CN111388568A - Composition for delaying aging and enhancing immunity and application - Google Patents
Composition for delaying aging and enhancing immunity and application Download PDFInfo
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- CN111388568A CN111388568A CN202010286490.1A CN202010286490A CN111388568A CN 111388568 A CN111388568 A CN 111388568A CN 202010286490 A CN202010286490 A CN 202010286490A CN 111388568 A CN111388568 A CN 111388568A
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Abstract
The invention relates to a composition for delaying senility and enhancing immunity and application thereof, wherein the composition for delaying senility and enhancing immunity comprises the following raw material components: 550 parts of mixed algae, 10-20 parts of ginkgo leaf extract, 5-10 parts of trigonella foenum graecum seed extract, 0.1-0.3 part of tea polyphenol, 10-15 parts of selenium-enriched yeast and 7-9 parts of vitamin; the mixed algae comprises sargassum fusiforme, Codium spinulosum and Antarctic ice algae according to the weight ratio of 2-6:0.5-1: 0.5-1. The invention has the beneficial effects that: the composition can effectively remove redundant free radicals in a human body, and simultaneously enhances the capability of the human body to remove the free radicals, namely the composition can provide three-dimensional protection for diseases and aging caused by the free radicals from a cell layer. The invention is developed from the root cause of aging and diseases, has scientific compatibility, obvious curative effect, natural raw material source, safety and no toxic or side effect.
Description
Technical Field
The invention belongs to the technical field of health care products, and particularly relates to a composition for delaying senescence and enhancing immunity and application thereof.
Background
When a person breathes continuously, about 95% of oxygen enters the body from the respiratory system, is consumed by the combustion conversion into energy substances, and 1% to 4% of oxygen forms free radicals. Life cannot leave free radical activities. The human body can carry out various chemical reactions at every moment, can not stop at every moment, and can burn energy at every moment, and the 'mover' responsible for transferring energy is the free radical. The free radicals also play the role of messengers in human bodies to transmit signals, and the blood pressure is stable because NO free radicals are needed to participate, and the blood pressure is unstable when NO free radicals are lacked. The free radicals not only help to transmit energy and signals, but also can kill bacteria and parasites and eliminate toxins in human bodies. Under normal physiological conditions, the quantity of free radicals generated in a human body and the quantity of consumed free radicals are in a balanced state, and the free radicals do not harm cells of the human body. However, when the amount of the generated radicals increases or the consumption of the radicals decreases, excessive radicals are formed, and if the excessive radicals cannot be removed in time, the excessive radicals may cause damage to human cells, thus causing various diseases and aging of the human body.
The human body is provided with two systems for eliminating redundant free radicals, one is an enzyme system, the other is a non-enzyme system, and the two systems prevent the free radicals from damaging normal cells so as to maintain the balance of the free radicals, thereby protecting the safety of the cells and avoiding diseases. The enzyme system for eliminating free radicals mainly comprises four enzyme substances, namely superoxide dismutase (SOD), Catalase (CAT), glutathione enzyme (GSH-Px) and glutathione-S-transferase (GST). The two enzymes with the greatest effect are superoxide dismutase (SOD) and glutathione enzyme (GSH-Px), wherein the superoxide dismutase (SOD) mainly eliminates superoxide anion free radicals (0)2 -) The glutathione enzyme (GSH-Px) has strong capability of eliminating hydroxyl radicals (0H.), which are the most active and toxic radicals and the most harmful radicals to human cells and are most easy to cause canceration of cells.
Two antioxidant systems are available for the human body to scavenge unwanted free radicals in the body, why are aging and diseases? The four enzymes for scavenging free radicals are mainly derived from liver and are synthesized by liver, after the age of 50, the number of liver cells is reduced sharply, and the number of the four enzymes synthesized by liver is reduced, so that the capability of an enzyme system for scavenging free radicals is reduced continuously. Another reason is that under the action of certain factors, such as endogenous factors including depression, sadness, fear, overwork, diseases or exogenous factors including food additives, residual pesticides, medicines, radiation and bad living habits, a large amount of free radicals are locally generated in a human body in an explosive manner in a short time, and the free radical scavenging system of the human body cannot complete the task of scavenging the free radicals in a very short time. Because the ability of endogenous free radical scavengers to scavenge free radicals is continuously reduced along with the aging, more exogenous free radical scavengers are searched and discovered in order to reduce the damage of the free radicals to human cells, and the substances are used as a 'shield', so that the attack of the free radicals to the cells is blocked, and the purposes of delaying aging, enhancing immunity and avoiding the occurrence and development of diseases are achieved.
There are dozens of kinds of free radicals in human body, and there are mainly 9 kinds of free radicals having the greatest influence on human body, including: superoxide anion radical (O)2 -H), hydroxyl radical (OH), hydrogen peroxide radical (H)2O2Hydrogen peroxide, peroxy radicals (HOO-), alkoxy Radicals (RO), peroxy alkyl radicals (ROO-), hydroperoxidesSubstance (ROOH), nitroxyl radical (NO) and singlet oxygen1O2. Among them, the alkane radical (R. cndot.), the alkoxy radical (R0. cndot.) and the peroxy alkane radical (R00. cndot.) are called fatty radicals, which are the products after the oxygen radicals attack the unsaturated fatty acids. The other radicals belong to the non-fatty ones. Each free radical has different characteristics, long and short survival time, strong and weak toxicity (attack ability to other substances), and large and small distribution range. Compared with the prior art, the number of superoxide anion free radicals is the largest, and the distribution range is the widest; hydroxyl radical is the most toxic and most likely to cause cellular variation; the singlet oxygen free radicals exist in the human body for the shortest time, only 100 ten-thousandth of a second. At present, the existing exogenous free radical scavengers in the market are mainly single or compound vitamin products and the like, have very limited free radical scavenging capacity, narrow range of free radicals capable of scavenging, and cannot play an effective scavenging role on all free radicals, so that the problem of self free radical scavenging capacity reduction and local free radical outbreak caused by age increase are difficult to truly compensate.
Disclosure of Invention
The composition is used as an exogenous free radical scavenger, and through scientific compatibility, multiple antioxidant mechanisms are complemented and matched for synergism, so that harmful free radicals of all types of human bodies can be eliminated, and the antioxidant effect is remarkable. The composition can supplement an antioxidant system of a human body, and effectively makes up for the deficiency of the capability of removing free radicals caused by factors such as aging, diseases and the like.
The technical scheme of the invention is as follows:
a composition for delaying senility and enhancing immunity comprises the following raw material components: 550 parts of mixed algae, 10-20 parts of ginkgo leaf extract, 5-10 parts of trigonella foenum graecum seed extract, 0.1-0.3 part of tea polyphenol, 10-15 parts of selenium-enriched yeast and 7-9 parts of vitamin; the mixed algae comprises sargassum fusiforme, Codium spinulosum and Antarctic ice algae according to the weight ratio of 2-6:0.5-1: 0.5-1.
Mixed algae, 1, sargassum fusiforme is one of algae plants of sargassum of brown algae phylum, and the edible value and the medicinal value of the sargassum fusiforme are recorded in Shen nong Ben Cao Jing. Nowadays, a great number of reports have been made that the sargassum fusiforme containing sulfated polysaccharide has biological functions of anticoagulation, blood fat reduction, oxidation resistance, immunity regulation and the like. The sulfated polysaccharide can improve the protection effect of antioxidant system of organism, and relieve diseases. Studies show that the sulfated hizikia fusiforme polysaccharide can reduce the level of lipid oxidation products and active oxygen by increasing the enzyme activity of antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the like in vivo. 2. The sea spiny algae is tender algae of sea spiny algae (Codium fragile, soft vegetable, etc.) Hariot, namely, sea spiny algae, black green algae, spongy mass, white villi, 10-30 cm high and upward multi-branch branches, grows on low-tide rock, Bohai sea and yellow sea, and is harvested for fresh eating, each hectogram of the sea spiny algae contains 13.3 g of protein, 0.45 g of fat and 16.79 g of carbohydrate, and also contains various vitamins, minerals, etc. the sea spiny algae has sweet and salty taste, has the effects of clearing away heat and toxic materials, relieving swelling and promoting diuresis and expelling parasites, is used for edema, dysuresia, etc. the main component of the sea spiny algae is carbohydrate, accounts for 66.86%, wherein the insoluble dietary fiber accounts for 43.04%, the protein content is higher, accounts for 21.67%, the total amino acid accounts for 42.2%, the amino acid constitutes high-quality protein, the fat content is low, 3.47%, wherein the unsaturated fatty acid is abundant and 47.46%, especially the polyunsaturated fatty acid such as EPA with remarkable health care function is high in content; the mineral elements are rich, especially the trace elements Cu, Fe and Zn are high. Therefore, the codium spinulosum is a natural ideal health food raw material with high dietary fiber, high protein, low fat and low calorie and rich in trace mineral elements such as iron, zinc and the like. 3. Antarctic ice algae is an algae plant growing in a fixed ice area and a floating ice area of an Antarctic sea area, mainly refers to large-scale micro algae growing in the Antarctic extreme environment sea ice, contains higher algae essence than common algae due to long-term cold absorption, and is pure natural algae food.
Ginkgo biloba extract, ginkgo tree is one of ancient tree species in China, is called as "activating stone" in the plant kingdom, has extremely strong vitality, Japanese Guangdong island and Nagasaki were attacked by atomic bomb, and the first plant to grow in the Guangdong island and the Nagasaki after war is the ginkgo tree. The chemical components of ginkgo leaf include more than 160 kinds of bilobalide, chelidonine, kaempferol and the like, and the ginkgo leaf serving as a traditional Chinese medicine has a history of more than one thousand years and is mainly used for treating asthma and bronchitis. In the sixties of the last century, German scientists extracted ginkgetin and bilobalide from ginkgo leaves, and the production process is named as EGb 761. Thereafter, ginkgo biloba extract (EGb) is used for treating various diseases such as cardiovascular and cerebrovascular diseases worldwide. The ginkgo leaf extract (EGb761) is a mixture of a group of flavonoids, glucose oxib and limonene, has unique pharmacological activity, and has three main active ingredients: the ginkgo flavone containing trihydroxy flavonoids accounts for about 24 percent, the ginkgo mushroom lactone accounts for about 6 percent, and the organic acid accounts for about 5 to 10 percent. The ginkgo leaf extract has obvious antioxidation, and the antioxidation and the free radical removal function of the ginkgo leaf extract are realized by three modes: directly inactivating various oxygen free radicals; increasing the activity of antioxidant enzymes including superoxide dismutase (SOD) and glutathione enzyme (GSH-Px); inhibiting lipid peroxidation. The folium Ginkgo extract can not only remove free radicals in cell membrane and biomembrane, but also remove free radicals distributed on membrane surface. The invention uses the ginkgo leaf extract and the vitamin C in a matching way, under the condition of equal concentration, the effect of removing free radicals is greatly improved, and the ginkgo leaf extract and the vitamin C have a matching synergistic effect in the aspects of removing superoxide anions and hydroxyl free radicals. On the other hand, the ginkgo leaf extract contains hydroxyl and phenolic hydroxyl structures, while algal polysaccharides rich in the mixed algae also contain hydroxyl and phenolic hydroxyl structures, and the same chemical structure provides conditions for the combined use of two active substances to exert a synergistic and synergistic effect.
Semen Trigonellae extract, semen Trigonellae is an annual herbaceous plant of Leguminosae, and has fragrance in whole plant, flowering phase of 4-6 months, and fruit phase of 7-8 months. It is mainly distributed in Anhui, Sichuan and Henan, and is thought to have the functions of invigorating kidney yang and dispelling cold and dampness by traditional Chinese medicine. Fenugreek seeds contain abundant proteins and vitamins and generally consist of: 10.30% of water, 3.15% of ash, 7.61% of fat, 16.97% of total protein, 3.68% of total sugar, 6.4% of cellulose and 11.98% of hemicellulose. The fatty oil contains solid fatty acid 92.9%, volatile fatty acid 1.5%, and unsaponifiable component 0.9%. The fatty acids are mainly linoleic acid and palmitic acid, and have a small amount of oleic acid and linolenic acid. The main total sugar is galactomannan and vitamin B1. The main active ingredients of the fenugreek seeds comprise alkaloids such as gentiinine, caricaine, choline and trigonelline, the alkaloids are nitrogen-containing alkaline organic compounds, most of the alkaloids have a ring structure, and nitrogen elements are contained in the ring, so that the fenugreek seeds have a remarkable effect of removing free radicals. Modern scientific research proves that the fenugreek can reduce cholesterol, control and treat diabetes, menopausal syndrome and the like.
Tea polyphenols, which are the general term for polyphenols in tea, are complexes of compounds such as theaflavins (flavanols), flavones, flavonols, anthocyanins, phenolic acids, depside acids, polymeric phenols, and mainly include four major classes of substances, catechins, flavones, anthocyanins, and phenolic acids. The content of catechins is the highest and accounts for 60-80% of the total amount. The catechins are mainly Epicatechin (EC), epicatechin gallate (ECG), Epigallocatechin (EGC) and epigallocatechin gallate (EGCG). Tea polyphenols have strong antioxidant effect, and have been widely used in food industry in addition to medical field. The antioxidation of tea polyphenol is realized by four ways: the oxygen free radical is directly eliminated; inhibiting lipid peroxidation; inducing metal ion complexation; activate the antioxidant defense system in the cell.
Selenium-enriched yeast, selenium is an indispensable trace element in human body, and the chemical property of selenium is between metal and nonmetal. The selenium content in adult is 14-21mg, the selenium content in liver, pancreas and kidney is high, and the selenium exists in the form of selenoprotein. Selenium scavenges free radicals about 10 times as effectively as vitamin E. Vitamin CWhile biotin functions primarily to scavenge free radicals at the cell membrane and biofilm surface, reduce lipid hydroperoxides, and protect cell membranes and biofilms from oxidative damage, selenium not only scavenges free radicals at the cell membrane surface, but also scavenges free radicals deep in cell membranes and biofilms, and scavenges hydroperoxides, lipid hydroperoxides, and other peroxides, thereby protecting cell membranes and biofilms. The free radical scavenging effect of selenium is mainly realized by glutathione enzyme (GSH-Px), and the selenium is positioned at the center of the activation reaction of the glutathione enzyme to endow the glutathione enzyme (GSH-Px) with activity, thereby enhancing the enzyme system of a human body. On the other hand, selenium and vitamin E have a synergistic effect in scavenging free radicals to protect cells. The vitamin E mainly inhibits free radicals from attacking unsaturated fatty acids, prevents cell membranes and biological membranes from undergoing lipid peroxidation chain reaction, and reduces peroxide generation. Although vitamin E has a strong ability to scavenge peroxides, it is not possible to completely prevent the generation of peroxides, which are very susceptible to generating more free radicals and are very harmful. Glutathione enzyme (GSH-Px) containing selenium can decompose peroxide and prevent peroxide from generating hydroxyl radical (OH) and singlet oxygen (G:)1O2) Playing a role in compensation. Selenium and vitamin E cooperate to cooperatively maintain the stability of cell membranes and biological membranes. Natural selenium is classified into two types, one is inorganic selenium including sodium selenate and sodium selenite, and the other is organic selenium. Inorganic selenium has no bioactivity and low absorption and utilization rate. The organic selenium is obtained by biotransformation of inorganic selenium to give bioactivity. The biological transformation is divided into two types, one is the transformation through plants, and the other is the transformation through a biological fermentation method, and the selenium-enriched yeast belongs to the microbial transformation. The organic selenium formed by microbial fermentation and conversion not only has high selenium concentration and stable content, but also has very high bioactivity, high absorption and utilization rate and stronger physiological functions including free radical removal, immunity enhancement, cancer resistance and the like.
The formula mechanism of the invention is that more than 60 free radicals exist in a human body, each free radical has different characteristics, and more antioxidant substances are found in the nature, but the ability of each antioxidant substance for scavenging free radicals is different.
Preferably, the raw material components comprise: 500 parts of mixed algae, 15 parts of ginkgo leaf extract, 8 parts of trigonella seed extract, 0.2 part of tea polyphenol, 12 parts of selenium-enriched yeast and 8.16 parts of vitamin; the mixed algae is composed of sargassum fusiforme, Codium spinulosum and Antarctic ice algae according to the weight ratio of 3:1: 1.
Preferably, the ginkgo biloba extract has a ginkgo flavone content of not less than 24 wt% and a ginkgolide content of not less than 6 wt%.
Preferably, the vitamin is a mixture of vitamin E and vitamin C in a weight ratio of 0.5-1.5: 5-8. Vitamin E and vitamin C are selected because the two substances have synergistic anti-oxidation effect, vitamin C reacts with vitamin E free radical (VE ·) to restore vitamin E free radical (VE ·) to reduced vitamin E, vitamin C itself becomes vitamin C free radical (VC ·), and vitamin C free radical (VC ·) can be reduced to vitamin C under the action of redox enzyme (NADH) system. The synergistic effect of vitamin C and vitamin E can improve the efficiency of scavenging free radicals. In addition, the vitamin E and the vitamin C can also be matched with other raw materials to realize a synergistic effect and further improve the antioxidant effect. In particular, the research shows that vitamin E, vitamin C and selenium have a coordination effect and form a triangular relationship. Although vitamin E has strong capacity of eliminating peroxide, the generation of the peroxide cannot be completely prevented, and the peroxide is very easy to generate chain reaction to generate more free radicals, so that the harm is very great. Glutathione enzyme (GSH-Px) containing selenium can decompose peroxide and prevent peroxide from generating hydroxyl radical (0H.) and singlet oxygen ((0H))1O2) Playing a role in compensation. In addition, selenium and vitamin E are matched, and the selenium-vitamin E compound also has the effect of cooperatively maintaining the safety of cell membranes and biological membranes.
Preferably, the fenugreek seed extract is prepared by the following method: (1) drying and crushing fenugreek seeds, and adding absolute ethyl alcohol to perform first ultrasonic extraction to obtain a first extracting solution; adding 50-80% ethanol into the reaction residue after the first ultrasonic extraction for second ultrasonic extraction to obtain a second extracting solution; mixing extractive solutions, filtering, and concentrating to obtain concentrated solution; (2) diluting the concentrated solution with water, sequentially purifying with D101 macroporous adsorbent resin column, eluting with 80% ethanol, collecting eluate, concentrating under reduced pressure, and spray drying.
Preferably, in the step (1), the material-liquid ratio of the first ultrasonic extraction is 1:10-20g/m L, the ultrasonic extraction power is 220- & lt 400 & gt, the ultrasonic extraction temperature is below 40 ℃, the extraction time is 25-30min, the material-liquid ratio of the second ultrasonic extraction is 1:40-60g/m L, the ultrasonic extraction power is 600- & lt 800 & gt, the extraction temperature is 40-60 ℃, the extraction time is 15-20min, and the concentration is carried out until the volume is 1/4-1/3.
Preferably, the composition for delaying senility and enhancing immunity is a preparation prepared by taking the raw materials as active ingredients and adding pharmaceutically acceptable auxiliary materials. Preferably, the preparation is any one of oral tablets, capsules, granules and solutions.
The composition for delaying senility and enhancing immunity is applied to the preparation of foods, health-care products and medicines for improving immunity and delaying senility.
The invention has the beneficial effects that:
1. according to the invention, the mixed algae consisting of sargassum fusiforme, spiny algae and Antarctic ice algae is combined with the ginkgo leaf extract, trigonella foenum-graecum seed extract, tea polyphenol, selenium-enriched yeast and vitamins and reasonably proportioned, so that a remarkable antioxidant synergistic effect is generated among the raw materials, on one hand, the algae is used as the supplement of an exogenous free radical scavenger to effectively remove redundant free radicals causing diseases and aging in a human body, not only can water-phase free radicals be removed, but also lipid-phase free radicals and free radicals at water-phase and lipid-phase interfaces can be removed, on the other hand, the activity of antioxidant enzymes of the human body is enhanced, and the capability of the endogenous free radical scavenger of the human body to remove free radicals is greatly improved. Through the two action mechanisms, the invention can provide three-dimensional protection for diseases and aging caused by free radicals from a cellular level. The invention develops from the root cause of aging and diseases, has scientific compatibility, obvious curative effect, natural raw material source, safety and no toxic or side effect
2. The composition has double health-care functions of enhancing immunity and delaying senility.
3. The composition has the advantages of easily available raw materials, high safety and simple preparation method, and is suitable for popularization and utilization.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention. In the following examples, 1 part by weight represents 1 g.
Example 1
The embodiment provides a composition for delaying senescence and enhancing immunity, which comprises the following raw materials in parts by weight:
the method comprises the following steps of (1) drying and crushing fenugreek seeds, adding absolute ethyl alcohol to carry out first ultrasonic extraction (material-liquid ratio: 1:10g/m L, 40 ℃, 600w, 20min) to obtain a first extracting solution, adding 50% ethyl alcohol to reaction residues obtained after the first ultrasonic extraction to carry out second ultrasonic extraction (material-liquid ratio: 1:10g/m L, 40 ℃, 600w, 20min) to obtain a second extracting solution, combining the extracting solutions, filtering to obtain an extracting solution, concentrating the extracting solution into a concentrated solution, adding water, concentrating the concentrated solution into a concentrated solution, and carrying out macroporous adsorption and purification to obtain a macroporous resin eluent, wherein the concentrated solution is prepared by adding water, concentrating the concentrated solution into a column containing 80% of pure water, and concentrating the macroporous resin to obtain a concentrated solution, and concentrating the macroporous resin.
Example 2
The embodiment provides a composition for delaying senescence and enhancing immunity, which comprises the following raw materials in parts by weight:
550 parts of mixed algae, 10 parts of ginkgo biloba extract (24 wt% of ginkgetin and 6 wt% of ginkgolide), 10 parts of fenugreek seed extract, 0.1 part of tea polyphenol, 15 parts of selenium-enriched yeast and 7 parts of vitamin, wherein the mixed algae comprises sargassum fusiforme, acanthomonas spinosa and antarctic ice algae in a weight ratio of 6:1:0.5, the vitamin is a mixture of vitamin E and vitamin C in a weight ratio of 1.5:5, the fenugreek seed extract is prepared by (1) drying and crushing fenugreek seeds, adding absolute ethyl alcohol to perform first ultrasonic extraction (material-liquid ratio: 1:20g/m L, 40 ℃, 400w and 25min) to obtain a first extract, adding 80% ethyl alcohol to reaction residues subjected to the first ultrasonic extraction to perform second ultrasonic extraction (material-liquid ratio: 1:60g/m 8632, 60 ℃, 800w and 15min), mixing the extracts, filtering the extracts until the volume of the extracts is 52%, concentrating by adding water, concentrating, sequentially, concentrating, and concentrating by using a macroporous resin to obtain a concentrated eluent, and concentrating and purifying by using a macroporous resin column (2) to obtain a concentrated eluent after the eluent is performed.
Example 3
The embodiment provides a composition for delaying senescence and enhancing immunity, which comprises the following raw materials in parts by weight:
500 parts of mixed algae, 15 parts of ginkgo leaf extract, 8 parts of trigonella foenum-graecum seed extract, 0.2 part of tea polyphenol, 12 parts of selenium-enriched yeast and 8.16 parts of vitamin, wherein the mixed algae is prepared by drying and crushing trigonella foenum-graecum, garcinia spinifera and antarctic ice algae in a weight ratio of 3:1:1, the vitamin is a mixture of vitamin E and vitamin C in a weight ratio of 0.96:7.2, the trigonella foenum-graecum seed extract is prepared by (1) drying and crushing trigonella foenum-graecum seeds, adding absolute ethyl alcohol to perform first ultrasonic extraction (material-liquid ratio: 1:15g/m L, 40 ℃, 300w and 25min) to obtain a first extract, adding 70% ethyl alcohol to reaction residues subjected to first ultrasonic extraction to perform second ultrasonic extraction (material-liquid ratio: 1:50g/m L, 50 ℃, 600w and 20min) to obtain a second extract, combining, filtering and concentrating to obtain a concentrated solution with the volume of the original 1/3, adding water to perform reduced pressure adsorption, concentrating, performing concentration, performing spray drying, and collecting macroporous elution, and purifying by using a macroporous resin column to obtain an eluent, and purifying eluent.
Further, a method for preparing the tablet A by the composition for delaying senility and enhancing immunity is provided:
(1) diluting tea polyphenol: uniformly mixing tea polyphenol and 19 parts by weight of starch to obtain a mixture A;
(2) primary mixing: adding 20 parts by weight of lactose, 25 parts by weight of microcrystalline cellulose, a ginkgo biloba extract, a fenugreek seed extract, selenium-enriched yeast and vitamins into the mixture A in sequence, and uniformly mixing to obtain a mixture B;
(3) and (3) secondary mixing: fully and uniformly mixing the mixture B with the mixed algae to obtain a mixture C;
(4) sieving: sieving the mixture C with a 40-mesh sieve;
(5) tabletting, wherein the weight of the tablet is 0.6 g;
(6) and (5) spraying a film coat. The use equipment comprises the following steps: a suspension coating machine; coating materials: hydroxypropyl methylcellulose (HPMC) with concentration of 2% (at 20 deg.C), viscosity of 6-7cP, and pH of 6-8; the weight increase of the coated tablets is controlled to be 1-2 percent;
(7) content determination: measuring according to the enterprise standard;
(8) subpackaging in plastic bottles, and labeling;
(9) radiation sterilization; cobalt 60 irradiation with a dose of 6-8KGY for 180 minutes;
(10) and (5) hygiene examination.
Comparative example 1
This comparative example provides a composition for delaying aging and enhancing immunity, which differs from the composition described in example 3 only in that the mixed algae consists of codium spinulosum and antarctic ice algae in a weight ratio of 1: 1. Further, tablet B including the composition described in this comparative example was prepared according to the tablet preparation method of example 3.
Comparative example 2
This comparative example provides a composition for delaying aging and enhancing immunity, which is different from the composition described in example 3 only in that the mixed algae is composed of Hizikia fusiforme and Antarctic ice algae in a weight ratio of 3: 1. Further, tablet C including the composition described in this comparative example was prepared according to the tablet preparation method of example 3.
Comparative example 3
This comparative example provides a composition for delaying aging and enhancing immunity, which is different from the composition described in example 3 only in that the mixed algae is composed of Hizikia fusiforme and Codium spinulosum in a weight ratio of 3: 1. Further, tablet D including the composition described in this comparative example was prepared according to the tablet preparation method of example 3.
Comparative example 4
This comparative example provides a composition for delaying aging and enhancing immunity, which is different from the composition described in example 3 only in that mixed algae is replaced with a single Hizikia fusiforme. Further, tablet E including the composition described in this comparative example was prepared according to the tablet preparation method of example 3.
Comparative example 5
This comparative example provides a composition for delaying aging and enhancing immunity, which differs from the composition described in example 3 only in the preparation method of fenugreek seed extract, eliminating the step of second ultrasound extraction. Further, a tablet F including the composition described in this comparative example was prepared according to the tablet preparation method of example 3.
Experimental study of immunomodulatory Effect of the invention
1. Test samples: example 3 and comparative examples 1-5.
2. Test animal
70 female second-class mice with female age of Kunming 1 provided by the experimental animal center of Chinese medical academy of sciences, the weight of which is 18-22g (license number: SCXK11-00-0006), are randomly divided into 7 groups of a control group and tablets A-F according to the weight, and each group comprises 10 mice. Body weights were not significantly different between groups by T-test (p > 0.05).
3. Dose selection
And (3) according to the recommended dose of a human body, performing intragastric administration on the mice 30 times of 4.8g/60kg.bw every day, namely performing intragastric administration on the mice once every day by adopting the dose of 2.4g/kg.bw, performing intragastric administration on the mice of a control group by using distilled water with the same volume, and measuring each index after each group of mice continuously administer the medicine for 28 days.
4. Instruments and reagents
752 spectrophotometer, AE100 electronic balance, electric heating three-purpose water tank, water-proof constant temperature incubator, screw micrometer, KA-1000 type desk centrifuge, 80-2 type desk centrifuge, slide rack, carbon dioxide incubator, super clean bench, 24-hole culture plate, microscope, 96-hole culture plate, CF-plate type enzyme labeling instrument, desk type multifunctional high speed freezing centrifuge;
sheep Red Blood Cells (SRBC), Hank's solution, India ink, sodium carbonate, guinea pig serum, normal saline, SA buffer solution, RPM1640 culture solution, calf serum, 2-mercaptoethanol, penicillin, streptomycin, concanavalin A (ConA), hydrochloric acid, isopropanol, MTT, PBS buffer solution (pH7.2-7.4), chicken red blood cells, acetone, methanol, Giemsa staining solution, sodium lactate, tetrazole nitrochloride, phenazine dimethyl sulfate and oxidized coenzyme I.
5. Detection method
5.1 ConA-induced mouse lymphocyte transformation assay (MTT method)
Sterile collecting spleen of mouse, placing in a small plate containing appropriate amount of sterile Hank's solution, gently grinding into single cell suspension, filtering with 200 mesh sieve, washing with Hank's solution for 3 times, centrifuging for 10min (1000r/min) each time, suspending spleen cells in 1640 complete culture solution of 2.00m L, counting by microscopic examination, and adjusting cell concentration to 2 × 106Adding the cell suspension into a 24-well culture plate in two wells, adding 0.050m L (0.1mg/m L) ConA solution into one well, placing the well in a carbon dioxide incubator at 37.0 ℃ for culturing for 72h, wherein the concentration of carbon dioxide is 5%, adding MTT (5mg/m L) 4h before the culture is finished, adding 1.00m L acid isopropanol after the culture is finished, uniformly blowing, uniformly mixing, measuring the optical density value at 570nm, and adding ConAThe optical density of the wells minus the optical density of the wells without ConA represents the proliferative capacity of the lymphocytes.
5.2 delayed hypersensitivity (DTH) (thickening of the plantar aspect)
Each mouse was immunized by intraperitoneal injection of 0.2m L2% (V/V) SRBC 4d after immunization, left and right toe thickness was measured by a micrometer screw, then 0.02m L20% (V/V) SRBC was subcutaneously injected at the measurement site, left and right toe thickness was measured 24h after, the same site was measured 3 times, and the degree of DTH was expressed as the difference in toe thickness before and after the challenge by taking the mean value.
5.3 antibody producing cell (PFC) assay
Intraperitoneal injection of 0.2m L2% (V/V) SRBC for immunizing each mouse for 5 days, and preparing the spleen of the mouse into the cell concentration of 5 × 106Spleen cell fluid of L/m, agarose was dissolved to make a 1% solution, incubated in a 48 ℃ water bath, mixed with an equal amount of 2-fold concentration of Hank's solution, dispensed into small tubes, 0.5m L per tube, 0.05m L10% SRBC and 0.01m L spleen cell suspension were added, mixed well rapidly, poured onto slides, incubated for 1h in a 37 ℃ incubator, complement was added to the slide bath, and the plaques were counted after 1.5h incubation.
5.4 measurement of serum hemolysin
Injecting 0.2m L2% (V/V) SRBC into the abdominal cavity to immunize each mouse, taking blood from the eyeball and placing the blood in a centrifugal tube after 5d, placing the blood for 1h, centrifuging for 10min at 2000r/min, collecting serum, diluting the serum of the mouse by 400 times by using SA liquid, sequentially adding 1.00m L of the diluted serum of the mouse, 0.5m L of SRBC0.5m of 10% (V/V) and complement (diluted by using SA liquid 1: 10) 1.00m L, additionally arranging a control tube without adding serum (replacing by using SA liquid), placing the control tube in a water bath at 37 ℃ for 20min, stopping the reaction by using an ice water bath, centrifuging for 10min at 2000r/min, taking 1.00m L of supernatant, placing 3.00m L of all reagents in the test tube, simultaneously taking 10% (V/V) SRBC0.25m L and 4.00m L of all reagents as half dissolved blood vessels, placing the test result of the optical density value at the wavelength of 540nm is expressed as a half-hemolysis value (50).
5.5 mouse carbon clearance test
The mouse was administered a 4-fold dilution of India ink (0.1m L/10 g by intravenous injection) into the tail of each mouse, and when the ink was recorded immediately after the injection, 0.02m L blood was added to a 2.00m L sodium carbonate solution 1 and 10min after the injection, respectively, and the optical density was measured at a wavelength of 600nm, using the sodium carbonate solution as a control, and the liver and spleen were weighed to express the mouse's ability to clear carbon by phagocytosis index (α).
5.6 experiment of phagocytosis of chicken red blood cells by macrophages in abdominal cavity of mouse
Injecting 20% chicken erythrocyte suspension into abdominal cavity of mouse at 1.0m L at intervals of 30min, dislocating cervical vertebra, cutting abdominal wall skin, injecting Hank's solution into abdominal cavity at 2.0m L, massaging abdominal cavity, sucking out abdominal cavity washing liquid at 1.0m L, dripping onto two glass slides respectively, incubating in 37.0 deg.C incubator for 30min, fixing with 1:1 acetone methanol solution, staining with 4% (V/V) Giesma-phosphoric acid buffer solution, rinsing with distilled water, air drying, counting macrophages under oil microscope, counting 100 per slide, and expressing phagocytic capacity of mouse macrophages with macrophage phagocytic percentage and macrophage phagocytic index.
6. Test results
TABLE 1 Effect on ConA-induced lymphocyte transformation in mice (Unit: optical Density values)
Group of | Animal number (only) | Proliferative capacity of lymphocytes |
Control group | 10 | 0.205±0.067 |
Tablet A group | 10 | 0.257±0.042* |
Tablet B group | 10 | 0.213±0.051 |
Tablet C group | 10 | 0.221±0.049 |
Tablet D group | 10 | 0.228±0.073 |
Tablet E group | 10 | 0.219±0.038 |
Tablet F group | 10 | 0.233±0.035 |
Compared with the contrast group, the ratio of the composition,*p<0.05。
as can be seen from table 1, after 28 days of oral administration of each test substance to mice, the lymphocyte proliferation ability of tablet a group was increased by 25% (p <0.05) compared to the control group.
TABLE 2 Effect on delayed allergy (DTH) in Normal mice
Group of | Animal number (only) | Toe swelling difference (mm) |
Control group | 10 | 0.62±0.15 |
Tablet A group | 10 | 0.87±0.20** |
Tablet B group | 10 | 0.65±0.14 |
Tablet C group | 10 | 0.73±0.17 |
Tablet D group | 10 | 0.75±0.25 |
Tablet E group | 10 | 0.71±0.11 |
Tablet F group | 10 | 0.76±0.08 |
Compared with the contrast group, the ratio of the composition,**p<0.01。
as can be seen from table 2, after 28 days of oral administration of each test substance to mice, the toe swelling degree of tablet a group was increased by 40% (p <0.01) compared to the control group.
TABLE 3 Effect on PFC in Normal mice (units: log)10Number of plaques/full spleen)
Group of | Animal number (only) | PFC |
Control group | 10 | 4.77±0.41 |
Tablet A group | 10 | 5.20±0.23** |
Tablet B group | 10 | 4.80±0.28 |
Tablet C group | 10 | 4.82±0.33 |
Tablet D group | 10 | 4.88±0.46 |
Tablet E group | 10 | 4.86±0.37 |
Tablet F group | 10 | 4.90±0.27 |
Compared with the control group,**p<0.01。
As can be seen from table 3, the PFC level in tablet a group was increased by 9% (p <0.01) compared to the control group 28 days after oral administration of each test subject to the mice.
TABLE 4 Effect on Normal mouse serum hemolysin
Group of | Animal number (only) | HC50 |
Control group | 10 | 90±70 |
Tablet A group | 10 | 214±79** |
Tablet B group | 10 | 135±94 |
Tablet C group | 10 | 144±93 |
Tablet D group | 10 | 150±88 |
Tablet E group | 10 | 155±85 |
Tablet F group | 10 | 163±72* |
Compared with the contrast group, the ratio of the composition,*p<0.05,**p<0.01。
as can be seen from table 4, HC50 in tablet a group was increased 1.4-fold (p <0.01) compared to the control group 28 days after oral administration of each test substance to mice.
TABLE 5 Effect on carbon clearance in Normal mice
Compared with the contrast group, the ratio of the composition,*p<0.05,**p<0.01。
as can be seen from table 5, the clearance phagocytosis index of tablet a group was increased by 13% (p <0.01) compared to the control group 28 days after oral administration of each test substance to mice.
TABLE 6 Effect on macrophage phagocytosis Rate in Normal mice
Group of | Animal number (only) | Phagocytosis rate/%) |
Control group | 10 | 26±8 |
Tablet A group | 10 | 36±10* |
Tablet B group | 10 | 27±9 |
Tablet C group | 10 | 29±6 |
Tablet D group | 10 | 30±8 |
Tablet E group | 10 | 31±5 |
Tablet F group | 10 | 33±6* |
Compared with the contrast group, the ratio of the composition,*p<0.05。
as can be seen from table 6, the macrophage phagocytosis rate of tablet a group was increased by 38% (p <0.05) compared to the control group 28 days after oral administration of each test substance to mice.
TABLE 7 Effect of test substances on macrophage phagocytosis index in Normal mice
Compared with the contrast group, the ratio of the composition,*p<0.05。
as can be seen from table 7, the macrophage phagocytosis index of tablet a group was increased by 49% (p <0.05) compared to the control group 28 days after oral administration of each test substance to mice.
And (4) conclusion: 1. after the tablets A-F samples of the invention are orally given to mice for 28 days, compared with a control group, the lymphocyte proliferation capacity of the tablet A group is improved by 25 percent (p is less than 0.05), the toe swelling degree is improved by 40 percent (p is less than 0.01), the PFC number is improved by 9 percent (p is less than 0.01), HC50 is improved by 1.4 times (p is less than 0.01), the carbon clearance phagocytosis index is improved by 13 percent (p is less than 0.01), the macrophage phagocytosis rate is improved by 38 percent (p is less than 0.05), and the macrophage phagocytosis index is improved by 49 percent (p is less than 0.05), and the result is combined, thereby the invention has the obvious effect of enhancing immunity. 2. The indexes of the group A of the tablet are obviously superior to those of other groups, namely the immunoregulation function of the composition prepared from the mixed algae consisting of 3 raw materials of sargassum fusiforme, spiny pine algae and antarctic ice algae is obviously superior to the immunoregulation function of the composition prepared from any one or two of the 3 raw materials of sargassum fusiforme, spiny pine algae and antarctic ice algae and the immunoregulation function of the composition containing the trigonella foenum graecum seed extract prepared by different extraction methods.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. The composition for delaying senility and enhancing immunity is characterized by comprising the following raw material components:
550 parts of mixed algae, 10-20 parts of ginkgo leaf extract, 5-10 parts of trigonella foenum graecum seed extract, 0.1-0.3 part of tea polyphenol, 10-15 parts of selenium-enriched yeast and 7-9 parts of vitamin; the mixed algae comprises sargassum fusiforme, Codium spinulosum and Antarctic ice algae according to the weight ratio of 2-6:0.5-1: 0.5-1.
2. The composition for delaying aging and enhancing immunity according to claim 1, wherein the raw material components comprise:
500 parts of mixed algae, 15 parts of ginkgo leaf extract, 8 parts of trigonella seed extract, 0.2 part of tea polyphenol, 12 parts of selenium-enriched yeast and 8.16 parts of vitamin; the mixed algae is composed of sargassum fusiforme, Codium spinulosum and Antarctic ice algae according to the weight ratio of 3:1: 1.
3. The composition for delaying aging and enhancing immunity according to claim 1, wherein the ginkgo biloba extract has a ginkgo flavone content of not less than 24 wt% and a ginkgolide content of not less than 6 wt%.
4. The composition for delaying aging and enhancing immunity as claimed in claim 1, wherein the vitamin is a mixture of vitamin E and vitamin C in a weight ratio of 0.5-1.5: 5-8.
5. The composition for delaying aging and enhancing immunity according to claim 1, wherein the fenugreek seed extract is prepared by the following method: (1) drying and crushing fenugreek seeds, and adding absolute ethyl alcohol to perform first ultrasonic extraction to obtain a first extracting solution; adding 50-80% ethanol into the reaction residue after the first ultrasonic extraction for second ultrasonic extraction to obtain a second extracting solution; mixing extractive solutions, filtering, and concentrating to obtain concentrated solution; (2) diluting the concentrated solution with water, sequentially purifying with D101 macroporous adsorbent resin column, eluting with 80% ethanol, collecting eluate, concentrating under reduced pressure, and spray drying.
6. The composition for delaying senescence and enhancing immunity according to claim 5, wherein in the step (1), the material-liquid ratio of the first ultrasonic extraction is 1:10-20g/m L, the ultrasonic extraction power is 220-400w, the ultrasonic extraction temperature is below 40 ℃, the extraction time is 25-30min, the material-liquid ratio of the second ultrasonic extraction is 1:40-60g/m L, the ultrasonic extraction power is 600-800w, the extraction temperature is 40-60 ℃, the extraction time is 15-20min, and the concentration is carried out until the volume is 1/4-1/3.
7. The composition for delaying aging and enhancing immunity as claimed in claim 1, which is a preparation prepared from the raw materials as active ingredients and pharmaceutically acceptable auxiliary materials.
8. The composition for delaying aging and enhancing immunity according to claim 7, wherein the preparation is an oral preparation.
9. The composition for delaying aging and enhancing immunity according to claim 8, wherein the oral preparation is any one of tablets, capsules, granules and solutions.
10. Use of the anti-aging and immunity-enhancing composition according to any one of claims 1 to 9 for the preparation of foods, health products and medicines for enhancing immunity and delaying aging.
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