CN115260333B - Active polysaccharide or active polysaccharide composition and application thereof in preparation of products for relieving and/or treating liver injury - Google Patents
Active polysaccharide or active polysaccharide composition and application thereof in preparation of products for relieving and/or treating liver injury Download PDFInfo
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- CN115260333B CN115260333B CN202211045712.6A CN202211045712A CN115260333B CN 115260333 B CN115260333 B CN 115260333B CN 202211045712 A CN202211045712 A CN 202211045712A CN 115260333 B CN115260333 B CN 115260333B
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 197
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 197
- 150000004676 glycans Chemical class 0.000 title claims abstract description 193
- 206010067125 Liver injury Diseases 0.000 title claims abstract description 44
- 231100000753 hepatic injury Toxicity 0.000 title claims abstract description 40
- 239000000203 mixture Substances 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims description 14
- 241001428166 Eucheuma Species 0.000 claims abstract description 140
- 238000000502 dialysis Methods 0.000 claims abstract description 89
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 53
- 235000009917 Crataegus X brevipes Nutrition 0.000 claims abstract description 49
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 claims abstract description 49
- 235000009685 Crataegus X maligna Nutrition 0.000 claims abstract description 49
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 claims abstract description 49
- 235000009486 Crataegus bullatus Nutrition 0.000 claims abstract description 49
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 claims abstract description 49
- 235000009682 Crataegus limnophila Nutrition 0.000 claims abstract description 49
- 235000004423 Crataegus monogyna Nutrition 0.000 claims abstract description 49
- 235000002313 Crataegus paludosa Nutrition 0.000 claims abstract description 49
- 235000009840 Crataegus x incaedua Nutrition 0.000 claims abstract description 49
- 238000000034 method Methods 0.000 claims abstract description 33
- 239000003814 drug Substances 0.000 claims abstract description 32
- 230000000694 effects Effects 0.000 claims abstract description 31
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000012530 fluid Substances 0.000 claims abstract description 23
- 238000001035 drying Methods 0.000 claims abstract description 10
- 238000001556 precipitation Methods 0.000 claims abstract description 7
- 238000003809 water extraction Methods 0.000 claims abstract description 7
- 241001092040 Crataegus Species 0.000 claims description 52
- 239000011148 porous material Substances 0.000 claims description 23
- 239000012465 retentate Substances 0.000 claims description 7
- 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
- 238000011049 filling Methods 0.000 claims description 2
- 235000013311 vegetables Nutrition 0.000 claims description 2
- 229940079593 drug Drugs 0.000 abstract description 17
- 238000011160 research Methods 0.000 abstract description 7
- 240000000171 Crataegus monogyna Species 0.000 abstract 1
- 239000012141 concentrate Substances 0.000 description 21
- 230000000052 comparative effect Effects 0.000 description 10
- 238000010438 heat treatment Methods 0.000 description 7
- 210000004185 liver Anatomy 0.000 description 7
- 235000020717 hawthorn extract Nutrition 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 125000003158 alcohol group Chemical group 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 238000010992 reflux Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 235000014493 Crataegus Nutrition 0.000 description 4
- -1 haw polysaccharide Chemical class 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 230000001476 alcoholic effect Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 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
- 230000006378 damage Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 235000007106 Crataegus suborbiculata Nutrition 0.000 description 1
- 241000073432 Crataegus suborbiculata Species 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- 235000013202 a hawthorn Nutrition 0.000 description 1
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000006909 anti-apoptosis Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
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- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
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- 239000012153 distilled water Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
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- 230000006870 function Effects 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
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- 238000007254 oxidation reaction Methods 0.000 description 1
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- 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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- 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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- 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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- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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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, extracting 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 with a first dialysis bag to obtain a first dialysate and a first trapped fluid; s13, the first dialysate is filled into a second dialysis bag for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide. The active polysaccharide composition comprises eucheuma polysaccharide and hawthorn polysaccharide. The 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 biological medicine, in particular to an active polysaccharide or an active polysaccharide composition and application thereof in preparing products for relieving and/or treating liver injury.
Background
Liver is the main place of alcohol metabolism in the body, and a great deal of alcohol consumption can increase the burden of the liver, and further gradually damage the liver. Alcohol enters the liver and is discharged out of the body through a series of oxidation, and is oxidized into acetaldehyde, acetic acid and carbon dioxide and water in the liver. Acetaldehyde causes degeneration, necrosis and fibrosis of liver cells, and has obvious toxic action on liver cells. Acute alcoholic liver injury can be caused by one-time drinking of a large amount of alcohol, and along with the gradual increase of the incidence rate of the alcoholic liver injury, viral hepatitis, liver cirrhosis, liver cancer and other liver diseases are also increased.
Research shows that the compound Chinese medicine preparation can raise the alcohol damage resisting capacity of liver, i.e. has excellent liver protecting effect. Many traditional Chinese medicines have the functions of antioxidation and anti-apoptosis activity, and have the advantage of treating both symptoms and root causes in improving liver functions. In the process of developing liver-protecting functional foods and medicines, the treatment of alcoholic liver injury by introducing natural products of traditional Chinese medicines is an important method.
The eucheuma polysaccharide is polysaccharide extracted from Chinese medicine eucheuma; research shows that it has certain curative effect on liver injury; however, the crude polysaccharide extracted from Eucheuma was not significant for the treatment of liver injury. Therefore, the eucheuma polysaccharide with better treatment effect on liver injury is developed by taking eucheuma as a raw material, and has important application value.
Disclosure of Invention
In order to solve at least one technical problem existing 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 compared with the crude eucheuma polysaccharide, the eucheuma polysaccharide prepared by the method can greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury.
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, extracting 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 with a first dialysis bag to obtain a first dialysate and a first trapped fluid;
s13, the first dialysate is filled into a second dialysis bag for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide.
Preferably, the specific method of the water extraction and alcohol precipitation method in the step S11 is as follows:
heating and reflux extracting traditional Chinese medicine eucheuma with water to obtain eucheuma extract; concentrating the eucheuma extract to obtain eucheuma concentrate, and adding ethanol into the eucheuma concentrate to make the final volume fraction of the ethanol in the eucheuma concentrate be 70-90%; finally, standing and taking out the precipitate to obtain the crude eucheuma polysaccharide.
Preferably, the ratio of Eucheuma to water is 1g: 5-15 mL.
Most preferably, the ratio of Eucheuma to water is 1g:10mL.
Preferably, the eucheuma extract is concentrated to obtain a eucheuma concentrate such that the volume of the eucheuma concentrate is 1/3 to 1/5 of the volume of the water.
Most preferably, the eucheuma extract is concentrated to obtain a eucheuma concentrate such that the volume of the eucheuma concentrate is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the eucheuma concentrate is made 80%.
Preferably, the first dialysis bag in step S12 has a pore size of 4.5-5.5 kD.
Further preferably, the first dialysis bag in step S12 is a dialysis bag with a pore size of 5 kD.
Preferably, the second dialysis bag in step S13 has a pore size of 2.5-3.5 kD.
Further preferably, the second dialysis bag in step S13 is a dialysis bag having a pore size of 3 kD.
The invention surprisingly discovers in a large number of experiments that the eucheuma polysaccharide prepared by dialyzing the eucheuma crude polysaccharide by adopting a dialysis bag with the aperture of 4.5-5.5 kD and then dialyzing the eucheuma crude polysaccharide by adopting a dialysis bag with the aperture of 2.5-3.5 kD can greatly improve the treatment effect of the eucheuma crude polysaccharide on liver injury compared with the eucheuma crude polysaccharide.
The inventors herein have emphasized that during the preparation of eucheuma polysaccharide, the selection of dialysis pore size by two dialysis bags plays a decisive role in greatly improving the therapeutic effect on liver injury for the prepared eucheuma polysaccharide. In the preparation process of eucheuma polysaccharide, the dialysis pore diameters of the two dialysis bags are different, and the difference of the treatment effect of the prepared eucheuma polysaccharide on liver injury is huge; the two dialysis bags are used for improper selection of dialysis pore diameter, and compared with the crude eucheuma polysaccharide, the method can not improve or greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury, and even reduce the treatment effect.
The invention also provides an active polysaccharide composition, which comprises eucheuma polysaccharide and hawthorn polysaccharide;
the eucheuma polysaccharide is prepared by the method.
Preferably, the weight ratio of eucheuma polysaccharide to hawthorn polysaccharide is 1-3:1.
Most preferably, the weight ratio of eucheuma polysaccharide to crataegus polysaccharide is 2:1.
Preferably, the hawthorn polysaccharide is prepared by the following method:
s21, taking traditional Chinese medicine hawthorn, and then preparing crude hawthorn polysaccharide by using a water extraction and alcohol precipitation method;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing with a first dialysis bag to obtain a first dialyzate and a first trapped fluid;
s23, loading the first dialysate into a second dialysis bag for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the hawthorn vegetable polysaccharide.
Preferably, the specific method of the water extraction and alcohol precipitation method in the step S21 is as follows:
heating and reflux extracting fructus crataegi with water to obtain fructus crataegi extractive solution; concentrating the haw extract to obtain haw concentrate, and adding ethanol into the haw concentrate to make the final volume fraction of ethanol in the haw concentrate be 70-80%; finally, standing and taking out sediment 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 hawthorn to water is 1g:10mL.
Preferably, the hawthorn extract is concentrated to obtain 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 produce a 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 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 is a dialysis bag having a pore size of 8 kD.
Preferably, the second dialysis bag in step S13 has a pore size of 5.5-6.5 kD.
Further preferably, the second dialysis bag in step S13 is a dialysis bag with a pore size of 6 kD.
The inventor surprisingly found in a great deal of researches that the treatment effect of eucheuma polysaccharide on liver injury can be further greatly improved after the combination of the eucheuma polysaccharide with the eucheuma polysaccharide provided by the invention, wherein the crataegus polysaccharide is prepared by dialyzing the crataegus crude polysaccharide by adopting a dialysis bag with the aperture of 7.5-8.5 kD, and then dialyzing the crataegus crude polysaccharide by adopting a dialysis bag with the aperture of 5.5-6.5 kD.
However, the inventors herein emphasized 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 prepared hawthorn polysaccharide is combined with the eucheuma polysaccharide. Researches show that only the hawthorn polysaccharide prepared by dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 7.5-8.5 kD, and then dialyzing by adopting a dialysis bag with the aperture of 5.5-6.5 kD, can further greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined with the eucheuma polysaccharide; the hawthorn polysaccharide prepared by other methods can not further and greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined with the eucheuma polysaccharide.
The invention also provides an application of the active polysaccharide or the active polysaccharide composition in preparing products 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 pharmaceutical.
The beneficial effects are 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 compared with the crude eucheuma polysaccharide, the eucheuma polysaccharide prepared by the method can greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury. 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 effect of treating and relieving the liver injury. Because the active polysaccharide and the active polysaccharide composition have excellent effects of treating and relieving liver injury; therefore, the active ingredient of the extract is used for developing functional foods, dietary supplements or medicines for relieving and/or treating liver injury, and has important application value.
Detailed Description
The technical scheme of the present invention will be clearly and completely described in the following examples. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
EXAMPLE 1 preparation of active polysaccharide
S11, taking 400g of traditional Chinese medicine eucheuma, then adding 4L of water, heating and refluxing for extraction for 1h at 100 ℃ to obtain eucheuma extract, and concentrating the eucheuma extract to 1L to obtain eucheuma concentrate; adding 95% ethanol into the concentrated eucheuma liquid to make the final volume fraction of ethanol in the concentrated eucheuma liquid 80%, standing for 6 hr, filtering, and collecting solid to obtain eucheuma crude polysaccharide;
s12, dissolving crude eucheuma polysaccharide in water, and dialyzing with a dialysis bag with a pore diameter of 5kD to obtain a first dialysate and a first retentate;
s13, placing the first dialysate into a dialysis bag with the aperture of 3kD for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide; i.e. the active polysaccharide.
EXAMPLE 2 active polysaccharide composition
Uniformly mixing eucheuma polysaccharide and hawthorn polysaccharide prepared by the method of the example 1 according to the weight ratio of 2:1 to obtain the active polysaccharide composition;
the hawthorn polysaccharide is prepared by the following steps:
s21, taking 400g of traditional Chinese medicine hawthorn, then adding 4L of water, heating and reflux-extracting for 1h at 100 ℃ to obtain hawthorn extract, and concentrating the hawthorn extract to 1L to obtain hawthorn concentrate; adding 95% ethanol into the concentrated solution to obtain final volume fraction of ethanol in the concentrated solution of fructus crataegi of 75%, standing for 6 hr, filtering, and collecting solid to obtain crude polysaccharide of fructus crataegi;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing with a dialysis bag with the aperture of 8kD to obtain a first dialysate and a first retentate;
s23, placing the first dialysate into a dialysis bag with the aperture of 6kD for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the hawthorn polysaccharide.
Comparative example 1 preparation of active polysaccharide
S11, taking 400g of traditional Chinese medicine eucheuma, then adding 4L of water, heating and refluxing for extraction for 1h at 100 ℃ to obtain eucheuma extract, and concentrating the eucheuma extract to 1L to obtain eucheuma concentrate; adding 95% ethanol into the concentrated eucheuma liquid to make the final volume fraction of ethanol in the concentrated eucheuma liquid 80%, standing for 6 hr, filtering, and collecting solid to obtain eucheuma crude polysaccharide;
s12, dissolving crude eucheuma polysaccharide in water, and dialyzing with a dialysis bag with the aperture of 8kD to obtain a first dialysate and a first retentate;
s13, placing the first dialysate into a dialysis bag with the aperture of 6kD for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide; i.e. the active polysaccharide.
Comparative example 1 differs from example 1 in that comparative example 1 was prepared by dialyzing crude eucheuma polysaccharide with a dialysis bag having a pore size of 8kD, and then dialyzing with a dialysis bag having a pore size of 6kD to obtain eucheuma polysaccharide; in the embodiment 1, crude eucheuma polysaccharide is dialyzed by a dialysis bag with the aperture of 5kD, and then the eucheuma polysaccharide is prepared by dialysis by a dialysis bag with the aperture of 3 kD.
Comparative example 2 preparation of active polysaccharide
S11, taking 400g of traditional Chinese medicine eucheuma, then adding 4L of water, heating and refluxing for extraction for 1h at 100 ℃ to obtain eucheuma extract, and concentrating the eucheuma extract to 1L to obtain eucheuma concentrate; adding 95% ethanol into the concentrated eucheuma liquid to make the final volume fraction of ethanol in the concentrated eucheuma liquid 80%, standing for 6 hr, filtering, and collecting solid to obtain eucheuma crude polysaccharide;
s12, dissolving crude eucheuma polysaccharide in water, and dialyzing with a dialysis bag with a pore diameter of 3kD to obtain a first dialysate and a first retentate;
s13, placing the first dialysate into a dialysis bag with the aperture of 1kD for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide; i.e. the active polysaccharide.
Comparative example 2 differs from example 1 in that comparative example 2 was prepared by dialyzing crude eucheuma polysaccharide using a dialysis bag having a pore size of 3kD, and then dialyzing using a dialysis bag having a pore size of 1kD to obtain eucheuma polysaccharide; in the embodiment 1, crude eucheuma polysaccharide is dialyzed by a dialysis bag with the aperture of 5kD, and then the eucheuma polysaccharide is prepared by dialysis by a dialysis bag with the aperture of 3 kD.
Comparative example 3 active polysaccharide composition
Uniformly mixing eucheuma polysaccharide and hawthorn polysaccharide prepared by the method of the example 1 according to the weight ratio of 2:1 to obtain the active polysaccharide composition;
the hawthorn polysaccharide is prepared by the following steps:
s21, taking 400g of traditional Chinese medicine hawthorn, then adding 4L of water, heating and reflux-extracting for 1h at 100 ℃ to obtain hawthorn extract, and concentrating the hawthorn extract to 1L to obtain hawthorn concentrate; adding 95% ethanol into the concentrated solution to obtain final volume fraction of ethanol in the concentrated solution of fructus crataegi of 75%, standing for 6 hr, filtering, and collecting solid to obtain crude polysaccharide of fructus crataegi;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing with a dialysis bag with the aperture of 5kD to obtain a first dialysate and a first retentate;
s23, placing the first dialysate into a dialysis bag with the aperture of 3kD for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the hawthorn polysaccharide.
Comparative example 3 differs from example 2 in that comparative example 3 is a preparation of an active polysaccharide composition by combining a crude haw polysaccharide with eucheuma polysaccharide of the present invention, wherein the crude haw polysaccharide is dialyzed first with a dialysis bag having a pore size of 5kD and then with a dialysis bag having a pore size of 3 kD; in the embodiment 2, the crude haw polysaccharide is dialyzed by a dialysis bag with the aperture of 8kD, and then the haw polysaccharide prepared by dialysis by a dialysis bag with the aperture of 6kD is combined with the eucheuma polysaccharide to prepare the active polysaccharide composition.
Experimental example 1
The experimental mice with consistent weight are divided into a normal group, an alcohol group and a drug group, wherein 10 mice in each group are respectively male and female. The experiment is carried out as follows, except for the normal group, the mice in the alcohol group and the drug group are filled with 8mL/kg (about 3.5 g/kg) of white spirit (alcohol: water: v-53:47) once every day, and the corresponding drug (10 mg/kg/d) or equal volume distilled water is filled 3 hours before the white spirit is filled into the stomach, and the filling volume is 0.2mL/10g. Fasted for 12 hours on the 9 th day of molding, 4 hours after white wine is given on the 10 th day, and blood is taken after head breakage; serum was isolated and the ALT and AST contents in the serum were determined according to the kit instructions. The test results are shown in Table 1.
Pharmaceutical sets 1 to 6 were tested for crude eucheuma polysaccharide prepared according to the method of example 1, and for the active polysaccharide or active polysaccharide compositions prepared in examples 1, 2 and comparative examples 1 to 3, respectively.
Table 1.
From the experimental data in table 1, compared with the blank control group, the content of AST and ALT in the alcohol group is obviously increased, which proves that the construction of the mouse model of acute liver injury caused by ethanol is successful.
As can be seen from the experimental data in table 1, the AST and ALT contents of drug group 1 were lower than those of the alcohol group, but the magnitudes were not large; this shows that the crude eucheuma polysaccharide prepared by the method of example 1 has limited therapeutic effect on acute liver injury caused by ethanol. However, the AST and ALT content of drug group 2 was significantly reduced compared to the alcohol group and drug group 1; this illustrates: the eucheuma polysaccharide prepared by dialyzing the eucheuma crude polysaccharide by adopting a dialysis bag with the aperture of 4.5-5.5 kD, and then by adopting a dialysis bag with the aperture of 2.5-3.5 kD has excellent liver injury treatment effect; compared with the crude eucheuma polysaccharide, the method can greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury.
As can be seen from the experimental data in table 1, the AST and ALT contents of drug groups 4 and 5 were not significantly reduced compared to drug group 1, which was much less reduced than drug group 2; this illustrates: in the preparation process of eucheuma polysaccharide, the dialysis aperture is selected by two dialysis bags, and the method plays a decisive role in greatly improving the treatment effect on liver injury for the prepared eucheuma polysaccharide. In the preparation process of eucheuma polysaccharide, the dialysis pore diameters of the two dialysis bags are different, and the difference of the treatment effect of the prepared eucheuma polysaccharide on liver injury is huge; the two dialysis bags are used for improper selection of dialysis pore diameter, and compared with the crude eucheuma polysaccharide, the method can not improve or greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury, and even reduce the treatment effect. Only the eucheuma polysaccharide prepared by dialyzing the eucheuma crude polysaccharide by adopting a dialysis bag with the aperture of 4.5-5.5 kD, and then adopting a dialysis bag with the aperture of 2.5-3.5 kD has excellent liver injury treatment effect; compared with the crude eucheuma polysaccharide, the method can greatly improve the treatment effect of the crude eucheuma polysaccharide on liver injury.
As can be seen from the experimental data in table 1, the AST and ALT contents of the drug group 3 are further greatly reduced compared with those of the drug group 2; this is illustrated: the hawthorn polysaccharide is prepared by dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 7.5-8.5 kD, and then dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 5.5-6.5 kD, and the treatment effect of the eucheuma polysaccharide on liver injury can be further greatly improved after the hawthorn crude polysaccharide is combined with the eucheuma polysaccharide. However, the AST, ALT content of drug group 6 was not reduced compared to drug group 2 and was significantly higher than drug group 3; this further illustrates: the preparation method of the hawthorn polysaccharide has decisive effect on whether the prepared hawthorn polysaccharide and eucheuma polysaccharide can further greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined; only the hawthorn polysaccharide is prepared by dialyzing the hawthorn crude polysaccharide by adopting a dialysis bag with the aperture of 7.5-8.5 kD, and then dialyzing by adopting a dialysis bag with the aperture of 5.5-6.5 kD, and the treatment effect of the eucheuma polysaccharide on liver injury can be further greatly improved after the hawthorn polysaccharide is combined with the eucheuma polysaccharide; the hawthorn polysaccharide prepared by other methods can not further and greatly improve the treatment effect of the eucheuma polysaccharide on liver injury after being combined with the eucheuma polysaccharide.
Claims (6)
1. An active polysaccharide, characterized in that the active polysaccharide is eucheuma polysaccharide, the eucheuma polysaccharide is a natural polysaccharide
The preparation method of the eucheuma polysaccharide comprises the following steps:
s11, preparing crude polysaccharide of Chinese medicine eucheuma by using water extraction and alcohol precipitation method;
s12, dissolving the crude eucheuma polysaccharide in water, and dialyzing with a first dialysis bag to obtain a first dialysate
And a first retentate;
s13, the first dialysate is filled into a second dialysis bag for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the eucheuma polysaccharide;
in the step S12, the aperture of the first dialysis bag is 4.5-5.5 kD;
and in the step S13, the aperture of the second dialysis bag is 2.5-3.5 kD.
2. The active polysaccharide according to claim 1, wherein the first dialysis bag in step S12 has a pore size of 5 kD.
3. The active polysaccharide according to claim 1, wherein the second dialysis bag in step S13 has a pore size of 3 kD.
4. An active polysaccharide composition comprising eucheuma polysaccharide and hawkthorn polysaccharide; wherein, the weight ratio of eucheuma polysaccharide to hawthorn polysaccharide is 2:1;
the eucheuma polysaccharide is prepared by the method of any one of claims 1-3;
the hawthorn polysaccharide is prepared by the following steps:
s21, taking traditional Chinese medicine hawthorn, and then preparing crude hawthorn polysaccharide by using a water extraction and alcohol precipitation method;
s22, dissolving the hawthorn crude polysaccharide in water, and dialyzing with a first dialysis bag to obtain a first dialyzate and
a first retentate;
s23, filling the first dialysate into a second dialysis bag for dialysis to obtain second dialysate and second trapped fluid; concentrating and drying the second trapped fluid to obtain the hawthorn vegetable polysaccharide;
in the step S22, the aperture of the first dialysis bag is 7.5-8.5 kD;
and in the step S23, the aperture of the second dialysis bag is 5.5-6.5 kD.
5. Use of the active polysaccharide or the active polysaccharide composition according to any one of claims 1 to 4 for preparing a product having an effect of alleviating and/or treating liver injury;
the liver injury is acute liver injury caused by ethanol.
6. The use according to claim 5, wherein the product is a functional food, a dietary supplement or a medicament.
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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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