CN115260333A - Active polysaccharide or active polysaccharide composition and application thereof in preparation of product for relieving and/or treating liver injury - Google Patents
Active polysaccharide or active polysaccharide composition and application thereof in preparation of product for relieving and/or treating liver injury Download PDFInfo
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- CN115260333A CN115260333A CN202211045712.6A CN202211045712A CN115260333A CN 115260333 A CN115260333 A CN 115260333A CN 202211045712 A CN202211045712 A CN 202211045712A CN 115260333 A CN115260333 A CN 115260333A
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- 150000004676 glycans Chemical class 0.000 title claims abstract description 194
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 194
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 194
- 206010067125 Liver injury Diseases 0.000 title claims abstract description 46
- 231100000753 hepatic injury Toxicity 0.000 title claims abstract description 41
- 239000000203 mixture Substances 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title description 13
- 241001428166 Eucheuma Species 0.000 claims abstract description 142
- 238000000502 dialysis Methods 0.000 claims abstract description 81
- 235000009917 Crataegus X brevipes Nutrition 0.000 claims abstract description 64
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 claims abstract description 64
- 235000009685 Crataegus X maligna Nutrition 0.000 claims abstract description 64
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 claims abstract description 64
- 235000009486 Crataegus bullatus Nutrition 0.000 claims abstract description 64
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 claims abstract description 64
- 235000009682 Crataegus limnophila Nutrition 0.000 claims abstract description 64
- 235000004423 Crataegus monogyna Nutrition 0.000 claims abstract description 64
- 235000002313 Crataegus paludosa Nutrition 0.000 claims abstract description 64
- 235000009840 Crataegus x incaedua Nutrition 0.000 claims abstract description 64
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 50
- 239000003814 drug Substances 0.000 claims abstract description 34
- 230000000694 effects Effects 0.000 claims abstract description 31
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000012530 fluid Substances 0.000 claims abstract description 14
- 238000001035 drying Methods 0.000 claims abstract description 9
- 238000011049 filling Methods 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 238000001556 precipitation Methods 0.000 claims abstract description 6
- 238000003809 water extraction Methods 0.000 claims abstract description 6
- 241001092040 Crataegus Species 0.000 claims description 63
- 239000011148 porous material Substances 0.000 claims description 18
- 239000000047 product Substances 0.000 claims description 6
- 231100000439 acute liver injury Toxicity 0.000 claims description 4
- 235000013376 functional food Nutrition 0.000 claims description 4
- 235000015872 dietary supplement Nutrition 0.000 claims description 3
- 235000017159 Crataegus pinnatifida Nutrition 0.000 claims 1
- 241000657480 Crataegus pinnatifida Species 0.000 claims 1
- 231100000234 hepatic damage Toxicity 0.000 claims 1
- 230000008818 liver damage Effects 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 229940079593 drug Drugs 0.000 abstract description 15
- 238000011160 research Methods 0.000 abstract description 7
- 240000000171 Crataegus monogyna Species 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 10
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- 210000004185 liver Anatomy 0.000 description 7
- 238000010992 reflux Methods 0.000 description 7
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- 125000003158 alcohol group Chemical group 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
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- 241000699670 Mus sp. Species 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
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- 230000001476 alcoholic effect Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 235000020717 hawthorn extract Nutrition 0.000 description 3
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 230000035622 drinking Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000003304 gavage Methods 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
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- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 235000020927 12-h fasting Nutrition 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 241000123865 Gynura bicolor Species 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
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- 230000006378 damage Effects 0.000 description 1
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- 239000012153 distilled water Substances 0.000 description 1
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- 230000004761 fibrosis Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
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- 229930014626 natural product Natural products 0.000 description 1
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- 231100000331 toxic Toxicity 0.000 description 1
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- 201000001862 viral hepatitis Diseases 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/04—Rhodophycota or rhodophyta (red algae), e.g. Porphyra
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to the technical field of biological medicines, and particularly discloses an active polysaccharide or an active polysaccharide composition and application thereof in preparing a product for relieving and/or treating liver injury. The active polysaccharide is eucheuma polysaccharide, and the eucheuma polysaccharide is prepared by a method comprising the following steps: s11, taking the Chinese medicine eucheuma, and then preparing eucheuma crude polysaccharide by using a water extraction and alcohol precipitation method; s12, dissolving crude eucheuma polysaccharide in water, and dialyzing by using a first dialysis bag to obtain first dialysate and first cut-off liquid; s13, filling the first dialysate into a second dialysis bag for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped solution to obtain the eucheuma polysaccharide. The active polysaccharide composition comprises eucheuma polysaccharide and hawthorn polysaccharide. Research shows that the active polysaccharide and the active polysaccharide composition have excellent effects of treating and relieving liver injury.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to active polysaccharide or an active polysaccharide composition and application thereof in preparing a product for relieving and/or treating liver injury.
Background
The liver is the main site for alcohol metabolism in vivo, and the liver burden is increased by a large amount of alcohol drinking, so that the liver is gradually damaged. After entering the liver, alcohol is discharged out of the body through a series of oxidations, where it is first oxidized to acetaldehyde, further to acetic acid, and finally to carbon dioxide and water. Acetaldehyde causes degeneration, necrosis and fibrosis of liver cells, and has obvious toxic effect on liver cells. Acute alcoholic liver injury can be caused by one-time large-scale drinking, and as the incidence rate of alcoholic liver injury is gradually increased, liver diseases such as viral hepatitis, liver cirrhosis and even liver cancer are increased.
Research shows that the Chinese medicine compound preparation can improve the capacity of the liver to resist alcohol damage, namely, the Chinese medicine compound preparation has better protective effect on the liver. Many traditional Chinese medicines not only have the effects of resisting oxidation and resisting apoptosis activity, but also have the advantage of treating both principal and secondary aspect of disease in the aspect of improving liver function. In the process of developing functional food and medicine for protecting liver, the natural product of traditional Chinese medicine is introduced to treat alcoholic liver injury.
The Eucheuma polysaccharide is extracted from Chinese medicine Eucheuma; research shows that the traditional Chinese medicine composition has a certain treatment effect on liver injury; however, the simple crude polysaccharide from Eucheuma muricatum has no significant therapeutic effect on liver injury. Therefore, the eucheuma polysaccharide which has better treatment effect on liver injury and is developed by taking the eucheuma as the raw material has important application value.
Disclosure of Invention
In order to solve at least one technical problem in the prior art, the invention firstly provides an active polysaccharide. The active polysaccharide is eucheuma polysaccharide prepared by a brand-new method; research shows that the eucheuma polysaccharide prepared by the method can greatly improve the treatment effect of the eucheuma polysaccharide on liver injury compared with the eucheuma polysaccharide.
The technical scheme of the invention is as follows:
an active polysaccharide, wherein the active polysaccharide is eucheuma polysaccharide, and the eucheuma polysaccharide is prepared by a method comprising the following steps:
s11, taking Chinese medicine eucheuma, and then preparing crude eucheuma polysaccharide by a water extraction and alcohol precipitation method;
s12, dissolving crude eucheuma polysaccharide in water, and dialyzing by using a first dialysis bag to obtain first dialysate and first cut-off liquid;
s13, filling the first dialysate into a second dialysis bag for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped solution to obtain the eucheuma polysaccharide.
Preferably, the water extraction and alcohol precipitation method in step S11 specifically includes:
extracting Eucheuma Gelatinosum with water under heating and refluxing to obtain Eucheuma Gelatinosum extractive solution; concentrating the eucheuma extracting solution to obtain eucheuma concentrated solution, and adding ethanol into the eucheuma concentrated solution to ensure that the final volume fraction of the ethanol in the eucheuma concentrated solution is 70-90%; and finally, standing and taking the precipitate to obtain the crude polysaccharide of the eucheuma.
Preferably, the dosage ratio of the eucheuma to the water is 1g:5 to 15mL.
Most preferably, the dosage ratio of the eucheuma to the water is 1g:10mL.
Preferably, the eucheuma extracting solution is concentrated to obtain the eucheuma concentrated solution, so that the volume of the eucheuma concentrated solution is 1/3-1/5 of the volume of water.
Most preferably, the Eucheuma extract is concentrated to obtain the Eucheuma concentrated solution, so that the volume of the Eucheuma concentrated solution is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the Eucheuma concentrated solution is 80%.
Preferably, the first dialysis bag in step S12 has a pore size of 4.5 to 5.5 kD.
Further preferably, the first dialysis bag in step S12 has a dialysis bag with a pore size of 5 kD.
Preferably, the second dialysis bag in step S13 has a pore size of 2.5 to 3.5 kD.
Further preferably, the second dialysis bag in step S13 has a dialysis bag with a pore size of 3 kD.
The invention is surprisingly found in a large number of experiments, the eucheuma polysaccharide is prepared by dialyzing the eucheuma crude polysaccharide by a dialysis bag with the aperture of 4.5-5.5 kD and then dialyzing by a dialysis bag with the aperture of 2.5-3.5 kD, and compared with the eucheuma crude polysaccharide, the eucheuma crude polysaccharide can greatly improve the treatment effect of the eucheuma crude polysaccharide on liver injury.
The inventor needs to emphasize that in the preparation process of the eucheuma polysaccharide, the selection of the dialysis pore diameter by two dialysis bags plays a decisive role in greatly improving the treatment effect on liver injury for the prepared eucheuma polysaccharide. In the preparation process of the eucheuma polysaccharide, the dialysis pore diameters of the dialysis bags are selected differently, so that the difference of the treatment effect of the prepared eucheuma polysaccharide on liver injury is huge; the dialysis pore diameters of the two dialysis bags are not properly selected, so that the treatment effect of the crude eucheuma polysaccharide on liver injury cannot be improved or greatly improved, and even the treatment effect of the crude eucheuma polysaccharide on liver injury is reduced.
The invention also provides an active polysaccharide composition, which comprises eucheuma polysaccharide and hawthorn polysaccharide;
the Eucheuma polysaccharide is obtained by the above method.
Preferably, the weight ratio of the eucheuma polysaccharide to the hawthorn polysaccharide is 1-3:1.
Most preferably, the weight ratio of eucheuma polysaccharide to hawthorn polysaccharide is 2:1.
Preferably, the hawthorn polysaccharide is prepared by the following method:
s21, taking traditional Chinese medicine hawthorn, and then preparing hawthorn crude polysaccharide by using a water extraction and alcohol precipitation method;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing by using a first dialysis bag to obtain first dialysate and first cut-off liquid;
s23, filling the first dialysate into a second dialysis bag for dialysis to obtain a second dialysate and a second trapped fluid; and concentrating and drying the second trapped fluid to obtain the gynura bicolor polysaccharide.
Preferably, the water extraction and alcohol precipitation method in step S21 specifically includes:
extracting fructus crataegi with water under reflux to obtain fructus crataegi extractive solution; then concentrating the hawthorn extract to obtain hawthorn concentrate, and adding ethanol into the hawthorn concentrate to ensure that the final volume fraction of the ethanol in the hawthorn concentrate is 70-80%; and finally, standing and taking the precipitate to obtain the hawthorn crude polysaccharide.
Preferably, the dosage ratio of the hawthorn to the water is 1g: 5-15 mL.
Most preferably, the dosage ratio of the hawthorn to the water is 1g:10mL.
Preferably, the hawthorn extract is concentrated to obtain the hawthorn concentrate, so that the volume of the hawthorn concentrate is 1/3-1/5 of the volume of water.
Most preferably, the hawthorn extract is concentrated to obtain the hawthorn concentrate such that the volume of the hawthorn concentrate is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the hawthorn concentrate is made to be 75%.
Preferably, the first dialysis bag in step S22 has a pore size of 7.5-8.5 kD.
Further preferably, the first dialysis bag in step S12 has a dialysis bag with a pore size of 8 kD.
Preferably, the second dialysis bag in step S13 has a pore size of 5.5 to 6.5 kD.
Further preferably, the second dialysis bag in step S13 has a dialysis bag with a pore size of 6 kD.
The inventor surprisingly discovers in a great deal of research that after the hawthorn polysaccharide prepared by dialyzing the hawthorn crude polysaccharide by using a dialysis bag with the aperture of 7.5-8.5 kD and then dialyzing by using a dialysis bag with the aperture of 5.5-6.5 kD is combined with the eucheuma polysaccharide, the treatment effect of the eucheuma polysaccharide on liver injury can be further and greatly improved.
However, the inventor needs to emphasize here that the preparation method of the hawthorn polysaccharide plays a decisive role in further greatly improving the therapeutic effect of the eucheuma polysaccharide on liver injury after the hawthorn polysaccharide and the eucheuma polysaccharide are combined. Researches show that only after the crude hawthorn polysaccharide is dialyzed by a dialysis bag with the aperture of 7.5-8.5 kD and then is dialyzed by a dialysis bag with the aperture of 5.5-6.5 kD, the prepared hawthorn polysaccharide is combined with the eucheuma polysaccharide, so that the treatment effect of the eucheuma polysaccharide on liver injury can be further and greatly improved; the hawthorn polysaccharide prepared by other methods cannot further greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined with the eucheuma polysaccharide disclosed by the invention.
The invention also provides application of the active polysaccharide or the active polysaccharide composition in preparing a product with the effect of relieving and/or treating liver injury.
Preferably, the liver injury is acute liver injury caused by ethanol.
Preferably, the product is a functional food, a dietary supplement or a medicament.
Has the advantages that: the invention provides an active polysaccharide prepared by a brand new method; the active polysaccharide is eucheuma polysaccharide prepared by a brand-new method; research shows that the eucheuma polysaccharide prepared by the method can greatly improve the treatment effect of the eucheuma polysaccharide on liver injury compared with the eucheuma polysaccharide. Furthermore, the composition obtained by combining the hawthorn polysaccharide prepared by the method and the eucheuma polysaccharide can further greatly improve the treatment effect of the eucheuma polysaccharide on liver injury, and has very excellent effects of treating and relieving the liver injury. The active polysaccharide and the active polysaccharide composition have excellent effects of treating and relieving liver injury; therefore, the compound has important application value as an effective component for developing functional food, dietary supplement or medicament for relieving and/or treating liver injury.
Detailed Description
The technical solution of the present invention will be clearly and completely described with reference to the following examples. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 preparation of active polysaccharide
S11, taking 400g of Chinese medicine eucheuma, adding 4L of water, heating and refluxing for 1 hour at 100 ℃, obtaining eucheuma extracting solution, and concentrating the eucheuma extracting solution to 1L to obtain eucheuma concentrated solution; adding 95% ethanol by volume fraction into the eucheuma concentrated solution to make the final volume fraction of the ethanol in the eucheuma concentrated solution be 80%, standing for 6h, filtering, and taking the solid to obtain crude eucheuma polysaccharide;
s12, dissolving the crude eucheuma polysaccharide in water, and dialyzing by using a dialysis bag with the aperture of 5kD to obtain first dialysate and first cut-off liquid;
s13, filling the first dialysate into a dialysis bag with the aperture of 3kD for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped fluid to obtain eucheuma polysaccharide; i.e. the active polysaccharide.
Example 2 active polysaccharide composition
Uniformly mixing the eucheuma polysaccharide and the hawthorn polysaccharide which are prepared by the method in the embodiment 1 according to the weight ratio of 2:1 to obtain the active polysaccharide composition;
the hawthorn polysaccharide is prepared by the following method:
s21, taking 400g of traditional Chinese medicine hawthorn, adding 4L of water, heating and refluxing for extraction for 1 hour at 100 ℃ to obtain hawthorn extracting solution, and concentrating the hawthorn extracting solution to 1L to obtain hawthorn concentrated solution; adding 95% ethanol by volume into the concentrated hawthorn solution to make the final volume fraction of the ethanol in the concentrated hawthorn solution be 75%, standing for 6h, filtering, and taking the solid to obtain crude hawthorn polysaccharide;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing by using a dialysis bag with the aperture of 8kD to obtain first dialysate and first cut-off liquid;
s23, filling the first dialysate into a dialysis bag with the aperture of 6kD for dialysis to obtain a second dialysate and a second trapped fluid; and concentrating and drying the second trapped fluid to obtain the hawthorn polysaccharide.
Comparative example 1 preparation of active polysaccharide
S11, taking 400g of Chinese medicine eucheuma, adding 4L of water, heating and refluxing for 1 hour at 100 ℃, obtaining eucheuma extracting solution, and concentrating the eucheuma extracting solution to 1L to obtain eucheuma concentrated solution; adding 95% ethanol by volume fraction into the eucheuma concentrated solution to make the final volume fraction of the ethanol in the eucheuma concentrated solution be 80%, standing for 6h, filtering, and taking the solid to obtain crude eucheuma polysaccharide;
s12, dissolving the crude eucheuma polysaccharide in water, and dialyzing by using a dialysis bag with the aperture of 8kD to obtain first dialysate and first cut-off liquid;
s13, filling the first dialysate into a dialysis bag with the aperture of 6kD for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped solution to obtain Eucheuma polysaccharide; i.e. the active polysaccharide.
The difference between the comparative example 1 and the example 1 is that the eucheuma crude polysaccharide in the comparative example 1 is firstly dialyzed by a dialysis bag with the aperture of 8kD, and then is dialyzed by a dialysis bag with the aperture of 6kD to prepare the eucheuma polysaccharide; and in the embodiment 1, the eucheuma crude polysaccharide is dialyzed by a dialysis bag with the aperture of 5kD, and then is dialyzed by a dialysis bag with the aperture of 3kD to obtain the eucheuma polysaccharide.
Comparative example 2 preparation of active polysaccharide
S11, taking 400g of Chinese medicine eucheuma, adding 4L of water, heating and refluxing for 1 hour at 100 ℃, obtaining eucheuma extracting solution, and concentrating the eucheuma extracting solution to 1L to obtain eucheuma concentrated solution; adding 95% ethanol by volume fraction into the eucheuma concentrated solution to make the final volume fraction of the ethanol in the eucheuma concentrated solution be 80%, standing for 6h, filtering, and taking the solid to obtain crude eucheuma polysaccharide;
s12, dissolving the crude polysaccharide of the eucheuma in water, and dialyzing by using a dialysis bag with the aperture of 3kD to obtain first dialysate and first cut-off liquid;
s13, filling the first dialysate into a dialysis bag with the aperture of 1kD for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped fluid to obtain eucheuma polysaccharide; i.e. the active polysaccharide.
The difference between the comparative example 2 and the example 1 is that the comparative example 2 is that the eucheuma crude polysaccharide is firstly dialyzed by a dialysis bag with the aperture of 3kD, and then is dialyzed by a dialysis bag with the aperture of 1kD to prepare the eucheuma polysaccharide; in example 1, the crude Eucheuma polysaccharide is dialyzed by a dialysis bag with the aperture of 5kD, and then dialyzed by a dialysis bag with the aperture of 3kD to obtain the Eucheuma polysaccharide.
Comparative example 3 active polysaccharide composition
Mixing eucheuma polysaccharide and hawthorn polysaccharide prepared by the method of example 1 evenly according to the weight ratio of 2:1 to obtain the active polysaccharide composition;
the hawthorn polysaccharide is prepared by the following method:
s21, taking 400g of traditional Chinese medicine hawthorn, adding 4L of water, heating and refluxing for 1 hour at 100 ℃, obtaining hawthorn extracting solution, and concentrating the hawthorn extracting solution to 1L to obtain hawthorn concentrated solution; adding 95% ethanol by volume into the concentrated hawthorn solution to make the final volume fraction of the ethanol in the concentrated hawthorn solution be 75%, standing for 6h, filtering, and taking the solid to obtain crude hawthorn polysaccharide;
s22, dissolving the crude hawthorn polysaccharide in water, and dialyzing by using a dialysis bag with the aperture of 5kD to obtain first dialysate and first cut-off liquid;
s23, filling the first dialysate into a dialysis bag with the aperture of 3kD for dialysis to obtain a second dialysate and a second trapped fluid; and concentrating and drying the second trapped fluid to obtain the hawthorn polysaccharide.
The difference between the comparative example 3 and the example 2 is that in the comparative example 3, the hawthorn polysaccharide prepared by dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 5kD firstly and then dialyzing by adopting a dialysis bag with the aperture of 3kD is combined with the eucheuma polysaccharide to prepare the active polysaccharide composition; and in the embodiment 2, the hawthorn polysaccharide is prepared by firstly dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 8kD, and then dialyzing by adopting a dialysis bag with the aperture of 6kD, and the prepared hawthorn polysaccharide is combined with the eucheuma polysaccharide to prepare the active polysaccharide composition.
Experimental example 1
The experimental mice with consistent body weight are divided into a normal group, an alcohol group and a medicine group 1-6, wherein each group comprises 10 mice and each half of the mice is male and female. The experiment was carried out as follows, except for the normal group, mice in the alcohol group and the drug group were gavaged once every day at 8mL/kg (about 3.5 g/kg) with the corresponding drugs (10 mg/kg/d) or distilled water of equal volume for 3h before gavage, and the gavage volume was 0.2mL/10g. Fasting for 12h on the 9 th day of molding, and taking blood after 4h after white wine is given on the 10 th day; separating serum, and determining ALT and AST content in serum according to kit instruction method. The test results are shown in Table 1.
The crude Eucheuma polysaccharide prepared according to the method of example 1, and the active polysaccharides or active polysaccharide compositions prepared in examples 1 and 2 and comparative examples 1 to 3 were tested in drug groups 1 to 6, respectively.
Table 1.
As can be seen from the experimental data in Table 1, compared with the blank control group, the AST and ALT contents in the alcohol group are significantly increased, which indicates that the mouse model with acute liver injury caused by ethanol is successfully constructed.
As can be seen from the experimental data in Table 1, the AST and ALT contents of the drug group 1 are reduced but not much larger than that of the alcohol group; this indicates that the crude Eucheuma polysaccharide prepared by the method of example 1 has limited effect in treating acute liver injury caused by ethanol. However, the AST and ALT contents of drug group 2 were greatly reduced compared to those of the alcohol group and drug group 1; this indicates that: the eucheuma polysaccharide is dialyzed by a dialysis bag with the aperture of 4.5-5.5 kD, and then is dialyzed by a dialysis bag with the aperture of 2.5-3.5 kD, so that the eucheuma polysaccharide prepared has excellent effect of treating liver injury; compared with the crude polysaccharide of the eucheuma, the crude polysaccharide of the eucheuma can greatly improve the treatment effect of the crude polysaccharide of the eucheuma on liver injury.
As can be seen from the experimental data in Table 1, the AST and ALT contents of the drug groups 4 and 5 are not greatly reduced compared with the drug group 1, and the reduction range is far smaller than that of the drug group 2; this indicates that: in the preparation process of the eucheuma polysaccharide, the dialysis pore size is selected by two dialysis bags, and the decisive role is played on whether the treatment effect on liver injury can be greatly improved or not for the prepared eucheuma polysaccharide. In the preparation process of the eucheuma polysaccharide, the dialysis pore diameters of the dialysis bags are selected differently, so that the difference of the treatment effect of the prepared eucheuma polysaccharide on liver injury is huge; the dialysis pore diameters of the two dialysis bags are not properly selected, so that the treatment effect of the crude eucheuma polysaccharide on liver injury cannot be improved or greatly improved, and even the treatment effect of the crude eucheuma polysaccharide on liver injury is reduced. Only the eucheuma polysaccharide which is prepared by firstly dialyzing the eucheuma crude polysaccharide by a dialysis bag with the aperture of 4.5-5.5 kD and then dialyzing by a dialysis bag with the aperture of 2.5-3.5 kD has excellent effect of treating liver injury; compared with the crude polysaccharide of the eucheuma, the crude polysaccharide of the eucheuma can greatly improve the treatment effect of the crude polysaccharide of the eucheuma on liver injury.
As can be seen from the experimental data in Table 1, the AST and ALT contents in drug group 3 are further greatly reduced compared with those in drug group 2; so that the description is as follows: the crude polysaccharide of hawthorn is firstly dialyzed by a dialysis bag with the aperture of 7.5-8.5 kD, and then is dialyzed by a dialysis bag with the aperture of 5.5-6.5 kD to prepare the hawthorn polysaccharide, and after the combination with the eucheuma polysaccharide disclosed by the invention, the therapeutic effect of the eucheuma polysaccharide on liver injury can be further greatly improved. However, the AST and ALT contents of drug group 6 were not reduced compared to drug group 2 and were much higher than drug group 3; this is further illustrated: the preparation method of the hawthorn polysaccharide plays a decisive role in greatly improving the treatment effect of the eucheuma polysaccharide on liver injury after the hawthorn polysaccharide and the eucheuma polysaccharide are combined; after the crude hawthorn polysaccharide is combined with the eucheuma polysaccharide disclosed by the invention, the therapeutic effect of the eucheuma polysaccharide on liver injury can be further greatly improved only by dialyzing the crude hawthorn polysaccharide by using a dialysis bag with the aperture of 7.5-8.5 kD and then dialyzing the hawthorn polysaccharide by using a dialysis bag with the aperture of 5.5-6.5 kD; the hawthorn polysaccharide prepared by other methods cannot further greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined with the eucheuma polysaccharide disclosed by the invention.
Claims (10)
1. An active polysaccharide, wherein the active polysaccharide is eucheuma polysaccharide, and the eucheuma polysaccharide is prepared by a method comprising the following steps:
s11, taking Chinese medicine eucheuma, and then preparing crude eucheuma polysaccharide by a water extraction and alcohol precipitation method;
s12, dissolving crude eucheuma polysaccharide in water, and dialyzing by using a first dialysis bag to obtain first dialysate and first cut-off liquid;
s13, filling the first dialysate into a second dialysis bag for dialysis to obtain a second dialysate and a second trapped fluid; concentrating and drying the second trapped solution to obtain the eucheuma polysaccharide.
2. The active polysaccharide of claim 1, wherein the first dialysis bag in step S12 has a pore size of 4.5 to 5.5 kD.
3. The active polysaccharide of claim 1, wherein the first dialysis bag in step S12 has a pore size of 5 kD.
4. The active polysaccharide of claim 1, wherein the second dialysis bag in step S13 has a pore size of 2.5 to 3.5 kD.
5. The active polysaccharide of claim 1, wherein the second dialysis bag in step S13 has a 3kD pore size.
6. An active polysaccharide composition comprising eucheuma polysaccharide and hawthorn polysaccharide;
the eucheuma polysaccharide is prepared by the method of any one of claims 1 to 5.
7. The active polysaccharide composition of claim 6, wherein the weight ratio of Eucheuma polysaccharide to Crataegus pinnatifida polysaccharide is 1-3:1;
most preferably, the weight ratio of eucheuma polysaccharide to hawthorn polysaccharide is 2:1.
8. Use of an active polysaccharide or active polysaccharide composition according to any one of claims 1 to 7 in the manufacture of a product having the effect of alleviating and/or treating liver damage.
9. The use of claim 8, wherein the liver injury is acute liver injury caused by ethanol.
10. The use according to claim 8, wherein the product is a functional food, a dietary supplement or a medicament.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104721221A (en) * | 2015-01-29 | 2015-06-24 | 广州暨南生物医药研究开发基地有限公司 | Application of eucheuma gelatinae polysaccharide to preparation of medicines for inhibiting respiratory viruses |
| CN110464742A (en) * | 2019-09-08 | 2019-11-19 | 广东医科大学 | A kind of Extraction of Eucheuma gelatinae and its preparing the application in treating organs fibrosis medicine |
| CN114569641A (en) * | 2022-04-22 | 2022-06-03 | 上海朗泰凯尔生物技术有限公司 | Composition for relieving acute alcoholic liver injury and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104721221A (en) * | 2015-01-29 | 2015-06-24 | 广州暨南生物医药研究开发基地有限公司 | Application of eucheuma gelatinae polysaccharide to preparation of medicines for inhibiting respiratory viruses |
| CN110464742A (en) * | 2019-09-08 | 2019-11-19 | 广东医科大学 | A kind of Extraction of Eucheuma gelatinae and its preparing the application in treating organs fibrosis medicine |
| CN114569641A (en) * | 2022-04-22 | 2022-06-03 | 上海朗泰凯尔生物技术有限公司 | Composition for relieving acute alcoholic liver injury and preparation method and application thereof |
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