CN114557905A - Endogenous cell regulation system and application thereof - Google Patents

Abstract

The invention discloses an endogenous cell regulation system and application thereof. The cell regulation system provided by the invention carries out specific proportioning and combination by purifying nucleoside compounds, unsaturated fatty acids, saturated fatty acids, protein substances and the like in natural animals and plants, activates cells in a multi-target, multi-layer, multi-path and all-around way, optimizes the microecology of skin cells, restores the young state of the cells, maintains the healthy state of the cells, and reconstructs the endogenous function of the cells, thereby improving the anatomical structure and function of skin tissues. The special combination of various unsaturated fatty acids, saturated fatty acids, flavonoids, nucleosides and proteins is used as a main control cell metabolic pathway, the cell activity is improved, aging cells are reduced in a targeted manner, and oxidation is prevented, so that the young state of skin cells is recovered, the three-dimensional structure of the skin is repaired, and the health state of the cells is maintained; based on this regulatory system, an endogenous wisdom cell activation system (abbreviated ICEAS) was further invented.

Description

Endogenous cell regulation system and application thereof

Technical Field

The invention relates to the fields of cell biology, regenerative medicine and medicines, in particular to an endogenous cell regulation system and application thereof.

Background

Cellular senescence can be induced by a variety of factors such as DNA damage, epigenetic changes, telomere shortening, metabolic disorders, mitochondrial dysfunction, and inflammation. It is theoretically difficult to achieve resistance to cellular senescence or activation of cellular functions through a single pathway.

The human skin tissue mainly comprises two layers of dermis and epidermis, wherein the dermis provides solid support for the epidermis and provides nutrition for the epidermis; in addition, the nutritional support of skin tissue is accompanied by the provision of nutrients from the subcutaneous tissue and the exchange of nutrients by the microcirculation. The epidermal cells form a skin barrier on the outer layer of skin tissue, resist the entry of external harmful substances, irritants and sunlight, and have the effects of moisturizing, regulating and resisting inflammation. The dermis is composed mainly of fibroblasts and extracellular matrix, and fibroblasts proliferate and differentiate normally to maintain the normal structure and physiological functions of the skin. Collagen and elastin are synthesized and secreted by fibroblasts and are the main components that make up the three-dimensional structure of the dermal layer, forming the main extracellular matrix. Collagen can impart strength and toughness to the extracellular matrix; the elastic fiber provides tissue elasticity and enables skin to have the capability of stretching and folding; meanwhile, the extracellular matrix forms the microenvironment of cells, regulates the substance exchange of the cells, supports the skin structure and maintains the skin moisture. Cells in the subcutaneous tissue that provide trophic support are derived primarily from adipocytes in the subcutaneous tissue. The face skin of the infant is fine and glossy, and the main reason is that the subcutaneous fat cells are alive. The vascular endothelial cells, which form blood vessels, regulate the local capillary microcirculation.

In addition, the presence of excessive senescent cells in the tissue is detrimental to the survival and growth of normal cells. Senescent cells secrete a range of pro-inflammatory cytokines, chemokines, proteases and other factors, destroy surrounding tissues and inhibit normal cellular activity: for example, senescent cells secrete proinflammatory cytokines, create a chronic inflammatory environment around normal cells, accelerating cell aging; inflammatory factors, such as TNF- α, are negative regulators of collagen I.

The skin extracellular matrix is synthesized and secreted by cells and is composed mainly of collagen, elastin and glycoproteins, and proteoglycans. The extracellular matrix forms the microenvironment of the cells, regulates the exchange of substances by the cells, supports the skin structure, and maintains skin moisture. With age, the rate of collagen degradation by matrix metalloproteinases increases, and the collagen content decreases, manifesting in the formation of wrinkles in the skin.

Different types of cells in skin tissue exert their own cellular functions in a healthy and orderly manner, can maintain youthful skin tissue and resist aging of skin tissue. However, in the prior art, no product capable of comprehensively activating cells, optimizing the microecology of skin cells, recovering the young state of the cells, maintaining the healthy state of the cells and reconstructing the endogenous function of the cells by regulating the metabolic pathway of the cells and multiple targets, layers and paths is available, so that the anatomical structure and the function of skin tissues are improved.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides an endogenous cell regulation system which is used for promoting the self-regulation of skin cells to restore the young state and the healthy state of the cells and improving the physiological state of skin tissues.

In order to solve the technical problems, the technical scheme of the invention is as follows: an endogenous cell regulatory system comprising at least 2 of a nucleoside compound, an unsaturated fatty acid, a saturated fatty acid, and a proteinaceous substance;

the nucleoside compound is at least one of adenosine compounds, cytidine compounds and guanosine compounds;

the saturated fatty acid is at least one selected from stearic acid, palmitic acid ethyl palmitate, stearic acid derivatives, palmitic acid derivatives and palmitic acid derivatives;

the unsaturated fatty acid is at least one selected from oleic acid, linoleic acid, punicic acid, linolenic acid, octadecenoic acid and eicosenoic acid.

As a further description of the above technical solution: the protein material is selected from animal placenta protein, animal umbilical cord extract, oligopeptide (oligopeptide-1, oligopeptide-2, oligopeptide-3, oligopeptide-4, oligopeptide-5 or oligopeptide-6), polypeptide, silk amino acid, silk powder, silk sericin, silk extract, methionine, egg powder, egg yolk extract, egg shell membrane extract, egg white extract, dipeptide-1, dipeptide-2, dipeptide-4, glutamine, glutathione, citrulline, oligonucleotide, cystine, collagen amino acids, collagen extract, keratin amino acids, yeast, zymosan, yeast fermentation product extract, yeast/soy protein fermentation product, yeast/barley seed fermentation product filtrate, yeast extract, soy protein, soy, Yeast/barley seed fermentation filtrate, yeast/grape fermentation extract, yeast lysate extract, yeast/rice bran fermentation product, yeast polypeptides, yeast fermentation product, yeast fermentation filtrate, yeast fermentation lysate filtrate, yeast lysate extract, arginine, chitosamine, milk amino acids, milk protein extract, proline, glucosamine, lactic acid, milk protein, hydrolyzed silk, hydrolyzed soy protein, hydrolyzed rice protein, tyrosine, casein, tyrosine methyl ester of tyrosine HCL, potassium caseinate, sodium caseinate, casein extract, buttermilk powder, ornithine, nisin, whey protein, orotic acid, lactoglobulin, lactoferrin, hydrolyzed elastin, hydrolyzed eggshell membrane, hydrolyzed egg shell membrane, and the like, Hydrolyzed collagen, hydrolyzed keratin PG-propylmethylsilanediol, hydrolyzed keratin ethyl ester, hydrolyzed yeast protein, hydrolyzed yeast extract, hydrolyzed placenta extract of sheep, hydrolyzed placenta protein of pig, hydrolyzed placenta extract of pig, sericin, fibroin, tetrapeptide-1, tetrapeptide-4, aspartic acid, asparagine, serum albumin, serum protein, and histidine.

Preferably, the proteinaceous material is selected from one or more of milk protein, hydrolysed yeast extract, hydrolysed fibroin, oligopeptides, sericin, tyrosine, hydrolysed collagen, sericin, hydrolysed silk, fibroin, yeast polypeptides, histidine, proline, tyrosine, ornithine, aspartic acid, arginine, cystine, methionine, oligopeptides, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl tetrapeptide-10, xanthan gum, carnosine, filaggrin and astaxanthin yeast extract. Further preferably, the proteinaceous substance is selected from the group consisting of milk proteins, sericin, and mixtures of milk proteins and or sericin, and histidine, proline, tyrosine, ornithine, aspartic acid, citrulline, methionine, cystine, and arginine.

As a further description of the above technical solution: the nucleoside compounds, unsaturated fatty acids, saturated fatty acids and flavonoids are mainly extracted from natural products, and can also be artificially synthesized substances. More preferably, the nucleoside compound is derived from at least one of grape seed extract, pomegranate seed extract, rehmannia root extract, papaya fruit extract and ganoderma lucidum extract; more preferably, the nucleoside compounds are derived from one or both of grape seed extract and pomegranate seed extract.

As a further preference, the unsaturated fatty acids and saturated fatty acids are derived from at least one of grape seed extract, pomegranate seed extract, agaricus bisporus extract, coconut fruit extract, green tea extract, cocoa extract, and oat meal extract; more preferably, the unsaturated fatty acids and saturated fatty acids are derived from one or both of grape seed extract and pomegranate seed extract.

More preferably, the flavonoid is derived from at least one of selaginella tamariscina extract, soybean extract, sea buckthorn extract, cactus extract, changium smyrnioides, radix scrophulariae extract and angelica root extract. More preferably, the flavonoids are derived from selaginella tamariscina extract.

As a further description of the above technical solution: the molecular weight range of the nucleoside compound is 80-8000; the molecular weight range of the unsaturated fatty acid is 100-3000; the molecular weight range of the saturated fatty acid is 100-3000; the molecular weight range of the flavonoid is 100-2100. More preferably, the molecular weight of the nucleoside compound is 80-1200; the molecular weight range of the unsaturated fatty acid is 100-1000; the molecular weight range of the saturated fatty acid is 100-1000; the molecular weight range of the flavonoid is 100-1200, and the regulation effect of each component in the range on the young state and the healthy state of cells is found to be good.

In the system, the content of nucleoside compounds is 10 ng/mL-500 ug/mL, the content of unsaturated fatty acid is 10 ng/mL-500 ug/mL, and the content of saturated fatty acid is 10 ng/mL-500 ug/mL; the content of the protein substances is 100 mg/mL-10 g/mL.

As a further description of the above technical solution: the system also comprises flavonoids substances of 1ng-800 ng/mL.

Preferably, the flavonoid is one or more of amentoflavone, quercetin, wogonin, isoquercitrin, glycoside flavonoid, naringenin, soy isoflavone, quercetin, bioflavonoid, fisetin and 4', 6, 7-trihydroxyisoflavone.

As a further description of the above technical solution: the conditioning system further comprises a skin care ingredient a and/or a skin care ingredient b;

the skin care ingredient a is selected from one or more of antimicrobial ingredients, humectants, moisturizers, UV absorbers, epidermal barrier function modulators, skin rejuvenating and regenerating ingredients, skin tightening and anti-wrinkle agents, soothing and anti-inflammatory agents, anti-itch ingredients, antioxidants, free radical scavengers, UV-quenchers, sebum-regulating and anti-acne agents, stretch mark modulators, skin immune system modulators, skin lightening agents, skin microcirculation modulators, microfilaments and telangiectasia modulators, and tissue metalloproteinase inhibitors;

the skin care ingredient B is selected from the group consisting of proteins, ellagic acid, growth factors, coenzymes, enzyme inhibitors, peptides (e.g., di-, tri-, tetra-, penta-, and hexapeptides), carbohydrates (e.g., mono-, di-, tri-, and oligosaccharides), polysaccharides, glycosaminoglycans, glycosaminoglycan subunits, lipids, sphingosines, sphingolipids, glycosphingolipids, sulfatides, phospholipids, sterols, phytosterols, saponins, phenols, polyphenols, terpenes, alkaloids, provitamins, vitamin A group, vitamin B group, ascorbates (ascorbic acid, magnesium ascorbate, calcium ascorbate, ascorbyl polypeptides, ascorbyl glucosides, ascorbyl stearates, ascorbyl palmitate, ethyl ascorbic acid, isoascorbic acid, ascorbic acid/PCA magnesium, disodium ascorbyl sulfate, and other ascorbic acid derivatives), Vitamin D, vitamin E, vitamin K, folic acid, riboflavin, inositol, thiamine HCl, rosin oil, retinal, tretinoin, isotretinoin, alitretinoin, avilamate, avilamoids, chelators, creatinine, dimethylethanolamine, amino acids such as serine, glycine, aspartic acid, cysteine, glutamine, lysine, arginine, aspartic acid, glutamic acid, N-acetylcysteine, citrulline, collagen, gelatin, albumin, sericin, fibroin, anti-keratin microfilament aggregatin, transforming growth factor, insulin-like growth factor, epidermal growth factor, acidic and basic fibroblast growth factor, nerve growth factor, keratinocyte growth factor, hepatocyte growth factor, platelet-derived growth factor, granulocyte-macrophage colony stimulating factor, vascular endothelial growth factor, Trace elements (calcium, iron, magnesium), hydrogenated lecithin, hydrolyzed collagen, xanthan gum, sodium polyglutamate, carnosine, coenzyme Q10, nicotinamide adenine dinucleotide, glucose, resveratrol, fructose, mannose, dihydroxyacetone, erythrulose, sucrose, trehalose, maltose, galactomannan, glucomannan, beta-glucan, carrageenan, glycogen, chitosan, lentinan, lichenin, inulin, fucose, alginate, xyloglucan, dextran, amylose, levan, amylopectin, hyaluronic acid, chondroitin sulfate, heparin, dermatan sulfate, glucuronic acid, oat beta-glucan, N-acetylglucosamine, allantoin, ceramide, N-lipoyl sphingosine, cerebroside, ganglioside, sulfatide, phosphatidylcholine, Phosphatidylserine, phosphatidylethanolamine, lactic acid, citric acid, glycolic acid, azelaic acid, salicylic acid, lipoic acid, pyrrolidone carboxylic acid, uric acid, caffeic acid, conjugated linoleic acid, squalane, squalene, monoglycerides, diglycerides, triglycerides, petrolatum, lanolin, phytosphingosine, sitosterol, stigmasterol, brassicasterol, lupeol, glycyrrhizin, rutin, genistein, daidzein, fisetin, myricetin, luteolin, hesperetin, silybin, silymarin, apigenin, epigallocatechin gallate, nordihydroguaiaretic acid, ellagic acid resorcinol, glycyrrhetinic acid, farnesol, alpha-bisabolol, beta-bisabolol, theophylline, theobromine, usnic acid, zinc-, selenium-, manganese-salts, glycerol, propylene glycol, butylene glycol, alpha-bisabolol, catechin, zinc-, selenium-, manganese-salts, glycerol, propylene glycol, and mixtures thereof, Sorbitol, erythritol, hexylene glycol, phytantriol, carbomer, polysorbate-20, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, diglycerol, caprylhydroxamic acid, 1, 2-hexanediol, 1, 3-propylene glycol, cetyl palmitate, caprylic/capric triglyceride, palmitoyl tetrapeptide-10, polysorbate-60, sorbitan monostearate, polyethylene glycol-8, betaine, sclerostin, sodium hyaluronate, defensin, cathepsin, histatin, salicylate, cinnamate, caffeine, filaggrin, melatonin, urea, papain, bromelain, superoxide dismutase, lactoperoxidase, phospholipase, sphingomyelinase, transglutaminase, tranexamic acid, alpha 1-antitrypsin, leupeptin, Kinetin, anthranilate, alpha-carotene, beta-carotene, lycopene, luteine, zeaxanthin, palm oil, conventional cosmetic natural extracts (e.g., rehmannia root extract, papaya fruit, ganoderma lucidum, changium smyrnioides, cactus extract, angelica root extract, black ginseng root extract, cardamom seed extract, agaricus bisporus extract, tea extract, oat meal extract, coconut fruit extract, cocoa extract, grape seed extract, pomegranate seed extract, selaginella tamariscina extract, mustard extract, astaxanthin yeast extract, comfrey extract, ginkgo extract, centella, chamomile extract, aloe vera, calendula extract, licorice extract, witch hazel extract, sea buckthorn extract, evening primrose oil, green tea extract, nasturtium extract, thyme extract, red-scented water, black pepper extract, black pepper, One or more of centella asiatica extract, arnica montana extract, pearl extract, lalang grass rhizome extract, licorice extract, guarana extract, safflower extract, soybean extract, etc.), tea tree oil, oligomeric proanthocyanidin, and polyhydric alcohol.

As a further description of the above technical solution: the product forms of the conditioning system are solutions, emulsions (oil-in-water and water-in-oil), emulsions, lotions, dispersions, powders, ointments, gel forms and viscous surfactants, emulsifying polymers, pomades, hair growth stimulants, shampoos, soaps, gels, lyophilizates, powders, sticks, pens, sprays, body oils, masks or patch forms.

The invention also provides the application of the cell regulating system, which is used for endogenous repair and cell function activation of tissue cells; and reducing senescent cells and restoring the youthful state of the cells.

As a further description of the above technical solution: the regulatory system is used to promote the secretion of cytokines beneficial for repair and regeneration by tissue cells.

As a further description of the above technical solution: the regulation system is used for regulating and controlling the cell rhythm/biological clock of tissue cells; and is used for epigenetic regulation of tissue cells and reduction of DNA epigenetic clock age.

As a further description of the above technical solution: the regulation system is used for enhancing the mitochondrial oxidative metabolism function of tissue cells.

As a further description of the above technical solution: the tissue cell is one or more of epidermal cell, epidermal stem cell, fibroblast, fat cell, adipose mesenchymal stem cell and vascular endothelial cell; derived from human, monkey, rat, pig, cow, dog, sheep, rabbit, chicken or horse.

Further, the invention also provides application of the regulating system in preparing cosmetics.

Furthermore, the invention also provides application of the regulating system in preparing cell culture medium or serum-free cell culture medium of skin tissue cells.

The invention has the following characteristics: the harsher the natural environment the more persistent the life that is inoculated. The plants such as grapes, pomegranates, selaginella tamariscina and the like which are produced in the strong ultraviolet dry environment of the Yunnan plateau inflammation and strong ultraviolet rays are rich in natural anti-aging, anti-ultraviolet and anti-water-deficiency substances, are raw material sources of active ingredients of a plurality of skin care products, and are not excavated deeply and scientifically all the time, so that the effect is limited. The invention discloses an endogenous cell regulation system, which is characterized in that high-purity nucleoside compounds, unsaturated fatty acids, saturated fatty acids, flavonoids substances, protein hydrolysates, milk proteins, amino acids and the like from natural animals and plants are used for systematically regulating and controlling the metabolic activity, cell rhythm and fine epigenetic regulation and control of different types of cells in skin tissues, erasing aging blots, enhancing mitochondrial functions and the like, and reducing aging cells by a specific combination ratio, so that the repair and resistance capability of the skin tissues are enhanced, and skin aging is resisted.

Although most skin care products contain the components in the patent, such as nucleoside compounds, unsaturated fatty acids, saturated fatty acids, flavonoid substances and the like, the components are mostly used as mixed components in natural extracts in skin care products, and have low purity and high impurity content; the proportion of the components is not clear or the content is low; and are often used as adjunct ingredients rather than as primary active ingredients; the different effects of the combination partners disclosed in the present invention on different cells of the skin tissue are also not specifically defined in the prior art publications. The invention develops the effective combination and proportion of the substances as the main active ingredients of the skin care product for the first time to form a regulating system for promoting the self regulation of skin cells to restore the young state and the healthy state of the cells; furthermore, the regulating system can effectively regulate the metabolism and physiological state of different cells of skin tissues, activate the functions of different types of cells, play the self-regulating role of endogenous cells under the stimulation of no exogenous trophic factors, facilitate the cells of different differentiation types to recover to young and healthy states, and automatically regulate the functions of the cells according to the metabolic demand of the tissue microenvironment, thereby improving the physiological state of the skin tissues. Based on this regulatory System, an Endogenous smart Cell Activation System (ICEAS for short) has further been invented.

Compared with the prior art, the invention has the following technical effects: the cell regulation system provided by the invention can activate cells in a multi-target, multi-level, multi-path and all-round way, optimize the microecology of skin cells, recover the young state of the cells, maintain the healthy state of the cells and rebuild the endogenous function of the cells, thereby improving the anatomical structure and function of skin tissues. The special combination of various unsaturated fatty acids, saturated fatty acids, flavonoids, nucleosides and proteins is used for mainly regulating and controlling a cell metabolic pathway, improving the cell activity, reducing aged cells in a targeted manner and resisting oxidation, so that the young state of skin cells is restored, the three-dimensional structure of the skin is repaired, and the health state of the cells is maintained.

Drawings

FIG. 1 is a graph of the growth curve of skin fibroblasts (example 1 regulatory system);

FIG. 2 is a graph of epidermal cell growth curves (example 1 regulatory system);

FIG. 3 is a graph of a smooth muscle cell growth curve (example 1 regulatory system);

FIG. 4 is a graph of a vascular endothelial cell growth curve (example 1 regulatory system);

FIG. 5 is a graph of adipose mesenchymal stem cell growth curves (example 1 regulatory system);

FIG. 6 is a graph showing the result of the staining detection of sirius red, which is a skin fibroblast, with a detection wavelength of 540 nm;

FIG. 7 is a graph of the growth curve of skin fibroblasts (example 2 regulatory system);

FIG. 8 is a graph of epidermal cell growth curves (example 2 regulatory system);

FIG. 9 is a graph of a smooth muscle cell growth curve (example 2 regulatory system);

FIG. 10 is a graph of a vascular endothelial cell growth curve (example 2 regulatory system);

figure 11 is an adipose mesenchymal stem cell growth curve (example 2 regulatory system);

FIG. 12 shows the result of the staining test (detection wavelength 540nm) of sirius red of skin fibroblasts;

FIG. 13 shows the result of the staining test (detection wavelength 540nm) of sirius red of skin fibroblasts;

FIG. 14 shows the result of the staining test (detection wavelength 540nm) of sirius red of skin fibroblasts;

FIG. 15 shows procollagen type assays for secretion of dermal fibroblasts from individuals of different ages; a, quantitatively detecting procollagen type; wherein Y represents age "year of age", Average represents the amount of cell secretion before treatment, and DM represents a control group; b, a schematic diagram of the change of the secretion content of collagen in skin fibers at different ages after the essence water treatment;

FIG. 16 is a result of measuring the expression level of cytokine genes after 5 days of treatment with serum in adipose-derived mesenchymal stem cells;

FIG. 17 shows the results of measuring the expression levels of senescence-associated genes CDKNIA, CDKN2A and IL6 in different types of cells;

FIG. 18 is a graph showing the changes in gene expression levels before and after CBX1 treatment in skin fibroblasts from different ages;

FIG. 19 is a graph of the change in apparent genetic clock in skin fibroblasts following treatment by the regulatory system;

FIG. 20 is the expression of the SIRT3 gene in skin fibroblasts;

FIG. 21 shows the results of measuring the expression level of a gene (clock, BMAL1, RORA) involved in regulating cell rhythm in skin fibroblasts;

FIG. 22 shows the results of measuring the expression level of a gene (clock, BMAL1, RORA) involved in the regulation of cell rhythm in vascular endothelial cells;

FIG. 23 shows the results of measuring the expression level of a gene (clock, BMAL1, RORA) involved in regulation of cell rhythm in epidermal cells;

FIG. 24 shows the results of measurement of the expression levels of various cytokines and elastin, filaggrin, in human dermal fibroblasts;

FIG. 25 shows the results of measurement of the expression level of tissue inhibitor of metalloproteinase 1(TIMP1) in human epidermal cells;

FIG. 26 shows the results of the identification of β -galactosidase staining of skin fibroblasts;

FIG. 27 is a graph of the results of a skin moisture test on a population using a essence;

FIG. 28 is a measurement of the moisture content of the skin of a person staying up overnight after using the refined liquid;

FIG. 29 shows the results of pH measurements on the skin of a human subject;

figure 30 is a graph of the results of a test for skin moisture on a subject following discontinuation of use.

Detailed Description

The technical solutions of the present invention will be described in further detail below with reference to the drawings and the detailed description, but the present invention is not limited to the following technical solutions.

The protein materials used in the following examples were all commercially available. The nucleoside compounds, unsaturated fatty acids, saturated fatty acids and flavonoids used in the following examples are mainly plant sources and obtained by the existing extraction and separation method; the sources of the various active ingredients of the present invention are not limited to plant extracts, and can be obtained by artificial synthesis. When the nucleoside compounds, the fatty acids and the flavonoid substances are extracted from plants, the nucleoside compounds, the unsaturated fatty acids, the saturated fatty acids and the flavonoid substances are obtained by separation and purification, and the intercepted molecular weight ranges are as follows:

molecular weight range of nucleoside compounds: 80-1200;

molecular weight range of unsaturated fatty acids: 100 to 1000;

molecular weight range of saturated fatty acids: 100 to 1000;

the molecular weight range of the flavonoid is as follows: 100 to 1200.

The crude products of the nucleoside compounds comprise guanosine, adenosine and cytidine compounds, and the guanosine, adenosine and cytidine compounds can be respectively obtained by purification, wherein in the extraction process, the content of guanosine compounds in pomegranate seeds is lower, and the content of guanosine compounds in grape seeds is relatively higher.

Example 1

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 mug/mL adenosine compounds, 2 mug/mL cytidine compounds, 2.5 mug/mL guanosine compounds, 60 mug/mL unsaturated fatty acids, 30 mug/mL saturated fatty acids and 600mg/mL protein substances (various amino acids used in milk proteins and cosmetics, such as histidine, proline, tyrosine, ornithine, aspartic acid, cystine, citrulline, methionine and arginine, the concentration can be set arbitrarily) +350ng/mL flavonoid substances, trace elements (iron, magnesium, calcium) +83 mug/mL vitamins;

wherein the nucleoside compounds (adenosine compounds, cytidine compounds, and guanosine compounds), unsaturated fatty acids, and saturated fatty acids are derived from grape seed extract and pomegranate seed extract.

The flavonoids are derived from herba Selaginellae extract, and are prepared by pulverizing dried herba Selaginellae, sieving, extracting with organic solution, concentrating the extractive solution, purifying, and separating to obtain flavonoids mainly containing amentoflavone, quercetin, wogonin, isoamentoflavone, glycoside flavonoids, naringenin, soybean isoflavones, quercetin, bioflavonoids, fisetin and 4', 6, 7-trihydroxyisoflavone. The vitamins described in this example contain primarily vitamin E, ascorbic acid species.

Effect evaluation experiment

Respectively preparing human skin fibroblasts, human epidermal cells, human smooth muscle cells, human vascular endothelial cells and human adipose-derived mesenchymal stem cells; the day before the treatment by using the regulating system, the cell concentration is 5000-9500 cells/cm²Cell plating was performed.

The detection method comprises the following steps: on the first Day after plating (Day 1 or D1), cells were treated with the regulatory system prepared in the first step for each cell, and the cell culture medium was DMEM containing 20% (v/v) of the regulatory system, and the cell change was performed every 2 days. Culturing the cells in a carbon dioxide incubator with CO ₂5% concentration, and culturing at 37 ℃.

For the detection of cell proliferation activity, the method of MTT (MTT was performed according to the kit instructions of Biyun day) was used to detect the proliferation of cells for 9 consecutive days; the results showed that the cell proliferation activity was good for skin fibroblasts, epidermal cells, smooth muscle cells, vascular endothelial cells, and adipose mesenchymal stem cells treated with the regulatory system of this example (fig. 1 to 5).

For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. The skin fibroblasts were sampled before and after the conditioning system treatment (3 days for the cultured cells) in this example, the control group was a culture system (conventional cell culture system) containing 10% FBS in DMEM, and the absorbance value after sirius red staining was quantitatively measured using a wavelength of 540nm after 3 days for the cells were cultured; the results show that the regulation system in this example can effectively enhance the activity of skin fibroblasts, which is shown by the enhanced collagen secretion ability of skin fibroblasts (fig. 6), and the collagen expression level is increased by 2.06 times after 3 days of treatment, and is increased by 1.88 times in the control group.

Example 2

The cell regulation system (liquid; also named serum in subsequent experiments) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL protein (milk protein) +83 μ g/mL vitamin; the raw materials are from the same sources as in example 1.

Effect evaluation experiment

Respectively preparing human skin fibroblasts, human epidermal cells, human smooth muscle cells, human vascular endothelial cells and human adipose mesenchymal stem cells; make itTreating the previous day with the regulating system according to the ratio of 5000-9500 cells/cm²Cell plating was performed.

The detection method comprises the following steps: starting on the first Day after plating (time Day1 or D1), cells were treated with the regulatory system prepared in the first step for each cell, and the cell culture medium was DMEM containing 20% of the regulatory system, and the cell change was performed every 2 days. Culturing the cells in a carbon dioxide incubator with CO ₂5% concentration, and culturing at 37 ℃.

For the detection of cell proliferation activity, the method of MTT (MTT was performed according to the kit instructions of Biyun day) was used to detect the proliferation of cells for 9 consecutive days; the results showed that the cell proliferation activity was good for skin fibroblasts, epidermal cells, smooth muscle cells, vascular endothelial cells, and adipose mesenchymal stem cells treated with the regulatory system of this example (fig. 7 to 11).

For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. The skin fibroblasts were sampled before and after the conditioning system treatment (3 days for the cultured cells) in this example, the control group was a culture system containing 10% FBS in DMEM (cell conventional culture system), and the absorbance value after sirius red staining was quantitatively measured using a wavelength of 540nm after 3 days for the cells; the results show that the regulation system in this example can effectively enhance the activity of skin fibroblasts, which is shown by the enhanced collagen secretion ability of skin fibroblasts (fig. 12), and the collagen expression level is increased by 1.96 times after 3 days of treatment, and the collagen expression level is increased by 1.88 times in the control group.

Example 3

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 4.5. mu.g/mL adenosine compounds + 1. mu.g/mL cytidine compounds + 1. mu.g/mL unsaturated fatty acids +100ng/mL saturated fatty acids + 10. mu.g/mL guanosine compounds +150mg/mL protein compounds (milk proteins); wherein the nucleoside compounds (adenosine compounds, cytidine compounds, guanosine compounds), unsaturated fatty acids, and saturated fatty acids are derived from grape seed extract and pomegranate seed extract.

Preparing human skinThe fiber cells are treated by the regulating system for 5000-9500 cells/cm in the day before²Cell plating was performed.

For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. The skin fibroblasts were sampled before and after the conditioning system treatment (3 days for the cultured cells) in this example, the control group was a culture system (conventional cell culture system) containing 10% FBS in DMEM, and the absorbance value after sirius red staining was quantitatively measured using a wavelength of 540nm after 3 days for the cells were cultured; the results show that the regulation system in this example can effectively enhance the activity of skin fibroblasts, which is shown by the enhanced collagen secretion ability of skin fibroblasts (fig. 13), and the collagen expression level is increased by 1.82 times and 1.88 times after 3 days of treatment. Although the result of the experimental group is lower than that of the control group, the culture system containing 10% FBS in DMEM is a conventional, stable and good culture system for culturing skin fibroblasts, and the cells are cultured by adopting a solution of a natural product, so that the cells can be normally proliferated, the activity is good, and the experimental effect equivalent to the collagen expression amount of the control group can be achieved.

Example 4

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 100ng/mL adenosine compounds, 100ng/mL cytidine compounds, 10ng/mL guanosine substances, 100 mu g/mL unsaturated fatty acids, 100ng/mL saturated fatty acids, 1g/mL protein substances (hydrolyzed yeast extract) +10ng/mL flavonoid substances; wherein the nucleoside compounds (adenosine compounds, cytidine compounds, guanosine compounds), unsaturated fatty acids, saturated fatty acids are derived from grape seed extract and pomegranate seed extract, and the flavonoids are derived from herba Selaginellae extract.

Preparing human dermal fibroblast, treating with the above regulating system for 500 days0 to 9500 cells/cm²Cell plating was performed.

For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. For the secretion of collagen by skin fibroblasts, a sirius red stain (commercial kit) was used for detection. The skin fibroblasts were sampled before and after the conditioning system treatment (3 days for the cultured cells) in this example, the control group was a culture system (conventional cell culture system) containing 10% FBS in DMEM, and the absorbance value after sirius red staining was quantitatively measured using a wavelength of 540nm after 3 days for the cells were cultured; the results show that the regulation system in this example can effectively enhance the activity of skin fibroblasts, which is shown by the enhanced collagen secretion ability of skin fibroblasts (fig. 14), and the collagen expression level is increased by 1.70 times after 3 days of treatment, and the collagen expression level is increased by 1.88 times in the control group.

Example 5

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 35 μ g/mL adenosine compounds +3 μ g/mL cytidine compounds +20 μ g/mL guanosine compounds +400 μ g/mL unsaturated fatty acids +100 μ g/mL saturated fatty acids +5g/mL protein substances (hydrolyzed fibroin, milk protein, amino acids); wherein the nucleoside compounds (adenosine compounds, cytidine compounds, guanosine compounds), unsaturated fatty acids, and saturated fatty acids are derived from grape seed extract and pomegranate seed extract.

Example 6

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +4 μ g/mL cytidine +5 μ g/mL guanosine +60ng/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL proteinaceous material (milk protein, various amino acids, as in example 1) + trace elements (iron, magnesium, calcium) +83 μ g/mL vitamin; wherein the nucleoside compounds (adenosine compounds, cytidine compounds, and guanosine compounds), unsaturated fatty acids, saturated fatty acids are derived from grape seed extract and pomegranate seed extract, and the flavonoid is derived from herba Selaginellae extract. The cells which use the regulatory system to act are macaque skin fibroblasts.

Example 7

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 500ng/mL unsaturated fatty acid +500mg/mL protein material (oligopeptide, sericin, tyrosine, hydrolyzed collagen, sericin, histidine); wherein the unsaturated fatty acid is derived from grape seed extract and pomegranate seed extract.

Example 8

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 300 mug/mL adenosine compounds +50 mug/mL cytidine compounds +4g/mL protein substances (milk proteins); wherein the adenosine compound and cytidine compound are separated from semen Granati extract.

Example 9

The cell regulation system (liquid) was prepared according to the following combinations and ratios: adenosine compounds 50 mug/mL, cytidine compounds 20 mug/mL, guanosine compounds 2 mug/mL, unsaturated fatty acids 50 mug/mL (linoleic acid) + saturated fatty acids 10ng/mL (palmitic acid) + protein substances 1g/mL (milk protein) + flavonoids 600ng/mL (soy isoflavones, quercetin); wherein the nucleoside compounds are separated from grape seed extract and pomegranate seed extract, and the soybean isoflavone is separated from soybean extract. Quercetin, unsaturated fatty acid (linoleic acid) and saturated fatty acid (palmitic acid) are commercially available.

Example 10

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +4 μ g/mL cytidine +6 μ g/mL guanosine +100 μ g/mL unsaturated fatty acid (oleic acid, linoleic acid) +420 μ g/mL saturated fatty acid (stearate, ethyl palmitate, retinol palmitate, diglycerin (sebacic acid/isopalmitate) ester) +3g/mL proteinaceous material (hydrolyzed silk, yeast polypeptides, proline, tyrosine, ornithine, histidine, aspartic acid, arginine) +22 μ g/mL provitamin; wherein the nucleoside compounds (adenosine compounds, cytidine compounds, and guanosine compounds) are separated from grape seed extract and pomegranate seed extract.

Example 11

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 mug/mL adenosine compounds, 4 mug/mL cytidine compounds, 200 mug/mL unsaturated fatty acids (oleic acid and linoleic acid), 300mg/mL protein substances (milk protein) and trace elements (iron, magnesium and calcium); wherein the nucleoside compounds (adenosine compounds and cytidine compounds) are separated from semen Granati extract, and other raw materials are purchased from market.

Example 12

The cell regulation system (emulsion) was prepared according to the following combinations and ratios: 20 mug/mL adenosine compounds, 4 mug/mL cytidine compounds, 5 mug/mL guanosine compounds, 60 mug/mL unsaturated fatty acids, 30 mug/mL saturated fatty acids and 600mg/mL protein substances (milk proteins, various amino acids, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl tetrapeptide-10, xanthan gum and carnosine) +300ng/mL flavonoid compounds, sodium hyaluronate, 83 mug/mL vitamin, gallic acid, catechin, trace elements (iron, magnesium and calcium) + pearl extracts; wherein the nucleoside compounds (adenosine compounds, cytidine compounds, guanosine compounds), unsaturated fatty acids, and saturated fatty acids are separated from grape seed extract, the flavonoid compounds are separated from herba Selaginellae extract, and the gallic acid and catechin are separated from Hamamelis Virginiana extract.

Example 13

The cell regulation system (gel-like) was prepared according to the following combinations and ratios: 50 μ g/mL adenosine compounds +6 μ g/mL cytidine compounds +60 μ g/mL unsaturated fatty acids +30 μ g/mL saturated fatty acids +2g/mL protein substances (silky polyprens, astaxanthin yeast extract, fibroin) +50ng/mL flavonoids (amentoflavone and isocedar biflavone) + squalane + hyaluronic acid + ethyl palmitate + retinal + safflower extract + oligomeric procyanidins; wherein the nucleoside compounds, unsaturated fatty acids and saturated fatty acids are separated from grape seed extract and pomegranate seed extract, and the flavonoids are separated from herba Selaginellae extract.

Example 14

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +4 μ g/mL cytidine +5 μ g/mL guanosine +60ng/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL proteinaceous material (milk protein, various amino acids, as in example 1) + trace elements (iron, magnesium, calcium) +83 μ g/mL vitamin; wherein the nucleoside compounds (adenosine compounds, cytidine compounds, and guanosine compounds), unsaturated fatty acids, and saturated fatty acids are separated from grape seed extract and pomegranate seed extract, and the flavonoid compounds are separated from herba Selaginellae extract. The cells which use this regulatory system to act are mouse skin fibroblasts.

Example 15

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL proteinaceous (milk protein) +83 μ g/mL vitamin; wherein the nucleoside compounds are derived from Ganoderma extract, and are separated to obtain adenosine compounds and cytidine compounds, unsaturated fatty acids and saturated fatty acids are separated from herba Avenae Fatuae coarse powder extract, and flavonoids are separated from radix Changii extract.

Example 16

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL protein (milk protein) +83 μ g/mL vitamin; wherein the nucleoside compounds are derived from papaya fruits, and adenosine compounds, cytidine compounds and guanosine compounds are obtained by separation; the unsaturated fatty acids are derived from green tea extract, the saturated fatty acids are separated from cocoa extract, and the flavonoids are separated from cactus extract.

Example 17

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL protein (milk protein) +83 μ g/mL vitamin; wherein the nucleoside compounds are derived from radix rehmanniae extract, unsaturated fatty acids and saturated fatty acids are obtained by separating coconut fruit extract, and flavonoids are obtained by separating radix scrophulariae extract.

Example 18

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL protein (milk protein) +83 μ g/mL vitamin; wherein the nucleoside compounds and saturated fatty acids are separated from grape seed extract, the unsaturated fatty acids are separated from Agaricus bisporus extract, and the flavonoids are separated from fructus Amomi rotundus seed extract.

The vitamins described in this example contain primarily vitamin E, vitamin K, ascorbic acid species, folic acid, riboflavin, inositol, thiamine HCl.

Example 19

The cell regulation system (liquid) was prepared according to the following combinations and ratios: 20 μ g/mL adenosine +2.5 μ g/mL cytidine +3 μ g/mL guanosine +60 μ g/mL unsaturated fatty acid +30 μ g/mL saturated fatty acid +600mg/mL proteinaceous (milk protein) +83 μ g/mL vitamin; wherein the adenosine and cytidine compounds are separated from semen Granati extract, guanosine is purchased from market, unsaturated fatty acid is separated from tea extract, saturated fatty acid is separated from palm oil, and flavonoid is separated from radix Angelicae sinensis extract.

Examples 5 to 19 differ from the cell regulation system, and the methods for culturing cells and detecting collagen secretion in skin fibroblasts are the same as those of steps 2 and 3 of example 4; unless otherwise specified, the cells are of human origin. In examples 5 to 19, the collagen expression levels detected by sirius red staining are shown in table 1.

TABLE 1

Example 20

From the above experimental results, it was found that the regulation system in example 2 has the highest expression level of collagen in cells, and thus the skin fibroblasts derived from different ages of human origin were cultured and treated using the regulation system in example 2 (denoted as serum in this example): the cell culture solution is DMEM containing 20% of a regulating system (essence water); control group was DMEM with 10% fbs (dm). After 3 days of treatment, the amount of type1 procollagen secretion in the cell culture broth was measured using the procollagen type1 kit from Takara (FIG. 15A); the result shows that the collagen content of the cells is increased after the treatment by the essence water, and the collagen content of the middle-aged and old people (30-90 years old) is obviously increased; according to the content detection of type1 procollagen, after the treatment of essence water, the cell collagen synthesis and secretion capacity of an aged individual (80-90 years old) tends to the level of 50-60 years old individual, the cell collagen synthesis and secretion capacity of a 50-60 years old individual tends to the level of 30-40 years old individual, and the cell collagen synthesis and secretion capacity of a 30-40 years old individual tends to the level of 20-30 years old individual (fig. 15B).

The human adipose-derived mesenchymal stem cells were treated using the conditioning system (labeled serum in this example) of example 2, and the treated cell culture solution was a conditioning system (serum) containing 20% in DMEM. After 5 days of culture treatment, the mRNA expression levels of the relevant cytokines IGF1, VEGF and FGF2 in the cells were examined, and the results showed: the expression level of the relevant cytokine is increased, which is favorable for maintaining the cell activity (FIG. 16).

Example 21

Human dermal fibroblasts, epidermal cells, vascular endothelial cells, smooth muscle cells and adipose mesenchymal stem cells were treated with the regulatory system of example 2 in DMEM with 20% of the cell culture medium. After 6 days of culture treatment, the expression levels of senescence-associated genes CDKN1A, CDKN2A and IL6 in the cells were measured by a quantitative PCR method, and the results showed that: after the treatment with the regulatory system in example 2, the expression levels of senescence-associated genes CDKN1A, CDKN2A and IL6 were decreased (fig. 17).

Example 22

Using the regulatory system of example 2, human-derived dermal fibroblasts from different ages were treated and grouped by age as over 60 Years (60+ Years), 30 to 60 Years (30+ Years), 20 to 30 Years (20+ Years), 10 to 20 Years (10+ Years), and the cell culture medium used for the treatment was DMEM with 20% of the regulatory system. After 12 days of treatment, the expression level of CBX1 gene was measured. The results show that the epigenetic modifier CBX1 is highly expressed in young cells, and that the expression of CBX1 is significantly increased in individuals aged 60 (60+ Years), 30 to 60 (30+ Years) and 20 to 30 (20+ Years) after treatment with the regulatory system of this example (fig. 18).

Dermal fibroblasts from individuals of 20-40 years old of human origin were treated using the regulatory system of example 2. Culturing skin fibroblast from primary isolated to third generation in vitro with culture solution of control group (DMEM containing 10% FBS) and experiment group (DMEM containing 20% regulatory system), respectively, culturing in carbon dioxide incubator, and culturing in CO₂Culturing at 37 deg.C with concentration of 5%; two groups of cells were serially passaged simultaneously, and when the cells were passaged to the 10 th generation, the cells were collected, cell DNAs were extracted, methylation sequencing was performed, and epigenetic clock was calculated by analyzing the whole genome methylation level (DNA methylation, abbreviated as DNAm) (Mieko Matsuyama et al, 2019; labeled as DNAm Age). The results showed that, without treatment using the regulatory system of this example, DNAm age (DNAm age) increased significantly, while the increase of DNAm age by the regulatory system was smaller in magnitude, with no significant difference compared to the initial generation (fig. 19).

Example 23

Human dermal fibroblasts were treated using the conditioning system of example 2, with the cell culture medium being a 20% (v/v) conditioning system in DMEM and the control being a 10% FBS culture in DMEM. After 6 days of culture treatment, the expression level of SIRT3 gene in the cells was measured by quantitative PCR. The results showed that SIRT3, a gene involved in regulating the metabolic activity of mitochondria of cells, was highly expressed after treating the cells with the regulatory system of example 2, compared to the control group (fig. 20).

Example 24

Human dermal fibroblasts, epidermal cells and vascular endothelial cells were treated with the regulatory system of example 1, respectively, in DMEM containing 20% of the regulatory system. After 7 days of culture treatment, the change in the expression level of a gene (clock, BMAL1, RORA) involved in the regulation of cell and cell rhythm/biological clock was examined, and the results showed that: the expression level of the gene (clock, BMAL1, RORA) involved in the regulation of cell rhythm was increased after the treatment with the regulation system of example 1 (FIGS. 21 to 23): the expression of clock, BMAL1 and RORA is beneficial for maintaining normal biological clock regulation of cells, and is related to the state of cell rejuvenation.

Example 25

Human dermal fibroblasts and epidermal cells were treated with the regulatory system of example 1, respectively, in DMEM containing 20% of the regulatory system. After 6 days of culture treatment, the mRNA expression level of the relevant cell secretion factor or protein in the cells is detected, and the result shows that: in skin fibroblasts, after treatment, the expression level of cytokines related to collagen synthesis such as Epidermal Growth Factor (EGF), fibroblast growth factor 2(FGF2), Hepatocyte Growth Factor (HGF) and Keratinocyte Growth Factor (KGF) is increased; in addition, the expression level of elastin and filaggrin is also increased (fig. 24), which is beneficial to promoting the generation of reticular fibers and elastic fibers; in epidermal cells, the expression level of tissue inhibitor of metalloproteinase 1(TIMP1) that inhibits secretion of matrix metalloproteinase after treatment was increased (fig. 25), which is advantageous for preventing collagen degradation and protecting collagen.

Aged human dermal fibroblasts, which contained a large number of senescent cells (which appeared blue after staining with β -galactosidase), were treated using the regulatory system of example 1. The conditioned medium was DMEM containing 20% of the conditioning system, and after 3 days of conditioning, beta-galactosidase staining was identified, which showed a significant reduction in senescent cells (blue) in the sample (FIG. 26).

Example 26

Detection of skin moisture content: the conditioning system of example 1 (labeled as extract in this example) was tested on 80 volunteers. When in test, the environmental humidity is 20-30%, the subjects use the refined liquid on the facial skin after cleaning the face, and the skin moisture test is carried out after the refined liquid is used for 4 hours; the control group was 50 individuals, who used their own daily skin care products. The results showed that the skin moisture remained good and the moisture content increased after 4 hours of application of the extract to the volunteers (fig. 27).

Detecting the skin water content of the people staying up all night: each group of 25 persons (25-40 years old), one group used their own daily skin care product (control group), and one group used their own daily facial cream after the essence solution (adjustment system in example 1), and the subjects who entered the daily habit sleep after 12 am. When in test, the environmental humidity is 20-30%, the water content of the skin is detected after the skin is continuously used for 7 days, and the water content of the skin of people with the refined liquid group is increased (figure 28).

Detection of skin pH: and (4) bringing the recovery group into 38 persons, wherein the pH value of the facial skin of the person is more than or equal to 5.6, continuously using the refined liquid for 14 days, and then detecting the pH value of the facial skin of the subject again, wherein the results show that the pH value is reduced after using the refined liquid, and most of the pH value is recovered to be in a normal pH range. The normal group included 64 persons (facial skin PH within normal range), and after 14 days of continuous use of the essence, the facial skin PH of the subjects was checked again, and the results showed that the PH was still within normal range without abnormality after use (fig. 29).

Example 27

The volunteers used the cell-regulating system of example 1 in combination with their own cream for 24 months, and then stopped using the regulating system for 3 months and used only the subject's own cream. Detecting the moisture content of the face skin of the subject before use, 24 months after use, 1 month after use stop, 2 months after use stop and 3 months after use stop respectively; when in test, the environmental humidity is 20-30%. The detection result shows that the skin condition is good when the subject uses the regulating system for a long time, particularly the moisture content of the skin is improved and kept good; after cessation of use, the skin was well conditioned, in particular as a result of the skin maintaining a high moisture content (fig. 30). The regulation system is prompted to recover and maintain the healthy state of cells and does not generate side effects such as 'use dependence' and the like.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims (16)

1. An endogenous cellular regulatory system comprising at least 2 of a nucleoside compound, an unsaturated fatty acid, a saturated fatty acid, and a proteinaceous substance;

the nucleoside compound is at least one selected from adenosine compounds, cytidine compounds and guanosine compounds.

2. The regulatory system of claim 1, wherein the proteinaceous material is selected from the group consisting of animal placental protein, animal umbilical cord extract, oligopeptides, polypeptides, silk amino acids, silk meal, sericin, silk extract, methionine, egg meal, egg yolk extract, eggshell membrane extract, egg white extract, dipeptide-1, dipeptide-2, dipeptide-4, glutamine, glutathione, citrulline, oligonucleotides, cystine, collagen amino acids, collagen extract, keratin amino acids, yeast fermentation product extract, yeast/soy protein fermentation product, yeast/barley seed fermentation product filtrate, yeast/grape fermentation product extract, yeast/barley fermentation product extract, and combinations thereof, Yeast lysate extract, yeast/rice bran fermentation product, yeast polypeptides, yeast fermentation product filtrate, yeast fermentation lysate filtrate, yeast lysate extract, arginine, chitosamine, milk amino acids, milk protein extract, proline, glucosamine, lactic acid, milk protein, hydrolyzed silk, hydrolyzed soy protein, hydrolyzed rice protein, tyrosine, casein, tyrosine methyl ester HCL, potassium caseinate, sodium caseinate, casein extract, buttermilk powder, ornithine, nisin, whey protein, orotic acid, lactoglobulin, lactoferrin, hydrolyzed elastin, hydrolyzed eggshell membrane, hydrolyzed collagen, hydrolyzed keratin-propylmethylsilanediol, hydrolyzed keratin ethyl PG, hydrolyzed keratin, glucose, One or more of hydrolyzed yeast, hydrolyzed yeast protein, hydrolyzed yeast extract, hydrolyzed placenta caprae seu ovis protein, hydrolyzed placenta caprae seu ovis extract, sericin, fibroin, tetrapeptide-1, tetrapeptide-4, aspartic acid, asparagine, serum albumin, serum protein, and histidine.

3. The regulatory system of claim 1, wherein the proteinaceous material is selected from one or more of milk protein, hydrolyzed yeast extract, hydrolyzed fibroin, oligopeptides, sericin, tyrosine, hydrolyzed collagen, sericin, hydrolyzed silk, fibroin, yeast polypeptides, histidine, proline, tyrosine, ornithine, aspartic acid, arginine, cystine, methionine, oligopeptides, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, palmitoyl tetrapeptide-10, xanthan gum, carnosine, serine, and astaxanthin yeast extract.

4. The regulatory system of claim 1, wherein the nucleosides, unsaturated fatty acids, saturated fatty acids, and flavonoids are extracted from natural products; the molecular weight range of the nucleoside compound is 80-8000; the molecular weight range of the unsaturated fatty acid is 100-3000; the molecular weight range of the saturated fatty acid is 100-3000; the molecular weight range of the flavonoid is 100-2100.

5. The regulating system according to claim 1, wherein the content of nucleoside compounds in the regulating system is 10ng/mL to 500ug/mL, the content of unsaturated fatty acids is 10ng/mL to 500ug/mL, the content of saturated fatty acids is 10ng/mL to 500ug/mL, and the content of protein substances is 100mg/mL to 10 g/mL.

6. The conditioning system of claim 1 further comprising flavonoids in an amount of 1ng to 800 ng/mL.

7. The regulation system of claim 6 wherein the flavonoid is one or more of amentoflavone, quercetin, wogonin, isoquercitrin, glycoside flavonoid, naringenin, soy isoflavone, quercetin, bioflavonoid, sumac and 4', 6, 7-trihydroxyisoflavone.

8. The conditioning system according to claim 1, wherein the conditioning system further comprises a skin care ingredient a and/or a skin care ingredient b;

the skin care ingredient a is selected from one or more of antimicrobial ingredients, humectants, moisturizers, UV absorbers, epidermal barrier function modulators, skin rejuvenating and regenerating ingredients, skin tightening and anti-wrinkle agents, soothing and anti-inflammatory agents, anti-itch ingredients, antioxidants, free radical scavengers, UV-quenchers, sebum-regulating and anti-acne agents, stretch mark modulators, skin immune system modulators, skin lightening agents, skin microcirculation modulators, microfilaments and telangiectasia modulators, and tissue metalloproteinase inhibitors;

the skin care ingredient B is selected from the group consisting of proteins, ellagic acid, growth factors, coenzymes, enzyme inhibitors, peptides, carbohydrates, polysaccharides, glycosaminoglycans, glycosaminoglycan subunits, lipids, sphingosine, sphingolipids, glycosphingolipids, sulpholipids, phospholipids, sterols, phytosterols, saponins, phenols, polyphenols, terpenes, alkaloids, provitamins, vitamin A group, vitamin B group, ascorbates, vitamin D, vitamin E, vitamin K, folic acid, riboflavin, inositol, thiamine HCl, rosin oil, retinal, tretinoin, isotretinoin, alitretinoin, abamectin esters, avilamda, chelating agents, creatinine, dimethylethanolamine, serine, glycine, aspartic acid, cysteine, glutamine, lysine, arginine, aspartic acid, glutamic acid, N-acetylcysteine, citrulline, collagen, and mixtures thereof, Gelatin, albumin, sericin, fibroin, anti-keratin microfilament aggrecan, transforming growth factor, insulin-like growth factor, epidermal growth factor, acidic and basic fibroblast growth factor, nerve growth factor, keratinocyte growth factor, hepatocyte growth factor, platelet-derived growth factor, granulocyte-macrophage colony stimulating factor, vascular endothelial cell growth factor, trace elements, hydrogenated lecithin, hydrolyzed collagen, xanthan gum, sodium polyglutamate, carnosine, coenzyme Q10, nicotinamide adenine dinucleotide, glucose, resveratrol, fructose, mannose, dihydroxyacetone, erythrulose, sucrose, trehalose, maltose, galactomannan, glucomannan, beta-glucan, carrageenan, glycogen, chitosan, lentinan, lichenin, alpha-glucosidase, collagen, or a mixture thereof, Inulin, fucose, alginate, xyloglucan, dextran, amylose, levan, amylopectin, hyaluronic acid, chondroitin sulfate, heparin, dermatan sulfate, glucuronic acid, oat β -glucan, N-acetylglucosamine, allantoin, ceramide, N-lipoyl sphingosine glucoside, cerebroside, ganglioside, sulfatide, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, lactic acid, citric acid, glycolic acid, azelaic acid, salicylic acid, lipoic acid, pyrrolidone carboxylic acid, urinary acid, caffeic acid, conjugated linoleic acid, squalane, squalene, monoglycerides, diglycerides, triglycerides, petrolatum, lanolin, phytosphingosine, sitosterol, stigmasterol, brassicasterol, lupeol, glycyrrhizin, rutin, genistein, soybean, nonsetrone, nonsetretin, dextran, heparin, dermatan sulfate, heparin, dermatan sulfate, glucuronide, phosphatidyl-N-acetyl-glucosaminyl, salicylic acid, lipoic acid, pyrrolidone carboxylic acid, uric acid, caffeic, conjugated linoleic acid, squalane, squalene, monoglyceride, diglyceride, triglyceride, phytosterol, and phytolaccoline, Myricetin, luteolin, hesperetin, silibinin, silymarin, apigenin, epigallocatechin gallate, nordihydroguaiaretic acid, ellagic acid resorcinol, glycyrrhetinic acid, farnesol, alpha-bisabolol, beta-bisabolol, theophylline, theobromine, usnic acid, zinc-, selenium-, manganese-salt, glycerol, propylene glycol, butylene glycol, sorbitol, erythritol, hexylene glycol, phytantriol, carbomer, polysorbate-20, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, diglycerol, octanoyl hydroxamic acid, 1, 2-hexanediol, 1, 3-propylene glycol, cetyl palmitate, caprylic capric triglyceride, palmitoyl tetrapeptide-10, polysorbate-60, sorbitan monostearate, polyethylene glycol-8, polyethylene glycol, Betaine, sclerotium rolfsii, sodium hyaluronate, defensin, cathepsin, histatin, salicylate, cinnamate, caffeine, filaggrin, melatonin, urea, papain, bromelain, superoxide dismutase, lactoperoxidase, phospholipase, sphingomyelinase, transglutaminase, tranexamic acid, alpha 1-antitrypsin, leupeptin, kinetin, anthranilate, alpha-carotene, beta-carotene, lycopene, luteolin, zeaxanthin, palm oil, conventional cosmetic natural extracts, tea tree oil, oligomeric procyanidins, and polyhydric alcohols.

9. The conditioning system according to claim 1, wherein the product form of the conditioning system is in the form of a solution, emulsion, lotion, dispersion, powder, ointment, gel form, viscous surfactant, emulsifying polymer, pomade, hair tonic, shampoo, soap, gel, lyophilizate, powder, bar, pen, spray, body oil, mask or patch.

10. Use of the regulatory system according to any of claims 1 to 9 for endogenous repair of tissue cells and activation of cellular functions; and reducing senescent cells and restoring the youthful state of the cells.

11. Use of a regulatory system according to any of claims 1 to 9 for promoting tissue cell secretion of cytokines useful for repair and regeneration.

12. Use of the modulation system according to any one of claims 1 to 9 for the modulation of the cell rhythm/biological clock of tissue cells; and is used for epigenetic regulation of tissue cells and reduction of DNA epigenetic clock age.

13. Use of the regulatory system of any one of claims 1 to 9 for enhancing the oxidative metabolic function of mitochondria in tissue cells.

14. Use of the regulatory system of claim 10, wherein the tissue cells are one or more of epidermal cells, epidermal stem cells, fibroblasts, adipocytes, adipose-derived mesenchymal stem cells and vascular endothelial cells.

15. Use of a conditioning system according to any of claims 1 to 9 for the preparation of a cosmetic product.

16. Use of a regulatory system according to any of claims 1 to 9 for the preparation of a cell culture medium or a serum-free cell culture medium for skin tissue cells.

Images