CN107961366A - Application of the sunflower small-molecular peptides in anti-trioxypurine dissolves tophus repairing kidney functions - Google Patents
Application of the sunflower small-molecular peptides in anti-trioxypurine dissolves tophus repairing kidney functions Download PDFInfo
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Abstract
The invention discloses a kind of application of sunflower small-molecular peptides in anti-trioxypurine dissolving tophus repairing kidney functions medicine is prepared.Application of the sunflower small-molecular peptides of the present invention in anti-trioxypurine dissolving tophus repairing kidney functions medicine is prepared, sunflower small-molecular peptides can play anti-inflammatory, detumescence, analgesic and reduction blood uric acid, dissolve the effect of Monosodium urate, there is good repairing effect to compromised kidneys cell at the same time, treat both principal and secondary aspect of disease to high lithemia disease, from basic treatment high lithemia disease.
Description
Technical field
The present invention relates to technical field of pharmaceuticals, more particularly to a kind of sunflower small-molecular peptides are preparing anti-trioxypurine dissolving tophus
Application in repairing kidney functions medicine.
Background technology
Gout is a kind of metabolic disease caused by purine metabolic disturbance, and serum Uric Acid Concentration raises, and sodium urate crystals are deposited on
Lower limb Bones and joints, artery, vertebrae gap tissue and cause, show as joint it is red, it is swollen, hot, pain, joint motion is unfavorable, seriously
Person is in joint deformity and dysfunction, and has the characteristics that recurrent exerbation.The presence of gout can also result in coronary heart disease, diabetes,
The deterioration of the diseases such as hypertension.In terms for the treatment of, western modern medicine treatment gout, the method that there is no radical cure, colchicin, nonsteroidal
Though the medicines such as anti-inflammatory agent, allopurinol can mitigate symptom, blood uric acid is reduced, its effect is more single, it is difficult to fundamentally treat
Gout, and long-term use of such Western medicine injured caused by body it is more more than therapeutic effect.
Domestic and international number of the infected is in rising trend at present for hyperuricemia, is increased and kidney with uric acid generation in its pathogenesis
Based on dirty underexcretion, and the disease medication usually causes to damage and aggravates kidney and bears to kidney.
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, its 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, among the people to be also used as medical treatment atrophic gastritis herbal cuisine by the use of it.China is sunflower
Plant big country, the aboundresources of sunflower disk.But in tradition is applied, sunflower disk is as livestock after will typically removing sunflower seed
Forage feed livestock is used as burning material warming ＆ cooking, has not given play to the economic benefit and medical function of its bigger, has caused resource
Waste.
The content of the invention
In order to solve problem of the prior art, an embodiment of the present invention provides a kind of sunflower small-molecular peptides to prepare anti-trioxypurine
Dissolve the application in tophus repairing kidney functions medicine.The technical solution is as follows：
On the one hand, application of a kind of sunflower small-molecular peptides in anti-trioxypurine dissolving tophus repairing kidney functions medicine is prepared.
Further, thick protein ＞ 93% in the sunflower small-molecular peptides, the relative molecular mass distribution of 80% peptide fragment/
U ＜ 1000；Peptide content is 85.6%, moisture 3.5%, content of ashes 7.4%.
Further, the sunflower small-molecular peptides are prepared by the following method to obtain：
1) sunflower disk is crushed to 100-200 mesh, adds the deionized water of 6-10 times of quality, be sufficiently stirred, use 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 cumulative volume 0.1%-0.3%, digests 3-5h, and during reaction terminating, 85- is warming up in 5 minutes
90 DEG C, kept for 5-10 minutes, 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, obtain 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, up to sunflower small-molecular peptides.
The beneficial effect that technical solution provided in an embodiment of the present invention is brought is：Sunflower small-molecular peptides of the present invention are preparing drop
Application in uric acid dissolving tophus repairing kidney functions medicine, sunflower small-molecular peptides can play anti-inflammatory, subside a swelling, analgesic and reduction
Blood uric acid, dissolves the effect of Monosodium urate, while has good repairing effect to compromised kidneys cell, simultaneous to high lithemia disease sample
Control, from basic treatment high lithemia disease.
Brief 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 kidney cell figure in the embodiment of the present invention 2, and wherein A is 400 times of enlarged drawings, and B is 200 times of amplifications
Figure；
Fig. 2 is model group kidney cell figure in the embodiment of the present invention 2, and wherein A is 400 times of amplifications for 400 times of enlarged drawings a, B
Scheme b, C is 200 times of enlarged drawings；
Fig. 3 is sunflower small-molecular peptides group kidney cell figure in the embodiment of the present invention 2, and wherein A is 400 times of enlarged drawings, and B is
200 times of enlarged drawings；
Fig. 4 is other purine group kidney cell figure in the embodiment of the present invention 2, and wherein A is put for 400 times of enlarged drawings a, B for 400 times
Big figure b, C is 200 times of enlarged drawings.
Embodiment
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 small-molecular peptides of embodiment
1) sunflower disk 5kg is crushed to 100 mesh, adds the deionized water of 30kg, be sufficiently stirred, pH is adjusted with sodium hydroxide
It is worth for 8, is stirred 120 minutes at 45 DEG C, obtain 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 cumulative volume 0.1%:0.5:1
Mass ratio mixing), digest 3h, during reaction terminating, 85 DEG C be warming up in 5 minutes, is kept for 5 minutes, loses compound protease
Deactivation, gained enzymolysis liquid are spare.
3) enzymolysis liquid of step 2) is centrifuged 8 minutes under 5000 revs/min of rotating speed, collected supernatant, obtain sunflower egg
The thick liquid of white small-molecular peptides.
4) the thick liquid of sunflower protein small-molecular peptides of step 3) is removed through the hollow-fibre membrane ultrafiltration that molecular weight is 5000 of shutting off
High molecular weight protein is removed, obtains sunflower disk small-molecular peptides liquid.
5) the sunflower disk small-molecular peptides liquid of step 4) is concentrated, arrives its volume concentration to original a quarter, sprayed
It is dry, up to sunflower small-molecular peptides.
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 overall, add the phosphate buffer of 10 times of quality, homogenate, in 4 DEG C of undertissue's slurries, then with
The speed centrifugation of 3000rpm/min, takes supernatant to be heated to 40 DEG C and is rapidly cooled to 5 DEG C, be filtered to remove fat, dialysis removes
Salt, freeze-drying, to obtain the final product.
Thick protein ＞ 93% in the sunflower small-molecular peptides being prepared, relative molecular mass distribution/u ＜ of 80% peptide fragment
1000；Peptide content is 85.6%, moisture 3.5%, content of ashes 7.4%.
2 sunflower small-molecular peptides anti-trioxypurine of embodiment repairs compromised kidney test cell line
(1) material
Small-molecular peptides：Sunflower small-molecular peptides 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 mouse before carrying out, wherein 12 are only used as blank control group, remaining mouse is hyperuricemia
Model modeling group.During the experiment is weighed between daily morning.The method that yeast extract gavage is taken in this modeling experiment, it is a large amount of by giving
Purine substance increases its uric acid level, purine metabolic disturbance, so as to reach the effect of temporary rise blood uric acid value in the short time.
50 mouse are randomly divided into 4 groups, every group about 12, be respectively blank group, model group, sunflower disk small-molecular peptides group, Allopurinol
Group.During modeling, in addition to blank group (isometric water gavage), other each groups daily by 20g/kg yeast extracts 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.Start within 9th day, blank group isometric water daily
Gavage, randomly selects 12 according to blood uric acid value test result and is only used as model group, remainder is divided into 2 groups of administrations, and continue ferment in the morning
Female cream gavage, administration group gastric infusion in afternoon, successive administration after 7 days, 7 days night it is jejunitas, m seq take blood survey blood uric acid value.
Each group mouse is administered by table 1.
Gavage each group administration medicine is administered in 1 modeling of table
| Blank group | Model group | Sunflower small-molecular peptides group | Allopurinol group | |
| 1-8 days | Water | Yeast extract | Yeast extract | Yeast extract |
| 9-15 days | Water | Yeast extract | Yeast extract+small-molecular peptides | Yeast extract+Allopurinol |
(3) before mouse modeling, after modeling and after administration, measure blood uric acid value is horizontal
The serum uric acid level (μm ol/L) of each group mouse is measured respectively before mouse modeling, after modeling and after administration, as a result
It is shown in Table 2.
Before 2 each group modeling of table, after modeling and administration after blood uric acid value (μm ol/L)
| Before modeling | After modeling | After administration | |
| Blank group | 230.99 | 235.59 | 298.32 |
| Model group | 252.89 | 364.38 | 336.38 |
| Sunflower small-molecular peptides group | 285.58 | 497.83 | 149.51 |
| Allopurinol group | 239.38 | 409.68 | 91.75 |
Method by the way that hyperuricemia model intragastric administration on mice is administered, successive administration is after 7 days, finds Allopurinol group, small
Molecular peptide group has certain anti-trioxypurine ability, its P<0.05, there is significant difference.
From table 2 it can be seen that each group mouse serum uric acid level is close before modeling, model group, sunflower small-molecular peptides after modeling
Group and Allopurinol group mouse blood uric acid value significantly rise, and illustrate model success；Sunflower small-molecular peptides group and Allopurinol group after administration
Mouse blood uric acid value declines, or even also lower than before modeling.Blood uric acid value wherein after the administration of sunflower small-molecular peptides group mouse
Compared with have dropped 70% after modeling, the blood uric acid value after the administration of Allopurinol group mouse illustrates that sunflower is small compared with have dropped 78% after modeling
It is close with Allopurinol that molecular peptide reduces blood uric acid ability.This explanation sunflower disk powder small-molecular peptides can significantly reduce hyperuricemia
The blood uric acid value of model mice, can reach the purpose for the treatment of hyperuricemia.
(4) immunohistochemistry immunohistochemical experiment examines medicine to Influence on kidney
After 1. tissue immerses paraffin, tissue is moved into embedding with paraffin cup before embedding.Taken out from insulating box and be placed in triangle
Alcolhol burner is lighted on frame, under it, 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 just being inserted frame bottom towards decentralization and is gently being flattened, to protect
There is no bubble for card.
4. being affixed on label thereon before treating 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, to ensure that wax thoroughly solidifies, repair cut immediately.
6. taking out section, dyed using immunohistochemical kit, micro- sem observation is examined biopsy tissues cell state and clapped
According to.
Experimental result is shown in Fig. 1-4, and blank group (Fig. 1) mesonephric glomerulus is normal, and renal cells is full, marshalling,
Have no tubular ectasia, tube chamber is clear, without inflammation and fibrosis.Model group (Fig. 2) mesonephric glomerulus number has no difference, and kidney is small
Pipe lumen distention, epithelial cell cavitation, atrophy, necrosis, it is seen that cell infiltration, the visible protein cast of tube chamber and cellular cast.
Sunflower small-molecular peptides group (Fig. 3) coloration result no significant difference compared with blank group, illustrate small-molecular peptides administration group to kidney by
The repairing effect for damaging cell is best.The visible cortex renis cell infiltration of Allopurinol group (Fig. 4), obvious renal tubule lumen distention, kidney
Tubular epithelial cell cavitation, although illustrating that Allopurinol group anti-trioxypurine ability is best, it has compared with macrolesion kidney cell.
Immunohistochemical experiment shows, blood uric acid value of the sunflower small-molecular peptides except reducing model mice, at the same can to by
The kidney cell of damage has certain anti-inflammatory and repair, and sunflower small-molecular peptides are better than positive drug allopurinol, and positive
Medicine allopurinol can only reduce blood uric acid value, have certain damage to kidney cell.Illustrate that sunflower small-molecular peptides can not only
Uric acid level is reduced, and kidney can be repaired, treats both principal and secondary aspect of disease to high lithemia disease, can fundamentally treat high lithemia disease.
Application of the sunflower small-molecular peptides of the present invention in anti-trioxypurine dissolving tophus repairing kidney functions medicine is prepared, sunflower are small
Molecular peptide can play anti-inflammatory, detumescence, analgesic and reduction blood uric acid, dissolve the effect of Monosodium urate, while to compromised kidneys cell
There is good repairing effect, treat both principal and secondary aspect of disease to high lithemia disease, from basic treatment high lithemia disease.
The embodiments of the present invention are for illustration only, do 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. application of the sunflower small-molecular peptides in anti-trioxypurine dissolving tophus repairing kidney functions medicine is prepared.
2. application as claimed in claim 1, it is characterised in that thick protein ＞ 93% in the sunflower small-molecular peptides, 80%
The relative molecular mass distribution of peptide fragment/u ＜ 1000；Peptide content is 85.6%, moisture 3.5%, and content of ashes is
7.4%.
3. application as claimed in claim 1, it is characterised in that the sunflower small-molecular peptides are prepared by the following method to obtain：
1) sunflower disk is crushed to 100-200 mesh, adds the deionized water of 6-10 times of quality, be 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 cumulative volume 0.1%-0.3%, digests 3-5h, and during reaction terminating, 85-90 DEG C is warming up in 5 minutes,
Kept for 5-10 minutes, 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, obtain 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, obtains sunflower disk small-molecular peptides liquid；
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, up to sunflower small-molecular peptides.
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Cited By (5)
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| CN110066844A (en) * | 2019-04-20 | 2019-07-30 | 华南农业大学 | A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine |
| 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 |
| CN110521904A (en) * | 2018-05-24 | 2019-12-03 | 石家庄爱肽生物科技有限公司 | Sunflower disc protein peptides solid beverage process recipe and preparation process |
| CN116570640A (en) * | 2023-07-12 | 2023-08-11 | 清枫链食苏打饮品(吉林)有限公司 | Application of sunflower disc alkaloid and derivative in uric acid-reducing and tophus-dissolving product |
| 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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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110521904A (en) * | 2018-05-24 | 2019-12-03 | 石家庄爱肽生物科技有限公司 | Sunflower disc protein peptides solid beverage process recipe and preparation process |
| CN110066844A (en) * | 2019-04-20 | 2019-07-30 | 华南农业大学 | A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine |
| 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 |
| 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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| CN116570640A (en) * | 2023-07-12 | 2023-08-11 | 清枫链食苏打饮品(吉林)有限公司 | Application of sunflower disc alkaloid and derivative in uric acid-reducing and tophus-dissolving product |
| CN116570640B (en) * | 2023-07-12 | 2023-09-05 | 清枫链食苏打饮品(吉林)有限公司 | Application of sunflower disc alkaloid and derivative in uric acid-reducing and tophus-dissolving product |
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