CN104106763B - Polygonatum polysaccharide has the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation - Google Patents
Polygonatum polysaccharide has the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation Download PDFInfo
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Classifications
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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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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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- Sustainable Development (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention discloses polygonatum polysaccharide and has the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, wherein polygonatum polysaccharide prepares preferably by following method: take sealwort as raw material, through containing boiling water extraction, Deproteinization, dialysis and Ethanol Treatment operation prepare, described polygonatum polysaccharide has the effect regulating tumor microenvironment, polygonatum polysaccharide in the present invention can be generated by Tumor suppression new vessel, immunosuppression, anti-oxidant and suppress many approach such as colorectal carcinoma fibroblastic growth to regulate tumor microenvironment, show that it has good auxiliary suppression colorectal carcinoma function of tumor.
Description
Technical field
The invention belongs to polysaccharide technical field, it is specifically related to a kind of polygonatum polysaccharide, in preparation, there is the application in the auxiliary functional food suppressing colorectal carcinoma effect.
Background technology
Malignant tumour is the serious disease of current harm humans health, chemotherapy is one of main method being applied to malignant tumour whole body therapeutic, but major part anti-tumor medicine thing is while killing tumour cell, also major injury normal cell, the antitumor drug therefore finding high-efficiency low-toxicity is significant. And research in recent years finds, polysaccharide just have significantly anti-tumor activity and the toxic side effect of human body is relatively little, the bioactivity research of polysaccharide has been become at medical field the focus of domestic and international experts and scholars research. In recent years find that some polysaccharide and mixture thereof are to multiple disease, as immunologic derangement, cancer, diabetes, hypertension, hepatitis, thrombus, pneumonia, virus etc. all have significant curative effect, and participate in the adjustment of the various biological phenomena of cell. A large amount of pharmacology and clinical trial show that polysaccharide compound is excellent immunomodulator, phagocytic function, the induction that can increase scavenger cell and white corpuscle produce cell necrosis factor (TNF) and leukin, thus improve the immunologic function of human body, and to the almost non-toxic side effect of normal cell.
Sealwort is a kind of conventional Chinese medicine, and sealwort is used as medicine with rhizome. There is boosting qi and nourishing yin, strong spleen, moistening lung, kidney benefiting functions, being used for the treatment of weakness of the spleen and the stomach, fatigue and asthenia, dry food is few, deficiency syndrome of the lung cough caused by dryness, essence insufficiency of blood, interior heat such as is quenched one's thirst the disease, and there is no at present is that polysaccharide prepared by raw material taking sealwort, and the product development for a kind of auxiliary suppression colorectal carcinoma aspect.
Summary of the invention
It is an object of the invention to provide polygonatum polysaccharide, in preparation, there is the application in the auxiliary functional food suppressing colorectal carcinoma effect.
Wherein said auxiliary suppression colorectal carcinoma effect refers to by Tumor suppression new vessel generation, immunosuppression, anti-oxidant and suppress the effect such as colorectal carcinoma fibroblastic growth to regulate tumor microenvironment, reaches auxiliary effect suppressing colorectal carcinoma tumour.
As one embodiment of the present invention, polygonatum polysaccharide in the present invention prepares preferably by following method: take sealwort as raw material, through preparing containing boiling water extraction, Deproteinization, dialysis and Ethanol Treatment operation, described polygonatum polysaccharide has the effect regulating tumor microenvironment.
As the present invention one preferred embodiment, polygonatum polysaccharide of the present invention prepares especially by following method: take sealwort as raw material, after repeatedly extracting with boiling water, united extraction liquid, extracting solution is concentrated to obtain concentrated solution, concentrated solution adds trichloroacetic acid solution and carries out Deproteinization process, undertaken reaction solution after Deproteinization neutralizing, pricking a bag dialysis treatment, undertaken reaction solution in dialysis tubing concentrating, centrifugal treating, get supernatant liquor to add ethanol and carry out alcohol precipitation, drying treatment, obtain polygonatum polysaccharide.
The present invention is in the preparation process of polygonatum polysaccharide, and the concrete parameter of each operation is preferably as follows:
During boiling water extraction, extraction time is 1 ~ 5 time, and when extracting, the consumption of boiling water is 5 ~ 20 times of sealwort total mass every time, and each extraction time is 1.5 ~ 5h.
During boiling water extraction, the consumption of boiling water is preferably 5 ~ 20 times of sealwort total mass, and extraction time is preferably 3 times, and when wherein first time extracts, the consumption of boiling water is preferably 10 ~ 20 times of raw material sealwort total mass, extracts 2 ~ 5h; When second time is extracted, the consumption of boiling water is preferably 10 ~ 20 times of raw material sealwort total mass, extracts 1.5 ~ 5h; Third time, when extracting, the consumption of boiling water was preferably 10 ~ 20 times of raw material sealwort total mass, extracted 1.5 ~ 5h.
Extracting solution is preferably concentrated to former extracting solution cumulative volume about 1/10 concentrated solution.
The mass percentage of trichloroacetic acid solution of the present invention is preferably 35%, and the Deproteinization treatment time is preferably 3 ~ 6h, and the volume of trichloroacetic acid solution is preferably identical with the volume of concentrated solution.
Can adopt during neutralizing treatment mass percentage be 10% aqueous sodium hydroxide solution.
The dialysis tubing of molecular weight cut-off 3500Da is preferably adopted to carry out dialysis treatment when pricking bag dialysis treatment.
Ethanol carries out preferably adopting volumn concentration to be preferably the aqueous ethanolic solution of 70 ~ 95% when alcohol precipitation processes.
Concentrate and can adopt multiple condensing mode of the prior art, wherein best with vacuum concentration. During vacuum concentration, temperature is preferably 40 DEG C.
Polygonatum polysaccharide in the present invention, it is possible to directly oral, it is also possible to make said multiple formulation in pharmaceutics, such as capsule type, tablet, pulvis, granule or oral liquid etc., has auxiliary effect suppressing colorectal carcinoma tumour after edible.
Therefore, the formulation of functional food of the present invention is preferably capsule type, tablet, pulvis, granule or oral liquid etc.
Compared with prior art, tool of the present invention has the following advantages:
(1) polygonatum polysaccharide in the present invention, its purity and activity height, can passing through Tumor suppression new vessel generation, immunosuppression, anti-oxidant and suppress many approach such as colorectal carcinoma fibroblastic growth to regulate tumor microenvironment, it has good auxiliary suppression colorectal carcinoma function of tumor;
(2) preparation method of the polygonatum polysaccharide in the present invention, technique simple and stable is efficient, is applicable to suitability for industrialized production, and cost is low;
(3) polygonatum polysaccharide in the present invention, it is possible to for the preparation of having the auxiliary functional food suppressing colorectal carcinoma tumor function.
Accompanying drawing explanation
Fig. 1 is the activity of the luciferase with BRE reporter gene that polygonatum polysaccharide can suppress BMP2 to induce in the embodiment of the present invention 4;
Fig. 2 be in the embodiment of the present invention 5 polygonatum polysaccharide TGF can be suppressed beta induced with SMAD2/3/4 reporter gene activity;
Fig. 3 is the activity that in the embodiment of the present invention 6, polygonatum polysaccharide can suppress the luciferase with NF-�� B reporter gene;
Fig. 4 is the activity of the luciferase with HRE reporter gene that polygonatum polysaccharide can suppress Cocl2 to induce in the embodiment of the present invention 7;
Fig. 5 is that the embodiment of the present invention 8 polygonatum polysaccharide can suppress colorectal carcinoma fibroblastic growth.
Embodiment
Below in conjunction with drawings and Examples, the present invention will be further described, but does not limit the present invention in any form.
The each raw material adopted in following examples, if no special instructions, is commercially available prod.
Embodiment 1
Polygonatum polysaccharide in the present embodiment, prepares by the following method: get sealwort raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 20 times of raw material total mass, boils 5h and filters, when second time is extracted, the consumption of water is 20 times of raw material total mass, boils 3h and filters, third time, when extracting, the consumption of water was 10 times of raw material total mass, boils 2h and filters, merging filtrate, and vacuum concentration is to about the 1/10 of original volume, lower same. concentrated solution is carried out trichoroacetic acid(TCA) removing protein process, add 30% trichoroacetic acid(TCA) (mass percent) of same volume (lower same), treatment time is after 4h, use 10%NaOH(mass percent) neutralization, lower same, after neutralization, reaction solution is placed in dialysis tubing and pricks bag dialysis, during dialysis, the molecular weight cut-off of dialysis tubing is about 3500Da usually, lower same, then liquid in dialysis tubing is concentrated, centrifugal, get 85% aqueous ethanolic solution (volumn concentration) that supernatant liquor adds 3 ~ 4 times of volumes and carry out alcohol precipitation process, taking precipitate is through containing vacuum-drying (about 40 DEG C, lower same) process to obtain polygonatum polysaccharide component.
Embodiment 2
Polygonatum polysaccharide in the present embodiment, prepares by the following method: get sealwort raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 15 times of sealwort raw material total mass, boils 5h and filters; When second time is extracted, the consumption of water is 15 times of sealwort raw material total mass, boils 3h and filters; Third time, when extracting, the consumption of water was 10 times of sealwort raw material total mass, boils 3h and filters, merging filtrate, vacuum concentration. Concentrated solution is carried out trichoroacetic acid(TCA) removing protein process, add 25% trichoroacetic acid(TCA) (mass percent) of same volume, treatment time is after 3h, use 10%NaOH(mass percent) neutralization, after neutralization, reaction solution is placed in dialysis tubing and pricks bag dialysis, then liquid in dialysis tubing is concentrated, centrifugal, getting 70% aqueous ethanolic solution (volumn concentration) that supernatant liquor adds 3 ~ 4 times of volumes and carry out alcohol precipitation process, taking precipitate is through obtaining polygonatum polysaccharide component containing vacuum drying treatment.
Embodiment 3
Polygonatum polysaccharide in the present embodiment, prepares by the following method: get sealwort raw material, adopts boiling water extraction, and when first time extracts, the consumption of water is 10 times of sealwort raw material total mass, boils 3h and filters, when second time is extracted, the consumption of water is 20 times of sealwort raw material total mass, boils 3 filtration h, third time is when extracting, the consumption of water is 10 times of sealwort raw material total mass, boil 5h to filter, merging filtrate, vacuum concentration, concentrated solution is carried out trichoroacetic acid(TCA) removing protein process, add 15% trichoroacetic acid(TCA) (mass percent) of same volume, treatment time is after 5h, use 10%NaOH(mass percent) neutralization, after neutralization, reaction solution is placed in dialysis tubing and pricks bag dialysis, then liquid in dialysis tubing is concentrated, centrifugal, get 95% aqueous ethanolic solution (volumn concentration) that supernatant liquor adds 3 ~ 4 times of volumes and carry out alcohol precipitation process, taking precipitate is through obtaining polygonatum polysaccharide component containing vacuum drying treatment.
Embodiment 4 polygonatum polysaccharide can suppress the activity of the luciferase with BRE reporter gene that BMP2 induces
C2C12 cell strain is purchased from Chinese Academy of Sciences's cell bank, and C2C12-pGF1-BRE cell strain is built by Shanghai Pharmaceutical Inst., Chinese Academy of Sciences's fourth laboratory of chatting about, and is stored in liquid nitrogen. C2C12-pGF1-BRE cell, with containing 10%Gibco foetal calf serum DMEM culture medium culturing, is placed in 37 DEG C of saturated humidities, containing 5%CO2Thermostat container in cultivate, with 1 �� 104The density of cells/well is inoculated in 96 orifice plates, every hole 100 �� L. Sucking 60 �� L substratum after pasting wall 24h, add the sample 50mL (polygonatum polysaccharide in embodiment 1) configuring respective concentration, final concentration is respectively 0.5mg/mL, 0.1mg/mL, 0.02mg/mL, and 10 �� LBMP2 are adjusted to final concentration 200ng/mL. Establish blank group and the comparison of BMP2 group simultaneously. Suck substratum after administration 16h, the �� LReporterLysis1XBuffer that adds 20, the cell pyrolysis liquid 20 �� L of dissolving is transferred to white plate, adds 40 �� Lluciferase substrates, in 3 minutes, read plate, read RLU value. SPSS statistical software is adopted to carry out statistical procedures. Data all represent with all several �� standard deviations (.x �� s), compare and adopt independent sample t inspection, compare employing one-way analysis of variance between many groups between two groups.
Experimental result is as shown in Figure 1, polygonatum polysaccharide has the activity of the luciferase with BRE reporter gene suppressing BMP2 induction under 0.1mg/mL lower concentration, and this activity increases with polysaccharide concentration and significantly increases, show that this polysaccharide has dose-dependently in this activity. Known cancer new vessel is created in the generation development process of tumour and plays keying action, and BMP2 signal path plays keying action in tumor angiogenesis process. This polysaccharide can significantly suppress BMP2 signal path, shows that this polysaccharide has potential adjustment tumor microenvironment effect.
Embodiment 5 polygonatum polysaccharide can suppress TGF beta induced with SMAD2/3/4 reporter gene activity
HEK293T cell strain is purchased from Chinese Academy of Sciences's cell bank, and HEK293T-pGF1-SMAD2/3/4 cell strain is built by Shanghai Pharmaceutical Inst., Chinese Academy of Sciences's fourth laboratory of chatting about, and is stored in liquid nitrogen. HEK293T-pGF1-SMAD2/3/4 cell, with containing 10%Gibco foetal calf serum DMEM culture medium culturing, is placed in 37 DEG C of saturated humidities, containing 5%CO2Thermostat container in cultivate, with 2 �� 104The density of cells/well is inoculated in 96 orifice plates, every hole 100 �� L. Sucking 60 �� L substratum after pasting wall 24h, add the sample 50mL (polygonatum polysaccharide in embodiment 1) configuring respective concentration, final concentration is respectively 0.5mg/mL, 0.1mg/mL, 0.02mg/mL, and 10 �� LTGF �� 1 are adjusted to final concentration 50ng/mL. Establish blank group and TGF �� 1 group comparison simultaneously. Suck substratum after administration 16hr, the �� LReporterLysis1XBuffer that adds 20, the cell pyrolysis liquid 20 �� L of dissolving is transferred to white plate, adds 40 �� Lluciferase substrates, in 3 minutes, read plate, read RLU value. SPSS statistical software is adopted to carry out statistical procedures. Data all represent with all several �� standard deviations (.x �� s), compare and adopt independent sample t inspection, compare employing one-way analysis of variance between many groups between two groups.
Experimental result is as shown in Figure 2, polygonatum polysaccharide have under 0.5mg/mL lower concentration suppress TGF beta induced with SMAD2/3/4 signal path, and this activity increases with polygonatum polysaccharide concentration and significantly increases, show that this polysaccharide has dose-dependently in this activity. Known cancer new vessel is created in the generation development process of tumour and plays keying action, and the beta induced SMAD2/3/4 signal path of TGF plays keying action in tumor angiogenesis process. This polysaccharide can significantly suppress this signal path, shows that this polysaccharide has potential adjustment tumor microenvironment effect.
Embodiment 6 polygonatum polysaccharide can suppress the activity of the luciferase with NF-�� B reporter gene
THP-1 cell strain is purchased from Chinese Academy of Sciences's cell bank, and THP-1/pGF1-NF-�� B cell strain is built by Shanghai Pharmaceutical Inst., Chinese Academy of Sciences's fourth laboratory of chatting about, and is stored in liquid nitrogen. THP-1/pGF1-NF-�� B cell is with containing 10% foetal calf serum RPMI-1640 culture medium culturing. It is placed in 37 DEG C of saturated humidities, containing 5%CO2Thermostat container in cultivate, with 5 �� 104The density of cells/well is inoculated in 96 orifice plates, every hole 50 �� L volume. Add the polygonatum polysaccharide in given the test agent 50 �� L(embodiment 1), final concentration is respectively 0.5mg/mL, 0.Lmg/mL, 0.02mg/mL, sets up blank group and LPS control group simultaneously. Adding LPS solution 10 �� L, final concentration is 1 �� g/mL, is placed in 37 DEG C of saturated humidities, containing 5%CO2Thermostat container in overnight incubation. After overnight incubation, every sky adds 100 �� LBright-GloTMLuciferaseAssaySystem substrate, reads plate in microplate reader, reads RLU value.
SPSS statistical software is adopted to carry out statistical procedures. Data all represent with all several �� standard deviations (.x �� s), compare and adopt independent sample t inspection, compare employing one-way analysis of variance between many groups between two groups.
As shown in Figure 3, this polysaccharide has the activity suppressing NF-�� B signal path preferably to experimental result under 0.5mg/mL lower concentration. Known NF-�� B is the key nuclear factor participating in series of genes expression regulation, and it has vital role in the origin of cancer, development, angiogenesis and transfer, and closely related with the generation of cancer resistance. NF-�� B participates in these processes by different approach after activating, and the effect blocking NF-�� B can suppress the generation of cancer. Therefore, this polysaccharide is by suppressing NF-�� B signal path, thus it is active to play potential Tumor suppression.
Embodiment 7 polygonatum polysaccharide can suppress Cocl2The activity of the luciferase with HRE reporter gene of induction
HEK293T cell strain is purchased from Chinese Academy of Sciences's cell bank, and HEK293T-pGF1-HRE cell strain is built by Shanghai medicine institute of Chinese Academy of Sciences fourth laboratory of chatting about, and is stored in liquid nitrogen, with containing 10%Gibco foetal calf serum DMEM culture medium culturing. HEK293T-pGF1-HRE cell is with 2 �� 104The density of cells/well is inoculated in 96 orifice plates, every hole 100 �� L. Sucking 60 �� L substratum after pasting wall 24h, add sample (polygonatum polysaccharide in embodiment 1) the 50 �� L configuring respective concentration, final concentration is respectively 0.5mg/mL, 0.1mg/mL, 0.02mg/mL, 10 �� LCocl2It is adjusted to final concentration 250 ��Ms. Establish blank group and Cocl simultaneously2Group comparison. Substratum is sucked, the �� LReporterLysis1XBuffer that adds 20 after administration 16h, frozen to reading plate in-80 DEG C of refrigerators. Move out of plate to be detected from-80 DEG C of refrigerators, the cell pyrolysis liquid 20 �� L of dissolving is transferred to white plate, add 40 �� Lluciferase substrates, in 3 minutes, read plate, read RLU value. SPSS statistical software is adopted to carry out statistical procedures. Data all represent with all several �� standard deviations (.x �� s), compare and adopt independent sample t inspection, compare employing one-way analysis of variance between many groups between two groups.
As shown in Figure 4, polygonatum polysaccharide may suppress Cocl to polygonatum polysaccharide determination of activity result under the concentration of 0.5mg/mL2The activity of the luciferase with HRE reporter gene of induction, proves its anti-oxidant activity with potential quality.
Embodiment 8 polygonatum polysaccharide can suppress colorectal carcinoma fibroblastic growth
Colorectal carcinoma associated fibroblast clone CT26-CAF is built by Institute of Biophysics, Academia Sinica. Cell is inoculated in 96 orifice plates, and cell density is 1 �� 104/ hole. Polygonatum polysaccharide in embodiment 1 is dissolved in DMEM+ /+substratum so that it is concentration is 1.0mg/mL. Cultivate 4 days, adopt CCK detection kit (Beijing Zhuan Meng biotech firm) to measure cell growth state. Often kind of polysaccharide sample size is 3.
First data are carried out homogeneity test of variance by statistical analysis: compare employing variance analysis between continuous data many groups, calculate Levene statistical value, if variance is neat, then multiple comparisons between organizing, method is LSD and Dunnett, if heterogeneity of variance, then adopt correction method Dunnettt. Statistical software is SPSS17.0, and significance level is ��=0.05.
As shown in Figure 5, this polygonatum polysaccharide can significantly suppress the growth of colorectal carcinoma CT26-CAF to experimental result under the concentration of 0.5mg/mL. CT26-CAF plays important regulating and controlling effect in colorectal carcinoma, can promote the growth of colon cancer cell. This polysaccharide, by suppressing the growth of colorectal carcinoma CT26-CAF, plays the effect regulating colorectal carcinoma microenvironment.
To sum up experiment proves that the polygonatum polysaccharide in the present invention is by Tumor suppression new vessel generations, immunosuppression, anti-oxidant and suppress many approach adjustment tumor microenvironments such as colorectal carcinoma fibroblastic growth, and play potential inhibitor against colon carcinoma cells tumor promotion further, such that it is able to polygonatum polysaccharide is passed through common process, make said multiple formulation in pharmaceutics, for the preparation of having the auxiliary functional food suppressing colorectal carcinoma effect, the formulation of functional food can be capsule type, tablet, pulvis, granule or oral liquid etc.
Above-described embodiment is that the present invention preferably implements mode; but embodiments of the present invention are not restricted to the described embodiments; the change done under the spirit of other any the present invention of not deviating from and principle, modification, replacement, combination, simplification; all should be the substitute mode of equivalence, it is included in protection scope of the present invention.
Claims (6)
1. polygonatum polysaccharide is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation;
Described polygonatum polysaccharide prepares especially by following method: take sealwort as raw material, after repeatedly extracting with boiling water, united extraction liquid, extracting solution is concentrated to obtain concentrated solution, concentrated solution adds trichloroacetic acid solution and carries out Deproteinization process, undertaken reaction solution after Deproteinization neutralizing, pricking a bag dialysis treatment, undertaken reaction solution in dialysis tubing concentrating, centrifugal treating, get supernatant liquor to add ethanol and carry out alcohol precipitation, drying treatment, obtain polygonatum polysaccharide;
During dialysis, the molecular weight cut-off of dialysis tubing is 3500Da.
2. polygonatum polysaccharide according to claim 1 is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, it is characterized in that: described auxiliary suppression colorectal carcinoma effect refers to by suppressing colorectal carcinoma tumor angiogenesis, immunosuppression, anti-oxidant and suppression colorectal carcinoma fibroblastic growth effect, regulate colorectal carcinoma tumor microenvironment, thus play auxiliary suppression colorectal carcinoma effect.
3. polygonatum polysaccharide according to claim 1 is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, it is characterized in that: during boiling water extraction, extraction time is 1 ~ 5 time, when extracting, the consumption of boiling water is 5 ~ 20 times of sealwort total mass every time, and each extraction time is 1.5 ~ 5h.
4. polygonatum polysaccharide according to claim 1 is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, it is characterized in that: during Deproteinization, adopt trichloroacetic acid solution, the mass percentage of described trichloroacetic acid solution is 35%, and the treatment time is 3 ~ 6h.
5. polygonatum polysaccharide according to claim 1 is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, it is characterized in that: adopt volumn concentration to be the aqueous ethanolic solution of 70 ~ 95% during Ethanol Treatment.
6. polygonatum polysaccharide according to claim 1 is auxiliary as uniquely having suppresses the activeconstituents of colorectal carcinoma effect to have the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation, it is characterized in that: the formulation of described functional food is capsule type, tablet, pulvis, granule or oral liquid.
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CN106511278A (en) * | 2016-12-06 | 2017-03-22 | 四川农业大学 | Polygonatum polysaccharide granules and preparation method thereof |
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101837091A (en) * | 2010-03-17 | 2010-09-22 | 安徽师范大学 | Process for continuously extracting polygonatum saponin and polygonatum polysaccharose |
CN103638352A (en) * | 2013-12-02 | 2014-03-19 | 李玉艳 | Traditional Chinese medicine composition for treating colorectal cancer |
-
2014
- 2014-06-19 CN CN201410274076.3A patent/CN104106763B/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101837091A (en) * | 2010-03-17 | 2010-09-22 | 安徽师范大学 | Process for continuously extracting polygonatum saponin and polygonatum polysaccharose |
CN103638352A (en) * | 2013-12-02 | 2014-03-19 | 李玉艳 | Traditional Chinese medicine composition for treating colorectal cancer |
Non-Patent Citations (1)
Title |
---|
黄精、巴戟天、白芷有效成分体外抗肿瘤作用的研究;孙晓娟;《中国优秀硕士学位论文 医药卫生科技辑》;20121031;第2页第2.1.1小节、第4页第2.4.3小节、第5页第4节 * |
Cited By (1)
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---|---|---|---|---|
CN110423284A (en) * | 2019-08-14 | 2019-11-08 | 浙江万寿康药业有限公司 | Lucid ganoderma spore powder polysaccharide has effects that auxiliary inhibits the application in the functional food of colon cancer in preparation |
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