CN107156844A - A kind of preparation method of hidden sheath sheath silk algae ethanol extract and application - Google Patents
A kind of preparation method of hidden sheath sheath silk algae ethanol extract and application Download PDFInfo
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- 241000195493 Cryptophyta Species 0.000 title claims abstract description 106
- 239000000469 ethanolic extract Substances 0.000 title claims abstract description 53
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 238000001556 precipitation Methods 0.000 claims abstract description 21
- 230000003064 anti-oxidating effect Effects 0.000 claims abstract description 16
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 7
- 235000013402 health food Nutrition 0.000 claims abstract description 6
- 239000006228 supernatant Substances 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 5
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims 1
- 241000244203 Caenorhabditis elegans Species 0.000 abstract description 28
- 102000019197 Superoxide Dismutase Human genes 0.000 abstract description 17
- 108010012715 Superoxide dismutase Proteins 0.000 abstract description 17
- 230000036542 oxidative stress Effects 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 9
- 229910052760 oxygen Inorganic materials 0.000 abstract description 7
- 239000001301 oxygen Substances 0.000 abstract description 7
- 238000001727 in vivo Methods 0.000 abstract description 6
- 230000032683 aging Effects 0.000 abstract description 5
- 238000012360 testing method Methods 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 150000003254 radicals Chemical class 0.000 description 13
- 210000002977 intracellular fluid Anatomy 0.000 description 12
- 230000004083 survival effect Effects 0.000 description 12
- 239000000463 material Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 6
- 238000007254 oxidation reaction Methods 0.000 description 6
- 239000003963 antioxidant agent Substances 0.000 description 5
- 230000003078 antioxidant effect Effects 0.000 description 5
- 235000006708 antioxidants Nutrition 0.000 description 5
- 230000003647 oxidation Effects 0.000 description 5
- 230000003712 anti-aging effect Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- -1 oxygen radical Chemical class 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000155 melt Substances 0.000 description 3
- FIKAKWIAUPDISJ-UHFFFAOYSA-L paraquat dichloride Chemical compound [Cl-].[Cl-].C1=C[N+](C)=CC=C1C1=CC=[N+](C)C=C1 FIKAKWIAUPDISJ-UHFFFAOYSA-L 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- 241001134698 Lyngbya Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
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- 230000009758 senescence Effects 0.000 description 2
- 235000016425 Arthrospira platensis Nutrition 0.000 description 1
- 240000002900 Arthrospira platensis Species 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001501873 Isochrysis galbana Species 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
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- 239000004530 micro-emulsion Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 231100000590 oncogenic Toxicity 0.000 description 1
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- 239000003960 organic solvent Substances 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
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- 239000001508 potassium citrate Substances 0.000 description 1
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- QEEAPRPFLLJWCF-UHFFFAOYSA-K potassium citrate (anhydrous) Chemical compound [K+].[K+].[K+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O QEEAPRPFLLJWCF-UHFFFAOYSA-K 0.000 description 1
- 235000011082 potassium citrates Nutrition 0.000 description 1
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A61K2236/30—Extraction of the material
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention belongs to field of marine biotechnology, there is provided a kind of preparation method of hidden sheath sheath silk algae ethanol extract and application.Hidden sheath sheath that the present invention is provided silk algae ethanol extract is by by the fresh hidden sheath sheath silk wet algae multigelation of algae, centrifuging and taking precipitation plus ethanol carry out ultrasonic extraction and are made, obtained hidden sheath sheath silk algae ethanol extract can significantly improve the removing of the vigor of superoxide dismutase (SOD) and promotion activity in vivo oxygen radical (ROS) in Caenorhabditis elegans body, reduce the level of activity in vivo oxygen radical (ROS), it can alleviate the early ageing phenomenon as caused by oxidative stress simultaneously, with significant antioxidation, it can be applied to prepare health food and medicine with anti-oxidation function.
Description
Technical field
The invention belongs to field of marine biotechnology, more particularly to a kind of preparation method of hidden sheath sheath silk algae ethanol extract
With application.
Background technology
Free radical refers to the atom containing unpaired electronics, molecule or group, due to containing not paired electricity in free radical
Son, the tendency with pairing, situation is unstable, the activity with height.Under normal circumstances, the free radical in human body is to be in
Among the dynamic equilibrium for constantly producing and removing.Free radical produces excessive or removed slow, unnecessary free radical by attacking life
Macromolecular substances and various cells are ordered, body can be caused in the various damages of molecular level, cellular level and histoorgan level,
Accelerate the senescence process of body and induce various diseases, such as free radical causes DNA oxidative damages, the transformation of cytogenetics information is led
It is oncogenic to occur；Free radical makes lipid peroxidation, causes artery sclerosis etc..
The definition of polyphenoils is " any material that there is the oxidation reaction that just can effectively suppress free radical with low concentration ",
Its mechanism of action can act directly on free radical, or consume the material of easily generation free radical indirectly, prevent
Further reaction.Human body resists the anti-oxidant of free radical producing naturally while free radical is inevitably generated, also
Material, to offset oxidative attacks of the free radical to human body cell.Research has shown that, the antioxidant system of human body be one can with it is immune
System is comparable, system with perfect and complicated function, and the oxidation resistant ability of body is stronger, more healthy, life
It is longer.
Aid in improving by screening the material with anti-oxidation characteristics the oxidation resistance of body, anti-aging it is straight
Connect, effectively, one of convenient important method.Microalgae has the marine alga of important research value as a class, its inoxidizability
One of focus always studied.At present, reported that the microalgae with antioxidation activity mainly has following several：Spirulina, ball
Isochrysis galbana, Rhodella reticulata, purple ball algae, flat algae and chlorella.
Hidden sheath sheath silk algae (Lyngbya cryptovaginatus), also makes hidden sheath float silk algae sometimes, plant Dan Sheng, directly or
Slightly curved song, in addition to off rating, without hard sheath；Algal filament attenuates gradually from middle part to top, with emitting shape structure, it is impossible to transport
Move or substantially do not move, it is wide 3.5~10 μm；Cell cylinder, rare square, air bag is full of cell.Through retrieving, in the prior art
The report on hidden sheath sheath silk algae related application is there are no, especially with regard to the anti-oxidant application of hidden sheath sheath silk algae ethanol extract
Report.
The content of the invention
It is an object of the invention to provide a kind of preparation method of hidden sheath sheath silk algae ethanol extract.It is hidden that the present invention is provided
Sheath sheath silk algae ethanol extract has significant antioxidation, can be applied to prepare health food with anti-oxidation function and
Medicine.
Technical scheme will be further described in detail below reflect and description.
A kind of preparation method of hidden sheath sheath silk algae ethanol extract, comprises the following steps：
S1, the fresh hidden sheath sheath silk wet algae of algae is taken, be to freeze 12 hours under conditions of -20 DEG C~-40 DEG C in temperature, room temperature
Melt 12 hours, multigelation 3~5 times, 5~15min centrifuged under conditions of rotating speed is 5000r/min~10000r/min,
Supernatant and precipitation are separated, take precipitation standby；Supernatant is hidden sheath sheath silk algae intracellular fluid；
S2, the EtOH Sonicate that addition volume fraction is 70%~99% into the precipitation described in step S1 are extracted 2~4 times,
5~15min is centrifuged under conditions of rotating speed is 5000r/min~10000r/min, supernatant and precipitation are separated, supernatant is taken
It is dried in vacuo, the one-level water of the product after vacuum drying is dissolved, produces hidden sheath sheath silk algae ethanol extract.
Preferably, add ethanol by the solid-to-liquid ratio 1g/mL of wet algae and ethanol in step S2 and carry out ultrasonic extraction.
Preferably, the frequency of ultrasonic extraction is 35~40kHz in step S2, and temperature is 4 DEG C.
Preferably, the time of each ultrasonic extraction is 10~30min in step S2.
The health care with anti-oxidation function is being prepared it is a further object of the present invention to provide hidden sheath sheath silk algae ethanol extract
Application in food and medicine.
Compared with prior art, the invention has the advantages that：
(1) the hidden sheath sheath silk algae ethanol extract that the present invention is provided can significantly improve super oxygen in Caenorhabditis elegans body
Thing mutase (SOD) vigor and the removing for promoting activity in vivo oxygen radical (ROS), significantly reduce activity in vivo oxygen radical
(ROS) level, with significant antioxidation.
(2) the hidden sheath sheath silk algae ethanol extract that the present invention is provided can extend Caenorhabditis elegans under oxidative stress conditions
Average life span, its survival rate is improved, with significant anti-aging and antioxidation.
(3) preparation method for the hidden sheath sheath silk algae ethanol extract that the present invention is provided has the cycle short, organic solvent consumption
Few, technique is simple, safety non-toxic, the features such as stability and high efficiency, is adapted to industrialized production.
Brief description of the drawings
Influence of the hidden sheath sheath silk algae ethanol extracts of Fig. 1 to Caenorhabditis elegans survival rate under oxidative stress conditions.
Influence of the hidden sheath sheath silk algae ethanol extracts of Fig. 2 to ROS levels in Caenorhabditis elegans body.
Influence of the hidden sheath sheath silk algae ethanol extracts of Fig. 3 to SOD vigor in Caenorhabditis elegans body.
Influence of the hidden sheath sheath silk algae intracellular fluids of Fig. 4 to Caenorhabditis elegans survival rate under oxidative stress conditions.
Influence of the hidden sheath sheath silk algae intracellular fluids of Fig. 5 to SOD vigor in Caenorhabditis elegans body.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
Hidden sheath sheath silk algae (Lyngbya cryptovaginatus) of the present invention is studied by Chinese Academy of Sciences aquatile
There is provided, its numbering in algae kind storehouse is FACHB-890.
One-level water of the present invention meets the standard of one-level water in GB/T 6682-2008.
The present inventor has carried out substantial amounts of experiment during studying microalgae, has been surprisingly found that hidden sheath sheath silk algae
Intracellular fluid and the hidden sheath sheath silk algae that remaining residue progress EtOH Sonicate extraction after extraction hidden sheath sheath silk algae intracellular fluid is made
Ethanol extract is respectively provided with significant antioxidation.And the antioxidation of hidden sheath sheath silk algae ethanol extract is especially pronounced.
Tested in whole animal level with oxidative stress model and aging correlation model, result of the test shows this hair
The hidden sheath sheath silk algae ethanol extract of bright offer can significantly extend the average life span of Caenorhabditis elegans under oxidative stress conditions,
Improve its survival rate；Meanwhile, it can significantly improve the vigor and rush of the superoxide dismutase (SOD) in Caenorhabditis elegans body
Enter activity in vivo oxygen radical (ROS) removing, significantly reduce activity in vivo oxygen radical (ROS) level.The present invention is provided
Hidden sheath sheath silk algae ethanol extract there is significant antioxidation, process of the test does not find hidden sheath sheath silk algae ethanol extract
There is toxic action to Caenorhabditis elegans, illustrate the hidden sheath sheath silk algae ethanol extract safety non-toxic that the present invention is provided, can answer
For preparing health food and medicine with anti-oxidation function.
Antioxidation of the present invention refer to under pathology or physiological condition because occur response to oxidative stress produced
Raw change is the aging of principal character and the preventive and therapeutic effect of diseases associated with senescence.
The present invention hidden sheath sheath silk algae ethanol extract can be shared individually or with other active components, as solely or mainly living
Property composition be applied to prepare health food and medicine with anti-oxidation function, the formulation of the health food and medicine is preferably
Tablet, capsule, parenteral solution, oral liquid, granule, pill, powder and pill etc..
Test method used in the embodiment of the present invention, is conventional method unless otherwise specified；Used material
Material, reagent, are the reagent and material commercially obtained unless otherwise specified.Wherein, S Basal solution and S
The preparation method of Medium solution is as follows：
S Basal solution：Weigh 5.8gNaCl, 1.3g K2HPO4·3H2O and 6.0g KH2PO4It is placed in reagent bottle, plus
Enter 750mL deionized waters, be settled to 1L after being completely dissolved, sterilize 30min, be cooled to 40-50 DEG C it is standby.
S Medium solution：The S Basal solution after 1L sterilizings is taken, 1mL 5.0mg/mL cholesterol ethanol is added dropwise molten
Liquid, while add change to shake, makes it fully dissolve (solution switch to micro emulsion by water white transparency transparent in vain, no obvious sediment), then successively
Add 3mL 1M CaCl2Solution, 3mL 1M MgSO4, 10mL 1M potassium citrate solutions (pH=6.0) and 10mL trace elements are molten
Liquid, room temperature is standby.
The preparation of the hidden sheath sheath silk algae ethanol extract of embodiment 1
S1, the fresh hidden sheath sheath silk wet algae of algae is taken, be to freeze 12 hours under conditions of -30 DEG C in temperature, room temperature melts 12
Hour, multigelation 4 times centrifuges 10min under conditions of rotating speed is 8000r/min, supernatant and precipitation is separated, precipitation is taken
It is standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 85%
EtOH Sonicate is extracted 3 times, and the frequency of ultrasonic extraction is 40kHz, and temperature is 4 DEG C, and the time of ultrasound is 20min every time, in rotating speed
To centrifuge 10min under conditions of 8000r/min, supernatant and precipitation are separated, takes supernatant to be dried in vacuo, vacuum is done
Product after dry is dissolved with one-level water, and it is that (A represents hidden to 1gA/mL to adjust it relative to the concentration of hidden sheath sheath silk algae (wet algae) weight
The wet algae of sheath sheath silk algae), produce hidden sheath sheath silk algae ethanol extract.
In addition, take step S1 centrifuge after isolated supernatant, plus to adjust it (wet relative to hidden sheath sheath silk algae for one-level water
Algae) concentration of weight is 2gA/mL (A represents the wet algae of hidden sheath sheath silk algae), produces hidden sheath sheath silk algae intracellular fluid.
The preparation of the hidden sheath sheath silk algae ethanol extract of embodiment 2
S1, the fresh hidden sheath sheath silk wet algae of algae is taken, be to freeze 12 hours under conditions of -20 DEG C in temperature, room temperature melts 12
Hour, multigelation 5 times centrifuges 8min under conditions of rotating speed is 10000r/min, supernatant and precipitation is separated, precipitation is taken
It is standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 70%
EtOH Sonicate is extracted 3 times, and the frequency of ultrasonic extraction is 35kHz, and temperature is 4 DEG C, and the time of ultrasound is 30min every time, in rotating speed
To centrifuge 10min under conditions of 8000r/min, supernatant and precipitation are separated, takes supernatant to be dried in vacuo, vacuum is done
Product after dry is dissolved with one-level water, and it is that (A represents hidden to 1gA/mL to adjust it relative to the concentration of hidden sheath sheath silk algae (wet algae) weight
The wet algae of sheath sheath silk algae), produce hidden sheath sheath silk algae ethanol extract.
The preparation of the hidden sheath sheath silk algae ethanol extract of embodiment 3
S1, the fresh hidden sheath sheath silk wet algae of algae is taken, be to freeze 12 hours under conditions of -40 DEG C in temperature, room temperature melts 12
Hour, multigelation 3 times centrifuges 15min under conditions of rotating speed is 8000r/min, supernatant and precipitation is separated, precipitation is taken
It is standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 90%
EtOH Sonicate is extracted 4 times, and the frequency of ultrasonic extraction is 40kHz, and temperature is 4 DEG C, and the time of ultrasound is 15min every time, in rotating speed
To centrifuge 15min under conditions of 5000r/min, supernatant and precipitation are separated, takes supernatant to be dried in vacuo, vacuum is done
Product after dry is dissolved with one-level water, and it is that (A represents hidden to 1gA/mL to adjust it relative to the concentration of hidden sheath sheath silk algae (wet algae) weight
The wet algae of sheath sheath silk algae), produce hidden sheath sheath silk algae ethanol extract.
Influence of the hidden sheath sheath silk algae ethanol extract of embodiment 4 to Caenorhabditis elegans survival rate under oxidative stress conditions
Hidden sheath sheath silk algae ethanol extract made from Example 1, adult initial stage is added to by final concentration 300mgA/mL
In Wild-type C. elegans (N2) (being provided by Univ Minnesota-Twin Cities USA's Caenorhabditis elegans hereditary information collection),
Control group adds isometric S Medium solution, is placed in after 20 DEG C of culture 24h, the paraquat for adding final concentration of 50mM is carried out
Oxidative stress modeling, then counts the ratio of Caenorhabditis elegans survival every 12h, until it is all dead.Beautiful hidden bar line
The survival rate of worm is represented with survivorship curve, is as a result analyzed with Kaplan-Meier methods.As a result it is as shown in Figure 1.
Fig. 1 results show that hidden sheath sheath silk algae ethanol extract can extend Caenorhabditis elegans under oxidative stress conditions
Average life span, improves its survival rate.Result of the test show the present invention provide hidden sheath sheath silk algae ethanol extract can alleviate by
Early ageing phenomenon caused by oxidative stress, with significant anti-oxidant and anti-aging effects.
Influence of the hidden sheath sheath silk algae ethanol extract of embodiment 5 to ROS levels in Caenorhabditis elegans body
24 well culture plates are taken, if test group and control group, every group sets two holes, be 1mL per hole final volume, test group adds together
Hidden sheath sheath silk algae ethanol extract, hidden sheath sheath made from L4 phases Wild-type C. elegans (N2) and embodiment 1 after stepization
The final concentration 100mgA/mL of silk algae ethanol extract, control group adds the L4 phases Wild-type C. elegans (N2) after synchronizing
With the S Medium solution isometric with test group hidden sheath sheath silk algae ethanol extract, it is placed in after 20 DEG C of culture 24h, is added per hole
Paraquat continues to cultivate after 48h, collects each group Caenorhabditis elegans to final concentration of 2mM, by its respectively under condition of ice bath it is even
Slurry, 5min is centrifuged by homogenate under the conditions of 4 DEG C, 10000g, collects supernatant, final concentration of 50 μM are added into supernatant
DCFH-DA probes (are purchased from Sigma companies, No. CAS：2044-85-1) solution, using fluorescence microplate reader in 485nm excitation wavelengths
With fluorescence intensity under 538nm launch wavelengths, room temperature detection 2h is surveyed once per 10min.As a result F check analyses are used, as a result such as
Shown in Fig. 2.
Fig. 2 results are shown, compared with control group, after the silk algae ethanol extract feeding of hidden sheath sheath, Caenorhabditis elegans body
Interior ROS levels are remarkably decreased.Result of the test shows that the hidden sheath sheath silk algae ethanol extract that the present invention is provided can promote internal ROS
Removing, internal ROS levels are reduced, with significant oxidation resistance.
Influence of the hidden sheath sheath silk algae ethanol extract of embodiment 6 to SOD vigor in Caenorhabditis elegans body
24 well culture plates are taken, if test group and control group, every group sets two holes, be 1mL per hole final volume, test group adds together
Hidden sheath sheath silk algae ethanol extract, hidden sheath sheath made from L4 phases Wild-type C. elegans (N2) and embodiment 1 after stepization
The final concentration 100mgA/mL of silk algae ethanol extract, control group adds the L4 phases Wild-type C. elegans (N2) after synchronizing
With the S Medium solution isometric with test group hidden sheath sheath silk algae ethanol extract, it is placed in after 20 DEG C of culture 72h, collects each group
Caenorhabditis elegans, it is homogenized under condition of ice bath respectively, homogenate is centrifuged into 5min under the conditions of 4 DEG C, 10000g, is collected
Supernatant, (is purchased from green skies biotechnology and grinds make internal disorder or usurp institute, production code member using total SOD activity detection kits：S0101) determine elegant
SOD vigor in beautiful hidden rhabditida homogenate supernatant, as a result as shown in Figure 3.
Fig. 3 results are shown, compared with control group, and hidden sheath sheath silk algae ethanol extract can significantly improve Caenorhabditis elegans body
Interior SOD vigor.Result of the test shows that the hidden sheath sheath silk algae ethanol extract that the present invention is provided can significantly improve internal SOD's
Vigor, with significant oxidation resistance.
Influence of the hidden sheath sheath silk algae intracellular fluid of embodiment 7 to Caenorhabditis elegans survival rate under oxidative stress conditions
Hidden sheath sheath silk algae intracellular fluid made from Example 1, the wild of adult initial stage is added to by final concentration 300mgA/mL
In type Caenorhabditis elegans (N2), control group adds isometric S Medium solution, is placed in after 20 DEG C of culture 24h, adds eventually
Concentration carries out oxidative stress modeling for 50mM paraquat, and the ratio of Caenorhabditis elegans survival is then counted every 12h, until
It is all dead.The survival rate of Caenorhabditis elegans is represented with survivorship curve, is as a result analyzed with Kaplan-Meier methods.Knot
Fruit is as shown in Figure 4.
Fig. 4 results show that liquid energy enough extends the existence of Caenorhabditis elegans under oxidative stress conditions in hidden sheath sheath silk gonidium
Time, improve its survival rate.Result of the test shows that liquid energy is enough alleviated by oxidative stress in the hidden sheath sheath silk gonidium of the invention provided
Caused early ageing phenomenon, with significant anti-oxidant and anti-aging effects.
Influence of the hidden sheath sheath silk algae intracellular fluid of embodiment 8 to SOD vigor in Caenorhabditis elegans body
24 well culture plates are taken, if test group and control group, every group sets two holes, be 1mL per hole final volume, test group adds together
Hidden sheath sheath silk algae intracellular fluid made from L4 phases Wild-type C. elegans (N2) and embodiment 1 after stepization, hidden sheath sheath silk algae
The final concentration 100mgA/mL of intracellular fluid, control group add synchronize after L4 phases Wild-type C. elegans (N2) and with experiment
The isometric S Medium solution of the hidden sheath sheath silk algae intracellular fluid of group, is placed in after 20 DEG C of culture 72h, collects the beautiful hidden bar line of each group
Worm, it is homogenized under condition of ice bath respectively, homogenate is centrifuged into 5min under the conditions of 4 DEG C, 10000g, collects supernatant, profit
(it is purchased from green skies biotechnology with total SOD activity detection kits and grinds make internal disorder or usurp institute, production code member：S0101 beautiful hidden bar line) is determined
SOD vigor in worm homogenate supernatant.As a result it is as shown in Figure 5.
Fig. 5 results are shown, compared with control group, and liquid energy is significantly improved in Caenorhabditis elegans body in hidden sheath sheath silk gonidium
SOD vigor.Result of the test shows that liquid energy enough significantly improves internal SOD vigor in hidden sheath sheath silk gonidium that the present invention is provided,
With significant oxidation resistance.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to assert
The specific implementation of the present invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of not departing from present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention's
Protection domain.
Claims (5)
1. a kind of preparation method of hidden sheath sheath silk algae ethanol extract, it is characterised in that the preparation method comprises the following steps：
S1, the fresh hidden sheath sheath silk wet algae of algae is taken, be to freeze 12 hours under conditions of -20 DEG C~-40 DEG C in temperature, room temperature is melted
12 hours, multigelation 3~5 times centrifuged 5~15min under conditions of rotating speed is 5000r/min~10000r/min, will be upper
Clear liquid and precipitation are separated, and take precipitation standby；
S2, the EtOH Sonicate that addition volume fraction is 70%~99% into the precipitation described in step S1 are extracted 2~4 times, are being turned
Speed is 5~15min of centrifugation under conditions of 5000r/min~10000r/min, and supernatant and precipitation are separated, take supernatant to carry out
Vacuum drying, the one-level water of the product after vacuum drying is dissolved, and produces hidden sheath sheath silk algae ethanol extract.
2. the preparation method of hidden sheath sheath silk algae ethanol extract as claimed in claim 1, it is characterised in that in the step S2
Ethanol, which is added, by the solid-to-liquid ratio 1g/mL of wet algae and ethanol carries out ultrasonic extraction.
3. the preparation method of hidden sheath sheath silk algae ethanol extract as claimed in claim 1, it is characterised in that in the step S2
The frequency of ultrasonic extraction is 35~40kHz, and temperature is 4 DEG C.
4. the preparation method of hidden sheath sheath silk algae ethanol extract as claimed in claim 1, it is characterised in that in the step S2
The time of each ultrasonic extraction is 10~30min.
5. hidden sheath sheath silk algae ethanol made from the preparation method of hidden sheath sheath silk algae ethanol extract as claimed in claim 1 is extracted
Application of the thing in health food and medicine with anti-oxidation function is prepared.
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