CN118105441A - Composition containing unsaturated fatty acid, and preparation method and application thereof - Google Patents
Composition containing unsaturated fatty acid, and preparation method and application thereof Download PDFInfo
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- CN118105441A CN118105441A CN202410168026.0A CN202410168026A CN118105441A CN 118105441 A CN118105441 A CN 118105441A CN 202410168026 A CN202410168026 A CN 202410168026A CN 118105441 A CN118105441 A CN 118105441A
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- oil
- fatty acid
- omega
- polyunsaturated fatty
- unsaturated fatty
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Abstract
The invention provides a composition containing unsaturated fatty acid, a preparation method and application thereof. The compositions of the present invention include omega-3, omega-6 polyunsaturated fatty acids, omega-7, omega-9 monounsaturated fatty acids, and grifola frondosa polysaccharides. According to experimental research, omega-3, omega-6 polyunsaturated fatty acid, omega-7 and omega-9 monounsaturated fatty acid and grifola frondosa polysaccharide have a coordinated synergistic effect in reducing blood fat, and the composition can obviously reduce the level of triglyceride, cholesterol and low-density lipoprotein by compounding the same according to a specific proportion, so that an excellent blood fat reducing effect is achieved, the injury to the liver is reduced, and meanwhile, the effects of reducing blood sugar and blood pressure are achieved, so that the composition can be used for preparing medicines for reducing three highs and foods or food additives for assisting in reducing three highs.
Description
Technical Field
The invention relates to the technical field of biological medicine, in particular to a composition containing unsaturated fatty acid, a preparation method and application thereof.
Background
Hyperlipidemia refers to the abnormal metabolism or operation of fat that causes one or more lipids in the plasma to be higher than normal. It is a systemic disease that manifests itself in blood Total Cholesterol (TC) and/or in hypertriglyceridemia or in high density lipoprotein cholesterol (HDL-C) which are too low, and modern medicine is called dyslipidemia, also commonly referred to as hyperlipoproteinemia. Most of the hyperlipidemia in the early stage has no symptoms, and if the hyperlipidemia is not treated in time, damage to the body can be caused. Current data studies indicate that hyperlipidemia can cause the following diseases: coronary heart disease, myocardial infarction, sudden cardiac death, cerebral apoplexy, hypertension, diabetes, fatty liver, liver cirrhosis, cholelithiasis, pancreatitis, hyperuricemia, etc. Hyperglycemia is a phenomenon in which the sugar content in blood of a patient continuously exceeds a normal level for a long time, and hypertension is a phenomenon in which the pressure value caused by blood flowing in a blood vessel to the wall of the blood vessel is continuously higher than the normal value; hyperlipidemia, hyperglycemia, and hypertension are referred to as "three highs".
Polyunsaturated fatty acids (PUFA) are linear fatty acids having at least two double bonds and a carbon chain length of 18 to 22 carbon atoms, and are classified into omega-3 and omega-6 polyunsaturated fatty acids according to the position of the 1 st double bond from the methyl end. Common omega-3 polyunsaturated fatty acids include alpha-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, etc., and common omega-6 polyunsaturated fatty acids include linoleic acid, gamma-linolenic acid, arachidonic acid, etc. Most PUFAs are essential to the human body, as are vitamins and minerals, and insufficient intake can easily lead to important organ disorders such as heart and brain. Monounsaturated fatty acids (MUFA) refer to fatty acids that contain only one unsaturated bond in the carbon chain, and include omega-7 and omega-9 monounsaturated fatty acids. Common omega-7 monounsaturated fatty acids are palmitoleic acid and common omega-9 monounsaturated fatty acids are oleic acid.
Grifola frondosa (Griflolafrondosa), named Polyporus frondosus, chestnut mushroom, lotus mushroom, maitake Mushroom, etc., is called "edible fungus prince", called "king of immunity" by Japanese medical community, and is fungus of Polyporaceae Polyporus. The Maitake Mushroom polysaccharide has effects of resisting tumor, hypertension, reducing blood sugar, obesity and hepatitis. According to the clinical experiments of special treatment hospitals for some cancers in the United states, the effect of inhibiting cancer cells by using the grifola frondosa polysaccharide is better than that of pure chemotherapy when the cancers are treated chemically. However, the addition of Grifola frondosa polysaccharide to monounsaturated fatty acid and polyunsaturated fatty acid to enhance the "three high" lowering function of unsaturated fatty acid has not been reported.
Disclosure of Invention
The invention aims to provide a composition containing unsaturated fatty acid, wherein components in the composition can act synergistically to enhance the effect of reducing 'three highs'.
In order to achieve the above object, the present invention provides the following technical solutions:
10-20 parts of omega-3 polyunsaturated fatty acid, 30-40 parts of omega-7 monounsaturated fatty acid, 20-30 parts of omega-6 polyunsaturated fatty acid, 5-10 parts of omega-9 monounsaturated fatty acid and 15-25 parts of grifola frondosa polysaccharide.
Preferably, the omega-3 polyunsaturated fatty acid comprises DHA, EPA or ALA, and the omega-3 polyunsaturated fatty acid is one or a combination of several of fish oil, krill oil, linseed oil, merry go round robinia oil, perilla seed oil, pumpkin seed oil, hemp seed oil, algae oil and microbial oil;
The omega-6 polyunsaturated fatty acid comprises LA or ARA, and the omega-6 polyunsaturated fatty acid is one or a combination of several of peanut oil, corn oil, soybean oil, sesame oil, rapeseed oil, algae oil and microbial grease.
Preferably, the omega-7 monounsaturated fatty acid comprises palmitoleic acid, and the omega-7 monounsaturated fatty acid is one or a combination of several of torreya seed oil, sea buckthorn fruit oil and microbial oil;
The omega-9 monounsaturated fatty acid comprises oleic acid, and the omega-9 monounsaturated fatty acid is one or a combination of several of canola oil, sunflower oil, olive oil, walnut oil, avocado oil, algae oil and microbial oil.
Preferably, the grifola frondosa polysaccharide is extracted from a grifola frondosa fruiting body.
The invention also provides a preparation method of the composition containing unsaturated fatty acid, polyunsaturated fatty acid and grifola frondosa polysaccharide are mixed and stirred, and the obtained oiling agent is the composition.
Preferably, the mixing is performed under vacuum or inert gas environment, the stirring speed is 1000-3000 rpm, and the stirring time is 20-30 min.
The invention also provides application of the composition containing unsaturated fatty acid in preparation of medicines for reducing 'three highs'.
Preferably, the dosage form of the medicine for reducing the three highs comprises pills, granules, electuaries, tablets, emulsions or capsules.
The invention also provides application of the composition containing unsaturated fatty acid in preparing food or food additives for assisting in reducing blood fat.
By adopting the technical scheme, the invention has the following beneficial effects: according to experimental research, the omega-3, omega-6 polyunsaturated fatty acid, omega-7 and omega-9 monounsaturated fatty acid and the tree flower polysaccharide have a coordinated synergistic effect in reducing blood fat, and the compound of the omega-3, omega-6 polyunsaturated fatty acid, omega-7 and omega-9 monounsaturated fatty acid and the tree flower polysaccharide in a specific proportion can obviously reduce the level of triglyceride, cholesterol and low-density lipoprotein, achieve an excellent blood fat reducing effect, reduce the damage to livers, and simultaneously have the effects of reducing blood sugar and blood pressure, and can be used for preparing medicines for reducing three highs, foods for assisting in reducing three highs or food additives.
Drawings
FIG. 1 is a graph showing the effect of a composition of the present invention on the viability of a model of high glucose injured INS-1 islet cells.
FIG. 2 is a graph showing the effect of the composition of the present invention on blood pressure in rats.
Detailed Description
The invention provides a composition containing unsaturated fatty acid, which comprises the following components in parts by weight: 10-20 parts of omega-3 polyunsaturated fatty acid, 30-40 parts of omega-7 monounsaturated fatty acid, 20-30 parts of omega-6 polyunsaturated fatty acid, 5-10 parts of omega-9 monounsaturated fatty acid and 15-25 parts of grifola frondosa polysaccharide.
In the composition of the present invention, the unsaturated fatty acid is 30 to 60 parts by weight, more preferably 40 to 50 parts by weight, still more preferably 45 parts by weight;
15-25 parts by weight of grifola frondosa polysaccharide, more preferably 18-22 parts by weight, and even more preferably 20 parts by weight;
Polyunsaturated fatty acids according to the invention include omega-3 and omega-6 polyunsaturated fatty acids, the weight ratio of omega-3 and omega-6 polyunsaturated fatty acids being from 1:3 to 5, more preferably from 1:3.5 to 4.5, and even more preferably 1:4.
The omega-3 polyunsaturated fatty acid comprises DHA, EPA or ALA, and the structural formula of the DHA (docosahexaenoic acid) is shown in formula 1:
The structural formula of EPA (eicosapentaenoic acid) is shown in formula 2:
the structural formula of ALA (alpha-linolenic acid) is shown in formula 3:
The omega-3 polyunsaturated fatty acid is one or a combination of a plurality of fish oil, krill oil, linseed oil, merry go round robinia oil, perilla seed oil, pumpkin seed oil, hemp seed oil, algae oil and microbial oil; the omega-3 polyunsaturated fatty acid content in the fish oil, krill oil, linseed oil, merry go round robinia oil, perilla seed oil, pumpkin seed oil, hemp seed oil, algae oil and microbial oil is not less than 60%.
The omega-7 monounsaturated fatty acid comprises palmitoleic acid, and the structural formula of the palmitoleic acid is shown as formula 4:
The omega-7 monounsaturated fatty acid is one or a combination of more than one of sea buckthorn fruit oil, chinese torreya seed oil and microbial oil, and the content of omega-7 in the sea buckthorn fruit oil is not less than 40%.
The omega-6 polyunsaturated fatty acid comprises LA or ARA, and the structural formula of the LA (linoleic acid) is shown in the formula 5:
the structural formula of ARA (arachidonic acid) is shown in formula 6:
The omega-6 polyunsaturated fatty acid is from peanut oil, corn oil, soybean oil, sesame oil, rapeseed oil and microbial grease, and the content of the omega-6 polyunsaturated fatty acid in the peanut oil, the corn oil, the soybean oil, the sesame oil, the rapeseed oil and the microbial grease is not less than 50 percent.
The omega-9 monounsaturated fatty acid comprises oleic acid, and the structural formula of the oleic acid is shown in formula 7:
The omega-9 monounsaturated fatty acid is one or a combination of several of canola oil, sunflower seed, olive oil, walnut oil and butter, algae oil and microbial oil, and the content of omega-9 monounsaturated fatty acid in the canola oil, sunflower seed oil, olive oil, walnut oil, butter, algae oil and microbial oil is not less than 60%.
In the invention, the grifola frondosa polysaccharide is extracted from the fruiting body of grifola frondosa, and specifically comprises the following steps:
(1) Grinding Maitake Mushroom fruiting body into powder, adding ethanol solution, and stirring to obtain precipitate;
(2) Weighing the precipitate, adding distilled water, and extracting;
(3) Centrifuging, collecting supernatant, and adding absolute ethanol for precipitation;
(4) Centrifuging, collecting precipitate, and vacuum drying to obtain Maitake Mushroom polysaccharide.
According to the invention, after the grifola frondosa fruiting body is ground into powder, adding an ethanol solution for stirring, wherein the volume fraction of ethanol is 93-98%, more preferably 94-97%, more preferably 95%, the volume ratio of the weight of the grifola frondosa fruiting body powder to the ethanol is 1 g:2-5 mL, more preferably 1 g:3-4 mL, more preferably 1g:3.5mL, and the stirring time is 1.5-2.5 h, more preferably 2h; adding ethanol, stirring for degreasing, and vacuum filtering to obtain precipitate. Weighing the precipitate, adding distilled water for extraction, wherein the weight volume ratio of the precipitate to the distilled water is 1 g:8-12 mL, more preferably 1 g:9-11 mL, more preferably 1g:10mL, the extraction time is 20-40 min, more preferably 25-35 min, more preferably 30min, centrifuging after the extraction is finished, taking supernatant, adding absolute ethyl alcohol for precipitation, wherein the volume ratio of the supernatant to the absolute ethyl alcohol is 1:0.5-1.5, more preferably 1:0.8-1.2, more preferably 1:1, and the precipitation time is 11-13 h, more preferably 12h. And centrifuging, and drying the precipitate, wherein the drying temperature is 50-70 ℃, more preferably 55-65 ℃, and still more preferably 60 ℃, so as to obtain the grifola frondosa polysaccharide.
The invention also provides a preparation method of the composition, which comprises the steps of mixing unsaturated fatty acid and grifola frondosa polysaccharide, and stirring to obtain an oil or suspension agent which is the composition. The mixing is performed under vacuum or inert gas environment, the stirring speed is 1000-3000 rpm, more preferably 1500-2500 rpm, more preferably 2000rpm, the stirring time is 20-30 min, more preferably 22-28 min, more preferably 25min, and the mixture is a uniform and stable oil or suspension.
The invention also provides application of the composition in preparation of the medicine for reducing three highs. The dosage forms of the medicine for reducing the three highs comprise pills, granules, electuaries, tablets, emulsions or capsules.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The fish oil, krill oil and algae oil in the embodiment of the invention are purchased from Jiangsu Aoqi ocean bioengineering Co., ltd, peanut oil, corn oil, sesame oil and sea buckthorn fruit oil are purchased from Wuhank biological medicine technology Co., ltd, and the brand of canola oil and sunflower seed oil is Nissin and the brand of avocado oil is Chosen Foods.
Example 1
The composition of this example consisted of the following raw materials:
Fish oil containing 10g omega-3 polyunsaturated fatty acids, seabuckthorn fruit oil containing 30g omega-7 monounsaturated fatty acids, peanut oil containing 20g omega-6 polyunsaturated fatty acids, canola oil containing 5g monounsaturated fatty acids, and grifola frondosa polysaccharide 15g.
Mixing fish oil, peanut oil and grifola frondosa polysaccharide under vacuum, and stirring at 1000rpm for 20min to obtain composition.
The grifola frondosa polysaccharide is prepared by the following method: grinding Maitake mushroom fruiting body into powder, adding 93% ethanol solution at a ratio of 1g to 2mL, stirring for degreasing, filtering for 1.5h, and obtaining precipitate. Weighing a precipitate, and adding a feed liquid ratio of 1:8 (g/mL) of distilled water, and extracting for 20min. Centrifuging, collecting supernatant, and adding 1:0.5 volume of absolute ethanol for precipitation for 11h. Centrifuging, collecting precipitate, and vacuum drying at 50deg.C to obtain Maitake Mushroom polysaccharide powder.
The sea buckthorn fruit extract is prepared by the following method: pulverizing fructus Hippophae, extracting with 85% ethanol solution at a ratio of 1g to 3mL for 40min, and recovering solvent to obtain solid fructus Hippophae extract.
Example 2
The composition of this example consisted of the following raw materials:
Krill oil containing 20g omega-3 polyunsaturated fatty acids, torreya seed oil containing 40g omega-7 monounsaturated fatty acids, rapeseed oil containing 30g omega-6 polyunsaturated fatty acids, sunflower seed oil containing 10g omega-9 monounsaturated fatty acids, grifola frondosa polysaccharide 25g, omega-3 polyunsaturated fatty acids derived from krill oil, omega-6 polyunsaturated fatty acids derived from corn oil.
Mixing krill oil, corn oil and Maitake Mushroom polysaccharide under vacuum, and stirring at 3000rpm for 30min to obtain composition.
The grifola frondosa polysaccharide is prepared by the following method: grinding Maitake mushroom fruiting body into powder, adding 98% ethanol solution at a ratio of 1g to 5mL, stirring for degreasing, filtering for 2.5h, and obtaining precipitate. Weighing a precipitate, and adding a feed liquid ratio of 1:12 Distilled water (g/mL), 40min. Centrifuging, collecting supernatant, and adding 1:1.5 volume of absolute ethanol for precipitation for 13h. Centrifuging, collecting precipitate, and vacuum drying at 50deg.C to obtain Maitake Mushroom polysaccharide powder.
Example 3
The composition of this example consisted of the following raw materials:
algae oil containing 15g omega-3 polyunsaturated fatty acid, sea buckthorn oil containing 35g omega-7 monounsaturated fatty acid, sesame oil containing 25g omega-6 polyunsaturated fatty acid, butter oil containing 8g omega-9 monounsaturated fatty acid, and Maitake Mushroom polysaccharide 20g.
Mixing Perilla seed oil, oleum Sesami, and Maitake Mushroom polysaccharide under vacuum, and stirring at 2000rpm for 25min to obtain composition.
The grifola frondosa polysaccharide is prepared by the following method: grinding Maitake mushroom fruiting body into powder, adding 95% ethanol solution according to a feed liquid ratio of 1g to 3mL, stirring for degreasing, and filtering after 2h to obtain precipitate. Weighing a precipitate, and adding a feed liquid ratio of 1:10 Distilled water (g/mL), extracting for 30min. Centrifuging, collecting supernatant, and adding 1:1 volume of absolute ethanol for precipitation for 12h. Centrifuging, collecting precipitate, and vacuum drying at 60deg.C to obtain Maitake Mushroom polysaccharide powder.
Example 4
The oil preparation oil powder in example 1 was prepared as follows:
40g of maltodextrin and 10g of Arabic gum were dissolved in 100g of distilled water at 60℃and stirred well, 35g of the oil solution of example 1 was sheared at 3000rpm for 8min, and then homogenized at 40MPa for 2min to obtain an emulsion. And then spray drying, wherein the temperature of an air inlet for spraying dysphoria is 180 ℃, the temperature of an air outlet is 100 ℃, and the grease powder is obtained after the spray drying is completed.
Pulverizing 7g of coix seed, 10g of konjak, 3g of black sesame, 4g of pumpkin, 8g of oat, 5g of small red bean, 2g of mung bean, 6g of black bean and 10g of Chinese yam, and mixing with 5g of the grease powder to obtain meal replacement powder.
Example 5
The oil in example 2 was prepared as in example 4 to give a grease powder.
12G of mango juice powder, 10g of blueberry juice powder, 5g of haw powder, 8g of mulberry fruit, 20g of whole milk powder, 5g of grease powder prepared by the oiling agent of example 2 and 3g of xylitol are mixed to obtain a solid beverage.
Example 6
Beating 5 eggs, separating yolk from egg white, stirring yolk uniformly, adding 15g white sugar and 50mL milk, stirring uniformly, adding 30g oiling agent in example 3, stirring uniformly, adding 90g low-gluten flour, stirring uniformly to obtain yolk paste, and covering with preservative film and standing at 4deg.C for use. Mixing the egg white with the yolk paste after beating, stirring uniformly, putting into an oven, and baking at 145 ℃ for 50min to obtain the cake.
Comparative example 1
Unlike example 1, comparative example 1 has an omega-3 polyunsaturated fatty acid content of 30g.
Comparative example 2
Unlike example 1, comparative example 2 does not contain grifola frondosa polysaccharide.
Comparative example 3
Unlike example 1, the omega-7 monounsaturated fatty acid content in comparative example 3 was 20g.
Comparative example 4
Unlike example 1, comparative example 4 has an omega-6 monounsaturated fatty acid content of 10g.
Experimental example 1 hypolipidemic animal experiment of the composition of the invention
1. Experimental materials
Fish oil, krill and algae oil are purchased from Jiangsu Aoqi ocean bioengineering Co., ltd, peanut oil, corn oil, sesame oil and sea buckthorn fruit oil are purchased from Wuhank Mike biomedical technology Co., ltd, and the brand of canola oil and sunflower seed oil is Nissin and the brand of butter fruit oil is ChosenFoods.
2. Experimental animals: SD rats, male, SPF grade, body weight (180+ -20) g, offered by Chengdu laboratory animal Co.
3. The main reagent comprises: kits of TC (total cholesterol), TG (total triglycerides), HDL-C (high density lipoprotein cholesterol), LDL-C (low density lipoprotein cholesterol) were all purchased from solarbio functional nets.
4. Feed: the general feed and the high-fat feed formula are as follows: 72.7% of basic feed, 2% of cholesterol, 5% of yolk powder, 10% of lard, 0.2% of propylthiouracil, 10% of sucrose and 0.1% of sodium cholate.
(A) Effect of the inventive composition on rat blood lipid level
The experimental rats purchased were subjected to laboratory adaptation for 1 week and were randomly divided into a blank control group, a model control group, a composition (examples 1 to 3 and comparative examples 1 to 3) low dose group (0.1 mL/kg.d), a medium dose group (0.3 mL/kg.d), a high dose group (0.5 mL/kg.d), and a positive drug (simvastatin) control group (1.5 mg/kg), 10 animals per group. From the day of the experiment, the blank control group and the model control group were perfused with physiological saline (10 mL/kg), and the composition group was perfused with the corresponding composition 1 time/d, 1 time of body weight per week, and the administration amount was adjusted according to the body weight. The blank control group was fed with normal feed, the remaining groups were fed with high fat feed, and free feeding, free drinking water, continuous molding and preventive dosing were performed for 4 weeks during the test period.
5. And (3) observing the indexes: and the medicine is fasted for 12 hours after the last administration, and water is not forbidden. The rats were anesthetized with 3% sodium pentobarbital, blood was collected from abdominal aorta, and serum was centrifuged to measure the total serum cholesterol (TC), triacylglycerol (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C), and the results are shown in Table 1.
TABLE 1 blood lipid changes in rats of each groupn＝10)
As can be seen from Table 1, the serum TC, TG, LDL-C levels in rats in the model group were significantly elevated (P < 0.05) compared to the normal group, and exhibited typical lipid metabolism disorders, indicating successful replication of the experimental hyperlipidemic model. Compared with the model group, the examples 1-3 can obviously reduce serum TC, TG, LDL-C content (P < 0.05), can obviously raise serum HDL-C content (P < 0.01) and show dose dependence. The composition of comparative example 2 does not contain grifola frondosa polysaccharide, and the contents of omega-7 monounsaturated fatty acids and omega-6 monounsaturated fatty acids of comparative examples 3 and 4 are not within the scope of the present invention, and the therapeutic effect is inferior to examples 1 to 3.
(B) Effect of the composition of the present invention on liver function in rats
ALT and AST in serum can reflect liver function. ALT and AST levels in serum of rats of the present invention after 4 weeks of administration of the composition and positive drug control are shown in Table 2.
TABLE 2 liver function changes in rats of each groupn＝10)
As can be seen from Table 2, the fat diet groups ALT and AST were significantly higher than the basal diet group (P < 0.01), indicating that administration of the high fat diet had some damage to the liver of the rats. The serum ALT and AST levels of each administration group are lower than those of the model control group, and the serum ALT and AST levels of rats in the medium-dose group, the high-dose group and the low-dose group are lower than those of the positive drug control group, so that compared with simvastatin, the composition can reduce the damage to the liver.
Experimental example 2 Effect of the composition of the present invention on the level of lipoprotein in adipocytes
3T3-L1 preadipocytes are one of the most typical cell lines for studying lipid metabolism of adipocytes, and in this example, 3T3-L1 preadipocytes are used as subjects for studying the effect of the compositions of the present invention on the levels of lipoproteins in adipocytes.
1. Effect of the composition on cell viability
Cell culture: 3T3-L1 cells (purchased from ATCC cell bank) were cultured in DMEM medium containing 10% FBS, and the cells were placed in a cell culture incubator containing 5% CO 2 at 37 ℃. When the cells are grown on the wall until the cell coverage reaches more than 80%, the cells are digested with 0.25% trypsin solution for passage or the next test is carried out. 100. Mu.L of the cell suspension (1X 10 5/mL) was inoculated into a 96-well plate, a control group and a test group were set, and after 24 hours of cell attachment, 100. Mu.L of the culture solution was added to the blank control group and the model control group, and 100. Mu.L of the sample solutions prepared in example 1 of different concentrations (0.05, 0.1, 0.25, 0.5 mg/mL) were added to the test group, respectively. After 24h of incubation, the old broth was discarded and incubated with 100. Mu. LMTT (0.5 mg/mL) for 4h.
Cell induction differentiation: the blank control group did not undergo induced differentiation, the model control group and the test group inoculated cells in 24-well plates (3×10 4 cells/well), and the cells were first cultured in DMEM complete medium until 90% confluence was achieved. The primary induced differentiation medium was changed for 2d, and the primary induced differentiation medium was a complete medium containing 0.5mM IBMX, 1. Mu.M DEX and 10. Mu.g/mL insulin. Then culturing for 2-4 d with secondary induced differentiation liquid, wherein the culture liquid is changed every 2d, and the secondary induced differentiation liquid is complete culture liquid containing 10 mug/mL insulin. Until 80% of the cells had differentiated into mature adipocytes, and the differentiation was completed. During the induction differentiation, the culture medium of the model control group is not added with the sample solution, and the sample solutions prepared in the example 1 with different concentrations (0.05, 0.1, 0.25 and 0.5 mg/mL) are respectively added into the primary induction differentiation liquid and the secondary induction differentiation liquid in each experimental group.
After the cell induction differentiation is finished, cell sediment is collected, washed by PBS buffer solution and then subjected to ultrasonic disruption, and detection is carried out according to the instructions of high-density lipoprotein and low-density lipoprotein kit (purchased from Shanghai ELISA). The effect of the hypolipidemic composition on the levels of high density lipoprotein and low density lipoprotein in 3T3-L1 cells is shown in table 3.
TABLE 3 high Density lipoprotein (HDL-C) and Low Density lipoprotein (LDL-C) levels in different groups of cells
As can be seen from table 3, the cells treated with the composition of the present invention significantly decreased the low density lipoprotein level compared to the model group, and the more the low density lipoprotein level decreased with increasing concentration of the composition; the higher the high density lipoprotein level of the composition-treated cells compared to the model group, and the higher the high density lipoprotein level, the higher the concentration of the composition. At a concentration of 0.5mg/mL, the cells treated with the composition of the invention had both high density lipoprotein levels increased by 77.41% and low density lipoprotein levels decreased by 49.04% relative to the model group.
From the above examples, the composition of the present invention can significantly reduce the level of triglyceride, cholesterol and low density lipoprotein by compounding unsaturated fatty acid and Maitake Mushroom polysaccharide, achieve excellent hypolipidemic effect, and reduce liver injury.
Experimental example 3 Effect of the composition of the invention on the viability of the high sugar damaged INS-1 islet cell model INS-1 islet cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum) in an incubator at 37℃with 5% CO 2. Taking INS-1 islet cells in exponential growth phase after passage, regulating cell density to 2.5X10 4/mL, inoculating to 96-well plate, culturing in 100 μL per well at 37deg.C in 5% CO 2 incubator for 24 hr, setting control group, 25mM high sugar model group, composition treatment group (composition of example 1 is dissolved in 25mM high sugar culture solution, filtering with 0.22 μm filter to final concentration of 0.04,0.2,1,5 mg/mL), continuously culturing for 48 hr, sucking out culture solution, adding 100 μL10% CCK8, culturing in carbon dioxide incubator for 2 hr, detecting absorbance (OD value) at 450nm with multifunctional enzyme-labeled instrument, and finally calculating cell survival rate of each group. 3 replicates were set for each experimental group, and the control group was added with the same volume of PBS as the composition group.
Cell viability (%) = (OD experimental group/OD control group) ×100% and the results are shown in fig. 1.
As can be seen from FIG. 1, the cell viability of the 25mM high-sugar model group was significantly reduced compared with that of the control group, and the INS-1 islet cell high-sugar damage model was successfully constructed. Compared with the 25mM high sugar model group with 85.6 percent of cell activity, the cell activities of the treatment groups of the compositions of 0.04mg/mL, 0.2mg/mL, 1mg/mL and 5mg/mL are respectively 91.4 percent, 95.3 percent, 98.8 percent and 94.2 percent, and the composition has certain injury inhibition effect, wherein the action effect of the composition of 1mg/mL is the best. Therefore, the composition of the invention can be applied to hypoglycemic drugs or health-care foods for assisting in reducing blood sugar.
Experimental example 4 Effect of the composition of the present invention on rat blood pressure
(1) Spontaneous hypertensive rats (SHRs, 10 week old, male, weight 250-300 g) with blood pressure exceeding 180mmHg were obtained from Shanghai Laike animal laboratory company;
(2) 6 groups of hypertensive rats are fed, and are circularly illuminated for 12 hours at the temperature of 23 ℃ to supply food and drinking water;
(3) Each hypertensive rat of the experimental group was fed with the composition of examples 1 to 3 at a dose of 0.5mL/kg·d, the control group was perfused with physiological saline at a dose of 10mL/kg·d, and the positive control group was perfused with physiological saline at the same dose while feeding the antihypertensive drug lisinopril (1.5 mg/kg);
(4) After gastric lavage, blood pressure was measured in these hypertensive rats at 0h, 1h, 2h, 4h, 6h and 8h, respectively. All the results were measured in triplicate and averaged, and the results are shown in figure 2.
As can be seen from fig. 2, the compositions of examples 1 to 3 all had significantly lower blood pressure than the control group, indicating that they had a blood pressure lowering effect in vivo, while the three compositions had better blood pressure lowering effect than lisinopril within 6 hours of administration compared to lisinopril group. Therefore, the composition of the invention can be applied to antihypertensive drugs or health care foods for assisting in lowering blood pressure.
From the above examples, the present invention provides an unsaturated fatty acid-containing composition comprising omega-3, omega-6 polyunsaturated fatty acids, omega-7, omega-9 monounsaturated fatty acids and grifola frondosa polysaccharides. The composition has excellent blood lipid reducing effect, reduces the damage to the liver, and simultaneously has the effects of reducing blood sugar and blood pressure.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (9)
1. The composition containing unsaturated fatty acid is characterized by comprising the following components in parts by weight: 10-20 parts of omega-3 polyunsaturated fatty acid, 30-40 parts of omega-7 monounsaturated fatty acid, 20-30 parts of omega-6 polyunsaturated fatty acid, 5-10 parts of omega-9 monounsaturated fatty acid and 15-25 parts of grifola frondosa polysaccharide.
2. The unsaturated fatty acid-containing composition of claim 1, wherein the omega-3 polyunsaturated fatty acid comprises DHA, EPA, or ALA, and the omega-3 polyunsaturated fatty acid is derived from one or a combination of several of fish oil, krill oil, linseed oil, merry go round robinia oil, perilla seed oil, pumpkin seed oil, hemp oil, algae oil, and microbial oil;
The omega-6 polyunsaturated fatty acid comprises LA or ARA, and the omega-6 polyunsaturated fatty acid is one or a combination of several of peanut oil, corn oil, soybean oil, sesame oil, rapeseed oil, algae oil and microbial grease.
3. The unsaturated fatty acid-containing composition according to claim 2, wherein the omega-7 monounsaturated fatty acid comprises palmitoleic acid, and the omega-7 monounsaturated fatty acid is derived from one or a combination of several of torreya seed oil, sea buckthorn fruit oil, and microbial oil;
The omega-9 monounsaturated fatty acid comprises oleic acid, and the omega-9 monounsaturated fatty acid is one or a combination of several of canola oil, sunflower oil, olive oil, walnut oil, avocado oil, algae oil and microbial oil.
4. The unsaturated fatty acid-containing composition according to claim 1, wherein the grifola frondosa polysaccharide is extracted from a grifola frondosa fruiting body.
5. The method for producing an unsaturated fatty acid-containing composition according to any one of claims 1 to 4, wherein the polyunsaturated fatty acid and the grifola frondosa polysaccharide are mixed and stirred to obtain an oil agent, i.e., the unsaturated fatty acid-containing composition.
6. The method according to claim 5, wherein the mixing is performed under vacuum or an inert gas atmosphere, the stirring speed is 1000 to 3000rpm, and the stirring time is 20 to 30 minutes.
7. Use of a composition comprising an unsaturated fatty acid according to any one of claims 1 to 6 for the preparation of a medicament for reducing "three high" lipid.
8. The use according to claim 7, wherein the dosage form of the drug for reducing "three high" comprises a pill, a granule, a tablet, an emulsion or a capsule.
9. Use of the unsaturated fatty acid-containing composition according to any one of claims 1 to 4 for the preparation of a food or food additive for aiding in the reduction of "three high".
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