CN104857018B - Raspberry polysaccharide prepares the purposes of antineoplastic combination medication with chemotherapeutics - Google Patents
Raspberry polysaccharide prepares the purposes of antineoplastic combination medication with chemotherapeutics Download PDFInfo
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Abstract
The invention provides raspberry polysaccharide and purposes of the chemotherapeutics in antitumor combination medicine is prepared.Pharmacodynamic experiment result shows, raspberry polysaccharide, which has, improves immunity of organisms, strengthens chemicals antitumous effect, the advantages that reducing its toxic side effect.
Description
Technical field
The invention belongs to medical sci-tech field, and in particular to the drug combination of raspberry polysaccharide and chemotherapeutics.
Technical background
Raspberry (Rubus idaeus L.) also known as Rubus corchorifolius, raspberry, raspberry etc., belong to rose family rubus.Raspberry master
Northern Hemisphere temperate zone and frigid zone are distributed in, its fruit sweet slightly sour to starch small aggregate fruit more.Raspberry contains polysaccharide, organic acid, Huang
The various active composition such as ketone, anthocyanidin, polyphenol, vitamin, mineral element, volatile oil, tannin, have high nutritive value and
Medical care effect, it is described as " gold fruit ".
Polyoses content is high in raspberry, and polysaccharide yield may be up to 12%, is a kind of preferably Polyose extraction raw material.Research hair
Existing, polysaccharide has regulation immunity of organisms, antitumor, anti-oxidant, antibacterial, radiation proof, antifatigue, reducing blood sugar and blood fat etc. a variety of
Physiologically active.Raspberry product in the market is mainly food, fruit juice, jam, fruit wine such as using raspberry as main component.
The polysaccharide of great activity value in raspberry is extracted, and the example for being applied to health products is considerably less, and using raspberry polysaccharide as work
The lozenge researches of property composition.
Malignant tumour is still the major disease of serious threat human life.Nearest statistics shows that China is new every year
It was found that the people of cancer patient about 20,000,000, wherein 1,500,000 people die from cancer.The death toll of cancer accounts for the 1/5 of total death toll.
Currently, the most frequently used method for the treatment of cancer is still chemotherapy.But due to the nonspecific cell toxicant of chemicals
Property, the serious side effect such as bone marrow suppression, mucositis, dermatitis, diarrhoea can be produced.In addition, tumor patient is after multiple chemotherapy
Easily develop immunity to drugs, so that the effect of chemotherapy is had a greatly reduced quality.The maximum drawback for the embolic chemotherapy applied at present is it
The immune system of body is inhibited, cancer cell is easily escaped the immunosurveillance of body so as to become more menace.Cause
This, it is extremely urgent to find safer and more effective treatment method.In recent years, the immunotherapy of tumour has turned into the new heat of oncotherapy
Point.So-called immunization therapy, broadly refer to by transferring body self immune system come one kind side for the treatment of to anti-malignant tumor
Method.At present, existing such medicine enters clinical investigation phase.
Polysaccharide is that one kind is widely present in animals and plants, has the large biological molecule of a variety of physiologically actives.In recent years, due to
Naturally, nontoxic characteristic and it is good the effect of, polysaccharide has become the new midicinal resources of a kind of great exploitation potential.Immunological regulation
It is the most important physiologically active of polysaccharide.Research shows that lentinan, GL-B, sea grass polysaccharide, flat beautiful snail polysaccharide etc. all have
Good immunological enhancement.But polysaccharide is used with chemotherapy drugs in combination also fewer to treat the example of tumour.
The content of the invention
It is an object of the invention to provide the new application of raspberry polysaccharide.
Specifically, the invention provides raspberry polysaccharide and use of the chemotherapeutics in antitumor combination medicine is prepared
On the way.
Wherein, the chemotherapeutics is selected from taxol, cis-platinum or/and docetaxel.
Wherein, the tumour is selected from melanoma, lung cancer, liver cancer, stomach cancer, breast cancer, kidney, carcinoma of endometrium or colon
Cancer.
Wherein, the chemotherapeutics uses injection type.Wherein, the injection type can be selected from hypodermic injection, muscle
Usual way in the various tumor therapeutic procedures such as injection or intravenous injection.
Wherein, the raspberry polysaccharide uses peroral dosage form.Wherein, the peroral dosage form be selected from tablet, capsule, pill,
Granule, powder, extract or oral liquid.
Further, the dosage of raspberry polysaccharide and chemotherapeutics ratio is 20~40: 1w/w, and certainly, this ratio is
The confirmation carried out with zoopery, in human body in actual use it is possible that the deviation of dosage use ratio.
Wherein, in the raspberry polysaccharide, purity of polysaccharide is more than 50%；Further, purity of polysaccharide is more than 80%；More
Further, purity of polysaccharide is more than 85%；Preferably, purity of polysaccharide is 85~95%w/w, such as moderate purity of the present invention be 90~
92%.Above-mentioned purity is determined using Phenol-sulphate acid method.
Wherein, the raspberry polysaccharide is prepared using water extraction and alcohol precipitation method.
Further, the water extraction and alcohol precipitation method concrete operations are as follows：Take dry raspberry, after petroleum ether or ether defatting, use
75~95%v/v of concentration ethanol extracts, filtering, filter residue extracting in water, after water intaking extract concentration, adds ethanol to alcohol content to reach
75%~85%, cooling, stand, it is raspberry polysaccharide to take solid content.
Wherein, remove seed or non-remove seed before raspberry extraction.
Wherein, the raspberry is the fruit of rose family rubus plant.
Further, the raspberry is selected from raspberry or black raspberry.
Present invention research is found, raspberry polysaccharide and chemotherapy drugs in combination are used, and for the treatment of malignant tumour, is had and is carried
High immunity of organisms, strengthen chemicals antitumous effect, the advantages that reducing its toxic side effect.
Brief description of the drawings：
1 chemotherapeutics of accompanying drawing~raspberry polysaccharide combines the influence to tumor-bearing mice liver organization form；
2 chemotherapeutics of accompanying drawing~raspberry polysaccharide combines the influence to tumor-bearing mice renal tissue form；
3 chemotherapeutics of accompanying drawing~raspberry polysaccharide combines the influence to C57BL/6 mouse spleen indexes；
Influence of the raspberry polysaccharide of accompanying drawing 4 to tumor-bearing mice splenic lymphocytes in-vitro multiplication；
5 chemotherapeutics of accompanying drawing~raspberry polysaccharide combines the influence to Related Immune Enzyme Activity in tumor-bearing mice splenocyte；
6 chemotherapeutics of accompanying drawing~raspberry polysaccharide combine to CD4, CD8, IL in tumor-bearing mice serum~2, INF~γ and
The influence of TNF~alpha content.
Embodiment
The raspberry polysaccharide used in the specific embodiment of the invention is prepared into using conventional Polyose extraction, way of purification
Arrive, concrete operations are as follows in the present invention：
It is prepared by raspberry polysaccharide：Dry raspberry is taken, after crushing remove seed, adds 5 times of petroleum ethers (60~90 DEG C of boiling range) degreasing, eliminates stone
75%~95%v/v ethanol is added after oily ether in 60 DEG C of backflows, the filter residue after filtering is added into 10 times of pure water ultrasonic extraction (power
60W, temperature：80 DEG C, processing time：100min), centrifuging and taking supernatant, filter residue are extracted twice, merging filtrate, subtracted again by old terms
Pressure is concentrated into the 1/4 of original volume, and adding 95% ethanol makes solution alcohol content reach 75%v/v, and 24h is stood in 4 degrees Celsius, and filtering is received
Collect filter residue, filter residue dries to obtain raspberry polysaccharide.
After measured, the raspberry polysaccharide yield that prepared by the present invention is 10.4%~11.6%, detects to obtain through Phenol-sulphate acid method
Purity of polysaccharide is 90.8%~91.2%.
The present invention is in terms of tumor-inhibiting action, attenuation synergistic and immunological regulation three to the main component tree of raspberry polysaccharide buccal tablet
Certain kind of berries polysaccharide has carried out the pharmacodynamic study of adjuvant for chemotherapy of tumour, and raspberry polysaccharide dosage is in terms of polysaccharide in each embodiment.
Embodiment 1
Docetaxel~raspberry polysaccharide combines the inhibitory action to the growth of melanin tumour b16 F10 mouse xenograft tumors
Take the logarithm the melanoma cells B16F10 in growth period, is aseptically prepared into 5 × 107/ ml cell suspensions,
It is subcutaneous that C57BL/6 right side of mice armpits are inoculated in 0.1ml.Vernier caliper measurement mice-transplanted tumor diameter is used after 5 days, treats tumour
Grow to 60~90mm3Animal is grouped at random afterwards.Use the method in measurement knurl footpath, the antitumor effect of dynamic observation subject medicine
Fruit.The pendulous frequency of diameter of tumor is every 2 days 1 time, and measurement every time also needs to weigh mouse weight simultaneously.Packet and administering mode are as follows：
Docetaxel group 10mg/kg, is administered once for every three days using being subcutaneously injected.High administering drug combinations in docetaxel~raspberry polysaccharide
Group, docetaxel 10mg/kg, are administered once, raspberry polysaccharide uses gavage, and dosage is respectively for every three days using being subcutaneously injected
200mg/kg, 400mg/kg, it is administered once daily.Negative control group injects normal saline, 1 time a day.Gross tumor volume calculates
Formula：TV=0.52 × a × b2, wherein a, b represent length and width respectively.Relative tumour volume is calculated according to the result of measurement.It is anti-
The evaluation index of tumor promotion is Relative tumor proliferation rate T/C (%), calculation formula：T/C (%)=TRTV/CRTV× 100%,
TRTV：Treatment group RTV；CRTV：Negative control group RTV.
The inhibitory action that 1. docetaxels of table~raspberry polysaccharide drug combination grows to C57BL/6 mouse xenograft tumors
As a result：1 is shown in Table, docetaxel 10mg/kg groups are to the tumour inhibiting rate of melanin tumour b16 F10 mice-transplanted tumors
66.49% administering drug combinations group is respectively 70.90% and 74.30% to the tumour inhibiting rate of melanin tumour b16 F10 mice-transplanted tumors.Connection
Close and the inhibition of melanoma is used alone higher than docetaxel when using.And docetaxel toxicity is larger, animal body
Decline substantially again, animal has death in experimentation.After both are used in combination, experimental animal body weight is tested without being decreased obviously
During do not occur animal dead.
Therefore, melanin tumour b16 F10 mice-transplanted tumors growth inhibition test result shows, compared with negative control group, respectively
Growth inhibitory action (* P ＜ 0.05, * * P ＜ with conspicuousness of the individual administration group group to melanin tumour b16 F10 mice-transplanted tumors
0.01).Growth of the high administering drug combinations group to melanin tumour b16 F10 mice-transplanted tumors has aobvious in docetaxel~raspberry polysaccharide
The inhibitory action of work property, and the inhibition of the high administering drug combinations group of docetaxel~raspberry polysaccharide is higher than docetaxel independent medication
Group (ΔP ＜ 0.05).Illustrate that raspberry polysaccharide can strengthen the anti-tumor capacity of docetaxel.In addition, compared with docetaxel, raspberry
Polysaccharide has no significant effect to the body weight of experimental animal, has no obvious toxicity.
Embodiment 2
Cis-platinum~raspberry polysaccharide is combined to human lung cancer H460 nude mouse xenograft knurl growth inhibition tests
Take the logarithm the tumour cell in growth period, be aseptically prepared into 5 × 107/ ml cell suspensions, are connect with 0.1ml
Kind armpit on the right side of nude mice is subcutaneous.With vernier caliper measurement transplanted tumor in nude mice diameter, treat tumour growth to 100~200mm3After move
Thing is grouped at random.Using the method in measurement knurl footpath, dynamic observes antitumous effect.The pendulous frequency of diameter of tumor is every 2 days 1
Secondary, measurement every time also needs to weigh mouse weight simultaneously.Administering mode is as follows：Negative control group injects normal saline, 1 time a day；
Cis-platinum group 10mg/kg, weekly administration 1 time；High administering drug combinations group in cis-platinum~raspberry polysaccharide, cis-platinum is using hypodermic injection, 10mg/
Kg, weekly administration 1 time, raspberry polysaccharide use gavage, and dosage is respectively 200mg/kg, 400mg/kg, is administered once daily.
Gross tumor volume calculation formula：
TV=0.52 × a × b2
Wherein a, b represent length and width respectively.Relative tumour volume is calculated according to the result of measurement.The evaluation of antitumor activity
Index is Relative tumor proliferation rate T/C (%), and calculation formula is as follows：
T/C (%)=TRTV/CRTV× 100%
TRTV：Treatment group RTV；CRTV：Negative control group RTV
The inhibitory action that 2. cis-platinums of table~raspberry polysaccharide drug combination grows to human lung cancer H460 nude mouse xenograft tumors
As a result：2 are shown in Table, cis-platinum 10mg/kg groups are 61.49% to the tumour inhibiting rate of human lung cancer H460 transplanted tumor in nude mice；Cis-platinum
It is respectively 65.90%, 72.30% to the tumour inhibiting rate of human lung cancer H460 transplanted tumor in nude mice that~raspberry polysaccharide, which combines middle and high dosage group,.
Wherein, the inhibition of the high administering drug combinations group of cis-platinum~raspberry polysaccharide is higher than cis-platinum independent medication group.In addition, toxicity of cisplatin compared with
Greatly, the weight of animals declines obvious, and animal has death in experimentation.After both are used in combination, experimental animal body weight is without under obvious
Drop, and do not occur animal dead in experimentation.
Therefore, human lung cancer H460 transplanted tumor in nude mice growth inhibition test results are shown, compared with negative control group, cis-platinum
Growth of 10mg/kg groups, cis-platinum~the raspberry polysaccharide senior middle school administering drug combinations group to human lung cancer H460 transplantable tumors has pole conspicuousness
Inhibitory action (* P ＜ 0.05, * * P ＜ 0.01).Compared with positive control cis-platinum, the high administering drug combinations group pair of cis-platinum~raspberry polysaccharide
H460 transplantable tumors growth with conspicuousness inhibitory action (ΔP ＜ 0.05), and the body weight of experimental animal has no significant effect, not
See obvious toxicity.
Embodiment 3
Taxol~raspberry polysaccharide is combined the suppression to the growth of the nude mouse xenograft tumor of human liver cancer Bel~7402 and tested
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；High administering drug combinations group in taxol~raspberry polysaccharide, taxol is using subcutaneous
Injection, 10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, and dosage is respectively 200mg/kg, 400mg/kg, often
It is administered once.
Inhibitory action of 3. taxols of the table~raspberry polysaccharide to the growth of the nude mouse xenograft tumor of human liver cancer Bel~7402
As a result：It is shown in Table 3, taxol 10mg/kg groups are to the tumour inhibiting rate of the transplanted tumor in nude mice of human liver cancer Bel~7402
68.55%；Taxol~raspberry polysaccharide combines tumour inhibiting rate point of the middle and high dosage group to the transplanted tumor in nude mice of human liver cancer Bel~7402
Not up to 72.47%, 76.47%.Wherein, the inhibition of taxol~raspberry polysaccharide senior middle school administering drug combinations group is higher than taxol list
Private medicine group (ΔP ＜ 0.05).And taxol toxicity is larger, the weight of animals declines substantially, and animal has death in experimentation.Two
After person is used in combination, experimental animal body weight does not occur animal dead without being decreased obviously in experimentation.
Therefore, taxol~raspberry polysaccharide shows the transplanted tumor in nude mice growth inhibition test result of human liver cancer Bel~7402,
Compared with negative control group, taxol 10mg/kg groups, taxol~raspberry polysaccharide senior middle school administering drug combinations group to human liver cancer Bel~
The growth of 7402 transplantable tumors has the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.With positive controls Japanese yew alcohol phase
Than taxol~raspberry polysaccharide senior middle school's administering drug combinations have not significant impact to the body weight of experimental animal, have no that obvious poison is secondary anti-
Should.
Embodiment 4
Taxol~raspberry polysaccharide is combined to the nude mouse xenograft tumor growth inhibition test of human gastric cancer MGC~803
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；High administering drug combinations group in taxol~raspberry polysaccharide, taxol is using subcutaneous
Injection, 10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, and dosage is respectively 200mg/kg, 400mg/kg, often
It is administered once.
Inhibitory action of 4. taxols of the table~raspberry polysaccharide to the growth of the nude mouse xenograft tumor of human gastric cancer MGC~803
As a result：It is shown in Table 4, taxol 10mg/kg groups are to the tumour inhibiting rate of the transplanted tumor in nude mice of human gastric cancer MGC~803
65.26%；Taxol~raspberry polysaccharide combines middle and high dosage group and the tumour inhibiting rate of the transplanted tumor in nude mice of human gastric cancer MGC~803 is distinguished
Up to 70.35%, 72.48%.Wherein, the inhibition of taxol~raspberry polysaccharide senior middle school administering drug combinations group is independent higher than taxol
Medication group (ΔP ＜ 0.05).And taxol toxicity is larger, the weight of animals declines substantially, and animal has death in experimentation.Both
After being used in combination, experimental animal body weight does not occur animal dead without being decreased obviously in experimentation.
Therefore, taxol~raspberry polysaccharide shows the transplanted tumor in nude mice growth inhibition test result of human gastric cancer MGC~803,
Compared with negative control group, taxol 10mg/kg groups, taxol~raspberry polysaccharide senior middle school administering drug combinations group to human gastric cancer MGC~
The growth of 803 transplantable tumors has the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.With positive controls Japanese yew alcohol phase
Than taxol~raspberry polysaccharide senior middle school's administering drug combinations have not significant impact to the body weight of experimental animal, have no that obvious poison is secondary anti-
Should.
Embodiment 5
Taxol~raspberry polysaccharide is combined to the examination of the nude mouse xenograft tumor growth inhibition of human breast carcinoma MDA~MB~231
Test
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；Taxol~raspberry polysaccharide administering drug combinations group, taxol, which uses, to be subcutaneously injected,
10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, dosage 400mg/kg, are administered once daily.
Suppression of 5. taxols of the table~raspberry polysaccharide to the growth of the nude mouse xenograft tumor of human breast carcinoma MDA~MB~231
Effect
As a result：It is shown in Table 5, taxol 10mg/kg groups are to the tumour inhibiting rate of the transplanted tumor in nude mice of human breast carcinoma MDA~MB~231
70.87%；Inhibitory rate of the taxol~raspberry polysaccharide joint group to the transplanted tumor in nude mice of human breast carcinoma MDA~MB~231
75.36%.Wherein, the inhibition of taxol~raspberry polysaccharide administering drug combinations group higher than taxol independent medication group (ΔP ＜
0.05).And taxol toxicity is larger, the weight of animals declines substantially, and animal has death in experimentation.After both are used in combination,
Experimental animal body weight does not occur animal dead without being decreased obviously in experimentation.
Therefore, taxol~raspberry polysaccharide is to the transplanted tumor in nude mice growth inhibition test result of human breast carcinoma MDA~MB~231
Show, compared with negative control group, taxol 10mg/kg groups, taxol~raspberry polysaccharide administering drug combinations group are to human breast carcinoma MDA
The growth of the transplantable tumor of~MB~231 has the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.It is purple with positive controls
China fir alcohol is compared, and taxol~raspberry polysaccharide administering drug combinations have not significant impact to the body weight of experimental animal, has no that obvious poison is secondary
Reaction.
Embodiment 6
Taxol~raspberry polysaccharide is combined to people's kidney A498 nude mouse xenograft tumor growth inhibition tests
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；High administering drug combinations group in taxol~raspberry polysaccharide, taxol is using subcutaneous
Injection, 10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, and dosage is respectively 200mg/kg, 400mg/kg, often
It is administered once.
The inhibitory action that 6. taxols of table~raspberry polysaccharide grows to people kidney A498 nude mouse xenograft tumors
As a result：6 are shown in Table, taxol 10mg/kg groups are 60.21% to the tumour inhibiting rate of people's kidney A498 transplanted tumor in nude mice；It is purple
China fir alcohol~raspberry polysaccharide combines middle and high dosage group to the tumour inhibiting rate of people's kidney A498 transplanted tumor in nude mice respectively up to 69.55%,
72.19%.Wherein, in taxol~raspberry polysaccharide high administering drug combinations group inhibition higher than taxol independent medication group (ΔP
＜ 0.05).And taxol toxicity is larger, the weight of animals declines substantially, and animal has death in experimentation.Both are used in combination
Afterwards, experimental animal body weight is without being decreased obviously, and does not occur animal dead in experimentation.
Therefore, taxol~raspberry polysaccharide shows people's kidney A498 transplanted tumor in nude mice growth inhibition test results, with the moon
Property control group is compared, and taxol 10mg/kg groups, taxol~raspberry polysaccharide senior middle school administering drug combinations group are to people's kidney A498 transplantable tumors
Growth have the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.Compared with positive controls taxol, taxol
~raspberry polysaccharide senior middle school's administering drug combinations have not significant impact to the body weight of experimental animal, have no obvious toxicity.
Embodiment 7
Taxol~raspberry polysaccharide is combined the suppression to the growth of the nude mouse xenograft tumor of human colon carcinoma HT~29 and tested
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；Taxol~raspberry polysaccharide administering drug combinations group, taxol, which uses, to be subcutaneously injected,
10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, dosage 400mg/kg, are administered once daily.
Inhibitory action of 7. taxols of the table~raspberry polysaccharide to the growth of the nude mouse xenograft tumor of human colon carcinoma HT~29
As a result：It is shown in Table 7, taxol 10mg/kg groups are to the tumour inhibiting rate of the transplanted tumor in nude mice of human colon carcinoma HT~29
62.88%；Inhibitory rate 68.23% of the taxol~raspberry polysaccharide joint group to the transplanted tumor in nude mice of human colon carcinoma HT~29.Its
In, the inhibition of taxol~raspberry polysaccharide administering drug combinations group higher than taxol independent medication group (ΔP ＜ 0.05).And Japanese yew
Alcohol toxicity is larger, and the weight of animals declines substantially, and animal has death in experimentation.After both are used in combination, experimental animal body weight
Nothing is decreased obviously, and does not occur animal dead in experimentation.
Therefore, taxol~raspberry polysaccharide shows the transplanted tumor in nude mice growth inhibition test result of human colon carcinoma HT~29,
Compared with negative control group, taxol 10mg/kg groups, taxol~raspberry polysaccharide administering drug combinations group are moved to human colon carcinoma HT~29
Planting the growth of knurl has the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.It is purple compared with positive controls taxol
China fir alcohol~raspberry polysaccharide administering drug combinations have not significant impact to the body weight of experimental animal, have no obvious toxicity.
Embodiment 8
Taxol~raspberry polysaccharide is combined the suppression to the growth of people carcinoma of endometrium HHUA nude mouse xenograft tumors and tested
Specific embodiment is with reference to embodiment 2.Dosage regimen is as follows：Negative control group injection normal saline, daily 1
It is secondary；Taxol group 10mg/kg, weekly administration 1 time；High administering drug combinations group in taxol~raspberry polysaccharide, taxol is using subcutaneous
Injection, 10mg/kg, weekly administration 1 time, raspberry polysaccharide use gavage, and dosage is respectively 200mg/kg, 400mg/kg, often
It is administered once.
The inhibitory action that 8. taxols of table~raspberry polysaccharide grows to people carcinoma of endometrium HHUA nude mouse xenograft tumors
As a result：8 are shown in Table, taxol 10mg/kg groups are to the tumour inhibiting rate of people's carcinoma of endometrium HHUA transplanted tumor in nude mice
58.77%；Taxol~raspberry polysaccharide combines tumour inhibiting rate point of the middle and high dosage group to people's carcinoma of endometrium HHUA transplanted tumor in nude mice
Not up to 67.21%, 69.34%.Wherein, the inhibition of high administering drug combinations group is higher than taxol list in taxol~raspberry polysaccharide
Private medicine group (ΔP ＜ 0.05).And taxol toxicity is larger, the weight of animals declines substantially, and animal has death in experimentation.Two
After person is used in combination, experimental animal body weight does not occur animal dead without being decreased obviously in experimentation.
Therefore, taxol~raspberry polysaccharide is to people's carcinoma of endometrium HHUA transplanted tumor in nude mice growth inhibition test result tables
Bright, compared with negative control group, taxol 10mg/kg groups, taxol~raspberry polysaccharide senior middle school administering drug combinations group are to people's intrauterine
The growth of film cancer HHUA transplantable tumors has the inhibitory action (* P ＜ 0.05, * * P ＜ 0.01) of pole conspicuousness.With positive controls Japanese yew
Alcohol is compared, and high administering drug combinations have not significant impact to the body weight of experimental animal in taxol~raspberry polysaccharide, have no obvious poison
Side reaction.
Embodiment 9
Raspberry polysaccharide is to tumor-bearing mice liver, the influence of renal tissue form
After each group tumor-bearing mice anesthesia in the anti-tumor experiment of embodiment 1~8, liver and kidney are separated.By liver, kidney
Dirty be soaked in 10% formalin is fixed.By the tissue fixed through ethanol dehydration, dimethylbenzene is transparent, FFPE, cuts
Piece paster, dimethylbenzene dewaxing, HE dyeing, dehydration is transparent, and sealing is after optical microphotograph Microscopic observation and takes pictures.
The hepatic tissue section of each group mouse is as shown in figure 1, the liver cell edge clear of normal mouse, cytoplasm fill, carefully
Karyon is clear, and cell arrangement is neat.By contrast, the liver cell height swelling of chemotherapeutics group mouse, subregion hepatic tissue
Focal necrosis, interstitial stove cell infiltration.And the liver cell of chemotherapeutics~raspberry polysaccharide joint group mouse has obvious recovery,
The liver cell of wherein chemotherapeutics~raspberry polysaccharide high dose joint group mouse is similar to normal mouse liver cell.It can be seen that raspberry
Polysaccharide tumor-bearing mice hepatic injury to caused by chemotherapeutics has certain repair.
The nephridial tissue of each group mouse is cut into slices as shown in Fig. 2 the glomerulus regular shape of normal mouse, Bowman's capsule gap
Small, glomerular basement membrane is complete.By contrast, there is certain degeneration and damage in the glomerulus of chemotherapeutics group mouse, specifically
Show that glomerular basement membrane distorts, capsula glomeruli gap is substantially widened.The kidney of chemotherapeutics~raspberry polysaccharide joint group mouse is small
Ball recovers clearly, to be mainly manifested in capsula glomeruli gap and compared with chemotherapeutics group substantially diminish.It can be seen that raspberry polysaccharide is to chemotherapeutic
Tumor-bearing mice injury of kidney also has certain repair caused by thing.
It these results suggest that, raspberry polysaccharide can repair caused lesions of liver and kidney after chemotherapeutic drug therapy, chemotherapeutics and tree
Certain kind of berries polysaccharide is used in combination rear toxic side effect and reduced.
Embodiment 10
Influence of the raspberry polysaccharide to tumor-bearing mice index and spleen index
After the anti-tumor experiment of embodiment 1~8 terminates, by mouse anesthesia, tumor tissues and spleen are separated, is weighed.According to title
Weight result calculates index and spleen index.Calculation formula：Index and spleen index=spleen weight (g)/net body weight (kg), wherein, net body weight=body
Weight~knurl weight.Test obtained result to represent with mean ± SD, and count T inspections, P ＜ 0.05 are significant difference, P ＜
0.01 is pole significant difference (*：Compared with negative control group, # is compared with chemotherapeutics group).
As a result Fig. 3 is seen.Compared with chemotherapeutics group, the index and spleen index of chemotherapeutics~raspberry polysaccharide administering drug combinations group is aobvious
Write increase.
Embodiment 11
The influence of raspberry Polysaccharides on Mice splenic lymphocytes in-vitro multiplication
Cervical dislocation puts to death mouse, is soaked 1~2 minute in 75% ethanol, takes out spleen rapidly in aseptic operating platform,
Filled as 400 mesh nylon screens are equipped with advance in ice-cold PBS plate.Spleen tissue is gently twisted with the fingers with sterile nook closing member on screen cloth, is made
Individual cells filtration screen cloth enters plate.Cell suspension is collected after rinsing screen cloth with PBS, in 4 DEG C of 5 points of centrifugations (1000r/min)
Clock.Supernatant is abandoned, cell is resuspended with the culture medium of the RPMI containing 2%FBS~1640 and counts under the microscope, cell concentration is adjusted
For 1 × 107Individual/ml, by cell suspension inoculation into 96 orifice plates, 100 μ l/ holes, and 5 μ g/ml ConA is added into hole, in 37
DEG C, 5%CO2Overnight incubation in incubator.Next day, it is separately added into after raspberry polysaccharide is diluted into each predetermined concentration with nutrient solution
In 96 orifice plates (100 μ l/ holes).Using do not add ConA as blank control group, using only plus ConA do not add the group of raspberry polysaccharide as
Positive controls.At 37 DEG C, 5%CO2Incubator is incubated 48 hours.5mg/ml MTT is added into 96 orifice plates, per the μ l of hole 20,
Continue to cultivate 4h.Culture medium is sopped up, 150 μ l DMSO dissolvings are added per hole, shaking table gently mixes for 10 minutes.Surveyed with ELIASA
The a length of 570nm of standing wave, reference wavelength are that light absorption value is determined at 630nm.Test obtained result to represent with mean ± SD, and carry out
Count T to examine, P ＜ 0.05 are significant difference, and P ＜ 0.01 are pole significant difference (*：Compared with negative control group, # and sun
Property control group is compared).
As a result as shown in figure 4, the light absorption value of raspberry polysaccharide each group is all remarkably higher than negative control group, and dose-dependant is presented
Relation.In addition, since 64 μ g/ml, it is significantly higher than positive controls.Thus illustrate, raspberry polysaccharide can significantly stimulate mice spleen
The propagation of lymphocyte.
Embodiment 12
Influence of the raspberry polysaccharide to Related Immune Enzyme Activity in tumor-bearing mice splenocyte
By the spleen of each group tumor-bearing mice of embodiment 1~8 under condition of ice bath with cell crushing instrument it is broken (400 amperes, often
Secondary 5 seconds, it is repeated 3 times), crushing medium pH7.4,0.01mol/L Tris~HCl, 0.0001mol/L EDTA~2Na,
0.01mol/L sucrose, 0.8%NaCl.Supernatant is collected after centrifuging (5000r/min) 10 minutes in 4 DEG C, is examined respectively with testing cassete
Survey the activity of four kinds of immune relevant enzyme AKP, ACP, LDH and SOD in splenocyte.Obtained result is tested with mean ± SD tables
Show, and carry out counting T inspections, P ＜ 0.05 are significant difference, and P ＜ 0.01 are pole significant difference (*：With negative control group phase
Than # is compared with positive controls).
As a result as shown in figure 5, compared with negative control group, raspberry polysaccharide can be obviously improved the activity of 4 kinds of enzymes, and agent is presented
Measure dependence.Compared with chemotherapeutics group, the activity for improving ACP of chemotherapeutics~raspberry polysaccharide joint group conspicuousness.
Thus illustrate, after chemotherapeutics is used in combination with raspberry polysaccharide, its raising ability to ACP activity is strengthened.
Embodiment 13
Influence of the raspberry polysaccharide to CD4, CD8, IL in tumor-bearing mice serum~2, INF~γ and TNF~alpha content
Blood is taken by eyeball is extractd after each group tumor-bearing mice anesthesia in the anti-tumor experiment of embodiment 1~8.Blood stands a moment
Serum is collected after centrifuging (2500r/min) 10 minutes after 4 DEG C.Detected respectively with enzyme-linked immunologic detecting kit CD4 in serum,
CD8, IL~2, INF~γ and TNF~alpha content.Test obtained result to represent with mean ± SD, and carry out counting T inspections,
P ＜ 0.05 are significant difference, and P ＜ 0.01 are pole significant difference (*：Compared with negative control group, # and positive controls phase
Than).
As a result as shown in fig. 6, compared with positive controls, chemotherapeutics~raspberry polysaccharide joint group can be significantly or extremely aobvious
The content of 5 kinds of albumen of raising of work property.Thus illustrate, after docetaxel is used in combination with raspberry polysaccharide, it is to this five kinds of albumen
The raising ability of content is strengthened.
Each immunization experiment result further illustrates above, and raspberry polysaccharide can strengthen the immunity of tumor-bearing mice, and greatly
The big therapeutic effect for improving chemotherapeutics.
Claims (15)
1. raspberry polysaccharide and purposes of the chemotherapeutics in antitumor combination medicine is prepared.
2. purposes according to claim 1, it is characterised in that：The chemotherapeutics is selected from taxol, cis-platinum or/and more western
He matches.
3. purposes according to claim 1, it is characterised in that：The tumour be selected from melanoma, lung cancer, liver cancer, stomach cancer,
Breast cancer, kidney, carcinoma of endometrium or colon cancer.
4. purposes according to claim 1, it is characterised in that：The chemotherapeutics uses injection type.
5. purposes according to claim 1, it is characterised in that：The raspberry polysaccharide uses peroral dosage form.
6. according to the purposes described in Claims 1 to 5 any one, it is characterised in that：The use of raspberry polysaccharide and chemotherapeutics
Dose ratio is 20~40: 1w/w.
7. according to the purposes described in Claims 1 to 5 any one, it is characterised in that：In the raspberry polysaccharide, purity of polysaccharide exists
More than 50%.
8. purposes according to claim 7, it is characterised in that：Purity of polysaccharide is more than 80%.
9. purposes according to claim 8, it is characterised in that：Purity of polysaccharide is more than 85%.
10. purposes according to claim 9, it is characterised in that：Purity of polysaccharide is 85~95%w/w.
11. according to the purposes described in Claims 1 to 5 any one, it is characterised in that：The raspberry polysaccharide uses water extract-alcohol precipitation
Method is prepared.
12. purposes according to claim 11, it is characterised in that：The water extraction and alcohol precipitation method concrete operations are as follows：Take drying
After raspberry, petroleum ether or ether defatting, extracted with 75~95%v/v of concentration ethanol, filtering, filter residue extracting in water, extract of fetching water
After concentration, add ethanol to alcohol content to reach 75%~85%, cooling, stand, it is raspberry polysaccharide to take solid content.
13. purposes according to claim 12, it is characterised in that：Remove seed or non-remove seed before raspberry extraction.
14. according to the purposes described in Claims 1 to 5 any one, it is characterised in that：The raspberry is rose family rubus
The fruit of plant.
15. purposes according to claim 14, it is characterised in that：The raspberry is selected from raspberry or black raspberry.
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| CN105254773A (en) * | 2015-10-27 | 2016-01-20 | 齐齐哈尔医学院 | Preparation method of antitumor polysaccharide |
| CN112538121B (en) * | 2020-12-09 | 2021-10-22 | 青海大学 | Polysaccharide SM-0.4M and anti-tumor product prepared from same |
| CN112358553B (en) * | 2020-12-09 | 2021-10-01 | 青海大学 | Polysaccharide SM-0.2M and anti-tumor product prepared from same |
| CN115414465A (en) * | 2022-10-17 | 2022-12-02 | 苏州明人医药生物科技有限公司 | Application of JWA polypeptide in preparation of antitumor drug synergist |
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| CN102731668A (en) * | 2011-04-12 | 2012-10-17 | 谭智丽 | Method for extracting and purifying Codonopsis pilosula polysaccharide |
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| CN102731668A (en) * | 2011-04-12 | 2012-10-17 | 谭智丽 | Method for extracting and purifying Codonopsis pilosula polysaccharide |
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| 覆盆子多糖提取、结构分析及自由基清除作用研究;刘明学等;《食品科技》;20091231;第34卷(第7期);第163-167页 * |
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