CN106377533A - Trepang phospholipid and application thereof to increment of motor coordination ability of organism - Google Patents
Trepang phospholipid and application thereof to increment of motor coordination ability of organism Download PDFInfo
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- CN106377533A CN106377533A CN201610810275.0A CN201610810275A CN106377533A CN 106377533 A CN106377533 A CN 106377533A CN 201610810275 A CN201610810275 A CN 201610810275A CN 106377533 A CN106377533 A CN 106377533A
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- phospholipid
- stichopus japonicuss
- chloroform
- trepang
- methanol
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- 150000003904 phospholipids Chemical class 0.000 title claims abstract description 63
- 230000004973 motor coordination Effects 0.000 title abstract description 8
- 235000020673 eicosapentaenoic acid Nutrition 0.000 claims abstract description 7
- 229960005135 eicosapentaenoic acid Drugs 0.000 claims abstract description 5
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 claims abstract description 5
- -1 EPA phospholipid Chemical class 0.000 claims abstract 2
- 241000965254 Apostichopus japonicus Species 0.000 claims description 64
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 44
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 40
- 239000000284 extract Substances 0.000 claims description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 18
- 239000000741 silica gel Substances 0.000 claims description 18
- 229910002027 silica gel Inorganic materials 0.000 claims description 18
- 229960001866 silicon dioxide Drugs 0.000 claims description 18
- 238000002360 preparation method Methods 0.000 claims description 15
- 239000003480 eluent Substances 0.000 claims description 9
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000000287 crude extract Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 229910052710 silicon Inorganic materials 0.000 claims description 8
- 239000010703 silicon Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 7
- 238000004587 chromatography analysis Methods 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 4
- 235000013376 functional food Nutrition 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims description 3
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 claims description 3
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- 229910052740 iodine Inorganic materials 0.000 claims description 3
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- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 2
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- 241001465754 Metazoa Species 0.000 abstract description 5
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- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 3
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- 241000208340 Araliaceae Species 0.000 description 2
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- 235000003140 Panax quinquefolius Nutrition 0.000 description 2
- BGNXCDMCOKJUMV-UHFFFAOYSA-N Tert-Butylhydroquinone Chemical compound CC(C)(C)C1=CC(O)=CC=C1O BGNXCDMCOKJUMV-UHFFFAOYSA-N 0.000 description 2
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- 229920001353 Dextrin Polymers 0.000 description 1
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- 241000282894 Sus scrofa domesticus Species 0.000 description 1
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- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
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- 229920002678 cellulose Polymers 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
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- 239000000463 material Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- FFEARJCKVFRZRR-UHFFFAOYSA-N methionine Chemical compound CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 235000006109 methionine Nutrition 0.000 description 1
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Classifications
-
- 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
- A61K31/66—Phosphorus compounds
- A61K31/661—Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion or mevinphos
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/616—Echinodermata, e.g. starfish, sea cucumbers or sea urchins
-
- A—HUMAN NECESSITIES
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses trepang phospholipid and application thereof to the increment of the motor coordination ability of an organism. A main component of the trepang phospholipid is EPA (Eicosapentaenoic Acid) phospholipid with the content which is up to 50 to 65 percent. According to the trepang phospholipid and the application thereof to the increment of the motor coordination ability of the organism, an existing extraction method of the trepang phospholipid is improved; the purity of the trepang phospholipid and the content of the EPA phospholipid in the trepang phospholipid are obviously increased. According to the trepang phospholipid and the application thereof to the increment of the motor coordination ability of the organism, a corresponding animal test is designed; 0.5 percent of the trepang phospholipid is only added into normal feed, so that the latent period of a mouse in a rotarod test is obviously increased, and the abnormal behavior activity of the mouse in an open field test is corrected; by using the trepang phospholipid and the application thereof to the increment of the motor coordination ability of the organism, the dyskinesia of the mouse, which is induced by MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and is similar to parkinsonism, is obviously ameliorated; the trepang phospholipid has an obvious effect of ameliorating the motor coordination ability of the mouse; the trepang phospholipid can be applied to a product for ameliorating the motor coordination ability of the organism.
Description
Technical field
The invention belongs to screening bioactive compoundses technical field, more particularly, to a kind of Stichopus japonicuss phospholipid extract and preparation method,
And its application in prevention, improvement or treatment body movement disorder product.
Background technology
Movement disorder, includes the harmony dyskinesia at present and motion is unbalance, has multiple diseases can cause movement disorder, bag
Include：Parkinson disease, spasm disease, myocardial infarction disease, spinocerebellar degeneration disease etc..Clinically developing new improvement campaign mistake
When the medicine adjusted or treatment meanss, because of parkinson disease sickness rate height, the life and health to the mankind especially old people and life matter
The athletic injury performances such as the amount very big, movement disorder of impact substantially, and become the focus of researcher.With people's dietary structure and life
The change of custom of living and environmental pollution etc., movement disorder phenomenon is more and more universal, and increasingly tends to rejuvenation, it has also become
Serious threat one of human life and health and the principal disease of quality of life.Therefore, movement disorder already become by China or even
An important social problem and problem in science that the whole world pays high attention to, so with regard to researching and developing effective control patients symptomatic, and then
Improve the anti-movement disorder medicine of movement disorder life in patients or food becomes current focus of attention.
Although to a certain extent can be with patients in remission, because individual variation is big for ataxic medicine
Etc. reason, still there are the various problems such as the poor or different degrees of side effect of effect.People tend to selection take have prevention,
Slow down or treat the functional component of movement disorder effect or functional food slows down ataxic purpose to reach, therefore exploitation peace
Effectively there is prevention entirely, slow down or treat movement disorder effect functional component become an urgent demand.
Stichopus japonicuss belong to Echinodermata (Echinodermata) Holothuroidea (Holothurioider)
Handss mesh (Aspidochirota), is considered a kind of famous and precious nourishing food and medical material since ancient times.Phospholipid in Stichopus japonicuss is sea
One of active substance in ginseng, Recent study finds, the phospholipid in Stichopus japonicuss has blood fat reducing, blood pressure lowering, blood sugar lowering, improves immunity
The effects such as power, prevention alzheimer's disease, but up to the present, the body movement coordination ability is improved for it, then there is no correlation
Research report.
Content of the invention
It is an object of the invention to provide a kind of Stichopus japonicuss phospholipid and preparation method thereof, and study it and assist in the motion improving body
The application of tune ability aspect, to make up the deficiencies in the prior art.
For reaching above-mentioned purpose, the invention provides a kind of Stichopus japonicuss phospholipid, its key component is EPA（Eicosapentaenoic acid）
Phospholipid, content is up to 50-65%.
Present invention also offers a kind of preparation method of Stichopus japonicuss phospholipid, comprise the following steps：
1）Stichopus japonicuss make dry powder；
2）Take Stichopus japonicuss dry powder, in chloroform：Extract overnight in methanol mixed solution, after filtration, saline solution extracts, stratification, receives
Collection chloroform layer, then through being concentrated under reduced pressure to give Stichopus japonicuss lipid crude extract；
3）After activated for silica gel dress post, balance silicagel column with chloroform；Again by step 2）The Stichopus japonicuss phospholipid crude extract chlorine obtaining
After imitative dissolving, it is poured slowly in silicagel column, carries out eluting with chloroform, chloroform-methanol mixed liquor, acetone and methanol successively, collect
Meoh eluate, obtains Stichopus japonicuss phospholipid crude extract；
4）Purification：By the meoh eluate obtaining concentrate after, slowly pour in the silicagel column after activation, and with chloroform-methanol-
Water mixed liquid carries out eluting, and eluent carries out real-time thin-layer silicon offset plate chromatography again, silica gel plate top colour developing after centralized collection chromatography
Eluent, the concentrated drying of eluent be Stichopus japonicuss phospholipid.
Further, above-mentioned steps 1）Middle Stichopus japonicuss powder, crosses grit number and is at least 200 mesh.
Further, above-mentioned steps 2）Middle saline solution is preferably 0.88% Klorvess Liquid.
Further, above-mentioned steps 2）The addition of middle chloroform-methanol mixed liquor is 10-20 times of Stichopus japonicuss dry powder quality
（ml/g）.
Further, above-mentioned steps 3）In chloroform-methanol mixed liquor in, the volume ratio of chloroform and methanol is 9:1.
Further, above-mentioned steps 3）Used in silicagel column length and a diameter of 15cm*16cm, step 4）Middle use
The length of silicagel column and a diameter of 60cm*8cm.
Further, above-mentioned steps 4）Middle chloroform：Methanol：The volume ratio of water mixed liquid is 65:25:4.
Further, above-mentioned steps 4）Developing solvent used by middle thin-layer silicon offset plate chromatography is chloroform：Methanol：Water (80:15:1,
v/v).
Further, above-mentioned steps 4）Developer used by thin-layer silicon offset plate chromatography is iodine and Dittmer reagent.
Application in improving body movement coordination ability product for the above-mentioned Stichopus japonicuss phospholipid extract.
Further, described product is the compositionss improving the body movement coordination ability；It include functional food or
Medicine, and correlated product, such as feedstuff etc..
The beneficial technique effect of the present invention：The Stichopus japonicuss phospholipid purity that the present invention is obtained is high, and EPA phospholipid enriches, in the later stage
In chow diet, in zoopery, only add 0.5% Stichopus japonicuss phospholipid, just significantly improve mice latent in turn-club test
Fu Qi, and correct for irregularities activity in open field test for the mice；The present invention significantly improves the similar of MPTP induction
In the dyskinesia of parkinson mice, the sports coordination ability improving mice is had a significant effect, can be applied to change
The product of the kind body movement coordination ability.The preparation method low cost of the Stichopus japonicuss phospholipid extract that the present invention provides, step is simple,
The organic solvent amount using is few, and the organic solvent pollution that environment is produced is little, and the Stichopus japonicuss phospholipid extract of gained is through zoopery
Confirm significantly to improve the sports coordination ability of disease mice, can be used for exploitation and be safely and effectively used for preventing, improve or treating
Ataxic functional food or medicine.
Brief description
Fig. 1 is the result figure to the mouse movement coordination ability for the Stichopus japonicuss phospholipid in the embodiment of the present invention 2.
Fig. 2 is the result figure to mice autonomic movement ability for the Stichopus japonicuss phospholipid in the embodiment of the present invention 2.
Fig. 3 is movement locus figure in spacious field experiment for the mice in the embodiment of the present invention 2.
Specific embodiment
Below using embodiment and combine accompanying drawing to describe the present invention in detail.
Embodiment 1：The preparation of Stichopus japonicuss phospholipid extract
The first step, after commercially available North Atlantic Ocean melon ginseng is pulverized through vacuum lyophilization, clays into power（Cross 200 mesh sieves）.
Second step, takes Stichopus japonicuss dry powder 10kg, adds 100L chloroform/methanol (2:1, v/v), stir, filter, use
0.88% Klorvess Liquid extracting, stratification, collect chloroform layer, extract 3 times, merge concentrated solution, through being concentrated under reduced pressure to give
Stichopus japonicuss phospholipid crude extract；
3rd step, weighs silica gel 200g, adds 5% water, activates 4h at 120 DEG C, and room temperature cools down.Using wet method dress post
（10*15cm）, stand after silica gel sedimentation completely, carry out compression leg with 2 times of column volume chloroforms, make the silica gel in post more closely knit.
The Stichopus japonicuss crude extract sample taking 50g, with lacking after chloroform dissolving as far as possible, shifts this solution with dropper, uniform along chromatography column wall
Add.First suitably wash after 5 column volumes with chloroform, with chloroform-methanol 9:2 column volumes washed by 1 mixed liquor, then with 1, acetone
Column volume, finally uses methanol-eluted fractions, collects meoh eluate.
4th step, purification：Above-mentioned eluent is concentrated to after 500ml, slowly pours the silicon of the 60*8cm having activated into
Glue post（With the 3rd step）In, and with chloroform-methanol-water（65:25:4）Mixed liquor eluting, eluent carries out thin-layer silicon glue plate layer
Analysis (TLC), developing solvent is chloroform/methanol/water (80:15:1, v/v), and with iodine and Dittmer reagent colour development, collecting layer
The eluent of silica gel plate top colour developing after analysis, after confirmation, concentrating under reduced pressure removal residual organic solvent obtains the extraction of Stichopus japonicuss phospholipid
Thing.
Sample identification adopts TLC method, and silica gel plate is placed in 110 DEG C of baking ovens activation 20min, after point sample with chloroform/
Methanol/water (80:15:1, v/v) launch for developing solvent, after launching to finish, dry up, by developer (10% sulphuric acid-second
Alcoholic solution) uniformly it is sprayed on the lamellae being dried, it is placed in 110 DEG C of baking oven 5min, until presenting on TLC plate
Purple dot.
Prepared Stichopus japonicuss phospholipid yield is 50%；Purity is 95%；There is provided compared to prior art in terms of yield and purity
Method all significantly improves, and illustrates that the preparation method that the present invention provides is significantly improved.
Embodiment 2：Stichopus japonicuss phospholipid extract is tested to the improvement of the movement disorder ability with parkinson disease as representative
One. material and method
（1）Animal feeding
Laboratory animal adopts C57/BL6 mice, male, and body weight 22 ± 0.5g, purchased from Beijing dimension tonneau China laboratory animal technology
Company limited.Mice is randomly divided into 3 groups by body weight：Normal group, model control group, the Stichopus japonicuss of embodiment 1 preparation
Phospholipid extract, every group 8, the corresponding feedstuff of feeding respectively.Feedstuff with reference to AIN-93G rodent formula and adds
To improve（Formula is shown in Table 1）, the Stichopus japonicuss phospholipid extract of experimental group interpolation 0.5%, 3 weeks pneumoretroperitoneums of continuous nursing inject 1- first
Base -4- phenyl -1,2,3,6- tetrahydropyridine（MPTP）, once, continuously injection 7 days, cause similar to parkinson for daily injection
Movement disorder symptoms, be embodied in movement disorder, the last time inject after, sacrificed by decapitation mice.Rearing conditions：Animal
Single cage list is only fed, and freely ingests and drinks water, 23 ± 2 DEG C of room temperature, relative humidity 50-60%, and 12:12h light and shade replaces.
The composition of table 1 zoopery feedstuff
| Composition（g/Kg） | Normal group | Model group | Stichopus japonicuss phospholipid |
| Casein | 200 | 200 | 200 |
| Corn starch | 399.486 | 399.486 | 392.5 |
| Sucrose | 100 | 100 | 100 |
| Semen Maydis oil | 50 | 50 | 44.76 |
| Adeps Sus domestica | 150 | 150 | 134.29 |
| Mineral | 35 | 35 | 35 |
| Vitamin | 10 | 10 | 10 |
| Cellulose | 50 | 50 | 50 |
| Tartaric acid cholate | 2.5 | 2.5 | 2.5 |
| DL-methionine | 3 | 3 | 3 |
| TBHQ | 0.014 | 0.014 | 0.014 |
| Stichopus japonicuss lipid-soluble extract | —— | —— | 27.93 |
（2）Behavioral assessment-turn-club test
Mice participates in turn-club test to assess the sports coordination ability of mice.It is real that transfer rod equipment is purchased from Peking University's health science
Test room.This equipment has a live spindle（Diameter 7.3cm）The dividing plate independent with 5（Can 5 mices of disposable test）.?
Before injection MPTP, mice is trained to for three days on end in transfer rod instrument, and rotating speed is 18rpm/min, and each training time is
900s.After mice injects MPTP the last time, carry out formal turn-club test, rotating speed is 40rpm/min, and mice is on transfer rod
Residence time is recorded, and 900s is designated as the time grown most.For every mice, turn-club test is repeated 3 times, and result takes 3 times
Meansigma methodss.What data presented is mice residence time in transfer rod instrument.
（3）Mice autonomic movement capacity experimental-spacious field experiment
Spacious field device：Using computer-implemented monitoring system Morris software, this device is mainly by a spacious case（45*45*
35cm）It is divided into 25 grid（5*5）, case top central authorities are mounted with the video camera being connected with record system, are divided by system
Analysis mice real time imaging is to obtain behavioral data in spacious field for the mice.
Mice is participated in spacious field and tests to assess the autonomic movement ability of mice.Experiment starts front 30min and moves into mice
Test cabinet adapts to, to ensure that during experiment, mice activity tends towards stability.When experiment starts, mice is placed on the centre position of spacious case,
Allow its glitch-free motion 5min in spacious field, mice active procedure wherein monitored by video camera, Morris software records are little
The move distance of Mus.In experimentation, after every mouse experiment terminates, the alcohol wipe with 75% opens the bottom of case, to protect
The cleaning in the spacious bottom portion of card.
（4）Statistical procedures
Experimental data is represented with x ± SEM, carries out Student ' s t test and using SPSS 18.0 software
Tukey ' s test analyzes, with P ＜ 0.05 for there were significant differences.
Two. experimental result
Stichopus japonicuss phospholipid extract is to the mouse movement coordination ability（Turn-club test）Impact as shown in figure 1, representing mice incubation period
Residence time on transfer rod, for Normal group, mice significantly subtracts the incubation period in transfer rod instrument model group
Short, as it is desirable that, after Stichopus japonicuss phospholipid extract is intervened, compared with model group, incubation period on transfer rod for the mice
Significantly extend, this prompting Stichopus japonicuss phospholipid extract can significantly improve the motor coordination energy of the parkinson mice of MPTP induction
Power.
The autonomic movement ability to mice for the Stichopus japonicuss phospholipid extract（Spacious field is tested）Impact as shown in Fig. 2 and normal group
Mice is compared, and the distance that model group mice moves in spacious field significantly shortens, and Stichopus japonicuss phospholipid extract significantly increases
The distance that mice walks in spacious field, this prompting Stichopus japonicuss phospholipid extract can significantly improve the parkinson mice of MPTP induction
Autonomic movement ability and sports coordination ability.
Fig. 3 is movement locus in spacious field experiment for the mice, as illustrated, compared with matched group, model group mice is spacious
Movement locus in significantly decrease, and after Stichopus japonicuss phospholipid is intervened, the movement locus of mice significantly increase, and this carries
Show that Stichopus japonicuss phospholipid can significantly improve the motor capacity of mice.
Using similar parkinson disease as experimental subject, its movement disorder disease mechanism of production is more complicated, including dopamine god
Through the positive minimizing of unit, intracerebral oxidation stress wait, and the cell that in embodiment 2, main mechanism stress may lead to for intracerebral oxidation withers
The body movement causing of dying is lacked of proper care, autonomic activitieses ability reduces, that is, illustrate that Stichopus japonicuss phospholipid can significantly improve the motion of body
The coordination ability.
Embodiment 3：Capsule preparation with Stichopus japonicuss phospholipid as primary raw material
Stichopus japonicuss phosphatidase 1 00mg in embodiment 1, micropowder silica gel 20g, DEXTRIN 0g, starch 530g, mentioned component adds
Blender, after being sufficiently stirred for mixing, fills capsule, every 1g, can be made into capsule 1000.Wherein every capsule contains effectively
Composition Stichopus japonicuss lipid-soluble extract 0.1mg.
The above, be only presently preferred embodiments of the present invention, is not the restriction that the present invention is made with other forms, appoints
What those skilled in the art possibly also with the disclosure above technology contents changed or be modified as equivalent variations etc.
Effect embodiment.But every without departing from technical solution of the present invention content, according to the present invention technical spirit to above example institute
Any simple modification, equivalent variations and the remodeling made, still falls within the protection domain of technical solution of the present invention.
Claims (10)
1. a kind of Stichopus japonicuss phospholipid it is characterised in that in this Stichopus japonicuss phospholipid eicosapentaenoic acid phospholipid content be 50-65%.
2. application in improving body movement coordination ability product for the Stichopus japonicuss phospholipid described in claim 1.
3. the preparation method of the Stichopus japonicuss phospholipid described in claim 1 is it is characterised in that comprise the following steps：
1）Stichopus japonicuss make dry powder；
2）Take Stichopus japonicuss dry powder, in chloroform：Extract overnight in methanol mixed solution, after filtration, saline solution extracts, stratification, receives
Collection chloroform layer, then through being concentrated under reduced pressure to give Stichopus japonicuss lipid crude extract；
3）After activated for silica gel dress post, balance silicagel column with chloroform；Again by step 2）The Stichopus japonicuss phospholipid crude extract chlorine obtaining
After imitative dissolving, it is poured slowly in silicagel column, carries out eluting with chloroform, chloroform-methanol mixed liquor, acetone and methanol successively, collect
Meoh eluate, obtains Stichopus japonicuss phospholipid crude extract；
4）Purification：By the meoh eluate obtaining concentrate after, slowly pour in the silicagel column after activation, and with chloroform-methanol-
Water mixed liquid carries out eluting, and eluent carries out real-time thin-layer silicon offset plate chromatography again, silica gel plate top colour developing after centralized collection chromatography
Eluent, the concentrated drying of eluent be Stichopus japonicuss phospholipid.
4. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 2）Middle saline solution is
0.88% Klorvess Liquid.
5. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 2）Middle chloroform-methanol mixes
The solid-liquid ratio closing liquid with Stichopus japonicuss dry powder is 1：10-20.
6. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 3）In chloroform-methanol
In mixed liquor, the volume ratio of chloroform and methanol is 9:1.
7. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 4）Middle chloroform：Methanol：Water
The volume ratio of mixed liquor is 65:25:4.
8. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 4）Middle thin-layer silicon glue plate layer
Developing solvent used by analysis is chloroform：Methanol：Water (80:15:1, v/v).
9. the preparation method of Stichopus japonicuss phospholipid as claimed in claim 3 is it is characterised in that above-mentioned steps 4）Thin-layer silicon offset plate chromatographs
Developer used is iodine and Dittmer reagent.
10. Stichopus japonicuss phospholipid as claimed in claim 2 application in improving body movement coordination ability product, its feature exists
In this product includes functional food or medicine.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106880049A (en) * | 2017-03-07 | 2017-06-23 | 中国海洋大学 | Application of the n 3PUFA phosphatide in chronic fatigue syndrome product is improved |
| CN109662802A (en) * | 2018-12-26 | 2019-04-23 | 南京大学 | The method and application of motor coordination performance during a kind of detection pain measurement |
| CN110327358A (en) * | 2019-08-21 | 2019-10-15 | 广东海洋大学深圳研究院 | Sea cucumber phosphatide inhibits the application in neuroinflamation drug in preparation |
| CN110478374A (en) * | 2019-08-31 | 2019-11-22 | 广东海洋大学深圳研究院 | Sea cucumber phosphatide is preparing the application in neuronal cell protection drug |
| CN110538185A (en) * | 2019-10-09 | 2019-12-06 | 广东海洋大学深圳研究院 | Application of sea cucumber phospholipid in preparation of medicine for preventing and treating inflammation |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060068004A1 (en) * | 2004-09-28 | 2006-03-30 | Aum Nutrapharm Inc. | Buccal delivery of sea cucumber tablets |
| CN1933828A (en) * | 2004-01-19 | 2007-03-21 | 马泰克生物科学公司 | Reelin deficiency or dysfunction and methods related thereto |
| CN103263406A (en) * | 2005-12-21 | 2013-08-28 | 布鲁迪科技有限公司 | Use of DHA, epa or DHA-derived EPA for treating a pathology associated with cellular oxidative damage |
| CN103417579A (en) * | 2013-09-05 | 2013-12-04 | 中国海洋大学 | Sea cucumber extract and application thereof |
| WO2015084158A1 (en) * | 2013-12-06 | 2015-06-11 | N.V. Nutricia | A pyrimidine derivative and a fatty acid source for use in the treatment of constipation |
-
2016
- 2016-09-08 CN CN201610810275.0A patent/CN106377533A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1933828A (en) * | 2004-01-19 | 2007-03-21 | 马泰克生物科学公司 | Reelin deficiency or dysfunction and methods related thereto |
| US20060068004A1 (en) * | 2004-09-28 | 2006-03-30 | Aum Nutrapharm Inc. | Buccal delivery of sea cucumber tablets |
| CN103263406A (en) * | 2005-12-21 | 2013-08-28 | 布鲁迪科技有限公司 | Use of DHA, epa or DHA-derived EPA for treating a pathology associated with cellular oxidative damage |
| CN103417579A (en) * | 2013-09-05 | 2013-12-04 | 中国海洋大学 | Sea cucumber extract and application thereof |
| WO2015084158A1 (en) * | 2013-12-06 | 2015-06-11 | N.V. Nutricia | A pyrimidine derivative and a fatty acid source for use in the treatment of constipation |
Non-Patent Citations (1)
| Title |
|---|
| 刘志国等: "多不饱和脂肪酸对大脑功能影响研究进展", 《食品科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106880049A (en) * | 2017-03-07 | 2017-06-23 | 中国海洋大学 | Application of the n 3PUFA phosphatide in chronic fatigue syndrome product is improved |
| CN109662802A (en) * | 2018-12-26 | 2019-04-23 | 南京大学 | The method and application of motor coordination performance during a kind of detection pain measurement |
| CN110327358A (en) * | 2019-08-21 | 2019-10-15 | 广东海洋大学深圳研究院 | Sea cucumber phosphatide inhibits the application in neuroinflamation drug in preparation |
| CN110327358B (en) * | 2019-08-21 | 2022-11-22 | 广东海洋大学深圳研究院 | Application of sea cucumber phospholipid in preparation of medicine for inhibiting neuroinflammation |
| CN110478374A (en) * | 2019-08-31 | 2019-11-22 | 广东海洋大学深圳研究院 | Sea cucumber phosphatide is preparing the application in neuronal cell protection drug |
| CN110538185A (en) * | 2019-10-09 | 2019-12-06 | 广东海洋大学深圳研究院 | Application of sea cucumber phospholipid in preparation of medicine for preventing and treating inflammation |
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Application publication date: 20170208 |