CN106491488A - A kind of effect gel and preparation method thereof - Google Patents
A kind of effect gel and preparation method thereof Download PDFInfo
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- CN106491488A CN106491488A CN201611014389.0A CN201611014389A CN106491488A CN 106491488 A CN106491488 A CN 106491488A CN 201611014389 A CN201611014389 A CN 201611014389A CN 106491488 A CN106491488 A CN 106491488A
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- aqueous extracts
- pig placenta
- olive oil
- homogeneous
- protein
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Classifications
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- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
- A61K8/982—Reproductive organs; Embryos, Eggs
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- 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
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
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Abstract
A kind of effect gel and preparation method thereof, belongs to biomedicine technical field.By oviductus ranae with homogeneous after the immersion of pig placenta Aqueous extracts, after then sequentially adding olive oil and pig placenta Aqueous extracts homogeneous, olive oil emulsion is obtained；By water soluble chitosan with homogeneous after the immersion of pig placenta Aqueous extracts, olive oil emulsion is then added, then through homogeneous, obtain effect gel containing pig placenta Aqueous extracts, olive oil, water soluble chitosan and oviductus ranae.Product can be also oral with external application, with the ability for removing free radical, can mitigate injury of the free radical to biological tissue, with the function of delaying body aging.
Description
Technical field
The present invention relates to a kind of effect gel and preparation method thereof, belongs to biomedicine technical field.
Background technology
Placenta is the transition sexual organ that mammal grew up in the gestational period, and the material being mainly used between parent and fetus is handed over
Change.《Compendium of Materia Medica》Record：Human plactnta is warm in nature, sweet-salty, thoughts of returning home lung kidney channel.Human plactnta has anti-aging, enhance immunity, benefit
The effects such as blood beauty, it is referred to as the temperature compensation Chinese medicine of preciousness, but is difficult to obtain as medicine source, and there is ethics and cross-infection etc. asking
Topic；And the pig placenta equally from mammal is easy to get safely, it may have enrich blood well, improve learning and memory, resisting stress
The effects such as ability and enhance immunity, have to Human plactnta through the constituent analysis such as protein, trace element, amino acid confirmation similar
Effect material base.
Water soluble chitosan has the characteristics such as thickening, emulsification, moisturizing, film forming and gel, compared with shitosan, can be extensive
Dissolve in pH value water, nontoxic, tasteless, degradable, good biocompatibility, be suitable in all kinds of cream, breast, frost formula, in daily use chemicals, doctor
The numerous areas such as medicine are extensively applied.
In pig placenta, the separation identification of proteinaceous components has no report both at home and abroad at present；Former as effect with pig placenta Aqueous extracts
Material, water soluble chitosan are that the gel preparation product of matrix research and development even more be nobody shows any interest in.
Content of the invention
Present invention aim at proposing a kind of effect gel that makes using pig placenta Aqueous extracts for raw material.
The present invention is mainly made up of pig placenta Aqueous extracts, olive oil, water soluble chitosan and oviductus ranae；The water soluble shells
Glycan is any one in carboxymethyl chitosan, amber bat acyl shitosan, cross-linked chitosan, hydroxypropyl chitosan or thio chitosan
Kind.
It can also be Orally taken product that product of the present invention can be external application, be experimentally verified that effect gel has and remove certainly
By the ability of base, injury of the free radical to biological tissue can be mitigated, thus with delaying the function of body aging.Such as effect is coagulated
After glue therapeutic intervention mouse aging, internal SOD contents increase, and hinder infringement of the oxygen radical to cell, and repair in time
Damaging cells；The reduction of cellular damage degree has also been reacted in the minimizing of MDA contents in vivo.
The gel oil-containing non-greasy, easily applies exhibition, has comfort with external application, can further research and develop wrinkle, nti-freckle, moisturizing beautiful
White effect cosmetic；Orally in good taste, memory can be strengthened, enhance immunity can further research and develop preventing and treating Parkinson, A Erhai
Neurodegenerative disease functional food or the health medicines such as silent thatch disease；So effect gel of the present invention is eaten in household chemicals field, medicine
The aspect such as dual-purpose has a good application prospect.
The present invention also proposes the preparation method of above product, comprises the following steps：
1）Fresh pig placenta is removed to clean after crimson blood, umbilical cord, manadesma and is drained, crushed after freezing under the conditions of -20 DEG C, add water even
Slurry, is then centrifuged for, and by supernatant again with 0.45 μm of membrane filtration, the filter liquor of acquirement is pig placenta Aqueous extracts；
2）By oviductus ranae with homogeneous after the immersion of pig placenta Aqueous extracts, olive oil homogeneous is added, olive oil emulsifiable paste is obtained, is added again
After pig placenta Aqueous extracts homogeneous, olive oil emulsion is obtained；
3）By water soluble chitosan with homogeneous after the immersion of pig placenta Aqueous extracts, olive oil emulsion is then added, then through homogeneous, is taken
Obtain effect gel.
Process above normal-temperature operation, advantages of simple；Any organic solvent, environmental protection and energy saving are not used；Olive oil is dispersed in egg
Make the product stability that makes good in white matter and chitosan gel rubber space network, breast analysis, flocculation will not occur and settle.
Water soluble protein containing 0.1～10% mass fraction in the pig placenta Aqueous extracts.Water soluble protein is pig
Main effect component in placenta Aqueous extracts, in pig placenta Aqueous extracts, water-soluble protein content must reach 0.1～10% scope
Just there is meaning of the present invention.Pig placenta Aqueous extracts are to extract to obtain from normally mature fresh pig placenta, because pig placenta comes
Source（Porcine specy and the difference of feed）And water-soluble protein content difference caused by the difference of pig farrowing pig time, can be according to reality
Border needs water-soluble in pig placenta Aqueous extracts using adding pure water to dilute or add the method for the dense product of pig placenta Aqueous extracts freeze-dried powder to make
Protein content reaches a certain determination content in 0.1～10% scope, can be made as preventing and treating with the low side as content for health care
Can be with the high side with content.
Olive oil has lipid-loweringing, cosmetology multiple efficacies concurrently as gel NMF；Oviductus ranae protein content is higher,
There is good emulsification after being soaked to olive oil, also there are the health-care efficacies such as increase immunity；Water soluble chitosan is gel base
Matter, has stabilization to olive oil breast, has the functions such as moisturizing, activating cell concurrently.
The water soluble protein at least includes GRP78, heat shock protein 8, protein disulfide isomery
Enzyme A3 precursor proteins, prolyl 4 hydroxylase beta polypeptides, β actin cytoskeleton fragments, aldose reductase and β hemoglobins
Subunit.Including at least the protein component for having more than ten kinds of above seven kinds of water soluble proteins etc. in pig placenta Aqueous extracts, by
Other protein contents beyond above-mentioned seven kinds of albumen are relatively low, and can not differentiate its concrete species with technology today means.
In addition, when preparing, the mixing quality ratio of pig placenta Aqueous extracts, olive oil, water soluble chitosan and oviductus ranae is 34:
4∶1∶1.Pig placenta Aqueous extracts are water phases, and olive oil is oil phase, and the oviductus ranae being soaked is emulsifying agent, and shitosan is gel-type vehicle,
Whole system is really emulsion gel rubber system, is particularly advantageous for the absorption of functional component, the above amount ratio of experimentally determined each composition
The compound system physical stability can be kept, i.e., the phenomenons such as layering do not occur.
In addition, step 2 of the present invention）Middle soak time is 2～12h.In oviductus ranae, protein content is higher, protein
Belong to macromolecular compound, the speed being diffused in dissolving medium is more slowly compared with micromolecular compound.The present invention has been initially charged
Limit quantity of solvent makes oviductus ranae fully swelling, then emulsifies and dilutes.Swelling be crimp helically or Coiling-type macromolecule molten
The process that unfolds in agent completely, swelling need more than the 2h times completely, swelling not exclusively can not then dissolve and be dispersed in solvent；Molten
Swollen process by the way of heating or stirring can not promote to dissolve, and otherwise run counter to desire.
The step 3）Middle soak time is 2～12h.Shitosan belongs to macromolecular compound, is diffused in dissolving medium
Speed more slowly compared with micromolecular compound.The present invention is directly added into the fully swelling dissolving of sufficient solvent, beneficial to olive oil
Emulsifiable paste is sufficiently mixed.Swelling be crimp helically or Coiling-type the process unfolded completely in a solvent of macromolecule, swelling completely
Need more than the 2h times, swelling not exclusively can not then dissolve and be dispersed in solvent；Swelling process can not adopt heating or stir
Mode promote to dissolve, otherwise run counter to desire.
The rotating speed of the mixed system during homogeneous is 1300 r min-1, each action time be 1～5min, this be through
Determination is investigated in experiment, can reach preferable homogenizing effect.The air formation bubble that the too high vortex of rotating speed is brought into is difficult to catch up with and removes,
Mixing time is long to have breakdown of emulsion dangerous.
Description of the drawings
Fig. 1 is pig placenta Aqueous extracts sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE）Protein Separation
As a result.
Fig. 2 is the fingerprint mass spectrogram that the 1st electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 3 is the fingerprint mass spectrogram that the 2nd electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 4 is the fingerprint mass spectrogram that the 3rd electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 5 is the fingerprint mass spectrogram that the 4th electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 6 is the fingerprint mass spectrogram that the 5th electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 7 is the fingerprint mass spectrogram that the 6th electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 8 is the fingerprint mass spectrogram that the 7th electrophoretic band of pig placenta Aqueous extracts SDS-PAGE marks corresponds to albumen.
Fig. 9 is basis of microscopic observation normal group skin epidermis pathological change photo.
Figure 10 is basis of microscopic observation model group skin epidermis pathological change photo.
Figure 11 be basis of microscopic observation positive controls to mouse aging skin epidermis pathological change photo.
Figure 12 be basis of microscopic observation effect gel high dose group to mouse aging skin epidermis pathological change photo.
Figure 13 be basis of microscopic observation effect gel middle dose group to mouse aging skin epidermis pathological change photo.
Figure 14 be basis of microscopic observation effect gel low dose group to mouse aging skin epidermis pathological change photo.
Specific embodiment
First, the preparation of effect gel
1st, two, normally mature fresh pig placenta is taken, crimson blood, umbilical cord, manadesma is removed, is cleaned and is drained, weigh, gross weight is
166g；Crush after -20 DEG C of quick-frozen 48 h；Plus 250mL pure water, 12000 r min in tissue mashing machine-1, homogenate 30
min；8000 r min of homogenate-1, 4 DEG C, it is centrifuged 30 min；Supernatant through 0.45 μm of filter membrane leach filtrate 1..To centrifugation
Settlement section adds 250mL pure water, 12000 r min in tissue mashing machine-1, it is homogenized 30 min；8000 r of homogenate
min-1, 4 DEG C, it is centrifuged 30 min；Supernatant through 0.45 μm of filter membrane leach filtrate 2..Add 250mL pure again to centrifugal sedimentation part
Water purification, 12000 r min in tissue mashing machine-1, it is homogenized 30 min；8000 r min of homogenate-1, 4 DEG C, centrifugation 30
min；Supernatant through 0.45 μm of filter membrane leach filtrate 3..3. 2. 1. merging filtrate obtain pig placenta Aqueous extracts, is diluted with pure water
To 850g, the pig placenta Aqueous extracts of dilution are obtained.
2nd, 570g is taken from the pig placenta Aqueous extracts of dilution, then divide 2～12 h of 25g oviductus ranaes for taking that 250g immersions are cleaned
Afterwards, through 1300 r min-1Homogeneous 1min, is added dropwise 100g olive oil and continues 1～5min of homogeneous and obtain olive oil emulsifiable paste.
While remaining pig placenta Aqueous extracts are added dropwise in the olive oil emulsifiable paste that makes while continuing 1～5min of homogeneous obtains olive oil emulsion.
3rd, 280g immersion 250g water miscible carboxymethyl chitosan 2～12 h is taken from the pig placenta Aqueous extracts of 850g dilutions
Afterwards, through 1300 r min-1Homogeneous 1min, instills and continues 1～5min of homogeneous and obtain effect gel in olive oil breast
1000g, is divided in standby in plastic flexible pipe.
Above carboxymethyl chitosan can adopt amber to clap acyl shitosan, cross-linked chitosan, hydroxypropyl chitosan or sulfydryl shell
Any one replacement in glycan.
2nd, in the pig placenta Aqueous extracts of above 850g dilutions, water-soluble protein content is determined
Liquid is prepared with protein standard and is added to 30 mg BSA（Bovine serum albumin(BSA)）In, concentration is configured to after fully dissolving for 0.5
mg·ml-1Protein standard liquid.
Press 50 volumes BCA（Determination of protein concentration kit）Reagent A adds 1 volume BCA reagent B（50 1）Prepare appropriate BCA
Working solution is mixed, and is kept in dark place.
Standard items are added in 96 orifice plates by 0,1,2,4,8,12,16,20 μ l, plus 20 μ L are supplied in standard items dilution.
Plus 10 μ L samples in the sample well of 96 orifice plates, plus standard items are diluted to 20 μ L.
Each hole adds 200 μ L BCA working solutions, 37 DEG C of 20 min of placement.
Determine A562, measurement result see the table below：
With standard protein concentration as abscissa, with the absorbance values corresponding to each standard protein concentration as ordinate, paint
Calibration curve processed, carries out linear regression analysis.Equation of linear regression is A=0.3871C+0.0934, R2=0.9953 .
The pig placenta Aqueous extracts for taking above 850g dilution are appropriate, appropriate dilute after under 562nm wavelength mensuration absorbance A
Value, in the pig placenta Aqueous extracts of the 850g dilutions that thus absorbance and dilution factor are calculated, water-soluble protein content is 0.5084%.
3rd, in pig placenta Aqueous extracts, the SDS-PAGE of water-solubility protein is separated and molecular weight determination
Precision weighs the pig placenta Aqueous extracts of preparation, and deionized water is made into 5,10,15 mg ml respectively-1Solution, respectively
Take 20 μ L and add 5 μ L albumen buffer solutions（Contain 2%SDS, 0.1% bromophenol blue, the aqueous solution of 10% glycerine）, mix, 5 min of micro-boiling,
4 DEG C of preservations；Tris-base 15.1g, 94 g of glycine is weighed, adds 10%SDS 50mL, miscible rear plus deionized water to be settled to
1 L, determines pH and is electrophoretic buffer for 8.3；0.25g coomassie brilliant blue R250s are weighed, it is 45 to add methanol-water glacial acetic acid
45 10 mixed liquor to 100 mL is coomassie brilliant blue R250 dyeing liquor.
The preparation of running gel：Assembling electrophoresis glass plate, is clamped with clip, sealed bottom and both sides, after insertion comb away from
Carry out mark where 1 cm of comb bottom, order is prepared point successively from top to bottom according to 1 separation gel proportioning of table after leak detection 3 times
From glue, liquid-transfering gun comes and goes suction and mixes, is slowly added into mark along glass inner side, then moves back and forth light plus deionization along glass
Water, standing 30 min solidifies separation gel, and water is poured out；Order prepares concentration successively from top to bottom to press the concentration glue of table 1 proportioning again
Glue, is poured in glass plate crack immediately after mixing, is flushed with glass plate, be inserted into comb, is stood to gelling admittedly, is used preservative film
Parcel, 4 DEG C standby.PAGE gel formula see the table below：
Sample-adding：PAGE gel is moved into electrophoresis tank, electrophoretic buffer is filled it up with, comb, the smooth slotted eye of form slection state is slowly extracted
Sample-adding, sample applied sample amount are 20 μ l, and Marker is 5 μ l, carries out mark.
Electrophoresis：Connect electrode, enriching stage voltage：110 V, after bromophenol blue moves on to separation gel, voltage switchs to 140 V, when
Bromophenol blue move to apart from 0.5 cm of blob of viscose bottom when, deenergization, electrophoresis terminate.
Dyeing and decolouring：Blob of viscose in glass plate is taken out, coomassie brilliant blue R250 dyeing liquor shaking table is gone to after dyeing 30 min
Decolourize to background transparent in the aqueous solution containing 40% methyl alcohol and 10% glacial acetic acid, through fully automatic digital gel image analysis system
Tanon 2500 is scanned, and sees Fig. 1.
Fig. 1 shows that the electrophoretogram of pig placenta Aqueous extracts protein SDS-PAGE, wherein A, B, C are respectively sample protein solution
Concentration is the electrophoretic separation result of 5 mg/mL, 10 mg/mL and 15 mg/mL.
There is more than ten protein electrophoresis band as seen from Figure 1, choose separating degree preferably and 1 to 7 higher electrophoretic band of content
It is analyzed identification.
When molecular weight of albumen is separated in 15～200 KD scopes, the logarithm of its mobility and molecular weight is linearly closed
System：
In above formula, K, b are constant.
6 groups of mobilities that Maker protein electrophoresises are produced are mapped to obtain to its molecular weight calibration curve；According to calibration curve and
Sample protein mobility can try to achieve corresponding molecular weight.Most clearly 1～7 band in SDS-PAGE electrophoresis is chosen in this experiment（Corresponding
Protein content is higher）Be followed successively by 77.27 through 2500 fully automatic digital gel image analysis system validation its molecular weight of Tanon,
68.21st, 57.85,53.57,46.44,37.16,15 KD, specifically see the table below.
*Note：This experiment albumen Maker is only containing six different molecular weight albumen for having purified.
4th, in pig placenta extracting liquid water-solubility protein Mass Spectrometric Identification
1st, film dosim：
1. decolourize：1～7 band of SDS-PAGE electrophoresis in example 4 is oriented respectively and cuts 1 mm3It is placed in the EP pipes of 1.5 mL,
Add 200 μ L destainers（ACN（Acetonitrile）50 mM NH4HCO3Solution=1 1）, 60 DEG C shake up 15 min, are suctioned out with pipette tips and are washed
Liquid, repeats to decolourize 1 time, then deionized water is rinsed repeatedly and taken off to the greatest extent to blueness.
2. dehydrate：The ACN that 200 μ L are separately added into 1～7 band EP pipes after decolouring shakes up, and abandons after 10 min
Clear liquid, repeats the step to gel into milky, absorbs ACN, and room temperature air-dries micelle.
3. digest：15 μ L enzyme liquids are separately added into dried 1～7 band EP pipes（0.01 μ g/ μ L of trypsase）, 4
DEG C place 30 min, then plus 25 mM NH4HCO3To submergence blob of viscose, 37 DEG C are placed 15 h to 20 μ l of solution.
4. extract：20 μ l extracts are separately added into 1～7 band EP pipes after enzymolysis（Contain 50%ACN, 0.1%TFA（Three
Fluoroacetic acid）The aqueous solution）, 60 DEG C shake up 10 min, ultrasonic 1 min, suction out extract in new EP pipes, repeat extraction step,
Merge extract twice, 8 h of freeze-drying.
2nd, Mass Spectrometric Identification：
Add 3 μ L extracts to dissolve again in the albumen after above-mentioned being lyophilized, take 0.7 μ L, with the alpha-cyano -4- hydroxyls that is prepared with extract
Base cinnamic acid（CHCA）Matrix saturated solution equal-volume is sufficiently mixed, and puts on stainless steel MALDI target plates, is done under room temperature naturally
Dry, mass spectral analysis, Nd of the lasing light emitter for 200 Hz (355 nm wavelength) is carried out with 5800 time-of-flight mass spectrometry instrument:YAG swashs
Light device, accelerating potential are 20 kV, and MS is using reflection positive ion mode, mass range 800-4000 m/z, each collection of illustrative plates collection
4000 times.Each sample at most selects 20 MS peaks, minimum signal to noise ratio 20, resolution ratio 200.MS/MS adopts positive ion mode, touches
2 KV of energy is hit, CID is closed, and each collection of illustrative plates is accumulated 3000 times.Charge-mass ratio（m/z）In the range of 800～4000, pig placenta is gathered
1～7 electrophoretic band protein fingerprint spectrum of protein（PMF）, each collection of illustrative plates signal is stronger, and appearance situation preferably, distinguish by 1～7 band
Assume 132,849,189,790,848,886,496 signal peaks, see Fig. 2 to Fig. 8.
3rd, protein database search：
Carry out Protein hits with MASCOT softwares, database NCBInr, retrieval kind is mammal, maximum allowable leakage enzyme site
For 1, enzyme is trypsase, and quality error scope is arranged：MS100 ppm, MS/MS0.6 Da, proteic charge number are 1+, fix and repair
Decorations：Carbamidomethyl (C), variable modification：Oxidation (M), Oxidation (HW), Deamidated (NQ), retrieval
Protein Information.
After retrieval NCBInr databases, each band has a lot of coupling albumen results, i.e., look in Protein Data Bank
To in peptide fragment and the storehouse of pig placental protein matter endonuclease bamhi, the molecular weight information of each peptide fragment of same endonuclease bamhi is the most
The protein that matches somebody with somebody, between the albumen of multiple matching results, similarity is also very high, selects homologous with pig placenta wild boar in storehouse（Sus
scrofa）Coupling Protein Information, carry out Protein hits comparison with Mascot softwares.Albumen score is significant result more than 60
（P<0.05）, obtain mating the title of albumen, numbering, molecular weight, albumen and obtain grading information, see the table below.
Gi according to coupling albumen is numbered and is searched for Protein Information in ncbi database, is mated with 1 ~ 7 band of pig placental protein
Peptide fragment and its amino acid sequence coverage, sequential covering rate account for 16%, 16%, 20%, 9%, 34%, 33%, 59% respectively.Therefore, grape
Glucosal related protein 78, heat shock protein 8, protein disulfide bond isomerase A 3 precursor, prolyl 4 hydroxylase beta polypeptides, β cell bones
Frame actin, aldose reductase, β hemoglobin subunits are mated with band 1～7 respectively.
1～7 band of electrophoresis belongs to the albumen related to cell metabolism, immunity, cellular activity etc.：Glucose adjusts egg
White 78 tool immunocompetence, cell proliferation, anti-apoptotic play an important role；Heat shock protein 8 is heat-shock protein family a member, can
Cellular anti-oxidant anti-inflammatory is improved, after stress reaction, recovers cell home；Protein disulfide bond isomerase A 3 precursor protein energy
Regulation protein synthesis in endoplasmic reticulum, is catalyzed disulfide bond, forms native protein conformation；Prolyl 4 hydroxylase beta polypeptides are
A kind of endoplasmic reticulum molecular chaperone, its expression are in reciprocal relation with a kind of rna level in Tumor cells；β cytoskeletons flesh is moved
Albumen can maintain eucaryotic cell structure, and can regulate and control the contractile motion of internal muscle；Aldose reductase is a kind of metabolic enzyme, the enzyme
Activity relevant with diabetes and its complication；β hemoglobin subunits are present in red blood cell, with oxygen carrying and transport nutrients
The function of matter.
5th, the Efficacy experiments of effect gel
1st, DPPH.free radical removed by effect gel（DPPH）The measure of ability
After taking effect gel addition deionized water suitably dilution of 3mg, being gradually added to 95% ethanol solutions of DPPH makes which fade,
30 min are stored at room temperature, mensuration absorbance is designated as A1 at 519 nm of wavelength；Blank replaces effect gel to survey with 95% ethanol solution
Obtain light absorption value and be designated as A0；With 0.2 mg.ml-1Vitamin C is used as control.
The clearance rate formula of DPPH is as follows：
Now DPPH clearance rates mean value is 82%, with 0.2 mg.ml-1Vitamin c solution 2mL suitable.
2nd, the measure of superoxide anion ability removed by effect gel
According to superoxide anion resisitance determining free radicals kit specification step operation determine effect gel suppress super oxygen cloudy from
The ability of sub- free radical, using considerable amount of vitamin C as standard control, is averaged in triplicate.
Its formula is：
1g effect gel superoxide anion resisitances vigor is measured for 32.6U.L-1, suitable with the vitamin C of 1mg.
3rd, the measure of effect gel scavenging hydroxyl ability
The step of kit specification is determined according to hydroxy radical determines effect gel and suppresses hydroxyl radicals, with a great deal of
Vitamin C as standard control, average in triplicate.
Its formula is：
Measure 1g effects gel to suppress hydroxy radical vigor is 35.30 u.ml-1, with 0.001 mg.ml-1Vitamin C molten
Liquid phase is worked as.
4th, the experiment of effect gel anti-aging ability：
1）48 mouse are divided into aging group（n =40）And Normal group（n =8）, after adaptability is fed 3 days, aging group is little
20% D- galactolipins of mouse nape part hypodermic injection, 1 g.kg-1.d-1, Normal group 2 ml.kg of injection-1.d-1Physiological saline, makes
Totally 42 days mould time.
From modeling the 11st day, by aging group mouse be randomly divided into the high, medium and low dosage group of effect gel, positive controls,
Model group, 8 per group, percutaneous dosing dosage is respectively 30,20,10 mg.kg-1.d-1, 20 mg.kg of vitamin E-1.d-1And it is raw
Reason 20 ml.kg of salt solution-1.d-1；Normal group gives 20 ml.kg-1.d-1Physiological saline, successive administration 32 days.
2）Specimen collection and process：
1. mouse is weighed during being administered week about；Place after death takes its skin of abdomen alcohol disinfecting, removes subcutaneous fat, takes
0.5cm × 0.5cm sizes skin is preserved in -80 DEG C.Remaining animal skin weight is accurately weighed（g）Physiological saline volume=1 3,
Under the conditions of ice-water bath, mechanical homogenisation makes 33.3% homogenate.Take half homogenate 3000r.min-1, 10min is centrifuged to obtain supernatant
Liquid.
2. mouse orbit veniplex takes blood, and blood is placed in calparine pipe, is centrifuged 15 min in 3000 r/min, takes supernatant
300 μ l, are placed in 1.5 ml centrifuge tubes, add 0.6 ml of acetonitrile, 2 min of vortex mixed, are centrifuged 15 in 3000 r/min
Min, Aspirate supernatant are with 0.45 μm of filtering with microporous membrane, to be measured.
3）Indexs measure：
（1）Ordinary circumstance is observed：
The state of mind of observation each group mouse, active situation, hair luster degree, body weight etc..
（2）Skin biopsy：
The skin of 0.5cm × 0.5cm sizes is taken, AFF liquid after freezing microtome section, is used（40% formaldehyde, 15 ml, 95% ethanol, 80 ml, ice vinegar
5 ml of acid）1 min is fixed, HE is dyeed, mounting.
（3）SOD assays：
According to absorbance of the SOD kit specification step measurements skin supernatants at 550 nm of wavelength.Computing formula：
（4）Extinction of the MDA assays according to MDA kit specification step measurements skin homogenate at 532 nm of wavelength
Degree, computing formula：
4）Statistical procedures：
Metric results are usedRepresent, application SPSS17.0 softwares carry out statistical analysis, measurement data adopts single factor test variance
Analyze and do Multiple range test.With P<0.05, P<0.01 is that difference is statistically significant.
5）As a result：
（1）Ordinary circumstance compares：
Normal group appetite is good, in high spirits, hair color gloss, is quick on the draw, agile.
20 mg.kg-1.d-1、30 mg.kg-1.d-1Effect gel group, VE group appetite are good, in high spirits, hair color gloss, reaction
Sensitive.
10 mg.kg-1.d-1Effect gel group appetite is general, and dull is slow in reacting, does not like motion.
Aging group mouse appetite is not good enough, and cold, hair loss is slow in reacting, does not like motion, dead one.
（2）The comparison of each group Mouse Weight：
The continued weight of normal group mouse increases, and the body weight of other each group mouse has increased, and at the end of experiment, model group is little
Mouse body weight compares with normal group that there were significant differences（P<0.01, P<0.05）, the wherein high, medium and low dosage group of effect gel is in work(
After effect gel intervention, there were significant differences compared with model group（P<0.01, P<0.05）See the table below（,n=8）.
Note：Compare with normal group, a：P<0.05, b：P<0.01；Compare with model group, c：P<0.05, d：P<0.01.
（3）Each group mice skin tissue SOD contents, the comparison of MDA contents：
The mice skin tissue SOD contents of model group are significantly less than normal group, and MDA contents are apparently higher than normal group（P<0.01, P<
0.05）, after effect gel is intervened, there were significant differences compared with model group for the high, medium and low dosage group of effect gel（P<
0.01, P<0.05）See the table below.
Effect gel is to mouse aging SOD content, the impact of MDA contents（,n=8）
Note：Compare with normal group, a：P<0.05, b：P<0.01；Compare with model group, c：P<0.05, d：P<0.01.
（4）The comparison of each group mice serum SOD and MDA content：
In the mice serum of model group, SOD contents are significantly less than normal group, and MDA contents are apparently higher than normal group（P<0.01, P<
0.05）, after effect gel is intervened, the high, medium and low dosage group of effect gel and positive controls have aobvious compared with model group
Write difference（P<0.01, P<0.05）, see the table below（,n=8）
Note：Compare with normal group, a：P<0.05, b：P<0.01；Compare with model group, c：P<0.05, d：P<0.01.
（5）The section of each group mice skin tissue is compared：
The visible normal group mouse skin layer rule of HE pathological stainings, thickness are normal, and cell arrangement is neat.Compared with normal group, mould
Type group epidermis is irregular, lower thickness, and cell arrangement is disorderly；After effect gel is intervened, high, medium and low dose of effect gel
Amount group epidermal area rule, thickness increase, and cell arrangement is neat, as shown in accompanying drawing 9 to Figure 14.
According to free radical theory, in body, excessive free radical poisons sexual assault chromosome, mitochondria, cell membrane and connective
Tissue etc. biological tissue and cause body aging.This experiment determines removing DPPH.free radical respectively（DPPH）Energy
Power, superoxide anion ability, scavenging hydroxyl ability is removed evaluating effect gel antioxidant activity in vitro.SOD is in biology
Internal level height can be with visual evaluation aging and death, and it can resist and block the infringement that oxygen radical is caused to cell,
And repair damaging cells in time.MDA is the end-product that Free Radical is obtained after lipid occurs peroxidization, can be indirect
Reacting cells degree of injury.Therefore internal oxidation resistance can be evaluated by determining internal SOD contents, MDA contents.This reality
Test using vitamin E as positive control, effect gel is observed to mouse aging body weight, SOD contents, MDA contents in skin histology,
The impact of the indexs such as skin histology lesion degree is evaluating oxidation resistance in effect gel body.Test result indicate that：Application 20%
Mouse injected by D- galactolipins, and after 1 week, Mouse Weight does not increase, the body weight of model group mouse, SOD contents, MDA contents after 6 weeks
Compared with normal group, there were significant differences, and skin epidermis are substantially thinning, points out aging model to be successful.Mouse gives 30,
20,10 mg.kg-1.d-1After effect gel 6 weeks, although the body weight of low dose group mouse, SOD contents, MDA contents are not significantly
Difference, but height, the body weight of middle dose group mouse, SOD contents have substantially rising（P<0.05, P<0.01）, MDA contents have bright
Aobvious decline（P<0.05, P<0.01）；The skin epidermis of three groups of mouse have been thickened.
Shown by above experimental result：Effect gel has the ability for removing free radical in vitro, so as to mitigate free radical
Injury to biological tissue, also has the function of delaying body aging.After effect gel therapeutic intervention mouse aging, in vivo
SOD contents increase, and hinder infringement of the oxygen radical to cell, and repair damaging cells in time；The minimizing of MDA contents in vivo
Also the reduction of cellular damage degree has been reacted.
Claims (8)
1. a kind of effect gel, it is characterised in that main by pig placenta Aqueous extracts, olive oil, water soluble chitosan and oviductus ranae
Composition；The water soluble chitosan is carboxymethyl chitosan, amber bat acyl shitosan, cross-linked chitosan, hydroxypropyl chitosan or mercapto
Any one in base enclosure glycan.
2. the preparation method of effect gel as claimed in claim 1, it is characterised in that comprise the following steps：
1）Fresh pig placenta is removed to clean after crimson blood, umbilical cord, manadesma and is drained, crushed after freezing under the conditions of -20 DEG C, add water even
Slurry, is then centrifuged for, and by supernatant again with 0.45 μm of membrane filtration, the filter liquor of acquirement is pig placenta Aqueous extracts；
2）By oviductus ranae with homogeneous after the immersion of pig placenta Aqueous extracts, olive oil homogeneous is added, olive oil emulsifiable paste is obtained, is added again
After pig placenta Aqueous extracts homogeneous, olive oil emulsion is obtained；
3）By water soluble chitosan with homogeneous after the immersion of pig placenta Aqueous extracts, olive oil emulsion is then added, then through homogeneous, is taken
Obtain effect gel.
3. the preparation method of effect gel according to claim 2, it is characterised in that contain in the pig placenta Aqueous extracts
The water soluble protein of 0.1～10% mass fraction.
4. the preparation method of effect gel according to claim 3, it is characterised in that the water soluble protein is at least wrapped
Include GRP78, heat shock protein 8, protein disulfide bond isomerase A 3 precursor protein, prolyl 4 hydroxylase β many
Peptide, β actin cytoskeleton fragments, aldose reductase and β hemoglobin subunits.
5. according to claim 3 or 4 effect gel preparation method, it is characterised in that pig placenta Aqueous extracts, olive oil,
The mixing quality ratio of water soluble chitosan and oviductus ranae is 34: 4: 1: 1.
6. the preparation method of effect gel according to claim 2, it is characterised in that the step 2）Middle soak time is 2
～12h.
7. the preparation method of effect gel according to claim 2, it is characterised in that the step 3）Middle soak time is 2
～12h.
8. the preparation method of effect gel according to claim 2, it is characterised in that the mixed system during homogeneous
Rotating speed is 1300 r min-1, each action time is 1～5min.
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|---|---|---|---|---|
| CN107375333A (en) * | 2017-08-02 | 2017-11-24 | 广州准优生物科技有限公司 | Treat intacellin biogel preparation of premature ovarian failure and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1686177A (en) * | 2005-04-08 | 2005-10-26 | 广州拜迪生物医药有限公司 | Production method of zhutaisu (pig's born vegetarian) |
| CN101601688A (en) * | 2009-07-07 | 2009-12-16 | 王会强 | One boar placental article and preparation method thereof |
| CN104367623A (en) * | 2014-11-19 | 2015-02-25 | 杨公明 | Biological agent with function of treating skin burn wounds |
| CN103565839B (en) * | 2013-11-14 | 2015-08-26 | 江南大学 | The separating and extracting process of one boar Placenta extract |
-
2016
- 2016-11-18 CN CN201611014389.0A patent/CN106491488A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1686177A (en) * | 2005-04-08 | 2005-10-26 | 广州拜迪生物医药有限公司 | Production method of zhutaisu (pig's born vegetarian) |
| CN101601688A (en) * | 2009-07-07 | 2009-12-16 | 王会强 | One boar placental article and preparation method thereof |
| CN103565839B (en) * | 2013-11-14 | 2015-08-26 | 江南大学 | The separating and extracting process of one boar Placenta extract |
| CN104367623A (en) * | 2014-11-19 | 2015-02-25 | 杨公明 | Biological agent with function of treating skin burn wounds |
Non-Patent Citations (8)
| Title |
|---|
| 于晓燕: "《让你年轻十岁的美肌书》", 31 May 2014, 黑龙江科学技术出版社 * |
| 周国华等: "羊和猪胎盘肽的制备及其生物活性的初步研究", 《家畜生态学报》 * |
| 周国华等: "羊和猪胎盘肽的提取", 《动物医学进展》 * |
| 唐文林等: "猪胎盘综合利用工艺与应用研究", 《中国优秀硕士学位论文全文数据库》 * |
| 李小梅等: ""猪胎盘水提物对D-半乳糖诱导衰老小鼠抗氧化及学习记忆调节作用的影响"", 《中国新药杂志》 * |
| 温辉梁: "《生物化工产品生产技术》", 31 December 2004, 江西科学技术出版社 * |
| 邓明鲁: "《吉林省中药资源》", 30 November 2014, 中国中医药出版社 * |
| 骆小丽: "《女人这样做吃不胖、晒不黑、人不老》", 31 July 2016, 中国古籍出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107375333A (en) * | 2017-08-02 | 2017-11-24 | 广州准优生物科技有限公司 | Treat intacellin biogel preparation of premature ovarian failure and preparation method thereof |
| CN107375333B (en) * | 2017-08-02 | 2020-08-18 | 广州准优生物科技有限公司 | Placenta extract biological gel preparation for treating premature ovarian failure and preparation method thereof |
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