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CN108354956A - Sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug - Google Patents

Sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug Download PDF

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Publication number
CN108354956A
CN108354956A CN201810192046.6A CN201810192046A CN108354956A CN 108354956 A CN108354956 A CN 108354956A CN 201810192046 A CN201810192046 A CN 201810192046A CN 108354956 A CN108354956 A CN 108354956A
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China
Prior art keywords
sunflower disk
sunflower
liquid
active ingredient
protein
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Pending
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CN201810192046.6A
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Chinese (zh)
Inventor
李婉南
杨茂义
赵国安
宋海淼
葛振龙
康明
刘小波
高波
韩露
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Jilin University
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Jilin University
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Priority to CN201810192046.6A priority Critical patent/CN108354956A/en
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/28Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/01Hydrolysed proteins; Derivatives thereof
    • A61K38/011Hydrolysed proteins; Derivatives thereof from plants
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Botany (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biotechnology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Medical Informatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses a kind of sunflower disk powder and its active ingredient to prepare the application in repairing wounded hepatocytes drug.The sunflower disk powder and its active ingredient of the present invention has good anti-inflammatory and repair to the wounded hepatocytes caused by hyperuricemia.

Description

Sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug
Technical field
The present invention relates to technical field of pharmaceuticals, more particularly to a kind of sunflower disk powder and its active ingredient are repaired in preparation and are damaged Application in liver cell drug.
Background technology
Domestic and international number of the infected is in rising trend at present for hyperuricemia, is increased and kidney with uric acid generation in pathogenesis Based on dirty underexcretion, and the disease medication usually causes kidney liver to damage and aggravate liver kidney burden.
Floral disc of sunflower is the floral disc of feverfew sunflower (Helianthus annuuss L.), also known as sunflower flower Support, sunflower disc and certain herbaceous plants with big flowers room, mild-natured, slightly sweet flavor, nontoxic, return liver warp, major function have The flat liver of heat-clearing, analgesic, hemostasis, are used for Treatment of arthritis, mastitis, dizziness, tinnitus, abdominal pain etc..Sunflower plate decocting liquid has treatment function of tumor, sunflower flower Disk ethanol extract has certain antihypertensive effect, civil also to use it as medical treatment atrophic gastritis herbal cuisine.China is sunflower Plant big country, sunflower disk it is resourceful.But in tradition is applied, sunflower disk is as livestock after will typically removing sunflower seed Forage feed livestock or as burn material warming & cooking, do not play the economic benefit and medical function of its bigger, cause resource Waste.
Invention content
In order to solve problems in the prior art, an embodiment of the present invention provides a kind of sunflower disk powder and its active ingredient to make The standby application repaired in wounded hepatocytes drug.The technical solution is as follows:
On the one hand, a kind of sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug.
Further, the wounded hepatocytes are caused by hyperuricemia.
Further, the sunflower disk powder active ingredient is prepared by the following method to obtain:
1) sunflower disk is crushed to 100-200 mesh, the deionized water of 6-10 times of quality is added, is sufficiently stirred, uses hydroxide It is 8-9 that sodium, which adjusts pH value, is stirred 120-240 minutes at 45-50 DEG C, obtains sunflower disk protein isolate liquid;
2) sunflower disk protein isolate liquid is heated to 85-90 DEG C, stirs 1-3h, be cooled to 45-55 DEG C, addition accounts for sunflower disk The compound protease of protein isolate liquid total volume 0.1%-0.3% digests 3-5h, and when reaction terminating, 85- is warming up in 5 minutes It 90 DEG C, is kept for 5-10 minutes, so that compound protease is lost activity, gained enzymolysis liquid is spare;
3) enzymolysis liquid of step 2) is centrifuged 8-12 minutes under 5000 revs/min of rotating speed, collects supernatant, obtains sunflower The thick liquid of protein micromolecular peptide;
4) by the thick liquid of sunflower protein small-molecular peptides of step 3) through the hollow-fibre membrane that molecular weight is 5000-20000 that shuts off Ultrafiltration removes high molecular weight protein, obtains sunflower disk small-molecular peptides liquid;
5) the sunflower disk small-molecular peptides liquid of step 4) is concentrated, make its volume concentration to original a quarter to two/ One, spray drying to get.
The advantageous effect that technical solution provided in an embodiment of the present invention is brought is:The present invention sunflower disk powder and its effectively at Divide has good anti-inflammatory and repair to the wounded hepatocytes caused by hyperuricemia.
Description of the drawings
The technical solution in example is applied in order to illustrate more clearly of the present invention, required use in being described below to embodiment Attached drawing be briefly described.
Fig. 1 is blank group liver cell figure in the embodiment of the present invention 2;
Fig. 2 is model group liver cell figure in the embodiment of the present invention 2;
Fig. 3 is sunflower disk powder group of effective components liver cell figure in the embodiment of the present invention 2;
Fig. 4 is alkaloid group liver cell figure in the embodiment of the present invention 2;
Fig. 5 is general flavone group liver cell figure in the embodiment of the present invention 2;
Fig. 6 is polysaccharide group liver cell figure in the embodiment of the present invention 2;
Fig. 7 is sunflower disk powder group liver cell figure in the embodiment of the present invention 2;
Fig. 8 is other purine group liver cell figure in the embodiment of the present invention 2.
Specific implementation mode
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with attached drawing to embodiment party of the present invention Formula is described in further detail.
The preparation of 1 sunflower disk powder of embodiment and its active ingredient
1) sunflower disk 5kg is crushed to 100 mesh, the deionized water of 30kg is added, is sufficiently stirred, pH is adjusted with sodium hydroxide Value is 8, is stirred 120 minutes at 45 DEG C, obtains sunflower disk protein isolate liquid.
2) sunflower disk protein isolate liquid is heated to 85 DEG C, stirs 1h, be cooled to 45 DEG C, addition accounts for sunflower disk protein isolate (Ginger Protease, helicopepsin, bat moth Protein in Larvae digestive ferment press 1 to the compound protease of liquid total volume 0.1%:0.5:1 Mass ratio mixing), digest 3h, when reaction terminating, 85 DEG C be warming up in 5 minutes, is kept for 5 minutes, compound protease is made to lose Deactivation, gained enzymolysis liquid are spare.
3) enzymolysis liquid of step 2) is centrifuged 8 minutes under 5000 revs/min of rotating speed, collects supernatant, obtains sunflower egg The thick liquid of white small-molecular peptides.
4) the hollow-fibre membrane ultrafiltration for being 5000 through the molecular weight that shuts off by the thick liquid of sunflower protein small-molecular peptides of step 3), is removed High molecular weight protein is removed, sunflower disk small-molecular peptides liquid is obtained.
5) the sunflower disk small-molecular peptides liquid of step 4) is concentrated, its volume concentration to original a quarter is made to arrive, sprayed Drying is to get sunflower disk powder active ingredient.
The preparation method of bat moth Protein in Larvae digestive ferment includes the following steps:By the decaptitating of bat moth larvae and reject worm skin And fat, take alimentary canal whole, be added the phosphate buffer of 10 times of quality, homogenate, in 4 DEG C of undertissue's slurries, then with The speed of 3000rpm/min centrifuges, and takes supernatant to be heated to 40 DEG C and is rapidly cooled to 5 DEG C, is filtered to remove fat, dialysis removes Salt, freeze-drying to get.
2 sunflower disk powder of embodiment and its active ingredient anti-trioxypurine repair damaged liver test cell line
(1) material
Small-molecular peptides:Sunflower disk powder active ingredient prepared by embodiment 1.
Yeast extract:Oxiod companies of Britain.
Oxonic Acid sylvite:Sigma Co., USA, lot number:101310713.
Allopurinol tablet:The Place pharmacy (Jiangsu) Co., Ltd, lot number:201608301.
Mouse retention acid detection kit:Bioengineering Research Institute is built up in Nanjing.
SPF grades of KM mouse:Male, weight (22~25) g, is provided by No.1 Hospital of Jilin Univ.'s Experimental Animal Center.
(2) modeling is administered
Research weighs to mouse before carrying out, wherein 12 are only used as blank control group, remaining mouse is hyperuricemia Model modeling group.It weighs between daily morning during experiment.The method for taking yeast extract gavage is tested in this modeling, a large amount of by giving Purine substance makes its uric acid level increase, purine metabolic disturbance, to achieve the effect that temporary raising blood uric acid value in the short time, Then grouping is administered and achievees the effect that cause hepatic tissue different degrees of damage.100 mouse are randomly divided into 8 groups, often Group about 12, respectively blank group, model group, sunflower disk powder group of effective components, polysaccharide group, alkaloid group, general flavone group, sunflower Disk powder group and Allopurinol group.During modeling, in addition to blank group (isometric water gavage), other each groups press 20g/kg yeast extracts daily Carry out gavage, 1 time/d, continuous gavage 7 days, the 7th day and night between it is jejunitas, m seq take blood survey blood UA values, survey blood uric acid value result Show that blood uric acid value is increased with can be administered.Start within 9th day, blank group isometric water gavage daily, according to blood uric acid Value test result randomly selects 12 and is only used as model group, and remainder is divided into 6 groups of administrations, and continue yeast extract gavage, administration group in the morning Afternoon gastric infusion, night is jejunitas after successive administration 7 days, 7 days, and next day, which takes off neck and puts to death and liver is taken to carry out immunohistochemistry, divides Analysis.Gavage is carried out using the high purine diet containing yeast extract, mouse can be caused of short duration hyperuricemia phenomenon occur, and can Continue 15-20 days.
(3) immunohistochemistry immunohistochemical experiment examines drug to hepatic effects
1. after tissue immerses paraffin, tissue is moved into embedding in paraffin cup before embedding.It is placed in triangle from insulating box taking-up On frame, under light alcolhol burner, to keep the dissolved state of paraffin.
2. getting out embedding frame, after being heated on alcolhol burner by the tweezers of embedding and (prevent tweezers sticky wax), left hand holds wax Cup, right handgrip heating tweezers be placed on wax mouth (vertical) when falling wax allow paraffin along tweezers inject embedding frame along.
3. rapid tweezer tissue block is put into embedding frame paraffin, section is towards the positive merging frame bottom of decentralization and gently flattens, to protect There is no bubbles for card.
4. being affixed on label thereon before waiting for paraffin surface solidification, should be noted does not make label obscure with sample, when wax stone is cooled to When there is layer of transparent cere in wax face, immerses in cold water and be allowed to cool rapidly, crystallization is otherwise commonly formed in wax.
5. wax stone removes copper frame after consolidating firmly completely and repaiies cut immediately to ensure that wax thoroughly solidifies.
6. taking out slice, dyed using immunohistochemical kit, micro- sem observation is examined biopsy tissues cell state and clapped According to.
Experimental result is shown in Fig. 1-8, liver cell and hepatic cell cords are clear in lobuli hepatis in blank group (Fig. 1), liver cell cytoplasm It is full.Liver cell and hepatic cell cords are unintelligible in the lobuli hepatis of the part model group (Fig. 2), the visible dotted necrosis region of liver cell atrophy and Liver cell acidophile degeneration.Alkaloid group, general flavone group and the accidental spotty necrosis stove of polysaccharide group (Fig. 4-6) and acidophile degeneration.Sunflower Liver cell and hepatic cell cords are clear in disk powder group (Fig. 7) and sunflower disk powder group of effective components (Fig. 3) lobuli hepatis, and liver cell cytoplasm is full It is full, accidental spotty necrosis stove.There is good repair to liver cell.Liver cell in the lobuli hepatis of the part Allopurinol group (Fig. 8) And hepatic cell cords is unintelligible, liver cell atrophy is common to be dispersed in around cell infiltration stove, spotty necrosis stove and accidental central vein The light dye of liver cell cavitation, Allopurinol group to liver cell have compared with major injury.Microstructure prompt sunflower disk powder and its effectively at Dividing has impaired liver cell certain anti-inflammatory and repair, and what wherein effect was best is also sunflower disk powder active ingredient Group.
Sunflower disk powder of the present invention and its active ingredient are preparing the application in repairing wounded hepatocytes drug, sunflower disk powder and Its active ingredient can play anti-inflammatory, detumescence, analgesic and reduction blood uric acid, dissolve the effect of Monosodium urate, while to liver damage Cell has good repairing effect, to high lithemia disease treating both manifestation and root cause of disease, from basic treatment high lithemia disease.
The embodiments of the present invention are for illustration only, can not represent the quality of embodiment.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all the present invention spirit and Within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.

Claims (3)

1. sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug.
2. application as described in claim 1, which is characterized in that the wounded hepatocytes are caused by hyperuricemia.
3. application as described in claim 1, which is characterized in that the sunflower disk powder active ingredient is prepared by the following method It arrives:
1) sunflower disk is crushed to 100-200 mesh, the deionized water of 6-10 times of quality is added, is sufficiently stirred, with sodium hydroxide tune Section pH value is 8-9, is stirred 120-240 minutes at 45-50 DEG C, obtains sunflower disk protein isolate liquid;
2) sunflower disk protein isolate liquid is heated to 85-90 DEG C, stirs 1-3h, be cooled to 45-55 DEG C, addition accounts for sunflower disk separation The compound protease of protein liquid total volume 0.1%-0.3% digests 3-5h, and when reaction terminating, 85-90 DEG C is warming up in 5 minutes, It is kept for 5-10 minutes, so that compound protease is lost activity, gained enzymolysis liquid is spare;
3) enzymolysis liquid of step 2) is centrifuged 8-12 minutes under 5000 revs/min of rotating speed, collects supernatant, obtains sunflower protein The thick liquid of small-molecular peptides;
4) by the thick liquid of sunflower protein small-molecular peptides of step 3) through shut off molecular weight be 5000-20000 hollow-fibre membrane ultrafiltration, High molecular weight protein is removed, sunflower disk small-molecular peptides liquid is obtained;
5) the sunflower disk small-molecular peptides liquid of step 4) is concentrated, makes its volume concentration to original a quarter to half, Spray drying to get.
CN201810192046.6A 2018-03-08 2018-03-08 Sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug Pending CN108354956A (en)

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CN201810192046.6A CN108354956A (en) 2018-03-08 2018-03-08 Sunflower disk powder and its active ingredient are preparing the application in repairing wounded hepatocytes drug

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109045224A (en) * 2018-10-29 2018-12-21 吉林大学 A kind of sunflower disk semen coicis compound and the purposes in treatment hepatic injury
CN109260266A (en) * 2018-12-11 2019-01-25 吉林大学 The purposes of sunflower disk Gorgon fruit and pueraria lobata compound in anti-trioxypurine reparation lesions of liver and kidney
CN109288884A (en) * 2018-12-17 2019-02-01 广州诗美化妆品有限公司 A kind of preparation method and application of the Helianthi extract with repair of releiving
CN109517748A (en) * 2018-11-30 2019-03-26 吉林大学 Black food small-molecular peptides are repairing the medical application in hepatic injury
CN110478470A (en) * 2019-08-28 2019-11-22 长春中医药大学 A kind of anti-gout composition of the small-molecular peptides containing sunflower disk and preparation method thereof
CN112716990A (en) * 2020-12-25 2021-04-30 吉林大学 Sunflower disc extract, medicine containing sunflower disc extract, preparation method and application
CN116808172A (en) * 2023-01-07 2023-09-29 王晓娟 Sunflower disc liposome and application thereof in preparation of products for reducing uric acid and dissolving tophus

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CN105753935A (en) * 2016-05-06 2016-07-13 洮南市金塔生物科技有限公司 Small-molecule peptides of sunflower head as well as preparation method and medical application of small-molecule peptide
CN105878322A (en) * 2016-05-06 2016-08-24 洮南市金塔生物科技有限公司 Sunflower head extract prepared by crystallization enzymolysis method and preparation method of sunflower head extract

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CN105753935A (en) * 2016-05-06 2016-07-13 洮南市金塔生物科技有限公司 Small-molecule peptides of sunflower head as well as preparation method and medical application of small-molecule peptide
CN105878322A (en) * 2016-05-06 2016-08-24 洮南市金塔生物科技有限公司 Sunflower head extract prepared by crystallization enzymolysis method and preparation method of sunflower head extract

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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109045224A (en) * 2018-10-29 2018-12-21 吉林大学 A kind of sunflower disk semen coicis compound and the purposes in treatment hepatic injury
WO2020087784A1 (en) * 2018-10-29 2020-05-07 吉林大学 Sunflower head-job's tears seed compound composition and use thereof in preparing drugs for treating liver damage
CN113543795A (en) * 2018-10-29 2021-10-22 吉林大学 Sunflower head and coix seed compound composition and application thereof in preparation of medicine for treating liver injury
CN109517748A (en) * 2018-11-30 2019-03-26 吉林大学 Black food small-molecular peptides are repairing the medical application in hepatic injury
CN109260266A (en) * 2018-12-11 2019-01-25 吉林大学 The purposes of sunflower disk Gorgon fruit and pueraria lobata compound in anti-trioxypurine reparation lesions of liver and kidney
CN109288884A (en) * 2018-12-17 2019-02-01 广州诗美化妆品有限公司 A kind of preparation method and application of the Helianthi extract with repair of releiving
CN110478470A (en) * 2019-08-28 2019-11-22 长春中医药大学 A kind of anti-gout composition of the small-molecular peptides containing sunflower disk and preparation method thereof
CN112716990A (en) * 2020-12-25 2021-04-30 吉林大学 Sunflower disc extract, medicine containing sunflower disc extract, preparation method and application
CN116808172A (en) * 2023-01-07 2023-09-29 王晓娟 Sunflower disc liposome and application thereof in preparation of products for reducing uric acid and dissolving tophus
CN116808172B (en) * 2023-01-07 2024-02-13 龚曙初 Sunflower disc peptide, composite liposome and application thereof in preparation of products for reducing uric acid and dissolving tophus

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