Cell energy activator and application thereof

Abstract

The present application relates to the cosmetic technical field, specifically, a kind of preparation method of cell energy activator containing nicotinamide, VC ethyl ether and bacillus fermentation product and its application in skin antioxidant.The present application is prepared by preparing the first component by inoculating activated bacillus into the fermentation medium containing camellia nitidissima powder and glucose powder;The second component is prepared by inoculating activated bacillus into the culture medium;And the first component and the second component are mixed, then nicotinamide and VC ethyl ether are added, to prepare the cell energy activator.The cell energy activator of the present application can play a synergistic effect, effectively improve skin antioxidant activity, and improve skin barrier.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a method for preparing a cell energy activator containing microbial metabolites and its application in skin anti-oxidation and enhancing the skin barrier function. Background Technology

[0002] Free radicals are a natural byproduct of human life activities. All human activities, such as breathing, exercise, and eating, generate free radicals (for example, the human body continuously releases energy through oxidation during respiration, and mitochondria convert some oxygen into reactive oxygen species). In addition, external environmental influences also generate free radicals, such as smoking, alcohol consumption, ultraviolet radiation, electromagnetic waves, environmental pollutants, medications, and emotional stress. Cosmetic antioxidants primarily reduce oxidative stress-induced damage to cell membranes, DNA, lipids, or proteins by scavenging free radicals, lowering ROS levels, increasing antioxidant enzyme activity, and reducing lipid metabolites, thereby mitigating the damage caused by oxidation to human skin. Most antioxidant testing methods also use free radical scavenging, ROS level reduction, and antioxidant enzyme activity as evaluation criteria. With the continuous expansion of the skincare market, finding more effective and natural anti-aging ingredients in bio-based skincare formulations is a crucial direction for current research and development.

[0003] In the field of novel cosmetic development, postbiotics, as non-living microbial preparations, are often used to regulate the skin's microecology, improve skin condition, and thus combat aging. Postbiotics refer to soluble substances secreted by probiotics or released after the decomposition of bacterial cells, mainly consisting of bacterial components and metabolites. Postbiotics can be prepared using carbon-source culture media and fermentation with specific bacterial strains. They are composed of different metabolites and have important biological significance, such as antioxidant and anti-inflammatory effects.

[0004] The main objective of this invention is to develop an antioxidant bio-formulation containing post-biotics and to use it in cosmetic development. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing a skin antioxidant cell energy activator and its application.

[0006] Nicotinamide, also known as nicotinamide, is an amide compound of nicotinic acid (vitamin B3). It can be converted into NAD+ or NADP+ in the body, playing a role in transferring hydrogen ions and electrons in many redox reactions. It is mainly used to prevent and treat pellagra, stomatitis, glossitis, sick sinus syndrome, and atrioventricular block. Nicotinamide is a water-soluble vitamin and a member of the B vitamin group. It is an antioxidant that interacts with the skin, stimulating necessary coenzymes to better protect the epidermis from external damage. It enhances the healing ability of skin cells, improves the structure of different skin layers, ensures optimal skin hydration, and regulates sebum secretion.

[0007] VC ethyl ether, also known as 3-o-ethyl ascorbic acid (CAS number 86404-04-8), is increasingly favored by the domestic and international cosmetics industry due to its significant skincare effects and good compatibility. Reports indicate that VC ethyl ether derivatives are the best vitamin C derivatives to date, exhibiting not only excellent chemical stability but also easy biodegradation by enzymes after penetrating the skin to exert their vitamin C effects. Chemically, VC ethyl ether is an amphoteric vitamin C derivative with both lipophilic and hydrophilic properties. This makes it extremely convenient to use in formulations and allows it to easily penetrate the stratum corneum and reach the dermis. Once inside the skin, it is easily degraded by enzymes to exert its vitamin C effects, enhancing its bioavailability. It also possesses strong antioxidant effects, effectively removing free radicals.

[0008] Golden camellia (scientific name: *Camellia petelotii*) is an evergreen shrub belonging to the genus *Camellia* in the family Theaceae. Studies have shown that golden camellia is rich in various bioactive substances, including tea polysaccharides, tea polyphenols, saponins, and flavonoids. Among these, golden camellia polysaccharides have attracted considerable attention due to their unique bioactivity. However, the extraction of golden camellia polysaccharides from raw materials such as tea leaves, flowers, and seeds is easily interfered with by small molecule compounds such as polyphenols, pigments, proteins, and inorganic salts. These impurities not only affect the bioactivity of golden camellia polysaccharides but also interfere with their quantitative structural analysis and identification.

[0009] Prior art CN118512341A discloses a cell energy activating composition with anti-aging effects and its application in men's skincare. However, with further research, utilizing the antioxidant properties of microorganisms to protect the skin is also a popular research direction. Prior art CN119950377A discloses an emotional beauty composition that enhances cell vitality and reduces wrinkles, along with its preparation method and application, including Andrographis paniculata extract, Bacillus fermentation products, 4-tert-butylcyclohexanol, and Codonopsis pilosula extract. Prior art CN119632893A discloses a composition containing Camellia chrysantha extract for anti-pollution repair and acne-prone skin, along with its application. However, it does not disclose any content involving the use of post-biotics to enhance the skin's anti-aging effects. Microbial fermentation derivatives in skincare products are fermented under different external nutrient conditions to obtain skincare-value components. By breaking down cell walls and filtering the cell fluid, the effective components are extracted. Based on this, the inventors, building on previous research, further developed a combination based on Bacillus fermentation products and natural anti-aging substances, thus obtaining this invention. The antioxidant and anti-aging effects of this invention are demonstrated through the measurement of relevant indicators. SOD, CAT, and GSH are antioxidant enzyme groups that make up the cellular defense system. SOD is an important antioxidant enzyme in the body, which can scavenge superoxide anion free radicals produced by the body and can indirectly reflect the body's antioxidant capacity. CAT is an antioxidant enzyme present in all living tissues, which can degrade or reduce hydrogen peroxide into water and molecular oxygen. GSH is a peroxidase that maintains the normal function of the body's immune system, which can remove lipid peroxides and H2O2, protecting tissues and cells from free radical attack and damage. Total antioxidant capacity (T-AOC) refers to the total antioxidant level composed of various antioxidant enzyme systems and non-enzyme systems, such as antioxidant enzymes like SOD and CAT, and vitamins C and E, and is used to reflect the total antioxidant capacity of bioactive substances in the body.

[0010] One aspect of the present invention provides a method for preparing a cell energy activator, comprising the following steps:

[0011] 1) Dry the golden camellia and pulverize it, then pass it through a 100-150 mesh sieve to obtain golden camellia powder; add the golden camellia powder to 60℃ distilled water, mix and stir for 20 minutes, add glucose powder as a carbon source, stir until homogeneous, and then place it in a water bath and sterilize at 90-100℃ for 30 minutes to obtain fermentation medium; each liter of fermentation medium contains 5-15 parts of golden camellia powder and 2.5-5 parts of glucose powder, each part being 10g;

[0012] After activation, Bacillus was inoculated into the fermentation medium at an inoculation density of 5.0-6.0 × 10⁻⁶. 6CFU / mL, fermented at 28-33℃ for 18-24 hours, centrifuged at 6000rpm for 10min, the supernatant was collected, filtered through a 0.22μm filter membrane, the filtrate was collected, lyophilized and concentrated to prepare the first component;

[0013] 2) The activated Bacillus was inoculated into the culture medium for fermentation. After fermentation, the cells were homogenized and ultrasonically broken up. The ultrasonic power was 1000w for 20min, with 3s working and 3s intermittent. The supernatant was collected by centrifugation, freeze-dried and concentrated to prepare the second component.

[0014] 3) Preparation of cell energy activator: Mix the first component and the second component, add 3-5 parts of nicotinamide and 0.5-1.5 parts of VC ethyl ether to prepare the cell energy activator.

[0015] Furthermore, the content of golden camellia powder is 10 parts, and the content of glucose powder is 2.5 parts.

[0016] Furthermore, the inoculation density of Bacillus was 5.5 × 10⁻⁶. 6 cfu / mL.

[0017] Further, in step 3), add 3-5 parts of nicotinamide and 0.5-1.5 parts of VC ethyl ether.

[0018] One aspect of the present invention provides a cell energy activator prepared by the aforementioned method.

[0019] One aspect of the present invention provides a skin care product, characterized in that it includes the aforementioned cell energy activator.

[0020] Furthermore, it also includes at least one of the following: moisturizer, antioxidant, anti-inflammatory agent, and chelating agent.

[0021] One aspect of the present invention provides the use of the aforementioned cell energy activator in the preparation of skin antioxidant products.

[0022] Furthermore, it is administered to men.

[0023] One aspect of the present invention provides the use of the aforementioned cell energy activator in the preparation of products that enhance catalase activity.

[0024] The culture medium used to inoculate the activated Bacillus into the culture medium for fermentation includes components of conventional beef extract peptone medium. This is common knowledge in the field.

[0025] The beneficial effects of this invention are as follows:

[0026] (1) This invention uses a fermentation medium containing Camellia chrysantha powder and glucose powder to prepare a first component by activating Bacillus and inoculating it into the fermentation medium; the activated Bacillus is then inoculated into the medium to prepare a second component; the first and second components are mixed, and nicotinamide and VC ethyl ether are added to prepare a cell energy activator. The cell energy activator of this invention can effectively enhance the antioxidant activity of the skin.

[0027] (2) The Bacillus fermentation method of the present invention can obtain high-density fermentation products, and its antioxidant effect is obvious.

[0028] (3) The cell energy activator of the present invention can effectively increase the content of CAT, SOD and GSH in the skin after being prepared into skin care products. Detailed Implementation

[0029] This invention provides a method for preparing a cell energy activator containing microbial metabolites and its application. To better illustrate the purpose, technical solution, and advantages of this invention, specific embodiments will be used for further explanation below. The purpose is to provide a detailed understanding of the invention, not to limit it. All other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention. Unless otherwise specified, the experimental reagents and instruments designed in the embodiments and comparative examples of this invention are commonly used and commercially available. Unless otherwise specified, the experimental methods used in the embodiments and comparative examples are conventional methods; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch.

[0030] Example 1

[0031] A method for preparing a cell energy activator includes the following steps:

[0032] 1) Dry and pulverize the golden camellia, then pass it through a 140-mesh sieve to obtain golden camellia powder; add the golden camellia powder to 60℃ distilled water, mix and stir for 20 minutes, add glucose powder as a carbon source, stir and homogenize, then place in a water bath and sterilize at 90℃ for 30 minutes to obtain fermentation medium; each liter of fermentation medium contains 10 parts of golden camellia powder and 2.5 parts of glucose powder, each part being 10g;

[0033] After activation, Bacillus was inoculated into the fermentation medium at an inoculation density of 5.5 × 10⁻⁶. 6 CFU / mL, fermented at 30℃ for 20 hours, centrifuged at 6000 rpm for 10 min, the supernatant was collected, filtered through a 0.22 μm filter membrane, the filtrate was collected, lyophilized and concentrated to prepare the first component;

[0034] 2) The activated Bacillus was inoculated into the culture medium for fermentation at an inoculation density of 5.6 × 10⁶. 6 The concentration of cfu / mL was used to homogenize the cells after fermentation, followed by ultrasonic disruption at a power of 1000W for 20 minutes with 3 seconds of operation followed by 3 seconds of rest. The supernatant was then collected by centrifugation, freeze-dried, and concentrated to prepare the second component.

[0035] 3) Preparation of cell energy activator: Mix the first component and the second component, add 3-5 parts of nicotinamide and 0.5-1.5 parts of VC ethyl ether to prepare the cell energy activator.

[0036] Comparative Example 1-1

[0037] Based on Example 1, in step 1), 5 parts of Camellia chrysantha powder and 2.5 parts of glucose powder were used to obtain the fermentation culture medium, with the rest remaining unchanged.

[0038] Comparative Examples 1-2

[0039] Based on Example 1, in step 1), 15 parts of Camellia chrysantha powder and 2.5 parts of glucose powder were used to obtain the fermentation culture medium, with the rest remaining unchanged.

[0040] Comparative Examples 1-3

[0041] Based on Example 1, in step 1), 10 parts of Camellia chrysantha powder and 5 parts of glucose powder were used to obtain a fermentation culture medium, with the rest remaining unchanged.

[0042] Comparative Examples 1-4

[0043] Based on Example 1, in step 1), 15 parts of Camellia chrysantha powder and 5 parts of glucose powder were used to obtain a fermentation culture medium, with the rest remaining unchanged.

[0044] The antioxidant activity of Example 1 and Comparative Examples 1-4 was determined according to the instructions of the T-AOC assay kit (Nanjing Jiancheng Biotechnology Co., Ltd.). At 37°C, one unit of total antioxidant capacity (U) was defined as the increase in absorbance (OD) of the reaction system by 0.01 per minute per milliliter of serum (plasma). The results are shown in Table 1.

[0045] Table 1 Results of antioxidant activity assay

[0046]

[0047] As shown in Table 1, the total antioxidant activity of the fermentation medium obtained in Example 1 was better than that of Comparative Examples 1-1 to 1-4. This may be because, under a single carbon source, glucose serves as the carbon source for microbial fermentation, but excessive glucose can lead to substrate repression. Therefore, the preparation method of Example 1 was used to screen subsequent strain inoculation experiments.

[0048] Comparative Examples 1-5

[0049] Based on Example 1, in step 1), Bacillus thuringiensis is activated and inoculated into the fermentation medium at an inoculation density of 5.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0050] Comparative Examples 1-6

[0051] Based on Example 1, in step 1), Bacillus thuringiensis is activated and inoculated into the fermentation medium at an inoculation density of 6.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0052] Comparative Examples 1-7

[0053] Based on Example 1, in step 1), Bacillus was activated and inoculated into the fermentation medium at an inoculation density of 6.5 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0054] Comparative Examples 1-8

[0055] Based on Example 1, in step 1), Bacillus thuringiensis is activated and inoculated into the fermentation medium at an inoculation density of 7.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0056] The total antioxidant activity of the fermented components was further determined using the aforementioned method, and the results are shown in Table 2.

[0057] Table 2 Total antioxidant activity

[0058]

[0059] As shown in Table 2, the total antioxidant activity of the fermented components obtained from the fermentation medium prepared in Example 1 was superior to that of Comparative Examples 1-5 to 1-8. Therefore, the preparation method of Example 1 was screened for subsequent single-factor screening experiments on fermentation temperature and fermentation time, and the inoculation density of 5.5 × 10⁻⁶ was finally selected. 6 The optimal conditions are cfu / mL and fermentation at 30°C for 20 hours.

[0060] Example 2

[0061] A method for preparing a cell energy activator includes the following steps:

[0062] 1) Dry and pulverize the golden camellia, then pass it through a 140-mesh sieve to obtain golden camellia powder; add the golden camellia powder to 60℃ distilled water, mix and stir for 20 minutes, add glucose powder as a carbon source, stir and homogenize, then place in a water bath and sterilize at 90℃ for 30 minutes to obtain fermentation medium; each liter of fermentation medium contains 10 parts of golden camellia powder and 2.5 parts of glucose powder, each part being 10g;

[0063] After activation, Bacillus was inoculated into the fermentation medium at an inoculation density of 5.6 × 10⁶. 6 CFU / mL, fermented at 30℃ for 20 hours, centrifuged at 6000 rpm for 10 min, the supernatant was collected, filtered through a 0.22 μm filter membrane, the filtrate was collected, lyophilized and concentrated to prepare the first component;

[0064] 2) The activated Bacillus was inoculated into YPD medium for fermentation at an inoculation density of 5.6 × 10⁶. 6 The concentration of cfu / mL was used to homogenize the cells after fermentation, followed by ultrasonic disruption at a power of 1000W for 20 minutes with 3 seconds of operation followed by 3 seconds of rest. The supernatant was then collected by centrifugation, freeze-dried, and concentrated to prepare the second component.

[0065] 3) Preparation of cell energy activator: Mix the first component and the second component, add 3-5 parts of nicotinamide and 0.5-1.5 parts of VC ethyl ether to prepare the cell energy activator.

[0066] Comparative Example 2-1

[0067] Based on Example 2, in step 2), the inoculation density is 2.8 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0068] Comparative Example 2-2

[0069] Based on Example 2, in step 2), the inoculation density is 5.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0070] Comparative Examples 2-3

[0071] Based on Example 2, in step 2), the inoculation density is 6.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0072] Comparative Examples 2-4

[0073] Based on Example 2, in step 2), the inoculation density is 7.0 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0074] Comparative Examples 2-5

[0075] Based on Example 2, in step 2), the inoculation density is 8.4 × 10⁻⁶. 6 cfu / mL, all other values ​​remain unchanged.

[0076] The yield of postbiotics was expressed as the cell density in the fermentation broth at the end of fermentation. The specific operation method was carried out in accordance with the national standard GB4789.15-2016. The test results are shown in Table 3.

[0077] Table 3 Colony counts after Bacillus fermentation

[0078]

[0079] As shown in Table 3, the Bacillus inoculation density used in Example 1 resulted in better fermented cells than Comparative Examples 2-1 to 2-5. Therefore, a single-factor screening experiment was conducted to determine the inoculation dose for Example 1 to determine the cell lysis, and 5.6 × 10⁻⁶ was ultimately selected. 6 The optimal concentration of cfu / mL is achieved by homogenizing the cells after fermentation, followed by ultrasonic disruption at a power of 1000W for 20 minutes, with a working mode of 3 seconds on and 3 seconds off.

[0080] Example 3

[0081] The components obtained according to the steps described in Example 1 were used to conduct formulation tests on the compound composition.

[0082] A method for preparing a cell energy activator includes the following steps:

[0083] 1) Dry and pulverize the golden camellia, then pass it through a 140-mesh sieve to obtain golden camellia powder; add the golden camellia powder to 60℃ distilled water, mix and stir for 20 minutes, add glucose powder as a carbon source, stir and homogenize, then place in a water bath and sterilize at 90℃ for 30 minutes to obtain fermentation medium; each liter of fermentation medium contains 10 parts of golden camellia powder and 2.5 parts of glucose powder, each part being 10g;

[0084] After activation, Bacillus was inoculated into the fermentation medium at an inoculation density of 5.6 × 10⁶. 6 CFU / mL, fermented at 30℃ for 20 hours, centrifuged at 6000 rpm for 10 min, the supernatant was collected, filtered through a 0.22 μm filter membrane, the filtrate was collected, lyophilized and concentrated to prepare the first component;

[0085] 2) The activated Bacillus was inoculated into YPD medium for fermentation at an inoculation density of 5.6 × 10⁶. 6 The concentration of cfu / mL was used to homogenize the cells after fermentation, followed by ultrasonic disruption at a power of 1000W for 20 minutes with 3 seconds of operation followed by 3 seconds of rest. The supernatant was then collected by centrifugation, freeze-dried, and concentrated to prepare the second component.

[0086] 3) Preparation of cell energy activator: Mix the first component and the second component, add 3.5 parts of nicotinamide and 1.0 part of VC ethyl ether to prepare the cell energy activator.

[0087] Comparative Example 3-1

[0088] Based on Example 3, in step 3), 3 parts of nicotinamide and 0.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0089] Comparative Example 3-2

[0090] Based on Example 3, in step 3), 3 parts of nicotinamide and 1.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0091] Comparative Example 3-3

[0092] Based on Example 3, in step 3), 3.5 parts of nicotinamide and 0.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0093] Comparative Examples 3-4

[0094] Based on Example 3, in step 3), 3.5 parts of nicotinamide and 1.0 part of VC ethyl ether were added, while the rest remained unchanged.

[0095] Comparative Examples 3-5

[0096] Based on Example 3, in step 3), 3.5 parts of nicotinamide and 1.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0097] Comparative Examples 3-6

[0098] Based on Example 3, in step 3), 4.0 parts of nicotinamide and 0.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0099] Comparative Examples 3-7

[0100] Based on Example 3, in step 3), 4.0 parts of nicotinamide and 1.0 part of VC ethyl ether were added, while the rest remained unchanged.

[0101] Comparative Examples 3-8

[0102] Based on Example 3, in step 3), 4.0 parts of nicotinamide and 1.5 parts of VC ethyl ether were added, while the rest remained unchanged.

[0103] Comparative Examples 3-9

[0104] Based on Example 3, in step 3), 0 parts of nicotinamide and 1.0 part of VC ethyl ether were added, while the rest remained unchanged.

[0105] Comparative Examples 3-10

[0106] Based on Example 3, in step 3), 3.5 parts of nicotinamide and 0 parts of VC ethyl ether were added, while the rest remained unchanged.

[0107] Anti-glycation assay. High glucose is one of the factors that contributes to the accumulation of AGEs (Advanced Glycation End Products) in cells, thereby inducing cellular glycation damage. High glucose can trigger inflammation, exacerbate AGE-RACE binding, activate the glycation reaction, and promote cell apoptosis. This example is used to evaluate the effect of a compounded composition of the components obtained according to the steps described in Example 1 on the glycation of skin cells. The specific steps are as follows:

[0108] Cell preparation: Human skin fibroblast cells (HSF) were purchased from the Kunming Cell Bank, Chinese Academy of Sciences. HSFs were cultured using 89% DMEM medium + 10% fetal bovine serum + 1% penicillin-dextrose antibody, and passaged every 3 days at a density of 1.0 × 10⁶ cells / year. 5 Inoculate 150 μL / mL of the sample into 6-well plates and incubate at 37°C for 24 hours. Then, set up a control group, a model group, and an experimental group. The control group was inoculated with fresh medium, while the model and experimental groups were inoculated with high-glucose medium (containing 30 mM glucose). At the start of the experiment, PBS was added to the control group, high-glucose medium was added to the model group, and the test sample was added to the experimental group. Each group was inoculated with 150 μL of the sample, and this process continued for 14 days.

[0109] During testing, the in vitro AGEs formation inhibition rate was determined using the method published by Eze et al. PBS was used as a blank control. BSA and D-fructose were dissolved in 0.2 mol / L PBS (pH 7.4) to achieve a BSA concentration of 10 mg / mL and a D-fructose concentration of 0.5 mol / L. The mixture was then filtered and sterilized to obtain the reaction solution. The blank control sample was a mixture of PBS and the reaction solution, while the model and experimental groups were mixtures of the test sample and the reaction solution. All samples were incubated at 37°C in the dark for 14 days. The incubation endpoint was measured using a microplate reader (excitation wavelength 370 nm, emission wavelength 440 nm, slit width 5 nm). The inhibition rate was calculated using the following formula:

[0110] Inhibition rate % = 1 - (F1 - F2) / (F3 - F2), where F1 represents the fluorescence value of the experimental group, F2 represents the fluorescence value of the blank group, and F3 represents the fluorescence value of the model control group. The measurement results are shown in Table 4.

[0111] Table 4. Inhibition rate determined by AGEs

[0112] Group Inhibition rate (%) Example 3 75.2 Comparative Example 3-1 55.1 Comparative Example 3-2 64.0 Comparative Example 3-3 60.4 Comparative Examples 3-4 73.3 Comparative Examples 3-5 65.6 Comparative Examples 3-6 68.9 Comparative Examples 3-7 71.4 Comparative Examples 3-8 69.5 Comparative Examples 3-9 34.1 Comparative Examples 3-10 47.5

[0113] As shown in Table 4, the metabolites of Bacillus fermented Camellia chrysantha exhibit different anti-glycation abilities. Furthermore, the addition of nicotinamide and VC ethyl ether to the formulation resulted in a good synergistic effect. Except for Comparative Examples 3-1, 3-9, and 3-10, the in vitro synergistic inhibition rate of AGEs formation reached over 60%. The optimal in vitro synergistic inhibition of AGEs formation was achieved when 3.5 parts of nicotinamide and 1.0 part of VC ethyl ether were added.

[0114] Example 4

[0115] Numerous studies have shown that long-term, high-dose injections of D-galactose solution can generate excessive reactive oxygen species (ROS), which reduces the activity of antioxidant enzymes in various organs, forms more superoxide anions and various oxidation products, and causes cell damage, leading to functional decline in multiple organs and systems. Therefore, this study used a D-galactose-induced aging mouse model combined with UV light irradiation to determine the anti-aging effect of the composition.

[0116] Based on the cell energy activator (named M, solvent: glycerol) obtained in Example 3, propylene glycol and deionized water were added, and the mixture was prepared to obtain the experimental reagent. The preparation components are shown in Table 5.

[0117] Table 5 Reagent Preparation Table

[0118]

[0119] Twenty 7-week-old SPF male BALB / c mice (purchased from Guangdong Provincial Medical Laboratory Animal Center) were randomly divided into 5 groups (control group, aging group, high-dose experimental group, medium-dose experimental group, and low-dose experimental group), with 4 mice in each group. Veet hair removal cream was used to remove hair from the backs of the mice. The control group received a daily subcutaneous injection of 0.3 mL of physiological saline, while the aging group and experimental groups received a daily subcutaneous injection of D-galactose (100 mg / kg / bw) for 4 consecutive weeks. After this treatment, each group was then exposed to radiation at a dose of 80 mJ / cm². 2 Mice were irradiated with UVB lamps on their backs for one hour every other day for four weeks. During the experiment, the appearance of aging mice and mice in the experimental group was monitored, and changes in the skin on the backs of mice in each group were compared as the experiment progressed. Specific experimental groupings and treatments are shown in Table 6.

[0120] Table 6 Experimental Grouping and Treatment Methods

[0121]

[0122] After the experiment, the mice were euthanized. A 10% skin tissue homogenate was prepared from skin tissue. The sample extract was homogenized in a high-speed, low-temperature tissue homogenizer (homogenization frequency 100 Hz, temperature -10℃). The homogenate was centrifuged at 8000 rpm, 4℃, for 30 min, and the supernatant was used for subsequent measurements. The activities of catalase (CAT), superoxide dismutase (SOD), and trace amounts of reduced glutathione (GSH) in the skin tissue were measured according to the instructions of the commercially available assay kit. The results are shown in Table 7.

[0123] Table 7. Antioxidant assay of skin tissue

[0124]

[0125] As shown in Table 7, with the aging group as the control, the activities of CAT, SOD and GSH in the skin of medium and high dose model mice were significantly increased, and the various oxidation indicators were close to those of the control group.

[0126] The various embodiments described in this specification are presented in a progressive manner, with each embodiment focusing on its differences from other embodiments. Similar or identical parts between embodiments can be referred to interchangeably. The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a cell energy activator, characterized in that, The preparation method comprises the following steps: 1) Dry and pulverize the golden camellia, then pass it through a 100-150 mesh sieve to obtain golden camellia powder; add the golden camellia powder to 60℃ distilled water, mix and stir for 20 minutes, add glucose powder as a carbon source, stir until homogeneous, and then place it in a water bath and sterilize at 90-100℃ for 30 minutes to obtain fermentation medium; each liter of fermentation medium contains 10 parts of golden camellia powder and 2.5 parts of glucose powder, each part being 10g; After activation, Bacillus was inoculated into the fermentation medium at an inoculation density of 5.5 × 10⁻⁶. 6 CFU / mL, fermented at 28-33℃ for 18-24 hours, centrifuged at 6000rpm for 10min, the supernatant was collected, filtered through a 0.22μm filter membrane, the filtrate was collected, lyophilized and concentrated to prepare the first component; 2) The activated Bacillus was inoculated into YPD medium for fermentation. After fermentation, the cells were homogenized and ultrasonically broken up. The ultrasonic power was 800-1000w for 15-20min, with 3-5s working and 3-5s intermittent. The supernatant was collected by centrifugation, freeze-dried and concentrated to prepare the second component. 3) Composition preparation: Mix the first component and the second component, add nicotinamide and VC ethyl ether to prepare the cell energy activator.

2. The method according to claim 1, characterized in that, Add 3-5 parts of nicotinamide and 0.5-1.5 parts of VC ethyl ether to step 3).

3. The cell energy activator prepared according to the method of claim 1 or 2.

4. A skincare product, characterized in that, Includes the cell energy activator as described in claim 3.

5. The skincare product according to claim 4, characterized in that, It also includes at least one of the following: moisturizers, anti-inflammatory agents, and chelating agents.

6. Use of the cell energy activator according to claim 3 in the preparation of skin antioxidant products.

7. The use according to claim 6, characterized in that, It is administered to men.

8. Use of the cell energy activator according to claim 3 in the preparation of products that enhance catalase activity.

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