CN108969606A - Application of the composite plant nutrient in preparation cellular immunity preparation - Google Patents
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K35/64—Insects, e.g. bees, wasps or fleas
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Abstract
This application involves a kind of application of composite plant nutrient in preparation cellular immunity preparation, the composite plant nutrient, the vegetable oil of the grape seed extract of lycopene oleoresin, 19%~29% mass percentage, the beeswax of 3%~5% mass percentage and 24%~50% mass percentage including 28%~42% mass percentage, and the mass percentage of lycopene is 5%~8% in lycopene oleoresin.Above-mentioned composite plant nutrient, which can be prepared, not only has good antitumous effect, but also can effectively facilitate the cellular immunity preparation of cellular immune function.
Description
Technical field
The present invention relates to biological medicine health fields, more particularly to a kind of composite plant nutrient in preparation cellular immunity
Application in preparation.
Background technique
Tumour is one of the disease of most serious of being detrimental to health, and disease incidence tends to be young in rising year by year trend
Change.The treatment of tumour at present is main or uses traditional treatment method, such as operation, radiotherapy, chemotherapy and complex treatment.Chemotherapy begins
In in the 1940s, after decades of development, the sum of anti-tumor drug has reached hundreds of, and is matched with these material medicines
The various anti-tumor drugs of system are then up to more than 1000 kinds.Current market sales of anti-tumor drug is mostly the synthetic drug of structure determination
Object can also generate serious damage to other normal tissue cells while killing tumor tissues.
Summary of the invention
Based on this, it is necessary to provide a kind of application of composite plant nutrient in preparation cellular immunity preparation, can prepare
Not only there is good antitumous effect, but also the cellular immunity preparation of cellular immune function can be effectively facilitated.
A kind of application of composite plant nutrient in preparation cellular immunity preparation, the composite plant nutrient, with matter
Measure percentage composition meter, including following components:
The mass content of lycopene is 5%~8% in the lycopene oleoresin.
The composite plant nutrient in one of the embodiments, in terms of mass percentage, including following components:
In one of the embodiments, the vegetable oil be selected from medium chain triglyceride, soybean oil, sunflower oil, olive oil,
At least one of safflower oil, rice bran oil and maize germ oil.
The vegetable oil is the combination of medium chain triglyceride and soybean oil in one of the embodiments, and the middle chain is sweet
The mass ratio of oily three esters and soybean oil is 1:(1.7~1.9).
In one of the embodiments, the human body of the composite plant nutrient daily measure for 14.3mg/kg~
357.5mg/kg。
In one of the embodiments, the human body of the composite plant nutrient daily measure for 14.3mg/kg~
71.5mg/kg。
The cellular immunity preparation is that tumour is inhibited to increase and promote cellular immune function in one of the embodiments,
Preparation.
The tumour is melanoma tumors in one of the embodiments,.
The cellular immune function is that T cell is proliferated performance and NK cell activity in one of the embodiments,.
The dosage form of the cellular immunity preparation is soft capsule or oral solution in one of the embodiments,.
Application of the above-mentioned composite plant nutrient in preparation cellular immunity preparation can be prepared not only with good anti-swollen
Tumor effect, and the cellular immunity preparation of cellular immune function can be effectively facilitated.
Detailed description of the invention
Fig. 1 is influence diagram of the composite plant nutrient to tumor-bearing mice weight of an embodiment；
Fig. 2 is influence diagram of the composite plant nutrient to tumor-bearing mice gross tumor volume of an embodiment；
Fig. 3 be an embodiment composite plant nutrient to tumor-bearing mice tumour treated tissue picture；
Fig. 4 is influence diagram of the composite plant nutrient to tumor-bearing mice tumor quality of an embodiment；
Fig. 5 is influence diagram of the composite plant nutrient to tumor-bearing mice T cell proliferative capacity of an embodiment.
Specific embodiment
To facilitate the understanding of the present invention, below will to invention is more fully described, and give it is of the invention compared with
Good embodiment.But the invention can be realized in many different forms, however it is not limited to embodiment described herein.Phase
Instead, purpose of providing these embodiments is makes the disclosure of the present invention more thorough and comprehensive.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein "and/or" includes one or more phases
Any and all combinations of the listed item of pass.
Application of the composite plant nutrient of one embodiment in preparation cellular immunity preparation, the composite plant nutrient
The grape pip of lycopene oleoresin, 19%~29% mass percentage including 28%~42% mass percentage extracts
The vegetable oil of object, the beeswax of 3%~5% mass percentage and 24%~50% mass percentage, and lycopene oil
The mass percentage of lycopene is 5%~8% in resin.
Further, which includes the lycopene oil tree of 28%~38.5% mass percentage
Rouge, the grape seed extract of 19%~26% mass percentage, 3.5%~5% mass percentage beeswax and
The vegetable oil of 30.5%~48% mass percentage.
Wherein, vegetable oil is selected from medium chain triglyceride, soybean oil, sunflower oil, olive oil, safflower oil, rice bran oil and jade
At least one of rice germ oil.
Further, vegetable oil is the combination of medium chain triglyceride and soybean oil, the matter of medium chain triglyceride and soybean oil
Amount is than being 1:(1.7~1.9).
In the present embodiment, the human body of above-mentioned composite plant nutrient daily measure for 14.3mg/kg~
357.5mg/kg.Further, the human body of above-mentioned composite plant nutrient is daily measured as 14.3mg/kg~71.5mg/kg.
Further, cellular immunity preparation is that tumour is inhibited to increase and promote the preparation of cellular immune function.
Wherein, tumour is melanoma.Cellular immune function is that T cell is proliferated performance and NK cell activity.
Further, the dosage form of cellular immunity preparation is soft capsule or oral solution.
Why above-mentioned composite plant nutrient can be used for preparing cellular immunity preparation, and pharmacological action is as follows:
Lycopene: lycopene belongs to carotenoid, molecular formula C40H56, it is by 11 conjugated double bonds and 2
The height unsaturation straight chain type hydrocarbon compound that unconjugated carbon-to-carbon double bond is constituted is fat-soluble pigment, is mainly derived from kind
The plants such as eggplant, watermelon, guava, purple grape fruit, pawpaw and carrot, wherein content highest in tomato.The study found that tomato
Red pigment has anti-oxidant, anti-aging, inhibits tumour, reduce the diseases such as angiocarpy, improve the immunity of the human body, delaying sclerotin crisp
The effect of the diseases such as loose, anti-radiation, prevention prostate.
Lycopene is most one of powerful antioxidant for being found in the plant of current nature, its effect is vitamin
100 times of E, in carotenoid, lycopene understands superoxide radical (O2 -) speed be it is most fast, to DPPH's
Scavenging activity is 5.32 times of alpha-carotene respectively, is 22.73 times of lutein.In addition, lycopene also have it is very strong sudden
It goes out the ability of singlet oxygen.Lycopene can inhibit oxidation free radical, reduce tumorigenic danger by antioxidation
Property, it has proven to most effective to prevention prostate cancer, lung cancer, gastric cancer, also has to cancer of pancreas, cancer of the esophagus, carcinoma of mouth etc. preferable pre-
Anti- effect can be allowed to reduce, delay to spread, especially prostate cancer to established tumour.
Grape seed extract: the polyphenols in grape pip containing about 5%~8%, have catechin, epicatechin and
The various procyanidine (PA) that gallic acid is made into, or be glucosidase procyanidins (OPC).Procyanidine has stronger antioxygen
Change effect, can inhibit damage of the free radical to human body, have anti-agings, beauty and skin care, anti-radiation, anti-vision degeneration and resist
The effect of artery sclerosis and thrombosis can also prevent a variety of diseases such as arthritis, cancer, diabetes and heart disease.
However although lycopene and grape seed extract have significant antioxidation, due to by factors shadow
It rings, causes its bioavailability low.Such as alkali, temperature, illumination (especially ultraviolet light), oxidant, acid, reducing agent and certain gold
Belong to ion (Fe3+、Fe2+、Al3+、Cu2+) cis-trans isomerization and the oxidative degradation of lycopene can be promoted；PH, temperature, light, metal
The factors such as ion, additive can also produce bigger effect the stability of anthocyanidin, therefore can be absorbed into blood and play a role
Ratio very little, thus reduce the performance of antioxidation.
The composite plant nutrient for being used to prepare cellular immunity preparation of the application, lycopene therein use tomato red
The form of vegetable oil resin, and its stability is improved by the content of lycopene in control lycopene oleoresin, to mention
It is high its in the absorptivity of human body, then combine itself and grape seed extract, beeswax and vegetable oil, and use special proportion,
The composite plant nutrient is further increased in the absorptivity of human body, the performance of antioxidation is improved, can prepare and not only have
There is good antitumous effect, and the cellular immunity preparation of cellular immune function can be effectively facilitated.
The following are specific embodiments.
Embodiment 1
Composite plant nutrient is used to prepare cellular immunity preparation, which includes 28% mass percentage
The lycopene oleoresin of content, the grape seed extract of 19% mass percentage, 5% mass percentage beeswax,
The medium chain triglyceride of the soybean oil of 20% mass percentage and 28% mass percentage, and in lycopene oleoresin
The mass content of lycopene is 6%.
(1) its tumor-inhibiting action to B16 transplantable tumor mouse is tested:
1.1B16 subcutaneously inhibiting mouse model preparation
Conventional method recovery B16 melanoma cells, expand culture, and pancreatin digestion collects cell, is prepared into 1x107/mL
It is subcutaneous to be inoculated in C57BL/6 right side of mice armpit with 0.1mL for cell suspension.
1.2 animal packets and processing.
The diameter that vernier caliper measurement mice-transplanted tumor is used after inoculation 5 days, to tumour growth to 0.2cm3After be randomly divided into 4
Group: lotus knurl control group, low dose group (0.26mg/g/day), middle dose group (1.3mg/g/day), high dose group (5.6mg/g/
Day), every group 6.It is administered by the way of stomach-filling, one time a day, lotus knurl control group is with the physiological saline stomach-filling of equivalent.Each group is small
After mouse successive administration 15 days.
Influence of 1.3 composite plant nutrients to tumor-bearing mice weight
It is administered by the way of described in 1.2, weighs within every 2 days the weight and record of mouse, detect various dose composite plant
Influence of the nutrient to tumor-bearing mice weight, experimental result is shown in Fig. 1.
As shown in Figure 1, it can be seen that the composite plant of high, medium and low three dosage from the change curve of tumor-bearing mice weight
Nutrient has the effect of promoting mouse weight to increase, but all without significant difference.
Influence of 1.4 composite plant nutrients to tumor-bearing mice gross tumor volume
After being administered 6 days by the way of described in 1.2, the every 2 days length and widths using vernier caliper measurement tumour, and using swollen
Knurl accumulates calculation formula: TV=0.52 × a × b2(a, b respectively indicate length and width), calculates relative tumour volume, experimental result is shown in
Fig. 2.
As shown in Figure 2, it can be seen that the compound of low dosage and middle dosage from the change curve of the gross tumor volume of tumor-bearing mice
Nutrient for plants can effectively inhibit the further growth of gross tumor volume, significant difference and be in dose dependent；High dose
Composite plant nutrient acts on the growth unrestraint of tumour.
Influence of 1.5 composite plant nutrients to tumor-bearing mice tumor quality
After being administered 14 days by the way of described in 1.2, the careful tumour for removing tumor-bearing mice takes pictures and weighs, experiment knot
Fruit sees Fig. 3 and Fig. 4.
As can be seen from figs. 3 and 4 can be seen that low dose from the tumour photo of tumor-bearing mice and the change curve of tumor quality
The composite plant nutrient of amount and middle dosage can effectively reduce tumor quality, significant difference and be in dose dependent；High agent
The composite plant nutrient of amount is on tumor quality without influence.
In conclusion low dosage and the composite plant nutrient of middle metering have the effect of preferably inhibiting tumour.
(2) its influence to tumor-bearing mice cellular immune function is tested:
Influence of 2.1 composite plant nutrients to tumor-bearing mice T cell proliferative capacity
In-vitro multiplication activity of the present embodiment using CCK-8 living cells detection kit detection mouse T cell, specific experiment
Step are as follows:
(I) neck that breaks puts to death mouse, impregnates 5min with 75% alcohol.It is sterile in super-clean bench to take spleen, it is put into sterilized petri dishes,
1mL RPMI-1640 is added to grind spleen tissue, 200 mesh copper mesh are crossed after grinding, moves into centrifuge tube, then adds 1mL
RPMI-1640 rinses plate, crosses 200 mesh copper mesh, moves into same centrifuge tube, abandons supernatant after 2000rpm centrifugation 5min.
(II) after 2mL erythrocyte cracked liquid resuspension cell is added, 2-3min is cracked, adds the fetal calf serum containing 4% in equal volume
RPMI-1640 culture medium terminate cracking reaction, 2000rpm is discarded supernatant after being centrifuged 5min, is resuspended with 1640 culture mediums thin
Born of the same parents.
(III) viable count is carried out by trypan blue staining, guarantees that viable count is not less than 95%, then adjustment is thin
Born of the same parents' concentration is to 2*106/mL。
(IV) B cell is removed: with 37 DEG C of mineral wool column slow filtration cell suspensions of sterilizing, to remove B cell, with containing
Cell is resuspended in 2% serum, 1640 culture medium.
(V) 96 cell plates, every 100 μ L of hole is added in cell suspension.Then ConA final concentration of 5 is added in Positive control wells
μ g/mL, in 37 DEG C, 5%CO268h is cultivated in incubator.
(VI) every hole is added the reagent WST-8 that CCK-8 kit is matched, and concentration is 5 μ g/mL, 20 holes μ L/, in 37 DEG C,
4h is cultivated in 5%CO2 incubator.
(VII) the OD value in 570nm is measured with microplate reader.
Experimental result is shown in Fig. 5.
As shown in Figure 5, it can be seen that the composite plant battalion of low dosage and high dose from the histogram of T cell proliferation activity
Feeding element can promote the intracorporal T cell proliferation of mouse, and difference is extremely significant, and has dose dependent.These results indicate that compound
For nutrient for plants while inhibiting tumour, not only without inhibiting lymphocyte immunologic function, but also it is swollen to enhance zoografting
Lymphocyte immunologic function in tumor model.
Influence of 2.2 composite plant nutrients to tumor-bearing mice NK cell activity
Isolated NK cell and target cell YAC-1 cell are co-cultured, the lactic acid by detection target cell release is de-
The amount of hydrogen enzyme (LDH) detects the killing ability of NK cell.Specific experimental procedure is as follows:
(1) preparation of LDH matrix liquid
5 × 10-2mol/L of lithium lactate
Nitro tetrazolium chloride (INT) 6.6 × 10-4mol/L
Phenazine Dimethyl Sulfate (PMS) 2.8 × 10-4mol/L
Oxidized coenzyme I (NAD) 1.3 × 10-3mol/L
Mentioned reagent is dissolved in the Tris-HCl buffer of 0.2mol/L (pH8.2)
(2) passage (YAC-1 cell) of target cell
Target cell is subjected to secondary culture for 24 hours before experiment.It is washed 3 times using preceding with Hank's liquid, is trained completely with RPMI1640
It is 4 × 10 that nutrient solution, which adjusts cell concentration,5A/mL.
(3) preparation (effector cell) of splenocyte suspension
It is sterile to take spleen, it is placed in the small plate for filling appropriate sterile Hank's liquid, gently spleen is ground with tweezers, list is made
Cell suspension.Spleen is ground through 200 mesh net filtrations, or with 4 layers of gauze, or is washed 2 times with Hank's liquid, is centrifuged 10min every time
(1000r/min).It abandons supernatant to bounce cytoplasm, is added 0.5mL aqua sterilisa 20 seconds, 0.5mL 2 is added after splitting erythrocyte
Times Hank ' s liquid and 8mL Hank ' s liquid, 1000rpm, 10min centrifugation, the RPMI1640 with 1mL containing 10% calf serum are complete
Culture solution is resuspended, and with platform phenol orchid dyeing counting viable count (should be 95% or more), finally uses RPMI1640 complete culture solution tune
Whole cell concentration is 2 × 107/mL。
(4) NK cytoactive detection
Target cell and each 100 μ L of effector cell (effect target ratio 50:1) are taken, is added in U-shaped 96 well culture plate；Target cell is natural
Relief hole adds target cell and each 100 μ L of culture solution, and target cell maximum relief hole adds target cell and 1%NP40 or 2.5%Triton
Each 100 μ L；Above-mentioned items are all provided with three multiple holes, cultivate 4h in 37 DEG C, 5%CO2 incubator, then by 96 well culture plates with
1500r/min is centrifuged 5min, and every hole is drawn in 100 μ L horizontalization bottom of supernatant, 96 well culture plate, while 100 μ L of LDH matrix liquid is added,
3min is reacted, the HCl30 μ L of 1mol/L is added in every hole, measures OD value (OD) at microplate reader 490nm, experimental result is shown in Table
1。
Table 1
| Composite plant nutrient dosage mg/g | Experiment mice NK cell activity (U/L) |
| Control | 24.12±5.63 |
| 6.5 | 25.16±0.79 |
| 1.3 | 41.14±8.56 |
| 0.26 | 35.09±5.87 |
As it can be seen from table 1 the composite plant nutrient of low dosage and high dose can enhance the intracorporal NK cell of mouse
Killing activity, significant difference and have dose dependent.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of application of composite plant nutrient in preparation cellular immunity preparation, which is characterized in that the composite plant battalion
Element is supported, in terms of mass percentage, including following components:
The mass content of lycopene is 5%~8% in the lycopene oleoresin.
2. application of the composite plant nutrient according to claim 1 in preparation cellular immunity preparation, which is characterized in that
The composite plant nutrient, in terms of mass percentage, including following components:
3. application of the composite plant nutrient according to claim 1 or 2 in preparation cellular immunity preparation, feature exist
In the vegetable oil is selected from medium chain triglyceride, soybean oil, sunflower oil, olive oil, safflower oil, rice bran oil and maize germ
At least one of oil,.
4. application of the composite plant nutrient according to claim 3 in preparation cellular immunity preparation, which is characterized in that
The vegetable oil is the combination of medium chain triglyceride and soybean oil, and the mass ratio of the medium chain triglyceride and soybean oil is 1:
(1.7~1.9).
5. application of the composite plant nutrient according to claim 1 in preparation cellular immunity preparation, which is characterized in that
The human body of the composite plant nutrient is daily measured as 14.3mg/kg~357.5mg/kg.
6. application of the composite plant nutrient according to claim 1 in preparation cellular immunity preparation, which is characterized in that
The human body of the composite plant nutrient is daily measured as 14.3mg/kg~71.5mg/kg.
7. application of the composite plant nutrient according to claim 1 in preparation cellular immunity preparation, which is characterized in that
The cellular immunity preparation is that tumour is inhibited to increase and promote the preparation of cellular immune function.
8. application of the composite plant nutrient according to claim 7 in preparation cellular immunity preparation, which is characterized in that
The tumour is melanoma tumors.
9. application of the composite plant nutrient according to claim 7 in preparation cellular immunity preparation, which is characterized in that
The cellular immune function is that T cell is proliferated performance and NK cell activity.
10. application of the composite plant nutrient according to claim 1 in preparation cellular immunity preparation, feature exist
In the dosage form of the cellular immunity preparation is soft capsule or oral solution.
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| CN110917333A (en) * | 2019-11-15 | 2020-03-27 | 上海瑞思德生物科技有限公司 | Application of nerve stem cell active factor compound nano liposome and preparation thereof |
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