CN107198702A - One kind floating quiver algae water extract and application - Google Patents
One kind floating quiver algae water extract and application Download PDFInfo
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- CN107198702A CN107198702A CN201710388260.4A CN201710388260A CN107198702A CN 107198702 A CN107198702 A CN 107198702A CN 201710388260 A CN201710388260 A CN 201710388260A CN 107198702 A CN107198702 A CN 107198702A
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- 241000195493 Cryptophyta Species 0.000 title claims abstract description 106
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 238000001556 precipitation Methods 0.000 claims abstract description 49
- 239000006228 supernatant Substances 0.000 claims abstract description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 30
- 230000003064 anti-oxidating effect Effects 0.000 claims abstract description 17
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 10
- 238000005119 centrifugation Methods 0.000 claims abstract description 8
- 235000013402 health food Nutrition 0.000 claims abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 3
- 238000010438 heat treatment Methods 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 16
- 238000002360 preparation method Methods 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- 230000036542 oxidative stress Effects 0.000 abstract description 15
- 230000032683 aging Effects 0.000 abstract description 8
- 229910052760 oxygen Inorganic materials 0.000 abstract description 6
- 239000001301 oxygen Substances 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 238000001727 in vivo Methods 0.000 abstract description 3
- 241000244203 Caenorhabditis elegans Species 0.000 description 24
- 230000004083 survival effect Effects 0.000 description 13
- 210000002977 intracellular fluid Anatomy 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 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 5
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 230000003712 anti-aging effect Effects 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000192494 Oscillatoriales Species 0.000 description 2
- 241000244200 Rhabditida Species 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000000975 bioactive effect Effects 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
- 235000013305 food Nutrition 0.000 description 2
- -1 hydroxyl radical free radical Chemical class 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 230000008569 process 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
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 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
- 241000293712 Rhodella reticulata Species 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
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- 239000002537 cosmetic Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 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
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
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- 238000009472 formulation Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000001990 heterocyst Anatomy 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-N hydroperoxyl Chemical compound O[O] OUUQCZGPVNCOIJ-UHFFFAOYSA-N 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000003182 parenteral nutrition solution Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000029264 phototaxis Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000001508 potassium citrate Substances 0.000 description 1
- 229960002635 potassium citrate Drugs 0.000 description 1
- 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
- 239000000843 powder Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000011506 response to oxidative stress Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 229940082787 spirulina Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
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- 210000001519 tissue Anatomy 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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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
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/748—Cyanobacteria, i.e. blue-green bacteria or blue-green algae, e.g. spirulina
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
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- 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
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Abstract
The invention belongs to field of marine biotechnology, there is provided one kind floating quiver algae water extract and its application.The floating algae water extract that quivers of the present invention is by the way that by the fresh wet algae multigelation of the floating algae that quivers, supernatant is removed in centrifugation；Precipitation plus ethanol is taken to carry out ultrasonic extraction, supernatant is removed in centrifugation；Precipitation is taken, add water carry out heating extraction after being washed with water, be made.The floating algae water extract that quivers that the present invention is provided can alleviate the early ageing phenomenon as caused by oxidative stress, activity in vivo oxygen radical (ROS) removing can be promoted, reduce the level of activity in vivo oxygen radical (ROS), 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 floating quiver algae water extract and its application.
Background technology
Microalgae is the unicellular microscopic algae of a group physiology and biological morphology isomery, and tellurian kind is more than 200,000
Kind.Microalgae can produce many bioactive substances, it is most of apply to medicine production, food or feed addictive, cosmetics and
In other application.
Active oxygen, such as superoxide anion, hydroxyl radical free radical, Both peroxyl radical, can attack lipid, protein, enzyme,
The bioactive molecules such as DNA, RNA, cause cell or tissue to damage aging even carcinogenic.Antioxidant can effective scavenging capacity
Oxygen, protects body from the infringement of active oxygen.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.
The floating algae (Planktothrixsp.) that quivers, is subordinate to Oscillatoriopsis, Oscillatoriales, without heterocyst and akinete,
It is different with other microalgaes of Oscillatoriales by cell division growth formation trichome with independent trichome and inflation vacuole,
These trichomes have phototaxis, the slight motion of energy.Through retrieval, the report of floating algae related application of quivering is had no.
The content of the invention
Of the invention is that providing a kind of float quivers algae water extract and its preparing the food of the health care with anti-oxidation function
Application in product and medicine.The floating algae water extract that quivers that the present invention is provided be by by the fresh wet algae multigelation of the floating algae that quivers,
Supernatant is removed in centrifugation；Precipitation plus ethanol is taken to carry out ultrasonic extraction, supernatant is removed in centrifugation；Precipitation is taken, adds water and is heated after being washed with water
Extract and be made, with significant antioxidation, can be applied to prepare health food and medicine with anti-oxidation function.
To realize the purpose of the present invention, technical scheme is as follows：
The floating algae water extract that quivers of one kind, its preparation method comprises the following steps：
S1, the fresh floating wet algae of the algae that quivers is taken, be to freeze 10~12 hours under conditions of -20 DEG C~-40 DEG C in temperature, room temperature
Melt 10~12 hours, multigelation 3~5 times, rotating speed be 5000r/min~10000r/min under conditions of centrifugation 5~
15min, supernatant and precipitation are separated, take precipitation standby；Supernatant is the floating algae intracellular fluid that quivers；
S2, the EtOH Sonicate that addition volume fraction is 70%~99% into the precipitation described in step S1 are extracted 2~4 times,
10~15min is centrifuged under conditions of rotating speed is 5000r/min~10000r/min, supernatant and precipitation are separated, precipitation is taken
It is standby；
S3, precipitation 1~3 time described in step S2 is washed with one-stage water wash, be 5000r/min~10000r/min's in rotating speed
Under the conditions of centrifuge 10~15min, by supernatant and precipitation separate, remove supernatant, into precipitation add one-level water temperature be 70
DEG C~100 DEG C under conditions of extract 3~5 times, 30~60min every time, in the condition that rotating speed is 5000r/min~10000r/min
5~15min of lower centrifugation, supernatant and precipitation are separated, supernatant is taken, and produce the floating algae water extract that quivers.
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.
Preferably, add one-level water by the solid-to-liquid ratio 1g/mL of wet algae and one-level water in step S3 and carry out heating extraction.
The result of the test tested in whole animal level with oxidative stress model and aging correlation model shows,
The 300mgA/mL floating algae water extract that quivers can significantly improve the survival rate of Caenorhabditis elegans under oxidative stress conditions, extension
Its average life span, with significant anti-oxidant, anti-aging effects.Therefore, the floating algae water extract that quivers that the present invention is provided has aobvious
The antioxidation of work, can be applied to prepare health food and medicine with anti-oxidation function.
It is elegant under oxidative stress conditions in process of the test, it was also found that floating through 100mgA/mL quivers after the feeding of algae water extract
The survival rate reduction of beautiful hidden rhabditida, average life span is shortened, but compared with control group, no significant difference.
Meanwhile, inventor is in further research, it has been found that during the floating algae water extract that quivers of above-mentioned preparation,
The floating algae intracellular fluid that quivers that step S1 is obtained equally has certain antioxidation, and can also be applied to prepare has anti-oxidation function
Health food and medicine.
Compared with prior art, the invention has the advantages that：
(1) present invention provides the new opplication of the floating algae that quivers, floating to quiver algae water extract and there is intracellular fluid significant antioxygen to be turned into
With can be applied to preparation has the health food and medicine of anti-oxidation function.
(2) the floating algae water extract that quivers that the present invention is provided can significantly improve Caenorhabditis elegans under oxidative stress conditions
Survival rate, alleviates Caenorhabditis elegans early ageing phenomenon as caused by oxidative stress, with significant antioxidation.
(2) the floating algae water extract that quivers that the present invention is provided can promote internal ROS removing, reduce internal ROS levels, tool
There is significant oxidation resistance.
Brief description of the drawings
Floating influences of the algae water extract to Caenorhabditis elegans survival rate under oxidative stress conditions of quivering of Fig. 1.
Floating influences of the algae water extract to ROS levels in Caenorhabditis elegans body of quivering of Fig. 2.
Floating influences of the algae intracellular fluid to Caenorhabditis elegans survival rate under oxidative stress conditions of quivering of Fig. 3.
Floating influences of the algae intracellular fluid to Caenorhabditis elegans average life span under natural conditions of quivering of Fig. 4.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
The floating algae (Planktothrixsp.) that quivers of the present invention is provided by Inst. of Hydrobiology, Chinese Academy of Sciences, its
Numbering in Chinese Academy of Sciences's algae kind storehouse is FACHB-1365.
One-level water of the present invention meets the standard of one-level water in GB/T 6682-2008.
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 floating algae water extract that quivers of the present invention can be shared individually or with other active components, be used as solely or mainly active component
Applied to preparing health food and medicine with anti-oxidation function, the formulation of the health food and medicine be 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.
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 floating algae water extract that quivers of embodiment 1
S1, the fresh floating wet algae of the algae that quivers is taken, is to freeze 12 hours under conditions of -20 DEG C in temperature, room temperature is melted 10 hours,
Multigelation 5 times, centrifuges 10min under conditions of rotating speed is 8000r/min, supernatant and precipitation is separated, take precipitation 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 80%
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 30min every time, in rotating speed
To centrifuge 15min under conditions of 8000r/min, supernatant and precipitation are separated, take precipitation standby；
S3, precipitation 2 times described in step S2 is washed with one-stage water wash, centrifuged under conditions of rotating speed is 8000r/min
15min, supernatant and precipitation are separated, supernatant is removed, and one-level is added by wet algae and the solid-to-liquid ratio 1g/mL of one-level water into precipitation
Water is extracted 4 times under conditions of temperature is 100 DEG C, each 45min, and 15min is centrifuged under conditions of rotating speed is 8000r/min,
Supernatant and precipitation are separated, supernatant is taken, plus it is 1gA/ that one-level water, which adjusts it relative to the concentration of floating algae (wet algae) weight of quivering,
ML (A represents the floating wet algae of algae that quivers), produces the floating algae water extract that quivers.
In addition, step S1 supernatants isolated after centrifuging are taken, plus one-level water adjusts it relative to the floating algae (wet algae) that quivers
The concentration of weight is 2gA/mL (A represents the floating wet algae of algae that quivers), produces the floating algae intracellular fluid that quivers.
The preparation of the floating algae water extract that quivers of embodiment 2
S1, the fresh floating wet algae of the algae that quivers is taken, is to freeze 12 hours under conditions of -30 DEG C in temperature, room temperature is melted 12 hours,
Multigelation 4 times, centrifuges 5min under conditions of rotating speed is 10000r/min, supernatant and precipitation is separated, take precipitation 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 3 times, and the frequency of ultrasonic extraction is 35kHz, and temperature is 4 DEG C, and the time of ultrasound is 20min every time, in rotating speed
To centrifuge 10min under conditions of 10000r/min, supernatant and precipitation are separated, take precipitation standby；
S3, precipitation 3 times described in step S2 is washed with one-stage water wash, centrifuged under conditions of rotating speed is 10000r/min
15min, supernatant and precipitation are separated, supernatant is removed, and one-level is added by wet algae and the solid-to-liquid ratio 1g/mL of one-level water into precipitation
Water is extracted 3 times under conditions of temperature is 90 DEG C, each 30min, and 15min is centrifuged under conditions of rotating speed is 10000r/min,
Supernatant and precipitation are separated, supernatant is taken, plus it is 1gA/ that one-level water, which adjusts it relative to the concentration of floating algae (wet algae) weight of quivering,
ML (A represents the floating wet algae of algae that quivers), produces the floating algae water extract that quivers.
The preparation of the floating algae water extract that quivers of embodiment 3
S1, the fresh floating wet algae of the algae that quivers is taken, is to freeze 10 hours under conditions of -40 DEG C in temperature, room temperature is melted 12 hours,
Multigelation 3 times, centrifuges 15min under conditions of rotating speed is 5000r/min, supernatant and precipitation is separated, take precipitation 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 4 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 15min under conditions of 5000r/min, supernatant and precipitation are separated, take precipitation standby；
S3, precipitation 2 times described in step S2 is washed with one-stage water wash, centrifuged under conditions of rotating speed is 10000r/min
15min, supernatant and precipitation are separated, supernatant is removed, and one-level is added by wet algae and the solid-to-liquid ratio 1g/mL of one-level water into precipitation
Water is extracted 5 times under conditions of temperature is 70 DEG C, each 60min, and 10min is centrifuged under conditions of rotating speed is 10000r/min,
Supernatant and precipitation are separated, supernatant is taken, plus it is 1gA/ that one-level water, which adjusts it relative to the concentration of floating algae (wet algae) weight of quivering,
ML (A represents the floating wet algae of algae that quivers), produces the floating algae water extract that quivers.
Floating influence of the algae water extract to Caenorhabditis elegans survival rate under oxidative stress conditions of quivering of embodiment 4
Paraquat, scientific name methyl viologen is a kind of strong oxidizer, can induce a large amount of generation active oxygen radicals in vivo
(ROS) body is in oxidative stress environment, and then early ageing phenomenon occur.
The algae water extract that quivers is floated made from Example 1, the wild type at adult initial stage is added to by final concentration 300mgA/mL
In Caenorhabditis elegans (N2) (being provided by Univ Minnesota-Twin Cities USA's Caenorhabditis elegans hereditary information collection), control group
Isometric S Medium solution is added, is placed in after 20 DEG C of culture 24h, the paraquat for adding final concentration of 50mM carries out the oxidation side of body
Compel modeling, the ratio of Caenorhabditis elegans survival is then counted every 12h, until it is all dead.Caenorhabditis elegans is deposited
Motility rate 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 the 300mgA/mL floating algae water extract that quivers can significantly extend beautiful hidden under oxidative stress conditions
The average life span of rhabditida, improves its survival rate, illustrates that the floating algae water extract that quivers that the present invention is provided can be alleviated by the oxidation side of body
Early ageing phenomenon caused by compeling, with significant antioxidation.
Floating influence of the algae water extract to ROS levels in Caenorhabditis elegans body of quivering of embodiment 5
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
The algae water extract that quivers, floating algae water extraction of quivering are floated made from L4 phases Wild-type C. elegans (N2) and embodiment 1 after stepization
The final concentration 100mgA/mL of thing, control group adds the L4 phases Wild-type C. elegans (N2) after synchronizing and floating with test group
The isometric S Medium solution of the algae water extract that quivers, is placed in after 20 DEG C of culture 24h, and paraquat is added per hole to final concentration of
2mM, continues to cultivate after 48h, collects each group Caenorhabditis elegans, it is homogenized under condition of ice bath respectively, by homogenate 4
DEG C, 5min is centrifuged under the conditions of 10000g, collect supernatant, final concentration of 50 μM of DCFH-DA probes (purchase added into supernatant
In Sigma companies, No. CAS：2044-85-1) solution, using fluorescence microplate reader in 485nm excitation wavelengths and 538nm launch wavelengths
Lower fluorescence intensity, room temperature detection 2h, is surveyed once, as a result as shown in Figure 2 per 10min.
Fig. 2 results are shown, compared with control group, after floating algae water extract feeding of quivering, ROS water in Caenorhabditis elegans body
It is flat to be remarkably decreased.Result of the test shows that the floating algae water extract that quivers that the present invention is provided can promote internal ROS removing, reduces body
Interior ROS levels, with significant oxidation resistance.
Floating influence of the algae intracellular fluid to Caenorhabditis elegans survival rate under oxidative stress conditions of quivering of embodiment 6
Float and quiver algae intracellular fluid made from Example 1, the wild of adult initial stage is added separately to by final concentration 100mgA/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 3.
Fig. 3 results show that liquid energy enough extends the life span of Caenorhabditis elegans under oxidative stress conditions in the floating gonidium that quivers,
Improve its survival rate.Result of the test shows that liquid energy enough alleviates the early ageing as caused by oxidative stress in the floating gonidium that quivers of the invention provided
Phenomenon, with significant antioxidation.
Floating influence of the algae intracellular fluid to Caenorhabditis elegans average life span under natural conditions of quivering of embodiment 7
Final concentration of 100mgA/mL embodiment is added into the L4 phases Wild-type C. elegans (N2) after synchronization
The algae intracellular fluid that quivers is floated made from 1, control group adds isometric S Medium solution, is placed in 20 DEG C of cultures, is then united every 1 day
The survival condition of Caenorhabditis elegans is counted, until it is all dead.The life-span of Caenorhabditis elegans represents with survivorship curve, as a result
Analyzed with Kaplan-Meier methods.As a result it is as shown in Figure 4.
Fig. 4 results are shown, compared with control group, and liquid energy enough extends Caenorhabditis elegans under natural conditions in the floating gonidium that quivers
Average life span.Result of the test shows that the floating algae intracellular fluid that quivers that the present invention is provided has the effect of anti-aging process, i.e., with anti-
Aging is acted on.
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 (6)
1. the floating algae water extract that quivers of one kind, it is characterised in that the preparation method of the floating algae water extract that quivers comprises the following steps：
S1, the fresh floating wet algae of the algae that quivers is taken, be to freeze 10~12 hours under conditions of -20 DEG C~-40 DEG C in temperature, room temperature is melted
10~12 hours, multigelation 3~5 times centrifuged 5~15min under conditions of rotating speed is 5000r/min~10000r/min,
Supernatant and precipitation are separated, 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 10~15min of centrifugation under conditions of 5000r/min~10000r/min, and supernatant and precipitation are separated, take precipitation standby；
S3, precipitation 1~3 time described in step S2 is washed with one-stage water wash, in the condition that rotating speed is 5000r/min~10000r/min
10~15min of lower centrifugation, by supernatant and precipitation separate, remove supernatant, into precipitation add one-level water temperature be 70 DEG C~
Extracted 3~5 times under conditions of 100 DEG C, 30~60min every time, under conditions of rotating speed is 5000r/min~10000r/min from
5~15min of the heart, supernatant and precipitation are separated, supernatant is taken, and produce the floating algae water extract that quivers.
2. the floating algae water extract that quivers according to claim 1, it is characterised in that the preparation side of the floating algae water extract that quivers
Ethanol is added by the solid-to-liquid ratio 1g/mL of wet algae and ethanol carry out ultrasonic extraction in method, in step S2.
3. the floating algae water extract that quivers according to claim 1, it is characterised in that the preparation side of the floating algae water extract that quivers
In method, the frequency of ultrasonic extraction is 35~40kHz in step S2, and temperature is 4 DEG C.
4. the floating algae water extract that quivers according to claim 1, it is characterised in that the preparation side of the floating algae water extract that quivers
In method, the time of each ultrasonic extraction is 10~30min in step S2.
5. the floating algae water extract that quivers according to claim 1, it is characterised in that the preparation side of the floating algae water extract that quivers
One-level water is added by the solid-to-liquid ratio 1g/mL of wet algae and one-level water carry out heating extraction in method, in step S3.
6. the floating algae water extract that quivers according to claim 1 is in health food and medicine with anti-oxidation function is prepared
Application.
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