CN115317399A - Anti-aging composition and preparation method and application thereof - Google Patents
Anti-aging composition and preparation method and application thereof Download PDFInfo
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- CN115317399A CN115317399A CN202210801295.7A CN202210801295A CN115317399A CN 115317399 A CN115317399 A CN 115317399A CN 202210801295 A CN202210801295 A CN 202210801295A CN 115317399 A CN115317399 A CN 115317399A
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- bacillus subtilis
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- aging
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- plasmin
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention provides an anti-aging composition and a preparation method and application thereof, belonging to the technical field of bioengineering. The health-care food is prepared from the following raw materials in parts by weight: 50-60 parts of bacillus subtilis fibrinolytic enzyme, 20-25 parts of resveratrol, 8-13 parts of polygonum multiflorum and 15-20 parts of ginseng. The bacillus subtilis plasmin combination with the anti-aging function can eliminate free radicals in vivo and slow down aging, takes the bacillus subtilis plasmin as a main component and is assisted with traditional Chinese medicine components, free radical components in a mouse body can be obviously reduced, and no adverse reaction is observed.
Description
Technical Field
The invention relates to the technical field of bioengineering, and particularly relates to an anti-aging composition and a preparation method and application thereof.
Background
With the aging of the population and the pressure of increasing social competition, aging diseases associated with the decline of body functions increasingly affect the quality of life of people. Theoretical studies at present suggest that aging is mainly divided into two categories: physiological aging and pathological aging. The former refers to a physiological degenerative process occurring after maturation and the latter is an aging change due to various external factors including various diseases. The two are practically indistinguishable. In summary, aging is a corollary of the combined effects of many pathological, physiological and psychological processes, the biological psychological process of the final stages of growth and development of an individual. The aging process is continuously generated from fertilized ovum to death, and the aging characteristics are obviously shown only by a certain stage. The physiological changes in the aging process of human body are mainly reflected in the loss of organism tissue cells and constitutional substances, the slowing of organism metabolic rate and the hypofunction of organism and organs. Senescence is inevitable, but delaying senescence is possible. Reasonable diet and balanced nutrition are one of important measures for delaying aging and prolonging life.
The current aging theory considers that aging is directly connected with free radical oxidation of human body. Degenerative changes during aging are due to the deleterious effects of free radicals produced during normal cellular metabolism. The aging process of an organism is the result of accumulation of free radicals continuously produced by tissue cells of the organism, and the free radicals can cause DNA damage to cause mutation and induce tumor formation. Free radicals are intermediate products of normal metabolism, have strong reaction capability, and can oxidize various substances in cells to damage biological membranes. But also cross-link macromolecules such as proteins and nucleic acids, and influence the normal functions of the proteins and nucleic acids.
The oxygen free radicals act on cell membranes to induce lipid peroxidation, which is a collective index of traditional Chinese medicine. At present, the traditional Chinese medicine and western medicine components which are just aging have the effects of prolonging life and slowing aging, and the action mechanism is mainly focused on the aspect of free radicals. However, at present, few anti-aging drugs available to people are few, and a few anti-aging drugs sold on the market are expensive and have insignificant effects, and all the drugs need to be taken for a long time. There is a lack of an anti-aging drug with low price and good effect.
The aging degree in China is continuously increased, and an anti-aging medicament with low price and small side effect is urgently needed.
Disclosure of Invention
The invention aims to provide an anti-aging composition, a preparation method and application thereof, which can remove free radicals in vivo and slow down aging. The composition takes bacillus subtilis plasmin as a main component and is assisted by traditional Chinese medicine components, so that the free radical components in a mouse body can be remarkably reduced, and no adverse reaction is observed.
The technical scheme of the invention is realized as follows:
the invention provides an anti-aging composition which is prepared from the following raw materials in parts by weight: 50-60 parts of bacillus subtilis fibrinolytic enzyme, 20-25 parts of resveratrol, 8-13 parts of polygonum multiflorum and 15-20 parts of ginseng.
As a further improvement of the invention, the Bacillus subtilis fibrinolytic enzyme is prepared by fermenting a strain Bacillus subtilis QK02, wherein the strain is preserved in China center for type culture collection with the preservation number of CCTCC NO: M203078.
As a further improvement of the invention, the Bacillus subtilis plasmin is subjected to centrifugation, microfiltration, ultrafiltration and spray drying on fermentation liquor obtained by fermenting the Bacillus subtilis QK02 strain to obtain spray-dried powder.
As a further improvement of the invention, the activity of the bacillus subtilis plasmin is not less than 80000IU/g.
The invention further provides a preparation method of the anti-aging composition, which comprises the following steps: grinding bacillus subtilis fibrinolytic enzyme, white quinoa alcohol, polygonum multiflorum and ginseng into powder, sieving and mixing in proportion to obtain the anti-aging composition.
As a further improvement of the invention, the screening mesh number is 80-100 meshes, and the mixing is carried out by using a mixer, preferably a three-dimensional mixer.
As a further improvement of the invention, the method comprises the following steps: grinding bacillus subtilis fibrinolytic enzyme, white quinoa alcohol, polygonum multiflorum and ginseng into powder, sieving the powder with a 80-100-mesh sieve, and mixing the powder in proportion by using a three-dimensional mixer to obtain the anti-aging composition.
The invention further protects the application of the anti-aging composition in preparing cosmetics, foods, health-care foods and medicines for eliminating in-vivo free radicals and increasing glutamylcysteine ligase and glutathione.
The invention further protects a medicament, which is characterized by comprising the bacillus subtilis fibrinolytic enzyme composition with the anti-aging function.
As a further improvement of the invention, the medicament further comprises any one or a combination of at least two of pharmaceutically acceptable carriers, excipients or diluents.
Bacillus subtilis fibrinolytic enzyme: is a protein, namely bacillus subtilis fibrinolytic enzyme, which is produced by fermenting bacillus subtilis naturally existing in nature through soybean peptone and the like. The bacillus subtilis fibrinolytic enzyme has the effects of protecting blood vessels, purifying blood, clearing free radicals in vivo and the like, and has various effects of clearing free radicals, relieving blood vessel injury, protecting blood vessels and the like. At present, no document reports about the application of the bacillus subtilis plasmin in the aspect of anti-aging are found, and according to the experimental data, the single bacillus subtilis plasmin has the effects of clearing free radicals in mice and slowing aging, and the effect is more obvious by adding some traditional Chinese medicine components.
Resveratrol: resveratrol, a non-flavonoid polyphenolAn organic compound is an antitoxin produced by many plants when stimulated, and has a chemical formula of C 14 H 12 O 3 . Can be synthesized in grape leaf and grape skin, and is bioactive component in wine and grape juice. Resveratrol can activate acetylase, slow down the damage of free radicals in organism to cell and prolong the life of organism. At present, research shows that resveratrol can be used as an activator of the strongest silent information factor (SIRT 1), can simulate Caloric Restriction (CR) anti-aging reaction and participates in the regulation and control of the average life cycle of organic organisms. CR is a strong inducer of SIRT1, can increase the expression of SIRT1 in organs and tissues such as brain, heart, intestine, kidney, muscle, fat and the like, and can cause physiological changes of delaying senility and prolonging life span, and can be prolonged by 50 percent most obviously. Research has proved that the resveratrol has the effect of prolonging the life of yeast, nematode, fruit fly and low-grade fish.
Polygonum multiflorum: polygoni Multiflori radix, also known as Polygonum multiflorum, radix Stephaniae Sinicae, caulis Polygoni Multiflori, etc. Is perennial winding liana of Polygonum of Polygonaceae, with thick root tuber, oblong shape, and dark brown color. The term "long-term taking of long bones and tendons, benefiting essence and marrow, and not aging" from Kaibao materia Medica of Song Dynasty "is recorded in Kaibao Ben Cao. Modern researches find that the polygonum multiflorum can promote the growth of nerve cells and has an auxiliary treatment effect on neurasthenia and other nervous system diseases. And can regulate serum cholesterol, lower blood sugar, and improve the capacity of liver cells to convert and metabolize cholesterol. The fleece-flower root also has good free radical clearing and antioxidation effects.
Ginseng: ginseng is a perennial herb of the genus Panax of the family Araliaceae. Ginseng is a precious medicinal plant, and is believed from Shen nong Ben Cao Jing that Ginseng can "tonify five internal organs, calm spirit, calm soul, stop palpitation, remove pathogenic factors, improve eyesight, and improve mentality. Can lighten the body and prolong the life after long-time taking. Modern researches find that the health-care tea also has the effects of resisting oxidation, resisting aging, resisting fatigue, protecting liver, regulating cardiovascular function, exciting hematopoietic system function and the like. The anti-aging research of people over 50 years old by using the ginseng fruit saponin, such as Jilin Chinese medicine research institute Huo Yushu, proves that the ginseng fruit saponin has the anti-aging effect.
Has the advantages that: the anti-aging composition combines bacillus subtilis fibrinolytic enzyme with a plurality of traditional Chinese medicine components, the traditional Chinese medicine composition which is mainly prepared from the bacillus subtilis fibrinolytic enzyme and is supplemented with resveratrol, polygonum multiflorum and ginseng is obtained by screening, and after the traditional Chinese medicine composition is mixed according to a specific proportion, the traditional Chinese medicine composition and the resveratrol are mutually synergistic, so that a greater sauce and vinegar inhibition curative effect is generated, and the composition is safe and has no side effect.
The bacillus subtilis fibrinolytic enzyme is produced by fermenting bacillus subtilis, and the main component in the bacillus subtilis fibrinolytic enzyme is the bacillus subtilis fibrinolytic enzyme. The bacillus subtilis fibrinolytic enzyme and the resveratrol, the polygonum multiflorum and the ginseng are synergistically acted by some digestive enzymes of a human body in a certain ratio in a digestive tract, so that main components in the resveratrol, the polygonum multiflorum and the ginseng can be degraded into small molecular compounds, and the small molecular compounds can be more smoothly absorbed by intestinal tracts, enter blood and enter the liver, the brain and the heart in a body along with the blood, remove free radicals in the body and protect the liver, the brain and the heart. The components are mutually promoted, and the effect is better than that of a single component. Meanwhile, the proportional relation among the components is optimized, and a better effect is exerted.
Wherein, the composition takes bacillus subtilis fibrinolytic enzyme as a main component. Too little bacillus subtilis plasmin component is not beneficial to achieving the best effect by cooperating with the traditional Chinese medicines, and too much bacillus subtilis plasmin increases the manufacturing cost and the effect is not obviously improved.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Polygonum multiflorum and ginseng are purchased from pharmacies of Tongji Tang, hubei.
The resveratrol is available from Riemerald Biotech, inc. of Henan.
The Bacillus subtilis QK02 strain is preserved in China center for type culture collection, the preservation place is Wuhan university in Wuhan, china, the preservation number is M203078, and the preservation date is 11/17/2003.
Example 1
The raw materials comprise the following components in parts by weight: 30 parts of bacillus subtilis fibrinolytic enzyme, 35 parts of resveratrol, 15 parts of polygonum multiflorum and 20 parts of ginseng.
The bacillus subtilis fibrinolytic enzyme is prepared by the following fermentation process.
(1) Preparing a peptone culture medium, and adding the following components in percentage by mass into the culture medium: 2% of peptone, 3% of glucose, 1% of disodium hydrogen phosphate, 0.1% of sodium dihydrogen phosphate, 0.05% of magnesium sulfate, 0.02% of anhydrous calcium chloride, 0.1% of defoaming agent and the balance of water, and sterilizing at 115 ℃ for 30min for later use;
(2) Inoculating 3% of Bacillus subtilis QK02 strain by weight into a peptone culture medium, and fermenting for 24h at 37 ℃;
(3) And centrifuging, microfiltering, concentrating and spray drying the fermentation liquor to obtain the bacillus subtilis fibrinolytic enzyme, wherein the activity of the bacillus subtilis fibrinolytic enzyme is more than 80000IU/g.
The preparation method comprises the following steps: grinding bacillus subtilis fibrinolytic enzyme, white quinoa alcohol, polygonum multiflorum and ginseng into powder, sieving the powder by a 100-mesh sieve, and mixing the powder in proportion by a three-dimensional mixer to obtain the anti-aging composition.
Example 2
The difference from the example 1 is that the raw material ratio is different.
The raw materials comprise the following components in parts by weight: 50 parts of bacillus subtilis fibrinolytic enzyme, 23 parts of resveratrol, 10 parts of polygonum multiflorum and 17 parts of ginseng.
Example 3
The difference from the example 1 is that the raw material ratio is different.
The raw materials comprise the following components in parts by weight: 60 parts of bacillus subtilis fibrinolytic enzyme, 20 parts of resveratrol, 8 parts of polygonum multiflorum and 12 parts of ginseng.
Example 4
The difference from the example 1 is that the raw material ratio is different.
The raw materials comprise the following components in parts by weight: 75 parts of bacillus subtilis fibrinolytic enzyme, 12 parts of resveratrol, 5 parts of polygonum multiflorum and 8 parts of ginseng.
Test example 1
The embodiment of the invention takes the treatment of a D-galactose-induced aging mouse as an example, and the specific process is as follows:
glutathione (GSH) is an important antioxidant substance in the body, and can scavenge lipid peroxide or oxygen free radicals in the body. 5363A method for preparing GSH from organism comprises catalyzing the synthesis of GSH in vivo by Gu Guangxian Cysteine Ligase (GCL), increasing the content of GCL to increase the content of GSH in organism, thereby ensuring the antioxidation and free radical scavenging effects of organism. The experimental study verifies the antioxidation and the anti-aging effect of various compositions by carrying out the gavage treatment of mice by using D-galactose and then assisting the anti-aging compositions of various components, measuring the GCL and GSH content in liver, brain, heart and other tissues of the mice.
200 SPF-grade healthy Kunming mice with the age of 6 months are selected, the male and female are half of each other, and the weight is 20 +/-2 g. After one week of laboratory acclimation of purchased mice, the mice were randomly divided into 10 groups of 20 mice each, each half male and female. 10 groups of rats were divided into ten groups in sequence. Wherein the first group is a normal group; the second group is a senescence model group; the third group is a bacillus subtilis fibrinolytic enzyme single treatment group; the fourth group is a resveratrol single treatment group; the fifth group is the fleece-flower root single treatment group; the sixth group is a ginseng monotherapy group; the seventh group was the group of example 1, using the antiaging composition prepared in example 1, 10 in total. The eighth group was the group of example 2, and 10 anti-aging compositions prepared in example 2 were used. The ninth group was the group of example 3, and 10 of the anti-aging compositions prepared in example 3 were used. The tenth group was the group of example 4, and 10 of the anti-aging compositions prepared in example 4 were used. Wherein the second to tenth groups established aging mouse models, each mouse was injected with D-galactose at a dose of 200mg/Kg per day on the neck and back; first group-control group was injected with equal amount of physiological saline in the neck and back at the same dose. Injections were continued for 42 days. Meanwhile, the third group to the tenth group are subjected to intragastric administration according to the treatment combination formula, and the intragastric administration treatment is carried out according to the dose of 20mg/Kg every day; the first and second groups were gavaged daily with the same dose of saline and also were gavaged for 42 consecutive days.
After day 43, the mice were anesthetized and treated with cryosurgery to remove decibels from the liver, brain and heart, and after washing with 4 ℃ saline, the organs were weighed and homogenized, the supernatant was removed and the GCL and GSH content of each tissue was determined separately.
As can be seen from the comparison of the data in tables 1 and 2, the liver, brain and heart of the mice in the normal group (first group) were significantly reduced in GCL and GSH compared with those in the aged group (second group), and the differences were statistically significant (p < 0.01). The success of model construction is shown.
TABLE 1 GCL content (ug/(min. Times mg), x. + -. S) of different tissues of mice
| Group of | n | Liver disease | Brain | Heart |
| First group | 20 | 2.51±0.24 | 3.80±0.31 | 4.00±0.33 |
| Second group | 20 | 1.39±0.08 ＊＊ | 2.78±0.15 ＊＊ | 2.79±0.25 ＊＊ |
And (3) annotation:
it is a new method for preparing the compound: it was shown that the GCL of liver, brain and heart in the senescent group was very significantly different from that in the normal group.
TABLE 2 mouse different tissue GSH content (ug/(min. Times.mg), x. + -.s)
| Group of | n | Liver disease | Brain | Heart and heart |
| First group | 20 | 0.37±0.04 | 0.51±0.05 | 0.54±0.06 |
| Second group | 20 | 0.12±0.03 ＊＊ | 0.26±0.04 ＊＊ | 0.25±0.04 ＊＊ |
Note that:
it is, zhu: indicating that GSH of the liver, brain and heart of the aging group was significantly different compared to the normal group.
TABLE 3 GCL content (ug/(min. Times mg), x. + -. S) of different tissues of mice
| Group of | n | Liver disease | Brain | Heart and heart |
| Third group | 20 | 2.00±0.33 ＊ | 3.51±0.35 ＊ | 3.55±0.70 ＊ |
| Fourth group | 20 | 2.11±0.45 ＊ | 3.44±0.56 ＊ | 3.15±0.50 ＊ |
| Fifth group | 20 | 1.89±0.11 ＊ | 2.99±0.41 | 2.90±0.51 |
| Sixth group | 20 | 1.77±0.21 | 2.84±0.33 | 2.67±0.67 |
| Seventh group | 20 | 2.20±0.61 ＊ | 3.55±0.75 | 3.61±0.53 |
| Eighth group | 20 | 2.45±0.27 | 3.66±0.47 | 3.88±0.56 |
| Ninth group | 20 | 2.49±0.31 | 3.59±0.50 | 3.92±0.66 |
| Tenth group | 20 | 2.30±0.41 | 3.21±0.56 | 3.88±0.49 |
Note that:
it is: the GCL of liver, brain and heart showed significant differences compared to the control group in the formula group.
TABLE 4 GSH content (ug/(min. Times mg), x. + -. S) in different tissues of mice
Note that:
it is: indicating that the GSH of liver, brain and heart of the formula group was significantly different compared to the control group.
By comparing the data in tables 1, 2, 3 and 4, it can be seen that, when the preparations are treated alone in several formulations, GSH and GCL of the liver, brain and heart of the mice have significant effects respectively (compared with the control group, GCL and GSH in three tissues with the effect of treating alone with bacillus subtilis plasmin and resveratrol have significant difference compared with the model group, and p is less than 0.05); wherein, the contents of GCL and GSH are improved by fleece-flower root and salvia miltiorrhiza, and a plurality of data have significant difference compared with a model group (the fifth group of liver and the seventh group of liver, p is less than 0.05), and most data have no significant difference. However, after the substances are prepared according to a certain proportion, the effect of the intragastric administration treatment is better than that of a single component, which shows that the components have synergistic effect. And when the formula is proper, the content of GCL and GSH in the aged mice can be improved to a normal level (compared with the normal group, p is more than 0.05, and no significant difference exists).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. The anti-aging composition is characterized by being prepared from the following raw materials in parts by weight: 50-60 parts of bacillus subtilis fibrinolytic enzyme, 20-25 parts of resveratrol, 8-13 parts of polygonum multiflorum and 15-20 parts of ginseng.
2. The anti-aging composition according to claim 1, wherein the Bacillus subtilis plasmin is prepared by fermentation of a strain Bacillus subtilis QK02 deposited in the chinese type culture collection with the preservation number of CCTCC No. M203078.
3. The anti-aging composition according to claim 2, wherein the Bacillus subtilis plasmin is prepared into spray-dried powder by performing centrifugation, microfiltration, ultrafiltration and spray drying on fermentation liquor obtained by fermenting a strain Bacillus subtilis QK 02.
4. The anti-aging composition according to claim 1, wherein the bacillus subtilis plasmin activity is not less than 80000IU/g.
5. A method for preparing the antiaging composition according to any one of claims 1-4, characterized by comprising the steps of: grinding bacillus subtilis fibrinolytic enzyme, glauberol, polygonum multiflorum and ginseng into powder, sieving and mixing according to a proportion to obtain the anti-aging composition.
6. The method of claim 5, wherein the sieving is performed with a mesh size of 80-100 mesh, and the mixing is performed with a mixer, preferably a three-dimensional mixer.
7. The method of claim 5, comprising the steps of: grinding bacillus subtilis fibrinolytic enzyme, white quinoa alcohol, polygonum multiflorum and ginseng into powder, sieving the powder with a 80-100-mesh sieve, and mixing the powder in proportion by using a three-dimensional mixer to obtain the anti-aging composition.
8. Use of the antiaging composition according to any one of claims 1 to 4 for producing cosmetics, foods, health foods and medicines for scavenging in vivo free radicals and elevating glutamylcysteine ligase and glutathione.
9. A medicament comprising the anti-senescence Bacillus subtilis plasmin composition of any one of claims 1-4.
10. The medicament of claim 9, further comprising any one or a combination of at least two of a pharmaceutically acceptable carrier, excipient, or diluent.
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