay well - OD value of the blank control) / (OD value of the cell control group - OD value of the blank control) × 100%.

[0107] 2. Results Analysis

[0108] 2.1 Effects of Fruit Wine Components on Free Radicals

[0109] Free radicals are considered a key factor in cellular and biological aging. As we age, the balance between free radical production and clearance in the body is disrupted. Excessive free radicals can attack biomacromolecules within cells, leading to cell dysfunction and aging.

[0110] The experimental results are as follows Figure 3 As shown in the figure, with the increase of the concentration of wine components, the effects of DPPH·, O2 - The clearance rate of DPPH and OH· also increased, and was positively correlated with the dosage concentration. When the dosage concentration was greater than 4 mg / ml, the scavenging ability of DPPH· was slightly stronger than that of Vc, with a clearance rate of up to 89%. As the concentration continued to increase, it could reach a maximum of 93%. - It has a certain effect on the scavenging of DPPH free radicals, hydroxyl free radicals, and superoxide anion free radicals, and its ability increases with the increase of drug concentration, and is slightly weaker than that of Vc. Free radical scavenging experiments show that fruit wine components can consume DPPH free radicals, hydroxyl free radicals, and superoxide anion free radicals, playing a certain protective role on the body.

[0111] 2.2 Effects of fruit wine components on elastase activity

[0112] Elastase primarily targets elastin, a component of the extracellular matrix. During cellular aging, the extracellular matrix undergoes significant changes. Altered elastase activity may affect the structure and function of the extracellular matrix. Normally, the extracellular matrix provides physical support for cells and regulates intercellular signaling, among other functions. However, with cellular aging, components of the extracellular matrix, such as elastin, may be abnormally degraded or remodeled.

[0113] The experimental results are as follows Figure 4As shown in the results, the inhibition rate of elastase gradually and significantly increased with the increase in the concentration of the fruit wine component at concentrations between 0.25 and 32 mg / mL, reaching 45.2% at 32 mg / mL. The inhibition rate of the positive drug Vc was 61.0%. Therefore, the fruit wine component has the ability to inhibit elastase activity, playing a role in delaying aging.

[0114] 2.3 Determination of the effects of fruit wine components on HSF cell viability

[0115] The results are as follows Figure 5 As shown in the figure, after treatment with different concentrations of fruit wine ingredients, the cell viability of 20, 40, and 80 mg / mL fruit wine ingredients decreased to less than 80% compared with the blank group. Therefore, the fruit wine ingredient at a concentration of 10 mg / mL was selected for subsequent experiments.

[0116] 2.4 Effects of fruit wine components on UVB-induced HSF cell senescence

[0117] β-galactosidase is a commonly used marker of aging. As cells age, β-galactosidase activity increases significantly, particularly in the aging-associated β-galactosidase (SA-β-Gal) assay. This enzyme is stable at pH 6.0, whereas β-galactosidase in young cells is typically active at neutral pH. By measuring SA-β-Gal activity in cells, it is possible to visually determine whether a cell is in a senescent state.

[0118] The treatment group received the drug after irradiation, while the prevention group received the drug before irradiation. Figure 6 As shown in the results, compared with the control group, the number of blue-stained cells in the model group was significantly increased under the microscope. Compared with the model group, the number of senescent cells in the treatment and prevention groups was significantly reduced, with the effect being most pronounced in the high-dose group. Therefore, fruit wine ingredients can reduce UVB-induced HSF cell senescence.

[0119] 2.5 Effects of blended alcohol ingredients on oxidase activity in HSF cells

[0120] MDA interacts with various intracellular molecules and signaling pathways, and changes in its content can reflect some key mechanisms in the cellular aging process. For example, MDA can activate intracellular inflammatory signaling pathways, such as the NF-κB pathway, triggering an inflammatory response, which is a key part of "inflammatory aging" during cellular aging. By measuring MDA content and the expression of related inflammatory factors, we can further study the inflammatory mechanisms in the cellular aging process. At the same time, MDA is also closely related to the intracellular antioxidant enzyme system (such as superoxide dismutase and glutathione peroxidase). Studying changes in MDA can help reveal the dynamic changes in the oxidation-antioxidation balance during cellular aging.

[0121] The experimental results are as follows Figure 7 As shown, compared with the blank group, the MDA content in the model group increased significantly (P < 0.01). Compared with the model control group, the wine components in the treatment-L, treatment-H, prevention-L, and prevention-H groups were able to significantly inhibit and prevent the increase in UVB-induced MDA (P < 0.05 or P < 0.01). Therefore, fruit wine components can inhibit and prevent UVB-induced oxidative stress in HSF cells and enhance cellular antioxidant capacity.

[0122] 2.6 Effects of fruit wine components on SASP content in UVB-induced HSF cells

[0123] The senescence-associated secretory phenotype (SASP) refers to a phenotype composed of a series of biologically active proteins, cytokines, chemokines, growth factors and other components secreted by senescent cells. When cells are damaged or stressed, such as oxidative stress or DNA damage, they release IL-6. IL-6 can activate a series of downstream signaling pathways. For example, by binding to the IL-6 receptor on the cell membrane, it activates the JAK-STAT3 signaling pathway, leading to cell cycle arrest and prompting cells to enter a senescent state. MMP-1 is a member of the matrix metalloproteinase family (MMPs), and its main function is to degrade collagen in the extracellular matrix (ECM). During the process of cellular aging, the activity and expression of MMP-1 usually increase, breaking down collagen and causing structural changes in the extracellular matrix.

[0124] The experimental results are as follows Figure 8 As shown in the results, compared with the blank group, the IL-6 and MMP-1 levels in the model group were significantly increased (P < 0.05). The wine ingredients in the treatment-L, treatment-H, and prevention-H groups were able to improve the UVB-induced increase in IL-6 and MMP-1 levels (P < 0.05, or P < 0.01). The prevention-L group had no significant effect on IL-6 and MMP-1. In summary, it shows that fruit wine ingredients can inhibit the increase in IL-6 and MMP-1 levels during UVB-induced cell senescence.

[0125] 2.7 Effects of fruit wine components on the content of type I collagen in UVB-induced HSF cells

[0126] COL-1 is the most abundant type of collagen in the extracellular matrix. It possesses high tensile strength and provides structural support to tissues. It is widely present in various tissues, including skin, bone, tendon, and blood vessel walls. During cellular aging, the content and structure of COL-1 change. As previously mentioned, matrix metalloproteinases such as MMP-1 degrade COL-1, leading to a decrease in its content.

[0127] The experimental results are as follows Figure 9 As shown, compared with the blank group, the COL-1 content in the model group was significantly decreased (P < 0.05); compared with the model control group, the COL-1 content in the treatment-H group and the prevention-H group was significantly increased (P < 0.05), while there was no significant difference in the treatment-L group and the prevention-L group. This experiment shows that high concentrations of fruit wine ingredients can improve UVB-induced decreases in COL-1 content and delay the aging process of cells.

[0128] 2.8. Determination of the Effect of Fruit Wine Components on B16 Cell Viability

[0129] The experimental results are as follows Figure 10 As shown in the figure, after treatment with different concentrations of fruit wine ingredients, the cell viability of the 0.64-10.96 mg / mL fruit wine ingredients was significantly changed compared with the blank group. Therefore, the fruit wine ingredient at a concentration of 0.32 mg / mL was selected for subsequent experiments.

[0130] 2.9 Effects of fruit wine on melanin synthesis and tyrosinase activity in B16 cells

[0131] B16 cells, a mouse melanoma cell line, are highly capable of melanin synthesis. These cells hold significant application value in studying the mechanisms of melanogenesis and screening for skin-whitening agents. Tyrosinase (TYR) is a key enzyme in melanin synthesis. It catalyzes the conversion of tyrosine to dopa (DOPA), which is then further oxidized to dopaquinone. Dopaquinone undergoes a series of non-enzymatic reactions to ultimately form melanin. In B16 cells, tyrosinase activity and expression levels directly influence melanin synthesis.

[0132] The experimental results are as follows Figure 11 As shown in the results, compared with the blank group, the melanin and tyrosinase activities in the model group were significantly increased (P < 0.05); compared with the model control group, the melanin and tyrosinase activities in the low-dose group were not significantly different, but significantly decreased in the high-dose group. The results show that high-dose fruit wine ingredients can effectively inhibit tyrosinase activity and melanin synthesis in B16 cells.

[0133] The above is only an embodiment of the present invention, and common knowledge such as the specific technical solutions or characteristics in the solution is not described in detail here. For those skilled in the art, without departing from the technical solution of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the description can be used to interpret the content of the claims.

Claims

1. A low-alcohol fruit wine with the effects of beautifying the face and improving sleep, characterized in that: The fruit wine is composed of the following components in parts by weight: 1-7 parts of collagen tripeptide, 0.1-1 parts of walnut peptide, 1-3 parts of golden flower, 0.1-0.5 parts of sodium hyaluronate, and 500 parts of low-alcohol fruit wine with a content of 10-15 degrees.

2. The low-alcohol fruit wine with the effects of beautifying the face and improving sleep according to claim 1, characterized in that: The fruit wine is composed of the following components in parts by weight: 1-5 parts of collagen tripeptide, 0.5-1 part of walnut peptide, 1-2 parts of golden flower, 0.1-0.4 part of sodium hyaluronate, and 500 parts of low-alcohol fruit wine with a content of 10-15 degrees.

3. The low-alcohol fruit wine with the effects of beautifying the face and improving sleep according to claim 1, characterized in that: The fruit wine is composed of the following components in parts by weight: 3 parts of collagen tripeptide, 0.7 parts of walnut peptide, 1.1 parts of golden flower, 0.2 parts of sodium hyaluronate, and 500 parts of low-alcohol fruit wine with a content of 10-15 degrees.

4. The method for preparing a low-alcohol fruit wine having the effects of beautifying the face and improving sleep according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1. Mix 150 parts of 40-45 vol% liquor, 320 parts of water, and 30 parts of concentrated fruit juice to prepare a low-alcohol fruit wine; S2, collagen tripeptide is prepared by enzymatic hydrolysis, filtration, concentration and drying; S3, walnut peptide is prepared by enzymatic hydrolysis, filtration, concentration, drying, ethanol polarization treatment, filtration, and drying; S4, golden flower is prepared by crushing, enzymatic hydrolysis, ethanol polarization treatment, filtration, concentration and drying; S5. Add sodium hyaluronate and the collagen tripeptide, walnut peptide and golden flower in steps S2-S4 into the low-alcohol fruit wine in proportion.

5. The method for preparing a low-alcohol fruit wine having the effects of beautifying the face and improving sleep according to claim 3, characterized in that: The alcohol content of the liquor in step S1 is 42 vol%.

6. The low-alcohol fruit wine having the effects of beautifying the face and improving sleep according to any one of claims 1 to 3, characterized in that: The fruit wine is used in the fields of food and medicine.

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