CN116650380B - Anti-aging tea fermentation product for improving skin microcirculation and preparation method and application thereof - Google Patents
Anti-aging tea fermentation product for improving skin microcirculation and preparation method and application thereof Download PDFInfo
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- CN116650380B CN116650380B CN202310746182.6A CN202310746182A CN116650380B CN 116650380 B CN116650380 B CN 116650380B CN 202310746182 A CN202310746182 A CN 202310746182A CN 116650380 B CN116650380 B CN 116650380B
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
- A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
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- Cosmetics (AREA)
Abstract
The invention discloses an anti-aging tea fermentation product for improving skin microcirculation, and a preparation method and application thereof. The tea fermentation product provided by the invention is a fermentation product obtained by fermenting tea extract, pipecolic acid and zymophyte; mixing the folium Camelliae sinensis extract containing pipecolic acid with zymocyte, and dynamically fermenting to obtain the final product. The tea fermentation product provided by the invention can regulate and control TRPV4 signal pathway, increase endothelial Ca 2+ inflow, activate myosin light chain kinase, stimulate nitric oxide synthesis, activate a negative feedback loop mediated by nitric oxide, and increase vascular permeability through cGMP dependent attenuation, thereby protecting vascular barrier and achieving the effect of improving skin microcirculation. Meanwhile, the tea fermentation product can also improve the synthesis of type I collagen and hyaluronic acid of skin cells, and inhibit cells from secreting inflammatory factors by regulating and controlling transient TRPV3 and TRPM8 signal paths so as to achieve the effect of resisting cell aging.
Description
Technical Field
The invention belongs to the technical field of plant microorganism fermentation. More particularly, relates to an anti-aging tea fermentation product for improving skin microcirculation, a preparation method and application thereof.
Background
With age, the environmental problems are becoming more serious and the pressure from life and work is increasing gradually, and skin aging is unavoidable. Skin aging is the result of the combined action of the physiological mechanism of human body aging solid-liquid and the damage of external wind, sun, environmental pollution and the like. The clinical symptoms of skin aging are that skin is easy to dry, skin elasticity is poor, skin is thin, and wrinkles appear; the activity of sebaceous glands is reduced, and secretion is reduced; pigment spots grow out; the recovery of the skin after injury becomes slow and the skin becomes more prone to bruise. How to effectively resist skin aging and keep youth and charm is a difficult problem that people loving beauty have to face.
Skin microcirculation is a system of mutual exchange of substance, energy and biological information between cells and tissues, and comprises blood vessels with the diameter of less than 150 mu m, namely arteries, venules, lymphatic vessels and arteriovenous anastomosis. The microcirculation can provide oxygen and nutrients for keratinocytes and fibroblasts of the skin, promote the epidermal repair (accelerating the turnover and metabolism of the keratinocytes) and the turnover and metabolism of the fibroblasts, promote the formation of more collagen and slow down the skin aging. And when microcirculation damage occurs: (1) Blood stasis can not provide nutrition for cells, so that each layer of epidermis is damaged due to the fact that nutrition can not be obtained, and red blood streaks can be generated on the skin; (2) The pressure of the vein in the blood vessel is increased, so that the water in the blood vessel flows out of the blood vessel along the pressure difference and then enters the tissue to be accumulated in the tissue, and edema is caused; (3) Ischemia damage occurs on tissue cells, oxidation and phosphorylation in mitochondria are blocked, ATP is decomposed in a large amount to generate a large amount of free radicals, the free radicals attack healthy cells and aggravate cell damage, so that the change of cell structure and function can accelerate the aging of skin to a certain extent, thereby effectively avoiding and improving skin microcirculation, keeping skin in a 'healthy state', and effectively relieving skin aging.
Currently, methods for improving skin microcirculation are: the medicine improvement, the food therapy improvement and the movement improvement mainly comprise the steps of dilating micro blood vessels and accelerating blood flow, thereby increasing blood flow; the food therapy and the exercise improvement are adopted, the period is long, and the effect is very little. The skin care products in the current market are various, but a plurality of skin care products need to be added with preservative or essence components, and the chemical reagents have certain irritation to human bodies, are easy to cause skin allergy and have potential safety hazards. The prior art discloses a composition for promoting skin microcirculation, which takes pollen pini extract, dunaliella extract, sparassis crispa extract and oat kernel extract as active ingredients, adopts natural plant extract, has safe ingredients, can effectively promote skin blood microcirculation, accelerate skin self-renewal and start anti-aging repair, but has complex ingredients and high preparation ingredients, and the effect is still to be improved. Therefore, there is a need to develop more natural plant components which have better effects and natural non-irritation and can improve skin microcirculation and have anti-aging function, replace the traditional cosmeceutical products, fundamentally adjust and improve skin surface microcirculation, and have mild and non-irritation effects, thereby solving the skin problem.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings of the problems and provide an anti-aging tea fermentation product for improving skin microcirculation, and a preparation method and application thereof.
The invention aims to provide an anti-aging tea fermentation product for improving skin microcirculation.
It is another object of the present invention to provide a process for the preparation of the tea fermentation product.
It is a further object of the present invention to provide the use of the tea fermentation product.
It is still another object of the present invention to provide a cosmetic or skin care product that is anti-aging, anti-inflammatory, regulates vascular contractions, promotes skin barrier repair and/or improves skin microcirculation.
The above object of the present invention is achieved by the following technical scheme:
the invention provides an anti-aging tea fermentation product for improving skin microcirculation, which is a fermentation product obtained by fermenting tea extract, pipecolic acid and zymophyte.
The tea adopted by the invention belongs to a green natural product, has lower toxicity and is suitable for being applied to skin care products. A large number of researches prove that the tea contains theabrownin, polyphenol and other substances, and has remarkable antioxidant, anti-inflammatory and anti-wrinkle effects.
The pipecolic acid (Pipecolic acid, PA) is an organic compound of the formula C 5H11NO2, belonging to the class of amides, belonging to the class of organic chemicals like amino acids, consisting of amino acids and alpha-hydroxy acids, located at terminal positions in the amino acid spectrum. PA has various physiological functions of regulating blood circulation, resisting oxidation, protecting nervous system and the like, and is an important bioactive substance.
The invention adopts the microbial fermentation technology, and utilizes microbial cells or an intracellular enzyme catalytic reaction system to carry out structural modification and reformation on active substances in plant raw materials, thereby obtaining valuable fermentation liquor. The active substance obtained after the fermentation of the plant components has higher content, smaller molecular weight after microbial transformation, and better regulating effect on the skin microecology of people.
The invention prepares the tea fermentation product by adding the Pipecolic Acid (PA) into the natural plant tea extract and mixing, inoculating zymocyte for fermentation; the tea fermentation product not only can improve skin microcirculation, but also has good anti-aging effect. The tea fermentation product provided by the invention well reserves the effective components in tea, realizes the bidirectional fermentation of probiotics and tea after being fermented by the synergistic fermentation bacteria of the pimaric acid, and promotes the tea fermentation product to have better skin care effects of moisturizing, anti-aging, anti-inflammatory, skin barrier repair and the like; the PA and the tea extract are compounded and used, so that the effect is further improved, the TRPV4 signal pathway is regulated and controlled, the endothelial Ca 2+ inflow is increased, meanwhile, the myosin light chain kinase is activated, the nitric oxide synthesis is stimulated, and a negative feedback loop mediated by nitric oxide is activated, and the loop increases the vascular permeability through cGMP dependent attenuation, so that the vascular barrier is protected, and the effect of improving the skin microcirculation is achieved; can also obviously improve the synthesis of the type I collagen and hyaluronic acid of skin cells, and the secretion of inflammatory factors by the cells, and has the skin care effects of resisting inflammation, promoting skin barrier repair and the like. In addition, the tea fermentation product provided by the invention has mild components, no stimulation and high safety, and can be used as an efficacy additive to be added into skin care products and cosmetics.
Further, the tea leaf extract is obtained by drying and crushing tea leaves, extracting with water, and extracting under heating and reflux.
Further, the pipecolic acid is pipecolic acid aqueous solution with the concentration of 0.7 mol/L-1 mol/L, and the addition amount is 14% -34% of the volume of the tea extract.
Further, the zymocyte is bacillus subtilis, the concentration of the zymocyte is 10 6～108 CFU/mL, and the inoculation amount is 2-4% of the volume of the mixed solution of the tea extract and the pipecolic acid.
More preferably, the inoculum size of the fermentation tubes is 2%.
The invention provides a preparation method of a tea fermentation product, which comprises the following steps:
S1, cleaning fresh tea leaves, removing impurities, drying, crushing, sieving, and adding a feed liquid with a ratio of 1:8-10 pure water, soaking, sterilizing, heating and reflux extracting to obtain tea leaf extract;
s2, adding a pipecolic acid solution into the tea extract and mixing to obtain the tea extract containing PA;
S3, inoculating fermentation bacteria into the tea extract containing PA, performing dynamic fermentation, sterilizing after fermentation, and filtering to obtain a tea fermentation product.
Preferably, the drying in step S1 is a drying treatment at 80-100℃for 1-3 hours.
Preferably, the tea leaves in step S1 are fresh tea leaves, including but not limited to black tea.
Preferably, the dynamic fermentation conditions in step S3 are: stirring at constant temperature of 35-40 ℃ for 36-90 h at stirring speed of 100-150 rpm.
More preferably, the dynamic fermentation conditions in step S3 are: stirring was carried out at 37℃for 72h with a stirring speed of 120 rpm.
The invention provides an application of a tea fermentation product in preparing cosmetics or skin care products for resisting aging, resisting inflammation, regulating vasoconstriction, promoting skin barrier repair and/or improving skin microcirculation.
The invention also provides a cosmetic or skin care product which has the functions of resisting aging, resisting inflammation, regulating vasoconstriction, promoting skin barrier repair and/or improving skin microcirculation, and the tea fermentation product.
Preferably, the cosmetic or skin care product contains 2-5% of tea fermentation product by weight percent.
More preferably, the cosmetic or skin care product is an essence, cream, emulsion, water or paste product, further contains other basic components commonly used in the art, such as moisturizer, antioxidant, repairing agent, antibacterial agent, moistening agent, etc., and specifically can be added with: p-hydroxyacetophenone, 1, 2-hexanediol, carbomer 941, triethanolamine, disodium EDTA, and the like.
Further preferably, the cosmetic or skin care product contains 0.4% of hydroxyacetophenone, 0.6% of 1, 2-hexanediol, 0.1% of carbomer 941.1%, 0.1% of triethanolamine, 0.02% of EDTA disodium and the balance of deionized water in percentage by weight, and the black tea essence is prepared after uniform mixing.
The invention has the following beneficial effects:
The invention provides an anti-aging tea fermentation product for improving skin microcirculation and a preparation method thereof, and the tea fermentation product provided by the invention is a fermentation product obtained by fermenting tea extract, pipecolic acid and zymophyte; the method has the advantages that the effective components of the tea are extracted, and the pipecolic acid is further added, so that the prepared tea fermentation product has better skin care effects of improving microcirculation, resisting aging, resisting inflammation, promoting skin barrier repair and the like under the fermentation of probiotics. The invention combines PA and tea extract to have synergistic effect, can increase endothelial Ca 2+ inflow by regulating TRPV4 signal path, activate myosin light chain kinase, stimulate nitric oxide synthesis, and activate nitric oxide-mediated negative feedback loop, which increases vascular permeability by cGMP dependent attenuation, thereby protecting vascular barrier and achieving the effect of improving skin microcirculation. The tea fermentation product can also improve the synthesis of type I collagen and hyaluronic acid of skin cells, and the cells secrete inflammatory factors, and has the effects of resisting inflammation, promoting skin barrier repair and the like; in addition, the tea fermentation product provided by the invention has mild components, no irritation and high safety, and can be used as an efficacy additive to be added into skin care products and cosmetics.
The invention has the advantages of effectively simplifying the production process and leading the reaction process to be carried out under mild conditions. Meanwhile, the fermentation extraction method can also change the molecular structure of the raw material components, improve the efficacy of the raw materials, reduce toxicity and irritation, optimize color and taste, and the like, thereby being an ideal novel process for extracting the traditional Chinese medicine and being beneficial to popularization and application.
Drawings
FIG. 1 is a diagram showing the results of detecting the cell expressing TRPV4, TRPV3, TRPM8 genes;
FIG. 2 is a graph showing the expression level of myosin light chain kinase;
FIG. 3 is a graph showing the expression level of inflammatory factor COX-2 in keratinocytes;
FIG. 4 is a graph showing the results of the secretion test of type I collagen;
FIG. 5 is a graph showing the results of hyaluronic acid secretion test.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
The Bacillus subtilis (accession number MGNA-B207) and the Bacillus bifidus (accession number ATCC-15696) used in the following examples are laboratory deposited strains;
the HaCaT cells, keratinocytes, human fibroblast HSF used are all cell lines used in the laboratory preservation of the present invention.
Example 1 preparation of tea fermentation product
1. Preparation of PA-containing tea extract
Cleaning tea leaves, removing sludge, dust and impurities, drying at 90 ℃ for 1h, and removing water to obtain dried tea leaves treated matters; crushing the dried tea leaves, and sieving with a No. 2 sieve (850+/-29 um); 1, the method comprises the following steps: 8-10, respectively adding pure water, soaking for 2h, sterilizing with steam at 121deg.C, cooling to room temperature, and reflux extracting for several times to obtain tea extractive solution.
A proper amount of Pipecolic Acid (PA) was dissolved in pure water to prepare a PA solution having a concentration of 0.7 mmol/L.
Subsequently, a PA solution accounting for 34% of the volume of the tea extract was added to the tea extract to obtain a PA-containing tea extract.
2. Preparation of fermentation broth
(1) Activating strains: picking a colony of bacillus subtilis, placing the colony in a liquid culture medium, and placing the colony in a shaking table to activate strains to obtain activated bacterial liquid;
(2) And (3) strain purification: the activated bacterial liquid is subjected to gradient dilution and plating so as to obtain single bacterial colonies, and purified bacterial strains are obtained;
(3) And (3) strain expansion culture: inoculating the purified strain into a BS culture medium (pH value is 7.0), standing and culturing in an incubator at 37 ℃, when the OD value=0.6-1.0, preparing fermentation broth with the concentration of 10 6～108 CFU/mL when the strain is in a logarithmic phase, and adjusting the pH value of the fermentation broth to 6.5-7.5 by using NaOH and HCl for later use.
3. Preparation of tea fermentation product
Taking a proper amount of tea leaf extract containing PA, inoculating fermentation bacteria liquid accounting for 2% of the volume of the tea leaf extract containing PA, and stirring at a constant temperature of 37 ℃ for 72 hours at a stirring speed of 120 revolutions per minute; and then sterilizing by steam at 121 ℃, cooling to room temperature, and filtering by adopting a filter plate with the diameter of 8-12 mu m to obtain the tea fermentation product.
Example 2 preparation of tea fermentation product
1. Preparation of PA-containing tea extract
Cleaning tea leaves, removing sludge, dust and impurities, drying at 90 ℃ for 1h, and removing water to obtain dried tea leaves treated matters; crushing the dried tea leaves, and sieving with a No. 2 sieve (850+/-29 um); 1, the method comprises the following steps: 8-10, respectively adding pure water, soaking for 2 hr, sterilizing with steam at 121deg.C, cooling to room temperature, and reflux extracting for several times to obtain tea extractive solution.
An appropriate amount of PA was dissolved in pure water to prepare a PA solution having a concentration of 0.7 mmol/L.
And adding a PA solution with the volume ratio of 20% to the tea extract into the tea extract. Obtaining the tea extract containing PA.
3. Preparation of fermentation broth
(1) Activating strains: picking a colony-ring in a liquid culture medium, and placing the colony-ring in a shaking table to activate strains to obtain activated bacterial liquid;
(2) And (3) strain purification: the activated bacterial liquid is subjected to gradient dilution and plating so as to obtain single bacterial colonies, and purified bacterial strains are obtained;
(3) And (3) strain expansion culture: inoculating the purified strain into a BS culture medium (pH value is 7.0), standing and culturing in an incubator at 37 ℃, when the OD value=0.6-1.0, preparing fermentation broth with the concentration of 10 6～108 CFU/mL when the strain is in a logarithmic phase, and adjusting the pH value of the fermentation broth to 6.5-7.5 by using NaOH and HCl for later use.
3. Preparation of tea fermentation product
Taking a proper amount of tea leaf extract containing PA, inoculating fermentation broth with volume of 3% of the mixed solution, and stirring at 37 ℃ for 72h at a stirring speed of 120 rpm; and then sterilizing by steam at 121 ℃, cooling to room temperature, and filtering by adopting a filter plate with the diameter of 8-12 mu m to obtain the tea fermentation product.
Example 3 preparation of tea fermentation product
1. Preparation of PA-containing tea extract
Cleaning tea leaves, removing sludge, dust and impurities, drying at 90 ℃ for 1h, and removing water to obtain dried tea leaves treated matters; crushing the dried tea leaves, and sieving with a No. 2 sieve (850+/-29 mu m); 1, the method comprises the following steps: 8-10, respectively adding pure water, soaking for 2 hr, sterilizing with steam at 121deg.C, cooling to room temperature, and reflux extracting for several times to obtain tea extractive solution.
An appropriate amount of PA was dissolved in pure water to prepare a PA solution having a concentration of 0.7 mmol/L.
And adding a PA solution with the volume ratio of 14% to the tea extract into the tea extract. Obtaining the tea extract containing PA.
4. Preparation of fermentation broth
(1) Activating strains: picking a colony-ring in a liquid culture medium, and placing the colony-ring in a shaking table to activate strains to obtain activated bacterial liquid;
(2) And (3) strain purification: the activated bacterial liquid is subjected to gradient dilution and plating so as to obtain single bacterial colonies, and purified bacterial strains are obtained;
(3) And (3) strain expansion culture: inoculating the purified strain into a BS culture medium (pH value is 7.0), standing and culturing in an incubator at 37 ℃, when the OD value=0.6-1.0, preparing fermentation broth with the concentration of 10 6～108 CFU/mL when the strain is in a logarithmic phase, and adjusting the pH value of the fermentation broth to 6.5-7.5 by using NaOH and HCl for later use.
3. Preparation of tea fermentation product
Taking a proper amount of tea leaf extract containing PA, inoculating fermentation broth with the volume of 4% of the mixed solution, and stirring at a constant temperature of 37 ℃ for 72h at a stirring speed of 120 rpm; and then sterilizing by steam at 121 ℃, cooling to room temperature, and filtering by adopting a filter plate with the diameter of 8-12 mu m to obtain the tea fermentation product.
Comparative example 1
The raw materials and the preparation method of this comparative example were the same as in example 3, except that the tea leaf extract was not added.
Comparative example 2
The raw materials and the preparation method of this comparative example were the same as in example 3, except that no PA solution was added.
Comparative example 3
The raw materials and extraction method of this comparative example were the same as in example 3, except that bifidobacteria were used as the fermentation species.
Comparative example 4
The raw materials and extraction method of this comparative example were the same as those of example 3, except that the tea leaf extract was prepared by using pure water of the same volume instead of the bacterial liquid without performing the bacterial additional fermentation.
Test example 1 real-time fluorescent RT-PCR detection of the mRNA expression level of an indicator Gene
Inoculating equal amount of HaCaT cells (purchased from Shanghai cell bank of China academy of sciences) into a 6-hole plate, respectively taking equal amount of tea fermentation products or extracting solutions prepared in the above examples 1-3 and comparative examples 1-4, adding pure water for dissolution, preparing tea fermentation products or extracting solution dissolution solutions with concentration of 2g/mL, adding into serum-free culture medium, co-culturing with the HaCaT cells, and extracting cell total RNA with TRIZOL reagent after 24 hours. Cells were lysed with TRIZOL reagent, and 500. Mu.L chloroform was added to the cell lysate, and the heart was separated for 10min at 12000 rpm. The RNA-containing supernatant was collected, precipitated with an equal volume of isopropanol and incubated for 30min. The RNA pellet was centrifuged at 12000 rpm for 10 minutes, the nucleic acid was collected, washed with 70% ethanol and air-dried, and then dissolved in 20. Mu.L of sterile double distilled water, followed by quantification of RNA using a micro ultraviolet-visible spectrophotometer. And converting the RNA into cDNA by using a cDNA conversion kit for quantifying polymerase chain reaction. Quantitative polymerase chain reaction technology is adopted to detect mRNA expression levels of TRPV4, TRPV3 and TRPM8 genes expressed by cells, and PBS is used for replacing the equivalent solution by a blank group.
Wherein, the detection cell expresses TRPV4, TRPV3 and TRPM8 genes specifically comprises the following steps: the subtype TRPV3 protein of transient receptor vanilloid (TRANSIENT RECEPTOR POTENTIAL VANILLOID, TRPV) can play a role in regulating inflammation production by regulating cell participation in inflammation and nociception; TRPV4 proteins are expressed in different vascular endothelial cells and are involved in a variety of vascular functions including regulation of vascular tone, mechanical signaling, etc., activation of TRPV4 channels increases endothelial Ca 2+ influx, while activating myosin light chain kinase, stimulating nitric oxide synthesis, and activating a nitric oxide-mediated negative feedback loop that increases vascular permeability through cyclic guanosine monophosphate (CYCLIC GMP, CGMP) -dependent attenuation, thereby protecting vascular barriers and improving skin microcirculation. TRPM8 of transient receptor potential channel protein M type (TRANSIENT RECEPTOR POTENTIALMELASTATIN, TRPM) family is also a main receptor, and exists in neurons, keratinocytes, vascular endothelial cells and other non-neuronal cells, and the activation of TRPM8 can inhibit skin inflammation, regulate vascular contraction, promote skin barrier repair and the like.
As shown in figure 1, compared with a blank group, the tea fermentation products or the extracting solutions prepared by the examples and the comparative examples can effectively up-regulate the mRNA levels of TRPV4 and TRPM8 and also down-regulate the mRNA levels of TRPV3, but the effects of the tea fermentation products prepared by the examples 1-3 are obviously better than those of the comparative examples 1-4, the effects of increasing the inner flow of endothelial Ca 2+ by up-regulating TRPV4 and TRPM8, and the effects of regulating vascular tension and mechanical signal transduction, increasing vascular permeability, well inhibiting skin inflammation, regulating vascular contraction and promoting skin barrier repair; simultaneously, TRPV3 is regulated downwards to regulate inflammation, thereby protecting vascular barrier, improving skin microcirculation, and the synergistic effect is achieved when PA and tea extract are compounded.
Test example 2Elisa analysis of the expression level of myosin light chain kinase
Taking HaCaT cells as a model, respectively taking equal amounts of tea fermentation products or extracting solutions prepared in the examples 1-3 and the comparative examples 1-4, adding pure water for dissolution, preparing tea fermentation products or extracting solution dissolution solutions with the concentration of 2g/mL, adding the tea fermentation products or extracting solution dissolution solutions into a serum-free culture medium for co-culture with the HaCaT cells (the using amount and the concentration of the extracting solution adopted in each example and the comparative example are the same), collecting the cells after 24 hours, and taking PBS instead of the solution with the same amount in a blank group. ELISA method is adopted to detect the influence of each group of tea fermentation products or extracting solutions on myosin light chain kinase, and the experimental result is analyzed by data processing software.
The ELISA method detection results are shown in FIG. 2, and show that compared with comparative examples 1-4, the examples 1-3 of the invention improve the level of myosin light chain kinase in HaCaT cells, thereby participating in the regulation of TRPV4 signal channels, protecting vascular barriers and achieving the effect of improving skin microcirculation.
Test example 3Elisa analysis of the effect of tea fermentation products on secretion of inflammatory mediators by human keratinocytes
In this example, the effect of the tea fermentation extract on the secretion of inflammatory mediators by human keratinocytes was examined to evaluate the efficacy of skin microcirculation. HaCaT cells grown in culture dishes to a confluence of about 80-90% (purchased from Shanghai cell Bank of China academy of sciences) were collected and counted in a sterile operating table. According to the counting result, inoculating 2.0-8.0X10 4 cells/well into a 96-well cell culture plate, treating cells with Lipopolysaccharide (LPS) to generate inflammation, establishing a keratinocyte inflammation model, adding 10 mu L PBS into each blank hole, adding the tea fermentation products or extracting solutions prepared in examples 1-3 and comparative examples 1-4 into sample holes respectively, wherein the concentration is 2g/mL, and three concentrations are parallel to each other, and culturing for 24 hours. The effect of the samples of examples 1-3 and comparative examples 1-4 above on the inflammatory factor COX-2 of keratinocytes was then examined using the Elisa kit.
As shown in FIG. 3, the results of the tests show that the tea fermentation products provided by the examples 1-3 are better than those of the comparative examples 1-4 in that the level of the inflammatory factor COX-2 can be effectively reduced in a lipopolysaccharide LPS model, and the effects are better than those of the comparative examples 1-4, so that the effects of resisting cell aging through anti-inflammation are achieved. The effect of compounding the PA and the tea extract is obviously better than that of comparative examples 1-4, and the synergistic effect of compounding the PA and the tea extract is achieved.
Test example 4Elisa analysis of the effect of tea fermentation products on fibroblasts
In this example, the anti-aging effect of the tea fermentation extract was evaluated by detecting the effect of the tea fermentation extract on the secretion of type I collagen by human fibroblasts and the secretion of hyaluronic acid by human fibroblasts. In a sterile operating table, the human fibroblast HSF grown to 80-90% in the petri dish was collected and counted. According to the counting result, 1mL to 12-well cell culture plates are inoculated according to 1.0 to 5.0X10 4 cells/mL, 100 mu L of PBS is added into blank wells, the extracting solutions prepared in the examples 1 to 3 and the comparative examples 1 to 4 are respectively added into sample wells, the concentrations are 2g/mL, three of the extracting solutions are parallel to each other, and the extracting solutions are cultured for 24 hours. The well-grown cell supernatant was collected in sterile tubes. Samples of examples 1 to 3 and comparative examples 1 to 4 were tested for secretion of type I collagen from human fibroblasts and hyaluronic acid from human fibroblasts using an Elisa kit.
The results of the type I collagen secretion test are shown in FIG. 4, and show that the tea fermentation products provided by the embodiments 1-3 of the invention can effectively increase the type I collagen secretion level of fibroblasts in a lipopolysaccharide LPS model and have a positive effect on improving skin aging compared with the comparison examples 1-4.
The results of hyaluronic acid secretion detection are shown in fig. 5, and show that compared with comparative examples 1-4, the tea fermentation products provided in examples 1-3 of the present invention can effectively increase the hyaluronic acid secretion level of fibroblasts in Lipopolysaccharide (LPS) model, thereby delaying aging.
Test example 5 safety detection of tea fermentation products
According to the embodiment, through a human body patch test, the safety detection is carried out on the black tea essence added with the tea hair product prepared by the invention, and the method for preparing the essence is as follows: 2 to 5 percent of fermentation products in examples 1 to 3 and comparative examples 1 to 4, 0.4 percent of p-hydroxyacetophenone, 0.6 percent of 1, 2-hexanediol, 0.1 percent of carbomer 941, 0.1 percent of triethanolamine, 0.02 percent of EDTA disodium and the balance of deionized water are respectively taken, and the black tea essence is respectively prepared after uniform mixing.
And selecting qualified spot test equipment with the area of 40mm 2 and the depth of about 1 mm. 20 healthy subjects with no history of dermatological allergy between 20 and 50 years of age were selected. Test treatment: 0.025mL of the test cosmetic was placed in a patch tester chamber, and the patch tester with the test substance was applied to the forearm curved side of the subject with hypoallergenic tape, and applied to the skin uniformly by gentle palm pressure for 24 hours. A blank control treatment was set for each test, no material was placed in the control plaque tester wells, and the other procedures were the same as the test group treatment. Skin reactions were observed 30min after removal of the subject plaque tester (after disappearance of the indentation), 24h and 48h respectively, according to the criteria described in table 1 below (the skin reaction grade of the plaque test was tested for safety of cosmetics according to the standard record in the "cosmetic health Specification"), and the observations were recorded.
TABLE 1 skin Patch test skin response grading criteria
The skin reaction grading standards according to the skin patch test described in table 1 above test cases where the extracts of examples 1 to 3 and comparative examples 1 to 4 were applied to the skin, and the results are shown in table 2, and the use of the tea fermentation essence products of examples 1 to 3 was more remarkable in safety than those of comparative examples 1 to 4.
TABLE 2 skin Patch test skin reaction results
In conclusion, the tea fermentation product prepared by dynamically fermenting the tea active ingredient extract containing the pipecolic acid can regulate and control TRPV4 signal paths, increase endothelial Ca 2+ inflow, activate myosin light chain kinase, stimulate nitric oxide synthesis and activate a nitric oxide-mediated negative feedback loop, and the loop increases vascular permeability through cGMP dependent attenuation so as to protect vascular barriers and achieve the effect of improving skin microcirculation; meanwhile, the synthesis of the type I collagen and hyaluronic acid of skin cells can be improved, and the cell secretion inflammatory factors are inhibited by regulating and controlling the transient TRPV3 signal channel and the TRPM8 signal channel, so that the effect of resisting cell aging is achieved.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (7)
1. An anti-aging tea fermentation product for improving skin microcirculation is characterized by being a fermentation product obtained by fermenting tea extract, pipecolic acid and zymophyte; the zymophyte is bacillus subtilis;
the preparation method of the product comprises the following steps:
S1, cleaning fresh tea leaves, removing impurities, drying, crushing, sieving, and adding a feed liquid with the ratio of 1:8-10 pure water, soaking, sterilizing, heating and reflux extracting to obtain tea leaf extract;
S2, adding a pipecolic acid solution into the tea extract and mixing to obtain the PA-containing tea extract;
S3, inoculating zymophyte into the tea extract containing PA, and carrying out dynamic fermentation: stirring at constant temperature of 35-40 ℃ for 36-90 h, stirring at speed of 100-150 rpm, sterilizing after fermentation, and filtering to obtain tea fermentation product.
2. The product according to claim 1, wherein the tea extract is obtained by drying and pulverizing tea, extracting with water, and heating and reflux extracting.
3. The product according to claim 1, wherein the pipecolic acid is aqueous solution of pipecolic acid with concentration of 0.7mol/L to 1mol/L and the addition amount is 14% -34% of the volume of the tea extract.
4. The product according to claim 1, wherein the concentration of the fermentation bacteria is 10 6～108 CFU/mL, and the inoculation amount is 2-4% of the volume of the mixed solution of the tea extract and the pipecolic acid.
5. Use of a tea fermentation product according to any one of claims 1 to 4 in the manufacture of a cosmetic or skin care product for anti-ageing, anti-inflammatory and/or promoting skin barrier repair.
6. A cosmetic or skin care product which is anti-ageing, anti-inflammatory and/or promotes the repair of skin barriers, characterized in that it comprises a tea fermentation product according to any one of claims 1 to 4.
7. A cosmetic or skin care product according to claim 6, wherein the cosmetic or skin care product comprises 2% to 5% by weight of the tea fermentation product according to any one of claims 1 to 4.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20090085246A (en) * | 2008-02-04 | 2009-08-07 | 한불화장품주식회사 | A lip cosmetic composition |
| CN102517235A (en) * | 2011-12-27 | 2012-06-27 | 湖南农业大学 | Bacillus subtilis |
| CN102647913A (en) * | 2009-10-15 | 2012-08-22 | 株式会社爱茉莉太平洋 | Composition for improving blood circulation and alleviating cold hands and feet, containing fermented tea extract |
| CN107184953A (en) * | 2017-05-22 | 2017-09-22 | 山东省科学院生物研究所 | A kind of antibacterial lipopeptid sebum film remediation composition and its application |
| CN110496163A (en) * | 2019-09-26 | 2019-11-26 | 江西中医药大学 | The preparation method of citrous bud or flower or branch or leaf or fruit active component or ingredient and its application in medicine preparation |
| CN110997694A (en) * | 2017-07-04 | 2020-04-10 | 路博润先进材料公司 | Compounds useful for the treatment and/or care of the skin, hair, nails and/or mucous membranes |
| CN112245538A (en) * | 2020-10-28 | 2021-01-22 | 广州环亚化妆品科技有限公司 | Traditional Chinese medicine composition, traditional Chinese medicine fermentation product containing traditional Chinese medicine composition, and preparation method and application of traditional Chinese medicine fermentation product |
| CN112772657A (en) * | 2021-01-12 | 2021-05-11 | 青岛农业大学 | Application of pipecolic acid as plant disease-resistant activator in preventing and treating apple leaf diseases |
| WO2022081762A1 (en) * | 2020-10-14 | 2022-04-21 | Dermbiont, Inc. | Compositions and methods for improving skin health and for the treatment and prevention of diseases, disorders and conditions associated with fungi and other pathogenic microbes |
| CN114601761A (en) * | 2022-01-18 | 2022-06-10 | 科丝美诗(中国)化妆品有限公司 | Preparation method and application of plant fermentation liquor for repairing skin barrier |
| KR20220141191A (en) * | 2021-04-12 | 2022-10-19 | 전은숙 | Composition for Skin Whitening, Moisturizing, Strengthening Skin Barrier, Skin Cell Regeneration, and Anti-Wrinkle Property Comprising Fermented Extract of Broussonetia kazinoki as Active Ingredient |
-
2023
- 2023-06-21 CN CN202310746182.6A patent/CN116650380B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20090085246A (en) * | 2008-02-04 | 2009-08-07 | 한불화장품주식회사 | A lip cosmetic composition |
| CN102647913A (en) * | 2009-10-15 | 2012-08-22 | 株式会社爱茉莉太平洋 | Composition for improving blood circulation and alleviating cold hands and feet, containing fermented tea extract |
| CN102517235A (en) * | 2011-12-27 | 2012-06-27 | 湖南农业大学 | Bacillus subtilis |
| CN107184953A (en) * | 2017-05-22 | 2017-09-22 | 山东省科学院生物研究所 | A kind of antibacterial lipopeptid sebum film remediation composition and its application |
| CN110997694A (en) * | 2017-07-04 | 2020-04-10 | 路博润先进材料公司 | Compounds useful for the treatment and/or care of the skin, hair, nails and/or mucous membranes |
| CN110496163A (en) * | 2019-09-26 | 2019-11-26 | 江西中医药大学 | The preparation method of citrous bud or flower or branch or leaf or fruit active component or ingredient and its application in medicine preparation |
| WO2022081762A1 (en) * | 2020-10-14 | 2022-04-21 | Dermbiont, Inc. | Compositions and methods for improving skin health and for the treatment and prevention of diseases, disorders and conditions associated with fungi and other pathogenic microbes |
| CN112245538A (en) * | 2020-10-28 | 2021-01-22 | 广州环亚化妆品科技有限公司 | Traditional Chinese medicine composition, traditional Chinese medicine fermentation product containing traditional Chinese medicine composition, and preparation method and application of traditional Chinese medicine fermentation product |
| CN112772657A (en) * | 2021-01-12 | 2021-05-11 | 青岛农业大学 | Application of pipecolic acid as plant disease-resistant activator in preventing and treating apple leaf diseases |
| KR20220141191A (en) * | 2021-04-12 | 2022-10-19 | 전은숙 | Composition for Skin Whitening, Moisturizing, Strengthening Skin Barrier, Skin Cell Regeneration, and Anti-Wrinkle Property Comprising Fermented Extract of Broussonetia kazinoki as Active Ingredient |
| CN114601761A (en) * | 2022-01-18 | 2022-06-10 | 科丝美诗(中国)化妆品有限公司 | Preparation method and application of plant fermentation liquor for repairing skin barrier |
Non-Patent Citations (2)
| Title |
|---|
| "Recent advancements on the synthesis and biological significance of pipecolic acid and its derivatives";Munirah M. Al-Rooqi;《Journal of Molecular Structure》;20220616;133719 * |
| "透明质酸的生物合成及其代谢工程的研究进展";马艳琴;《生物技术通报》;20210816;第38卷(第02期);252-262 * |
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