CN112245345A - Use of carnosine for protecting the activity of plant exosomes - Google Patents

Abstract

The present invention provides the use of carnosine in protecting the activity of plant exosomes. The present invention also provides a composition for anti-aging, wherein the composition comprises carnosine and plant exosomes. The composition for resisting aging provided by the invention has a good anti-aging effect.

Description

Use of carnosine for protecting the activity of plant exosomes

Technical Field

The invention relates to the field of biomedicine. In particular, the invention relates to the use of carnosine in protecting the activity of plant exosomes.

Background

Skin aging can be divided into endogenous aging caused by intrinsic factors and exogenous aging secondary to the extrinsic factors, and is usually reflected in the reduction of skin elasticity and the generation of skin wrinkles, the reduction of collagen and elastin leads to the reduction of skin elasticity and the generation of skin wrinkles, and the collagen and elastin are regulated by collagenase and elastase, which can cause the degradation of collagen and elastin, thus leading to the reduction of collagen and elastin, leading to the generation of wrinkles and skin elasticity reduction.

Exosome is a small cup-shaped vesicle with a double-layer membrane structure and a diameter of about 30-100nm, secreted by various living cells, has a lipid bilayer structure, can carry a large amount of protein, lipid, mRNA and siRNA, is easy to fuse with receptor cells, and researches report that exosome can participate in processes such as cell communication, cell migration, angiogenesis, immunoreaction and the like.

Exosomes (exosomes) are small, two-layer membrane-structured, cup-shaped vesicles of approximately 30-100nm diameter that can be secreted by a variety of living cells. The exosome can transfer active substances such as protein, mRNA, microRNA and the like among cells, can participate in a plurality of important physiological and pathological processes such as cell communication, cell migration, angiogenesis, immune reaction and the like, has more and more attention on unique functions, can be secreted by cells under normal physiological conditions, has a lipid bilayer structure, contains membrane lipid, membrane protein, genetic substances and cytoplasm components, and indirectly reflects the properties and the state of the cells. Meanwhile, exosomes act as extracellular transport proteins mediating cell-cell communication by binding to other cells and tissues and transferring membrane components, mRNA, miRNA, proteins (growth hormone, cytokine, etc.) to recipient cells.

At present, animal-derived exosomes are researched and applied more, but the animal cell-derived exosomes are complex in extraction process, high in cost, low in yield, easy to cause side reactions in the application process and have certain safety problems. The exosome from plant sources is extracted and applied, the source is reliable and stable, the yield is relatively high, and the biocompatibility and the safety are more reliable. However, the activity of the plant exosome is difficult to ensure at normal temperature, which limits the application of the plant exosome and the efficacy of the product, so that the research and the application aiming at the plant exosome are less at present.

Disclosure of Invention

In order to solve the defects of the prior art, the invention provides the application of carnosine in protecting the activity of a plant exosome and also provides a composition for protecting the activity of the plant exosome.

The purpose of the invention is realized by the following technical scheme:

in a first aspect, the invention provides the use of carnosine in protecting the activity of a plant exosome.

In a preferred embodiment, the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

In a second aspect, the present invention provides a composition for anti-aging, wherein the composition comprises carnosine and plant exosomes.

In a preferred embodiment, the composition further comprises a humectant, a preservative and water.

In a preferred embodiment, the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

In a preferred embodiment, the humectants are glycerin and butylene glycol.

In a preferred embodiment, the preservative is phenoxyethanol.

In a preferred embodiment, the carnosine is present in an amount of 0.1 to 0.5 parts by weight. For example, the carnosine is present in an amount of 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45 or 0.5 parts by weight.

In a preferred embodiment, the weight fraction of the plant exosomes is 0.1-0.5. For example, the weight fraction of the plant exosomes is 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45 or 0.5.

In a preferred embodiment, the humectant is 1 to 6 parts by weight. For example, the humectant is 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 parts by weight.

In a preferred embodiment, the preservative is present in an amount of 0.1 to 0.4 parts by weight. For example, the preservative is present in an amount of 0.1, 0.15, 0.2, 0.25, 0.3, 0.35 or 0.4 parts by weight.

In a third aspect, the invention provides the use of the composition for resisting senility in preparing an anti-senility product.

In a fourth aspect, an anti-aging product is provided, which comprises the composition for resisting aging of the invention.

In a fifth aspect, there is provided a method for preparing a composition for anti-aging, wherein the method comprises the steps of:

s1, adding carnosine, plant exosome and preservative into a container in sequence and mixing to obtain phase A;

s2, sequentially adding water and a humectant into a container, mixing, heating to 80-85 ℃, and uniformly stirring to obtain a phase B;

and S3, cooling the phase B obtained in the step S2 to 40-45 ℃, adding the phase A, and stirring for 15-25 min.

In a preferred embodiment, the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

In a preferred embodiment, the plant exosomes are prepared as follows:

weighing 100-; centrifuging the solution in the middle layer at 10000-; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80min, then using 1mL sterile PBS to resuspend the precipitate, and ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80 min; after centrifugation, the pellet was resuspended in 1-2mL sterile PBS to obtain the plant exosomes.

In a preferred embodiment, the grape exosomes are prepared as follows:

weighing 100-; centrifuging the purple transparent solution in the middle layer at 10000-; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80min, then using 1mL sterile PBS to resuspend the precipitate, and ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80 min; after centrifugation, 1-2mL sterile PBS is used for resuspension and precipitation, and the grape exosome is obtained.

In the present invention, carnosine was purchased from south-fragrance perfumes (southwestern) limited by de with a lot number of 191001; the preservative is purchased from Shumei GmbH of Germany with the batch number of 191112; the moisturizer was purchased from sienna meicheng ltd under a lot number of 191101.

Compared with the prior art, the invention has the beneficial effects that:

the procyanidin and resveratrol contained in fructus Vitis Viniferae are powerful antioxidants, and have antiaging and free radical scavenging effects; the grape exosomes contain proteins, RNA, secondary metabolites and the like besides procyanidins and resveratrol, the components are evolved and selected from grapes, the components cooperate with each other to better play an anti-aging role, and the grape exosomes are of a phospholipid bilayer structure, have a small particle size ratio and are easier to react with cells, so that the anti-aging role is better played. The invention adopts a high-speed centrifugal extraction technology, ensures the extraction rate, simultaneously furthest retains the activity of the grape exosomes, prolongs the storage time of the grape exosomes, and ensures the effectiveness of the functional components so as to ensure the anti-aging effect of the product. However, procyanidin and resveratrol in the grape exosome are very easy to oxidize, and a protective agent is needed to protect the activity of the grape exosome.

The anti-aging raw materials on the market at present are a lot, the collagen accounts for the most, but the molecular weight of the anti-aging raw materials is large, and the collagen is difficult to absorb by the skin, while the grape exosome has the small molecular weight, not only can exert the anti-aging effect of the grape exosome, but also has the characteristic of a carrier, can carry other components into the deep layer of the skin to play the anti-aging effect, so that the anti-aging composition has a good anti-aging effect.

Drawings

FIG. 1 is a graph showing the results of a collagen content test;

FIG. 2 is a graph showing the results of measurement of MMP-1 release.

Detailed Description

The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly demonstrated. It will be understood by those skilled in the art that these specific embodiments and examples are for the purpose of illustrating the invention and are not to be construed as limiting the invention.

Example 1

The embodiment provides a composition for resisting aging, which comprises the following components in parts by weight:

0.1 part of grape exosome, 0.1 part of carnosine, 0.5 part of glycerol, 0.5 part of butanediol, 0.1 part of phenoxyethanol and 93 parts of water;

the preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Example 2

The embodiment provides a composition for resisting aging, which comprises the following components in parts by weight:

0.5 part of grape exosome, 0.5 part of carnosine, 3 parts of glycerol, 3 parts of butanediol, 0.4 part of phenoxyethanol and 96 parts of water;

the preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Example 3

The embodiment provides a composition for resisting aging, which comprises the following components in parts by weight:

0.2 part of grape exosome, 0.3 part of carnosine, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water;

the preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Example 4

This example provides a method for preparing a composition for anti-aging, wherein the method comprises the steps of:

s1, adding carnosine, grape exosomes and phenoxyethanol into a container in sequence, and mixing to obtain a phase A;

s2, sequentially adding water, glycerol and butanediol into a container, mixing, heating to 80-85 ℃, and uniformly stirring to obtain a phase B;

and S3, cooling the phase B obtained in the step S2 to 40-45 ℃, adding the phase A, and stirring for 15-25 min.

Example 5

This example provides the use of carnosine to protect the activity of plant exosomes.

Wherein the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

Wherein the preparation method of the plant exosome comprises the following steps:

weighing 100-; centrifuging the solution in the middle layer at 10000-; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80min, then using 1mL sterile PBS to resuspend the precipitate, and ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80 min; after centrifugation, the pellet was resuspended in 1-2mL sterile PBS to obtain the plant exosomes.

The preparation method of the grape exosome comprises the following steps:

weighing 100-; centrifuging the purple transparent solution in the middle layer at 10000-; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80min, then using 1mL sterile PBS to resuspend the precipitate, and ultracentrifuging at 120000-150000 Xg and 4 ℃ for 60-80 min; after centrifugation, 1-2mL sterile PBS is used for resuspension and precipitation, and the grape exosome is obtained.

Example 6

This example provides a composition for anti-aging, wherein the composition comprises carnosine and plant exosomes.

In some embodiments, the composition further comprises a humectant, a preservative, and water.

In some embodiments, the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes, or pomegranate exosomes.

In some embodiments, the humectants are glycerin and butylene glycol.

In some embodiments, the preservative is phenoxyethanol.

In some embodiments, the carnosine is present in an amount of 0.1 to 0.5 parts by weight.

In some embodiments, the weight fraction of the plant exosomes is 0.1-0.5.

In some embodiments, the humectant is 1 to 6 parts by weight.

In some embodiments, the preservative is present in an amount of 0.1 to 0.4 parts by weight.

Comparative example 1

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of onion exosome, 0.3 part of carnosine, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the onion exosome comprises the following steps:

weighing 100g of fresh onion, pouring into a crusher, adding 100mL of sterilized PBS, crushing, homogenizing, centrifuging at 3000 Xg at 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain onion exosomes.

Comparative example 2

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of grape exosome, 0.3 part of glutathione, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 3

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of grape exosome, 0.3 part of astaxanthin, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 4

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of grape exosome, 0.3 part of carnosine, 2 parts of glycerol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 5

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of grape exosome, 0.3 part of carnosine, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 6

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.2 part of grape exosome, 0.3 part of carnosine, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of potassium sorbate and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 7

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.05 part of grape exosome, 0.3 part of carnosine, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 8

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

1 part of grape exosome, 0.3 part of carnosine, 2 parts of glycerol, 2 parts of butanediol, 0.3 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 9

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

0.05 part of grape exosome, 0.05 part of carnosine, 0.4 part of glycerol, 0.4 part of butanediol, 0.05 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Comparative example 10

This comparative example provides a composition for anti-ageing comprising, in parts by weight:

1 part of grape exosome, 0.6 part of carnosine, 3.5 parts of glycerol, 3.5 parts of butanediol, 0.5 part of phenoxyethanol and 95 parts of water.

The preparation method of the grape exosome comprises the following steps:

weighing 100g of fresh grapes, pouring the fresh grapes into a crusher, adding 100mL of sterilized PBS into the grapes, crushing the grapes into uniform slurry, centrifuging the mixture at 3000 xg and 4 ℃ for 10min, and removing large residues; centrifuging the purple transparent solution in the middle layer at 10000 Xg and 4 deg.C for 40 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000 Xg and 4 ℃ for 60 min; after centrifugation, the precipitate was resuspended in 1mL sterile PBS to obtain the grape exosomes.

Experiment 1: collagen content test

And (3) testing a sample: samples obtained in examples 1 to 3 and comparative examples 1 to 10

The experimental method comprises the following steps:

after the samples were processed, they were tested using an ELISA kit (purchased from Shanghai Jianglai Biotech Co., Ltd., lot 1903) according to the kit instructions.

The method utilizes a detection model of Guangdong Boxi biological technology limited company for detection, and the model is as follows: fibroplast, batch number: 191001.

the results of the collagen content test are shown in FIG. 1.

Table 1 collagen content test experimental group

Also, to further illustrate the benefits of the present invention, the following examples are provided:

example 3.1 is provided, which example 3.1 differs from example 3 in that: replacing the grape exosomes with carrot exosomes;

example 3.2 is provided, which example 3.2 differs from example 3 in that: replacing the grape exosomes with tomato exosomes;

example 3.3 is provided, which example 3.3 differs from example 3 in that: replacing the grape exosomes with pomegranate exosomes;

the anti-aging composition prepared by combining the preparation method of the invention is tested for collagen content. The anti-aging effects of the compositions for anti-aging prepared in example 3.1, example 3.2, and example 3.3 were observed. The results were similar to those in example 3 above.

As can be seen from fig. 1:

collagen is a high molecular functional protein, which is the main component of skin and accounts for 80 percent of the proportion of the dermis of the skin; it is also a major component of the dermal extracellular matrix, and forms a fine elastic network in the skin, firmly retaining moisture, and maintaining skin elasticity. The loss of collagen directly causes the generation of skin wrinkles, while CollagenI is the most important collagen, the synthesis/content of CollagenI can directly reflect the collagen secretion capacity of fibroblasts, and the collagen synthesis promoting capacity of cosmetic active ingredients can be rapidly evaluated, so that the collaagen collagen is an effective index for rapidly evaluating the efficacy of anti-wrinkle.

The collagen content detection by using the Fibroplast model shows that the composition for protecting the activity of the plant exosomes has higher collagen I synthesis/content, which indicates that the plant exosomes can promote carnosine to enter the deep layer of the skin, so that the collagen is generated, and the composition for protecting the activity of the plant exosomes has a good anti-wrinkle effect, namely the composition for protecting the activity of the plant exosomes has a good anti-aging effect. And the composition for protecting the activity of the plant exosomes without using the formula system provided by the invention has obviously lower synthesis/content of Collageni, namely the anti-aging effect is not obvious.

Experiment 2: detection of MMP-1 Release amount

And (3) testing a sample: samples obtained in examples 1 to 3 and comparative examples 1 to 10

The experimental method comprises the following steps:

the assay was performed using an ELISA kit (purchased from Shanghai Jianglai Biotech Co., Ltd., lot No. 1903) according to the kit instructions.

The results of the measurement of the amount of MMP-1 released are shown in FIG. 2.

TABLE 2 experimental groups for the detection of the amount of MMP-1 released

Also, to further illustrate the benefits of the present invention, the following examples are provided:

example 3.1 is provided, which example 3.1 differs from example 3 in that: replacing the grape exosomes with carrot exosomes;

example 3.2 is provided, which example 3.2 differs from example 3 in that: replacing the grape exosomes with tomato exosomes;

example 3.3 is provided, which example 3.3 differs from example 3 in that: replacing the grape exosomes with pomegranate exosomes;

the composition for resisting aging is prepared by combining the preparation method of the invention, and the release amount of MMP-1 of the composition is detected. The anti-aging effects of the compositions for anti-aging prepared in example 3.1, example 3.2, and example 3.3 were observed. The results were similar to those in example 3 above.

As can be seen from fig. 2:

the detection of MMP-1 release is the main index for evaluating photoaging. MMP-1 is the most important enzyme for degrading type I and type III collagens, when skin cells excessively express MMP-1, the result of the dermal extracellular matrix is seriously damaged, particularly the normal structure of collagen fibers and elastic fibers, so that MMP-1 is the most important enzyme for causing aging symptoms such as wrinkles, fine lines and the like of the skin.

The detection of MMP-1 release amount by using a Fibroplast model shows that the MMP-1 release amount of the composition for protecting the activity of the plant exosomes is obviously lower, which indicates that the plant exosomes can promote carnosine to enter the deep layer of the skin, thereby avoiding the excessive expression of MMP-1 by skin cells, not only playing a corresponding anti-aging effect, but also carrying functional components to enter the deep layer of the skin to play an anti-aging effect, and indicates that the composition for protecting the activity of the plant exosomes has good anti-wrinkle, anti-fine line aging symptoms and the like, namely, the composition for protecting the activity of the plant exosomes has good anti-aging effect. The composition for protecting the activity of the plant exosome without using the formula system has higher MMP-1 release amount, namely the anti-aging effect is not obvious.

Experiment 3: human body efficacy testing

And (3) testing a sample: samples obtained in examples 1 to 3 and comparative examples 1 to 10

The experimental method comprises the following steps:

130 volunteers, aged between 28-55 years, were collected in groups of 10 persons, and the samples obtained in examples 1-3 and comparative examples 1-10 were used to test the anti-aging effect index, and the moisture content, elasticity and wrinkles in the anti-aging effect index were tested by a German CK apparatus, and then immediate efficacy test and long-term efficacy test were performed.

Wherein, the instant efficacy verification is as follows: the measured data before nursing and within one hour after nursing are the immediate effect data, the average value processing is carried out, and the improvement degree after one hour of nursing is calculated, so that the proportion of the number of improved people to the number of non-improved people is obtained.

Wherein, the long-acting efficacy is verified as follows: the original data before nursing and the data of one week to four weeks after the use of the test subjects are taken as long-term effect data to be subjected to average value processing, and the improvement degree of each volunteer after the use of the test subject for four weeks is calculated, so that the improved population ratio and the non-improved population ratio are obtained.

TABLE 3 instant efficacy test results

Table 4 long-acting efficacy verification results

As can be seen from tables 3 and 4:

the composition for protecting the activity of the plant exosomes provided by the invention is subjected to instant efficacy test and long-acting efficacy test, and volunteers have remarkable advantages in maintaining moisture content, keeping elasticity and resisting wrinkles, so that the composition for protecting the activity of the plant exosomes provided by the invention has a good anti-aging effect. The composition for protecting the activity of the plant exosomes without using the formula system of the invention is provided for instant efficacy verification and long-acting efficacy verification, and the volunteers have no obvious effects on the aspects of maintaining the water content, keeping the elasticity, resisting wrinkles and the like and resisting aging.

It is to be understood that the invention disclosed is not limited to the particular methodology, protocols, and materials described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

Those skilled in the art will also recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims (10)

1. Use of carnosine to protect the activity of a plant exosome.

2. The use according to claim 1, wherein the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

3. Composition for use in combating ageing, wherein the composition comprises carnosine and plant exosomes.

4. The composition of claim 3, wherein the composition further comprises a humectant, a preservative, and water.

5. The composition of claim 3, wherein the plant exosomes are grape exosomes, carrot exosomes, tomato exosomes or pomegranate exosomes.

6. The composition of claim 4, wherein the humectants are glycerin and butylene glycol.

7. The composition of claim 4, wherein the preservative is phenoxyethanol.

8. The composition according to claim 3, characterized in that said carnosine is present in a weight fraction of 0.1 to 0.5.

9. The composition of claim 3, wherein the weight fraction of the plant exosomes is 0.1-0.5.

10. A method for preparing a composition for combating ageing, said method comprising the steps of:

s1, adding carnosine, plant exosome and preservative into a container in sequence and mixing to obtain phase A;

s2, sequentially adding water and a humectant into a container, mixing, heating to 80-85 ℃, and uniformly stirring to obtain a phase B;

and S3, cooling the phase B obtained in the step S2 to 40-45 ℃, adding the phase A, and stirring for 15-25 min.

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