CN104523744A - Fucan sulfate and application of low-molecular-weight fucan sulfate in preparation of blood fat reduction and weight loss drugs and health care products - Google Patents
Fucan sulfate and application of low-molecular-weight fucan sulfate in preparation of blood fat reduction and weight loss drugs and health care products Download PDFInfo
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Abstract
The invention provides fucan sulfate and application of low-molecular-weight fucan sulfate in preparation of blood fat reduction and weight loss drugs and health care products. The fucan sulfate and the low-molecular-weight fucan sulfate can be used for remarkably suppressing aggregation of triglyceride and cholesterol of liver cells when concentration is larger than 50 micrograms per milliliter, and the fucan sulfate and the low-molecular-weight fucan sulfate can be used for effectively reducing the level of TG, TC and LDL in blood, increasing the level of HDL in the blood and obviously reducing weight and the content of abdominal fat. As is provided through experiments, FS and LFS have a good function of reducing blood fat and losing weight. The raw materials come from alga and have the advantages of being rich in resource, low in cost, high in safety performance and the like, the situation that the high blood fat and obesity state can be remarkably improved is proved both in a body and outside the body, and a new way is provided for developing the kind of drugs.
Description
Technical field
The invention belongs to medical art, be specifically related to fucoidan and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product.
Background technology
Hyperlipemia is mainly with cholesterol (cholesterol in blood plasma, TC), triglyceride (triglyceride, TG), low density lipoprotein, LDL (low density lipoprotein, LDL) raise, high density lipoprotein (high density lipoprotein, HDL) a kind of metabolism disorder of blood lipid state of performance is reduced to, be one of main predisposing factors of atherosclerosis, heart disease, apoplexy and fatty liver diseases, the health of people in serious threat.Effectively control the key that hyperlipemia is prevention cardiovascular and cerebrovascular disease, along with aged tendency of population rises and the increasing and momental minimizing of fat intake in people's diet year by year, the sickness rate of hyperlipemia and complication thereof is more and more higher.Show according to the Chinese residents nutrition and health survey that Ministry of Public Health carries out in China in recent years, within more than 18 years old, resident's hyperlipemia total prevalence rate is 18.6%, and national subject population reaches 1.6 hundred million.Though multiple Western medicine has the effect for reducing fat of affirmative at present, prolonged application has side effect in various degree, and easily has a rebound after drug withdrawal.Therefore, find out a kind of safety, effectively, and there is the natural health care of fat-reducing effect and medicine just seems extremely important.
Obesity refers to athero excessive in body.At present, there is the popular impetus in obesity in worldwide.Research shows, obesity is the disease of a series of feed regulation and control and the energy metabolism disorder caused by specific Some Circulating Factors, and pathogenic process is very complicated.At present, obesity instead of the disease caused by malnutrition and infection, suddenly becomes the primary killers of harm humans health.The diseases such as known obesity and diabetes, cardiovascular disease, some tumor have obvious dependency relation.Obesity has been not only the problem of the individual image of an impact, but becomes a kind of epidemic diseases, threatens the health of the whole mankind.
Marine polysaccharide is aboundresources not only, and containing the functional group such as carboxyl, sulfate group in molecule, pharmacologically active is extensive and toxicity is very low.Fucoidan was just found in 1913, was the electronegative biomacromolecule of the class be present in Brown algae, was mainly made up of fucose, galactose, mannose and sulfate etc.Existing many sections of documents disclose preparation method and the pharmaceutical applications thereof of fucoidan.Except anticoagulating active, studying more is the effect of fucoidan antitumor, antiviral, antioxidation, antiinflammatory, immunity moderation.In addition, 2009-2012 report Thallus Laminariae Undariapinnatifida originate fucoidan and lower-molecular-weight component there is hypoglycemic activity.But not yet there is fucoidan at blood fat reducing and the application report reduced in body weight, fat-reducing.
Summary of the invention
Fucoidan and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product are the object of this invention is to provide, through system experimentation, the present invention proves that fucoidan and low-molecular-weight fucoidan are in treatment hyperlipemia and reduction body weight, have beyond thought good therapeutic effect.
For achieving the above object, the present invention adopts following technical proposals to be achieved:
Fucoidan and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product.
The preparation method of described fucoidan and low-molecular-weight fucoidan is:
(1) by Brown algae powder organic solvent soak degreasing;
(2) then through water extraction, alcohol settling, the thick sulfated fucan of dry acquisition;
(3) the water solublity Algin in thick sulfated fucan is removed again;
(4) after dialysis, concentrating, then through alcohol settling, the described fucoidan of purification after drying, is obtained;
(5) by the obtained low-molecular-weight fucoidan of described fucoidan degraded.
Described fucoidan molecular weight is 200kD-2000kD, and the molecular weight of described low-molecular-weight fucoidan is 1000-200000 dalton.
Described Brown algae is one or more in Fucus Vesiculosus, yellow tang, Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, sargassum thunbergii, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae) or Thallus Laminariae (Thallus Eckloniae).
Described fucoidan and/or low-molecular-weight fucoidan are that more than 50ug/ml significantly suppresses liver cell triglyceride, cholesterol gathers in concentration.
Described fucoidan and/or low-molecular-weight fucoidan obviously reduce body weight and abdominal fat content under the dosage of dosage 20mg/kg/day.
Compared with prior art, advantage of the present invention and technique effect are: described fucoidan be by Brown algae powder after ethanol or methanol or the defat of acetone and other organic solvent soaked overnight, boiling water extraction precipitate with ethanol, dry thick sulfated fucan, then through CaCl
2or MgCl
2there is lower 20% alcohol settling and remove water solublity Algin, then through dialysis, concentrated after 75% alcohol settling, obtain the fucoidan of purification after drying; Fucoidan obtained in the present invention is degraded by suitable mode and can be obtained low-molecular-weight fucoidan, and its molecular weight more undegradable fucoidan molecular weight is low, and concrete molecular weight ranges is 1000-200000 dalton.
Described fucoidan and low-molecular-weight fucoidan are that more than 50ug/ml can significantly suppress liver cell triglyceride, cholesterol gathers in concentration respectively.Described fucoidan (FS) and low-molecular-weight fucoidan (LFS) significantly can alleviate the hold concurrently metabolism disorder of blood lipid of obesity mice ob/ob and body weight of hyperlipemia and raise symptom under the dosage of dosage 20mg/kg/day, namely effectively reduce TG, TC, LDL and raising HDL levels in blood, and obviously reduce body weight and abdominal fat content.Through experiment, the present invention proves that FS and LFS has effect of good blood fat reducing and fat-reducing.The present invention derives from Sargassum, has the plurality of advantages such as aboundresources, with low cost and safety be high, and in vivo with levels in vitro all proves significantly to improve hyperlipidemia and fat state, the exploitation for such medicine provides new approach.
Accompanying drawing explanation
Fig. 1 is the cytotoxic assay result of fucoidan and low-molecular-weight fucoidan.
Fig. 2 shows that fucoidan and low-molecular-weight fucoidan in the present invention are held concurrently for hyperlipemia the inhibitory action of obesity mice abdominal fat accumulation, and in figure, model control group is that hyperlipidemia is held concurrently obese model mice ob/ob; Simvastatin group is blood fat reducing positive drug group; Metformin is the heavy positive drug group of control volume; FS is fucoidan processed group; LFS is low-molecular-weight fucoidan group; Dosage is 20mg/kg/day; Model group gives the normal saline with same volume.* P<0.05, compared with comparison model group; #P<0.05, compared with positive drug group, n=12.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, technical scheme of the present invention is described in further detail.
Embodiment 1
The preparation method of fucoidan of the present invention comprises the following steps:
(1) Brown algae powder being placed in methanol, acetone or mass ratio is the organic solvents such as the ethanol of 80%, soaks 3h at every turn, repeat 3 times at 80 DEG C, with soak degreasing;
Described Brown algae takes from one or more in Fucus Vesiculosus, yellow tang, Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, sargassum thunbergii, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae) or Thallus Laminariae (Thallus Eckloniae), wherein preferred Fucus Vesiculosus.
(2) then water extraction at 80 DEG C, then be 80% alcohol settling through mass ratio, then dryly at 40 DEG C obtains thick sulfated fucan;
(3) then at CaCl
2or MgCl
2there is lower mass ratio is that 20% alcohol settling removes water solublity Algin;
(4) again through dialysing, concentrating, be 80% alcohol settling with mass ratio, then after 40 DEG C of dryings, obtain the fucoidan of purification; Obtained described fucoidan molecular weight is 200kD-2000kD (comprising endpoints thereof).
(5) the described fucoidan obtained is obtained low-molecular-weight fucoidan by the degraded of suitable mode (degraded mode includes but not limited to acid degradation method, alkaline degradation method, enzymatic degradation method, physical degradation methods, degradation of xylan with microwave, free-radical oxidation edman degradation Edman etc.), its molecular weight more undegradable fucoidan molecular weight is low, and the concrete molecular weight ranges of the low-molecular-weight fucoidan after degraded is 1000-200000 dalton (not comprising higher limit).
The Cytotoxicity tests of embodiment 2, fucoidan (FS) and low-molecular-weight fucoidan (LFS)
The present invention adopts the cell model of classical disorders of lipid metabolism, hyperlipidemia is held concurrently fat animal model, have rated the effect of the toxicity of described fucoidan on cell and animal level and blood fat reducing, reduction body weight.As shown in Figure 1, fucoidan (FS) and low-molecular-weight fucoidan (LFS) within the scope of 0-400 μ g/mL to cell without overt toxicity, when concentration reaches 500 μ g/mL and is above, side shows certain cytotoxicity.And experimental concentration of the present invention is up to 100 μ g/mL, therefore any toxicity can not be produced to cell.
Embodiment 3, fucoidan (FS) and the external reduction triglyceride of low-molecular-weight fucoidan (LFS) and cholesterol effect
The cell model utilizing human liver cell strain HepG2 to contact high fat and fat inducing factor culture medium to set up, have rated the inhibition of FS and LFS to triglyceride and cholesterol.Positive drug selects simvastatin and metformin.
Experimental result is as shown in table 1, FS and LFS can the content of substantial reduction in triglycerides and cholesterol, and compared with positive drug simvastatin, its triglyceride reducing effect is more obvious; And compared with positive drug metformin, its Lowering cholesterol effect is also better.Illustrate that FS with LFS compares with metformin with current clinical commonly used drug simvastatin, there is the effect better reducing triglyceride and cholesterol.
Table 1FS and LFS is on the impact of hepatocyte TG and TC
* P<0.05, compared with comparison model group;
The antiobesity action that in embodiment 4, fucoidan (FS) and low-molecular-weight fucoidan (LFS) body, blood fat reducing and suppression body weight raise
1), fucoidan (FS) and low-molecular-weight fucoidan (LFS) are held concurrently on Hyperlipidemia the impact of obesity mice body weight and food ration.
Model mice ob/ob is utilized to detect the effect of FS and LFS reduction body weight
Ob/ob mice 60, male, 5-7 week age, be divided into model group, positive control (metformin and simvastatin group) and FS and LFS group at random, often organize 12.Experimental session mice ad lib and drinking-water.Every day gastric infusion.Positive controls gives metformin 225mg/kgd and simvastatin 2.5mg/kgd, experimental group FS and LFS is respectively 10mg/kgd and 10mg/kgd, record food-intake every day and take body weight, 30 days are afterwards and measure final body weight and adopt nuclear-magnetism NMR to measure the content of overall fat, and concrete outcome is as follows:
Positive drug metformin and simvastatin have significant reducing effect (table 2) to food ration, show that they depend on the minimizing of food ration at least partly to the inhibitory action of body weight, and when every kg body weight administration 20mg every day, the food ration of FS and LFS process mice does not have significant change, but body weight relative model group obviously declines, and being significantly better than the inhibition of metformin and simvastatin, this illustrates that FS and LFS has the effect effectively suppressing body weight, fat-reducing under the condition not affecting food-intake.
Further employing desk-top employing nuclear magnetic resonance analyser (Minispec MQ10analyzer) measures the overall fat content of mice, and fatty slip reaches 24.8%, is better than the effect of metformin.Result shows (table 3 and Fig. 2), FS and LFS can fall low-fat content significantly, has effective antiobesity action.
Table 2FS and LFS is on the impact of body weight
* P<0.05, * * P<0.01 is compared with comparison model group; #P<0.05, compared with positive drug group, n=12.
Table 3FS and LFS is on the impact of overall fat content
* P<0.05, * * P<0.01 is compared with comparison model group; #P<0.05, compared with positive drug group, n=12.
2), fucoidan (FS) and low-molecular-weight fucoidan (LFS) are held concurrently on Hyperlipidemia the impact of obesity mice blood lipid metabolism.
Model mice ob/ob is utilized to detect the effect of FS and LFS adjusting blood lipid.
Ob/ob mice 60, male, 5-7 age in week, be divided at random blank group, model group, positive control (metformin and simvastatin group) and FS and LFS group, often organize 12.Experimental session mice ad lib and drinking-water.Every day gastric infusion.Positive controls gives metformin 225mg/kgd and simvastatin 2.5mg/kgd, experimental group FS and LFS is respectively 10mg/kgd and 10mg/kgd, extract eyeball of mouse after 30 days again and get blood, centrifuging and taking serum, measure mice serum free fatty acid (FFA), triglyceride (TG), T-CHOL (TC) content.
Concrete outcome is as table 4, experimental result shows, when every kg body weight administration 20mg every day, FS and LFS significantly can reduce the content of triglyceride TG and cholesterol TC and low density lipoprotein, LDL LDL-c, and the level of remarkable high density lipoprotein increasing HDL, in serum, the content of FFA also effectively reduces simultaneously.And suitable with the lipid-lowering effect of positive drug simvastatin and metformin, this shows that FS and LFS has the effect of obvious adjusting blood lipid.
Table 4FS and LFS is on the impact of blood fat
* P<0.05, * * P<0.01 is compared with comparison model group; N=12.
Experimental result of the present invention shows, fucoidan (FS) and low-molecular-weight fucoidan (LFS) under finite concentration, without any cytotoxicity; FS and LFS in vitro cellular level significantly can reduce the accumulation of TG and TC of liver cell, has the effect of good TG reducing and TC; In animal level can obviously inhibition mice ob/ob body weight rising and fat accumulation, and significantly reduce the content of blood fat TG, TC, LDL-c and free fatty FFA, and effectively raising the level of HDL, these parameter evaluating display FS and LFS has effect of good blood fat reducing and fat-reducing.
The present invention derives from Sargassum, has the plurality of advantages such as aboundresources, with low cost and safety be high, and in vivo with levels in vitro all proves significantly to improve hyperlipidemia and fat state, the exploitation for such medicine provides new approach.
Above embodiment only in order to technical scheme of the present invention to be described, but not is limited; Although with reference to previous embodiment to invention has been detailed description, for the person of ordinary skill of the art, still can modify to the technical scheme described in previous embodiment, or equivalent replacement is carried out to wherein portion of techniques feature; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the spirit and scope of the present invention's technical scheme required for protection.
Claims (10)
1. fucoidan and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product.
2. fucoidan according to claim 1 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that the preparation method of described fucoidan and low-molecular-weight fucoidan is:
(1) by Brown algae powder organic solvent soak degreasing;
(2) then through water extraction, alcohol settling, the thick sulfated fucan of dry acquisition;
(3) the water solublity Algin in thick sulfated fucan is removed again;
(4) after dialysis, concentrating, then through alcohol settling, the described fucoidan of purification after drying, is obtained;
(5) by the obtained low-molecular-weight fucoidan of described fucoidan degraded.
3. fucoidan according to claim 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, it is characterized in that: described fucoidan molecular weight is 200kD-2000kD, the molecular weight of described low-molecular-weight fucoidan is 1000-200000 dalton.
4. fucoidan according to claim 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that: described Brown algae is one or more in Fucus Vesiculosus, yellow tang, Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, sargassum thunbergii, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae) or Thallus Laminariae (Thallus Eckloniae).
5. fucoidan according to claim 4 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that: described Brown algae is Fucus Vesiculosus.
6. fucoidan according to claim 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, it is characterized in that: in described step (1), organic solvent is one or more in methanol, ethanol or acetone, and soak degreasing carries out at 80 DEG C.
7. fucoidan according to claim 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, it is characterized in that: in described step (2) at 80 DEG C water extraction, be the alcohol settling of 80% through mass ratio again, then dryly at 40 DEG C obtain thick sulfated fucan.
8. fucoidan according to claim 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that: at CaCl in described step (3)
2or MgCl
2under existent condition, adding mass ratio is that the ethanol of 20% is to precipitate removal water solublity Algin.
9. fucoidan according to claim 1 and 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that: described fucoidan and/or low-molecular-weight fucoidan are that more than 50ug/ml significantly suppresses liver cell triglyceride, cholesterol gathers in concentration.
10. fucoidan according to claim 1 and 2 and the application of low-molecular-weight fucoidan in the medicine preparing blood fat reducing and fat-reducing and health product, is characterized in that: described fucoidan and/or low-molecular-weight fucoidan obviously reduce body weight and abdominal fat content under the dosage of dosage 20mg/kg/day.
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Cited By (6)
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CN105273104A (en) * | 2015-10-24 | 2016-01-27 | 山东好当家海洋发展股份有限公司 | Method for extracting fucoidan by using Sargassum kjellmanianum |
CN106749439A (en) * | 2016-12-30 | 2017-05-31 | 山东省科学院生物研究所 | New fucoidan oligosaccharide and preparation method and application |
CN106866834A (en) * | 2017-03-16 | 2017-06-20 | 山东省食品发酵工业研究设计院 | It is a kind of to prepare the efficiently method of the fucoidin of customization molecular weight and its application |
CN108853150A (en) * | 2018-07-02 | 2018-11-23 | 集美大学 | Yellow tang extract and its preparation method and application |
CN109349473A (en) * | 2018-11-23 | 2019-02-19 | 盐城工学院 | A kind of Brazilian certain kind of berries fruit juice and preparation method thereof with fat-reducing effect |
CN115677870A (en) * | 2022-09-30 | 2023-02-03 | 广西大学 | Preparation method of sulfated fucogalactan with blood fat reducing effect |
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Cited By (8)
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CN105273104A (en) * | 2015-10-24 | 2016-01-27 | 山东好当家海洋发展股份有限公司 | Method for extracting fucoidan by using Sargassum kjellmanianum |
CN106749439A (en) * | 2016-12-30 | 2017-05-31 | 山东省科学院生物研究所 | New fucoidan oligosaccharide and preparation method and application |
CN106749439B (en) * | 2016-12-30 | 2019-09-17 | 山东省科学院生物研究所 | Fucoidan oligosaccharide and the preparation method and application thereof |
CN106866834A (en) * | 2017-03-16 | 2017-06-20 | 山东省食品发酵工业研究设计院 | It is a kind of to prepare the efficiently method of the fucoidin of customization molecular weight and its application |
CN106866834B (en) * | 2017-03-16 | 2018-01-30 | 山东省食品发酵工业研究设计院 | A kind of method and its application for preparing the efficiently fucoidin of customization molecular weight |
CN108853150A (en) * | 2018-07-02 | 2018-11-23 | 集美大学 | Yellow tang extract and its preparation method and application |
CN109349473A (en) * | 2018-11-23 | 2019-02-19 | 盐城工学院 | A kind of Brazilian certain kind of berries fruit juice and preparation method thereof with fat-reducing effect |
CN115677870A (en) * | 2022-09-30 | 2023-02-03 | 广西大学 | Preparation method of sulfated fucogalactan with blood fat reducing effect |
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Application publication date: 20150422 |