A protein carbonylation resistance composition and its application

Abstract

The present invention relates to an anti-protein carbonylation composition, which comprises a cornus extract, an eucommia leaf extract, a lily extract, a kelp extract and a collagen peptide. The present invention creatively combines the cornus extract, the eucommia leaf extract, the lily extract, the kelp extract and the collagen peptide, and the five substances have a potential mutual cooperation relationship and a synergistic effect in anti-protein carbonylation, and can be used in the fields of food, health food or medicine.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-protein carbonylation, and in particular relates to an anti-protein carbonylation composition and application thereof. Background Art

[0002] Protein carbonylation is regarded as a biomarker of oxidative stress and aging. It is a non-enzymatic and irreversible carbonyl modification of proteins. Proteins can react with active carbonyl substances from different sources to form protein carbonyl adducts, and can also self-oxidize to form protein carbonyl derivatives. With age, the body's ability to degrade carbonylated proteins weakens, and the level of protein carbonylation steadily increases, leading to the reduction and loss of protein and cell function, accelerating the accumulation of senile pigments, initiating or promoting the physiological and pathological processes of senile degenerative diseases, and ultimately accelerating the aging process.

[0003] The damage of carbonylated proteins to the skin is reflected in the following aspects: UV-exposed areas accumulate more carbonylated proteins than unexposed / light-protected areas; protein carbonylation is negatively correlated with skin water content, and carbonylated proteins can significantly reduce the content of bound water in the stratum corneum; corneum protein is composed of some cross-linked proteins. If the corneum cannot mature, it will seriously affect the barrier function, and the disorder of the corneum protein structure after carbonylation is not conducive to the maintenance of skin homeostasis; excessive accumulation of carbonylated proteins in the stratum corneum will significantly affect the refraction of light, reduce the transparency of the skin, and make the skin look dull and lifeless; oxidative stress (ultraviolet rays, lipid peroxidation, etc.) is the beginning of the formation of carbonylated proteins, and the formed carbonylated proteins can act as "photosensitizers" in the skin, which are activated by UVA and blue light, and produce more O through photosensitivity reactions. 2- , which in turn forms hydrogen peroxide, causing the production of new carbonylated proteins, and the cycle of damage continues.

[0004] At present, the main strategies for resisting protein carbonylation include: sun protection or using antioxidants, and some plant extracts have also been found to have excellent anti-carbonylation effects. For example, CN104173220A discloses a new use of green-calyx plum extract as an effective ingredient for anti-carbonylation reaction. Tests show that green-calyx plum extract has the effect of removing active carbonyl substances, and can be mixed with other cosmetic excipients and additives to prepare various skin care cosmetic compositions and various dosage forms.

[0005] However, there are few publicly reported strategies or products with significant anti-protein carbonylation effects. Therefore, it is very necessary to develop more products that can effectively resist protein carbonylation. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention aims to provide an anti-protein carbonylation composition and application thereof.

[0007] To achieve this object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides an anti-protein carbonylation composition, which comprises a cornus officinalis extract, an eucommia leaf extract, a lily extract, a kelp extract and collagen peptides.

[0009] The main components of Cornus officinalis extract include flavonoids such as cornus pyrogallol, various organic acids and other active ingredients such as cornel alcohol, etc. Cornus officinalis extract has anti-inflammatory, antibacterial and antioxidant effects. The main components of Eucommia ulmoides leaf extract are compounds such as chlorogenic acid and Eucommia ulmoides flavonoids. Eucommia ulmoides leaf extract has various biological activities, such as antioxidant, anti-inflammatory, antibacterial and antiviral effects. Lily extract has moisturizing, whitening and anti-aging effects. The main components of kelp extract include sodium alginate, etc. Kelp extract has various biological activities, such as anti-tumor, anti-inflammatory, antioxidant, anti-coagulation and other effects, and is widely used in medicine, health products, food and other fields. Collagen peptides are a type of product that is made from fresh animal tissues rich in collagen (including skin, bones, tendons, ribs, scales, etc.), which are extracted, hydrolyzed, and refined. After use, collagen peptides can increase skin moisture content and reduce wrinkles, effectively prevent the deepening of facial wrinkles, make the skin whiter and younger, slow down facial skin aging, and have a maintenance effect on the skin.

[0010] The present invention creatively combines cornus extract, eucommia leaf extract, lily extract, kelp extract and collagen peptide, and finds that the above five substances have a potential mutual cooperation relationship in anti-protein carbonylation and have a significant synergistic effect. The composition can be used in the field of food, health food or medicine.

[0011] Preferably, the anti-protein carbonylation composition comprises, by weight, 0.2-2 parts of Cornus officinalis extract, 0.5-2.5 parts of Eucommia ulmoides leaf extract, 1.0-3.5 parts of Lilium extract, 0.5-2.5 parts of Laminaria japonica extract and 2-6 parts of collagen peptide.

[0012] The weight proportions of the Cornus officinalis extract can be 0.2 parts, 0.3 parts, 0.5 parts, 0.8 parts, 0.9 parts, 1 parts, 1.3 parts, 1.5 parts, 1.8 parts, 1.9 parts, 2 parts, etc.

[0013] The weight proportions of the Eucommia leaf extract can be 0.5 parts, 0.8 parts, 0.9 parts, 1 parts, 1.3 parts, 1.5 parts, 1.8 parts, 1.9 parts, 2 parts, 2.2 parts, 2.4 parts, 2.5 parts, etc.

[0014] The weight proportions of the lily extract can be 1 part, 1.3 parts, 1.5 parts, 1.8 parts, 1.9 parts, 2 parts, 2.2 parts, 2.4 parts, 2.5 parts, 2.8 parts, 3 parts, 3.2 parts, 3.4 parts, 3.5 parts, etc.

[0015] The weight proportions of the kelp extract may be 0.5, 0.8, 0.9, 1, 1.3, 1.5, 1.8, 1.9, 2, 2.2, 2.4, 2.5, etc.

[0016] The weight proportions of the collagen peptide may be 3 parts, 4 parts, 5 parts, 7 parts, 8 parts, 9 parts, 10 parts, etc.

[0017] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0018] Preferably, the lily extract is prepared by a preparation method comprising the following steps: subjecting lily to hydrothermal extraction, followed by filtering and drying, to obtain the lily extract.

[0019] Preferably, the mass ratio of the lily to water is 1:(8-10), such as 1:8, 1:9, 1:10, etc.

[0020] Preferably, the temperature of the hydrothermal extraction is 60-80°C, for example, 60°C, 65°C, 70°C, 75°C, 78°C, 80°C, etc.

[0021] Preferably, the hydrothermal extraction is performed 2-4 times, for example 2 times, 3 times, 4 times, etc.; each time is independently 1-2 hours, for example 1 hour, 1.2 hours, 1.3 hours, 1.4 hours, 1.5 hours, 1.7 hours, 1.8 hours, 2 hours, etc.

[0022] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0023] Preferably, the kelp extract is prepared by a preparation method comprising the following steps: subjecting kelp to hydrothermal extraction, followed by filtering and drying, to obtain the kelp extract.

[0024] Preferably, the mass ratio of the kelp to water is 1:(10-15), for example, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, etc.

[0025] Preferably, the temperature of the water extraction is 70-90°C, for example, 70°C, 73°C, 75°C, 77°C, 78°C, 80°C, 83°C, 84°C, 85°C, 87°C, 88°C, 89°C, 90°C, etc.

[0026] Preferably, the hydrothermal extraction is performed 2-4 times, for example 2 times, 3 times, 4 times, etc.; each time is independently 1-2 hours, for example 1 hour, 1.2 hours, 1.3 hours, 1.4 hours, 1.5 hours, 1.7 hours, 1.8 hours, 2 hours, etc.

[0027] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0028] Preferably, the Cornus officinalis extract is prepared by the following preparation method: mixing Cornus officinalis with water, performing high-pressure pulse extraction, filtering, and drying to obtain the Cornus officinalis extract.

[0029] Preferably, the field strength of the high-voltage pulse extraction is 10-20 kV / cm, for example, 10 kV / cm, 12 kV / cm, 15 kV / cm, 18 kV / cm, 20 kV / cm, etc.; the pulse time is 500-800 μs, for example, 500 μs, 550 μs, 600 μs, 650 μs, 700 μs, 750 μs, 800 μs, etc.; the pulse frequency is 150-500 Hz, for example, 150 Hz, 200 Hz, 250 Hz, 300 Hz, 350 Hz, 400 Hz, 450 Hz, 500 Hz, etc.; the extraction time is 10-50 min, for example, 10 min, 20 min, 30 min, 40 min, 50 min, etc.

[0030] Preferably, the mass ratio of Cornus officinalis to water is 1:(5-15), for example, 1:5, 1:7, 1:8, 1:9, 1:10, 1:12, 1:13, 1:14, 1:15, etc.

[0031] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0032] Preferably, the Eucommia leaf extract is prepared by the following preparation method:

[0033] The eucommia leaf is mixed with a solution containing a deep eutectic solvent, and refluxed for extraction to obtain an extract, and the extract is filtered and dried to obtain the eucommia leaf extract.

[0034] Preferably, the ratio of the Eucommia ulmoides leaves to the solution containing the deep eutectic solvent is 1 g:(5-20) mL, for example, 1 g:5 mL, 1 g:10 mL, 1 g:15 mL, 1 g:20 mL, etc.

[0035] Preferably, the reflux extraction is performed 1-3 times, for example, 1 time, 2 times, 3 times; and the time of each extraction is independently 1-3 hours, for example, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, etc.

[0036] Preferably, the mass percentage of the deep eutectic solvent in the solution containing the deep eutectic solvent is 50-80%, for example, 50%, 60%, 70%, 78%, 80%, etc.

[0037] Preferably, the deep eutectic solvent is obtained by reacting a hydrogen bond donor and a hydrogen bond acceptor.

[0038] Preferably, the reaction temperature is 40-70°C (e.g., 40°C, 43°C, 45°C, 47°C, 48°C, 50°C, 53°C, 54°C, 55°C, 57°C, 58°C, 59°C, 60°C, 62°C, 64°C, 66°C, 68°C, 70°C, etc.), and the reaction time is 1-2h (e.g., 1h, 1.2h, 1.3h, 1.4h, 1.5h, 1.7h, 1.8h, 2h, etc.).

[0039] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0040] Preferably, the hydrogen bond donor is selected from any one of malic acid, D-sorbitol, fumaric acid or citric acid, or a combination of at least two thereof.

[0041] When malic acid, D-sorbitol, fumaric acid or citric acid were used as hydrogen bond donors of deep eutectic solvents in combination with specific hydrogen bond acceptors, the extracted Eucommia ulmoides leaf extracts had more excellent anti-protein carbonylation efficacy.

[0042] Preferably, the hydrogen bond acceptor is selected from any one or a combination of at least two of ammonium chloride, L-proline or betaine.

[0043] Preferably, the molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is (1-6):1, such as 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, etc.

[0044] Specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0045] In a second aspect, the present invention provides a use of the anti-protein carbonylation composition described in the first aspect in the preparation of an anti-protein carbonylation product.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] The present invention creatively combines cornus extract, eucommia leaf extract, lily extract, kelp extract and collagen peptide, and finds that the above five substances have a potential mutual cooperation relationship in anti-protein carbonylation and have a significant synergistic effect. The composition can be used in the field of food, health food or medicine. DETAILED DESCRIPTION

[0048] For the convenience of understanding the present invention, the present invention lists the following embodiments. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0049] The collagen peptide used in the following examples and comparative examples is a product with an average molecular weight of 1000 purchased from Hubei Ruibang Biotechnology Co., Ltd.

[0050] Preparation Example 1-1

[0051] This preparation example provides a Cornus officinalis extract, and the preparation method thereof is as follows:

[0052] 500 g of Cornus officinalis was used as raw material, coarsely crushed the raw material, mixed with 5 L of water for high-voltage pulse extraction, and the field strength was controlled to be 15 kV / cm, the pulse time was 600 μs, the pulse frequency was 300 Hz, and the extraction time was 20 min. After obtaining the extract, it was purified by macroporous resin D101, concentrated, and spray-dried to obtain the Cornus officinalis extract.

[0053] Preparation Example 1-2

[0054] This preparation example provides a Cornus officinalis extract, and the preparation method thereof is as follows:

[0055] 500 g of Cornus officinalis was used as raw material, coarsely crushed the raw material, mixed with 3 L of water for high-voltage pulse extraction, and the field strength was controlled to be 10 kV / cm, the pulse time was 500 μs, the pulse frequency was 200 Hz, and the extraction time was 40 min. After obtaining the extract, it was purified by macroporous resin D101, concentrated, and spray-dried to obtain the Cornus officinalis extract.

[0056] Preparation Example 1-3

[0057] This preparation example provides a Cornus officinalis extract, and the preparation method thereof is as follows:

[0058] 500 g of Cornus officinalis was used as raw material, coarsely crushed the raw material, mixed with 4 L of water for high-voltage pulse extraction, and the field strength was controlled to be 20 kV / cm, the pulse time was 800 μs, the pulse frequency was 300 Hz, and the extraction time was 10 min. After obtaining the extract, it was purified by macroporous resin D101, concentrated, and spray-dried to obtain the Cornus officinalis extract.

[0059] Preparation Example 1-4

[0060] This preparation example provides a cornus officinalis extract, and the preparation method thereof does not use high-pressure pulse extraction, but uses hydrothermal extraction, as follows:

[0061] 500 g of Cornus officinalis was used as raw material, the raw material was coarsely crushed, mixed with 5 L of water, and hydrothermally extracted at 60° C. for 40 min. The extract was purified by macroporous resin D101, concentrated, and spray-dried to obtain the Cornus officinalis extract.

[0062] Preparation Example 1-5

[0063] This preparation example provides a cornus officinalis extract, and the preparation method thereof is different from that of Preparation Example 1-1 only in that the high-voltage pulse extraction parameters used are different, as follows:

[0064] 500 g of Cornus officinalis was used as raw material, coarsely crushed the raw material, mixed with 5 L of water for high-voltage pulse extraction, and the field strength was controlled to be 25 kV / cm, the pulse time was 600 μs, the pulse frequency was 300 Hz, and the extraction time was 60 min. After obtaining the extract, it was purified by macroporous resin D101, concentrated, and spray-dried to obtain the Cornus officinalis extract.

[0065] Preparation Example 2-1

[0066] This preparation example provides an eucommia leaf extract, and the preparation method thereof is as follows:

[0067] (1) D-sorbitol and ammonium chloride were mixed at a molar ratio of 2:1, heated and stirred at 60° C. for 1 h, and then mixed with deionized water at a volume ratio of 3:2 to obtain a solution containing a deep eutectic solvent for later use.

[0068] (2) Mixing the Eucommia leaf powder with a solution containing a deep eutectic solvent at a ratio of 1:10 g / mL, reflux extraction for 2 h, extracting twice, and combining the obtained extracts; filtering the extracts, centrifuging, concentrating, and freeze-drying to obtain the Eucommia leaf extract.

[0069] Preparation Example 2-2

[0070] This preparation example provides an eucommia leaf extract, and the preparation method thereof is as follows:

[0071] (1) Fumaric acid and L-proline were mixed at a molar ratio of 4:1, heated and stirred at 50° C. for 2 h, and then mixed with deionized water at a volume ratio of 4:1 to obtain a solution containing a deep eutectic solvent for later use.

[0072] (2) The Eucommia leaf powder was mixed with a solution containing a deep eutectic solvent at a ratio of 1:15 g / mL, and refluxed for 1 hour, extracted three times, and the obtained extracts were combined; the extracts were filtered, centrifuged, concentrated, and freeze-dried to obtain the Eucommia leaf extract.

[0073] Preparation Example 2-3

[0074] This preparation example provides an eucommia leaf extract, and the preparation method thereof is as follows:

[0075] (1) Mix citric acid and betaine in a molar ratio of 6:1, heat and stir at 70° C. for 1 h, and then mix with deionized water in a volume ratio of 1:1 to obtain a solution containing a deep eutectic solvent for later use.

[0076] (2) Mixing the Eucommia leaf powder with a solution containing a deep eutectic solvent at a ratio of 1:5 g / mL, reflux extraction for 2 h, extracting twice, and combining the obtained extracts; filtering the extracts, centrifuging, concentrating, and freeze-drying to obtain the Eucommia leaf extract.

[0077] Preparation Example 2-4

[0078] This preparation example provides an eucommia leaf extract, and its preparation method is different from that of Preparation Example 2-1 only in that the composition of the hydrogen bond donor in the deep eutectic solvent is different, D-sorbitol is replaced by an equal amount of maleic acid, and other conditions remain unchanged. The other steps refer to Preparation Example 2-1.

[0079] Preparation Example 2-5

[0080] This preparation example provides an eucommia leaf extract, and its preparation method is different from that of Preparation Example 2-1 only in that the composition of the hydrogen bond donor in the deep eutectic solvent is different, D-sorbitol is replaced by an equal amount of ethylene glycol methyl ether, and other conditions remain unchanged. The other steps refer to Preparation Example 2-1.

[0081] Preparation Example 2-6

[0082] This preparation example provides an eucommia leaf extract, and its preparation method is different from that of Preparation Example 2-1 only in that the composition of the hydrogen bond donor in the deep eutectic solvent is different, D-sorbitol is replaced by an equal amount of malic acid, and other conditions remain unchanged. Other steps refer to Preparation Example 2-1.

[0083] Preparation Example 3-1

[0084] This preparation example provides a lily extract, and the preparation method thereof is as follows:

[0085] 500 g of lily was roughly chopped and then added to 4 L of water for extraction. The extraction was carried out by heating, the temperature was controlled at 70°C, and the extraction was performed twice, each time for 2 hours. The extracts were combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0086] Preparation Example 3-2

[0087] This preparation example provides a lily extract, and the preparation method thereof is as follows:

[0088] 500 g of lily was roughly chopped and then added with 5 L of water for extraction. The extraction was carried out by heating, the temperature was controlled at 65°C, and the extraction was performed 3 times, each time for 2 hours. The extracts were combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0089] Preparation Example 3-3

[0090] This preparation example provides a lily extract, and the preparation method thereof is as follows:

[0091] 500 g of lily was roughly chopped and then added to 4.5 L of water for extraction. The extraction was carried out by heating, the temperature was controlled at 80°C, and the extraction was performed 3 times, each time for 1 hour. The extracts were combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0092] Preparation Example 4-1

[0093] This preparation example provides a kelp extract, and the preparation method thereof is as follows:

[0094] The dried fronds of kelp are used as raw materials, the weight of the raw materials is 500g, the kelp is cut into pieces and 10 times the weight of water is added, and the extraction is heated at 80°C for 2 times, each time for 2 hours, the extracts are combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0095] Preparation Example 4-2

[0096] This preparation example provides a kelp extract, and the preparation method thereof is as follows:

[0097] The dried fronds of kelp are used as raw materials, the weight of the raw materials is 500g, the kelp is cut into pieces and 12.5 times the weight of water is added, and the extraction is heated at 70°C for 3 times, each time for 1.5 hours. The extracts are combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0098] Preparation Example 4-3

[0099] This preparation example provides a kelp extract, and the preparation method thereof is as follows:

[0100] The dried fronds of kelp are used as raw materials, the weight of the raw materials is 500g, the kelp is cut into pieces and 15 times the weight of water is added, and the extraction is heated at 90°C for 4 times, each time for 1 hour, the extracts are combined, filtered and centrifuged, concentrated and freeze-dried to obtain the product.

[0101] Example 1

[0102] This embodiment provides an anti-protein carbonylation composition, which is obtained by physically mixing the following components uniformly:

[0103] 2 parts of Cornus officinalis extract prepared in Preparation Example 1-1, 2.5 parts of Eucommia ulmoides leaf extract prepared in Preparation Example 2-1, 3.5 parts of Lilium sibiricum extract prepared in Preparation Example 3-1, 2.5 parts of Laminaria japonica extract prepared in Preparation Example 4-1, and 10 parts of collagen peptide

[0104] Example 2

[0105] This embodiment provides an anti-protein carbonylation composition, which is obtained by physically mixing the following components uniformly:

[0106] 1 part of Cornus officinalis extract was prepared in Preparation Example 1-2, 1.5 parts of Eucommia ulmoides leaf extract was prepared in Preparation Example 2-2, 2.5 parts of Lilium extract was prepared in Preparation Example 3-2, 1.5 parts of Kelp extract was prepared in Preparation Example 4-2, and 7 parts of collagen peptide were prepared.

[0107] Example 3

[0108] This embodiment provides an anti-protein carbonylation composition, which is obtained by physically mixing the following components uniformly:

[0109] 0.5 parts of Cornus officinalis extract prepared in Preparation Example 1-3, 1 part of Eucommia ulmoides leaf extract prepared in Preparation Example 2-3, 2 parts of Lilium extract prepared in Preparation Example 3-3, 1 part of Kelp extract prepared in Preparation Example 4-3 and 4 parts of collagen peptide.

[0110] Example 4

[0111] This embodiment provides an anti-protein carbonylation composition, which is different from Example 1 only in that an equal amount of the Cornus officinalis extract prepared in Preparation Example 1-1 is replaced by an equal amount of the Cornus officinalis extract prepared in Preparation Example 1-4, and other conditions remain unchanged.

[0112] Example 5

[0113] This embodiment provides an anti-protein carbonylation composition, which is different from Example 1 only in that an equal amount of the Cornus officinalis extract prepared in Preparation Example 1-1 is replaced by an equal amount of the Cornus officinalis extract prepared in Preparation Example 1-5, and other conditions remain unchanged.

[0114] Example 6

[0115] This embodiment provides an anti-protein carbonylation composition, which is different from Example 1 only in that an equal amount of the eucommia leaf extract prepared in Preparation Example 2-1 is replaced by an eucommia leaf extract prepared in Preparation Example 2-4, and other conditions remain unchanged.

[0116] Example 7

[0117] This embodiment provides an anti-protein carbonylation composition, which is different from Example 1 only in that an equal amount of the eucommia leaf extract prepared in Preparation Example 2-1 is replaced by an eucommia leaf extract prepared in Preparation Example 2-5, and other conditions remain unchanged.

[0118] Example 8

[0119] This embodiment provides an anti-protein carbonylation composition, which is different from Example 1 only in that an equal amount of the eucommia leaf extract prepared in Preparation Example 2-1 is replaced by the eucommia leaf extract prepared in Preparation Example 2-6, and other conditions remain unchanged.

[0120] Comparative Example 1

[0121] This comparative example provides an anti-protein carbonylation composition, which is different from Example 1 only in that it does not contain Cornus officinalis extract. The reduced portion of Cornus officinalis extract is proportionally distributed to Eucommia ulmoides leaf extract, lily extract, kelp extract and collagen peptide. The preparation method refers to Example 1.

[0122] Comparative Example 2

[0123] This comparative example provides an anti-protein carbonylation composition, which is different from Example 1 only in that it does not contain Eucommia leaf extract, and the reduced portion of Eucommia leaf extract is proportionally distributed to Cornus officinalis extract, Lilium extract, Laminaria extract and collagen peptide. The preparation method refers to Example 1.

[0124] Comparative Example 3

[0125] This comparative example provides an anti-protein carbonylation composition, which is different from Example 1 only in that it does not contain lily extract, and the reduced portion of lily extract is proportionally distributed to Cornus officinalis extract, Eucommia ulmoides leaf extract, Laminaria japonica extract and collagen peptide. The preparation method refers to Example 1.

[0126] Comparative Example 4

[0127] This comparative example provides an anti-protein carbonylation composition, which is different from Example 1 only in that it does not contain kelp extract. The reduced portion of kelp extract is proportionally distributed to Cornus officinalis extract, Eucommia ulmoides leaf extract, lily extract and collagen peptide. The preparation method refers to Example 1.

[0128] Comparative Example 5

[0129] This comparative example provides an anti-protein carbonylation composition, which is different from Example 1 only in that it does not contain collagen peptides. The reduced portion of collagen peptides is proportionally distributed to Cornus officinalis extract, Eucommia ulmoides leaf extract, Lilium extract and Laminaria japonica extract. The preparation method refers to Example 1.

[0130] Test Case

[0131] Weigh an appropriate amount of the anti-protein carbonylation composition prepared in Examples 1-8 and Comparative Examples 1-5, add it to purified water, stir until completely dissolved, and obtain a mother liquor with a concentration of 20 mg / mL. Take the mother liquor of the sample group (including Examples 1-8 and Comparative Examples 1-5) and the E.coil OP50 bacterial solution and mix them evenly in a ratio of 2:38. Use a pipette to draw 200 μL of bacterial solution and gently hit the center of each NGM plate. For the blank group, use a pipette to draw 200 μL of E.coil OP50 bacterial solution and gently hit the center of each NGM plate. Transfer the L4 nematode larvae to the blank group and the sample group (Examples 1-10 and Comparative Examples 1-5), the sample concentration is 1 mg / mL NGM, and the nematodes need to be picked to a new NGM plate every other day.

[0132] After 10 days of culture, the nematodes were washed three times with M9 buffer. A small amount of 6mmol / L levamisole was dripped on a 3% agarose pad slide. At least 30 nematodes were selected from each treatment group and transferred to the levamisole droplets to anesthetize the nematodes until there was no reaction. Fluorescence images and bright field images were taken and the fluorescence intensity of each nematode in the image was analyzed relatively quantitatively. Three parallel dishes were set up for each group to culture nematodes.

[0133] ImageJ software was used to perform relative quantitative analysis on the fluorescence intensity of lipofuscin in the nematode intestine in the image to obtain the fluorescence intensity value and relative fluorescence intensity value of each group. Fluorescence intensity inhibition rate (%) = (1-T / C) × 100%. (T-average fluorescence intensity value of the sample group; C-average fluorescence intensity value of the blank group).

[0134] The experimental results show that the fluorescence intensity inhibition rate of the sample group is greater than 0, and the fluorescence intensity value of the sample group is significantly different from that of the blank group (P < 0.05), indicating that the sample can inhibit the production of lipofuscin in the intestine of nematodes at this concentration, has the ability to resist the carbonylation of body proteins, and has anti-carbonylation activity. The experimental results are shown in Table 1.

[0135] Table 1

[0136]

[0137]

[0138] As shown in Table 1, compared with the comparative example, the composition of the present invention has excellent anti-carbonylation effect, and the extract of Cornus officinalis, the extract of Eucommia ulmoides leaves, the extract of Lilium sibiricum, the extract of Laminaria japonica and the collagen peptide synergize in the above effects. The extraction process of the plant extract also significantly affects the above effects.

[0139] The applicant declares that the present invention illustrates the detailed process equipment and process flow of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned detailed process equipment and process flow, that is, it does not mean that the present invention must rely on the above-mentioned detailed process equipment and process flow to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. An anti-protein carbonylation composition, characterized in that The anti-protein carbonylation composition comprises, by weight, 0.2-2 parts of cornus officinalis extract, 0.5-2.5 parts of eucommia leaf extract, 1.0-3.5 parts of lily extract, 0.5-2.5 parts of kelp extract and 2-6 parts of collagen peptide; The cornus officinalis extract is prepared by the following preparation method: mixing cornus officinalis and water at a mass ratio of 1: (5-15), performing high-voltage pulse extraction, with a field strength of 10-20 kV / cm, a pulse time of 500-800 μs, a pulse frequency of 150-500 Hz, an extraction time of 10-50 min, filtering, and drying to obtain the cornus officinalis extract; The eucommia leaf extract is prepared by the following preparation method: eucommia leaf is mixed with a solution containing 50-80% of a deep eutectic solvent at a ratio of 1 g: (5-20) mL, refluxed for 1-3 times, each extraction time is 1-3 hours, to obtain an extract, and the extract is filtered and dried to obtain the eucommia leaf extract; The deep eutectic solvent is obtained by reacting a hydrogen bond donor and a hydrogen bond acceptor at a molar ratio of (1-6):1 at 40-70°C for 1-2 h; the hydrogen bond donor is selected from any one of D-sorbitol, fumaric acid or citric acid or a combination of at least two of them; the hydrogen bond acceptor is selected from any one of ammonium chloride, L-proline or betaine or a combination of at least two of them.

2. The anti-protein carbonylation composition according to claim 1, characterized in that The lily extract is prepared by the following preparation method: performing hydrothermal extraction on the lily, and then filtering and drying to obtain the lily extract.

3. The anti-protein carbonylation composition according to claim 2, characterized in that: The mass ratio of the lily to water is 1:(8-10).

4. The anti-protein carbonylation composition according to claim 2, characterized in that The temperature of the hydrothermal extraction is 60-80°C.

5. The anti-protein carbonylation composition according to claim 2, characterized in that: The hydrothermal extraction is performed 2-4 times, and the time for each extraction is 1-2 h.

6. The anti-protein carbonylation composition according to claim 1, characterized in that: The kelp extract is prepared by the following preparation method: hydrothermal extraction is performed on kelp, followed by filtering and drying to obtain the kelp extract.

7. The anti-protein carbonylation composition according to claim 6, characterized in that: The mass ratio of the kelp to water is 1:(10-15).

8. The anti-protein carbonylation composition according to claim 6, characterized in that: The temperature of the hydrothermal extraction is 70-90°C.

9. The anti-protein carbonylation composition according to claim 6, characterized in that: The hydrothermal extraction is performed 2-4 times, and the time for each extraction is 1-2 h.

10. Use of the anti-protein carbonylation composition according to any one of claims 1 to 9 in the preparation of an anti-protein carbonylation product.

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