CN109329207A - A kind of experimental animal model for anticancer agents and its construction method - Google Patents
A kind of experimental animal model for anticancer agents and its construction method Download PDFInfo
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- 238000010171 animal model Methods 0.000 title claims abstract description 24
- 239000002246 antineoplastic agent Substances 0.000 title claims abstract description 16
- 238000010276 construction Methods 0.000 title claims abstract description 9
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 66
- 241001465754 Metazoa Species 0.000 claims abstract description 27
- 241000699666 Mus <mouse, genus> Species 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 16
- 239000012153 distilled water Substances 0.000 claims abstract description 14
- 238000011081 inoculation Methods 0.000 claims abstract description 14
- 241000699670 Mus sp. Species 0.000 claims abstract description 12
- 239000003085 diluting agent Substances 0.000 claims abstract description 12
- 238000005303 weighing Methods 0.000 claims abstract description 12
- 238000003304 gavage Methods 0.000 claims abstract description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 5
- 239000002504 physiological saline solution Substances 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 6
- 238000007920 subcutaneous administration Methods 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 5
- 230000000259 anti-tumor effect Effects 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 13
- 238000012795 verification Methods 0.000 abstract description 10
- 230000008569 process Effects 0.000 abstract description 8
- 241000251539 Vertebrata <Metazoa> Species 0.000 abstract description 2
- 241000700159 Rattus Species 0.000 description 19
- 239000002775 capsule Substances 0.000 description 8
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- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 5
- 201000005202 lung cancer Diseases 0.000 description 5
- 208000020816 lung neoplasm Diseases 0.000 description 5
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- 208000014018 liver neoplasm Diseases 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 206010039491 Sarcoma Diseases 0.000 description 3
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical group ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
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- 230000036541 health Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000003468 Ehrlich Tumor Carcinoma Diseases 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000700161 Rattus rattus Species 0.000 description 1
- 206010064390 Tumour invasion Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
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- 230000036737 immune function Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000037451 immune surveillance Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
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- 230000005760 tumorsuppression Effects 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/13—Tumour cells, irrespective of tissue of origin
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
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Abstract
The invention belongs to cultivate vertebrate technical field, a kind of experimental animal model for anticancer agents and its construction method are disclosed, laboratory mice or rat is taken to inoculate mouse or rat tumor sterile diluent by sterile working in its right axillary, grouping of weighing after 24 hours；It weighs after 24 hours, it is random to be grouped, every group 10, the daily 20mg/kgi.p of CTX, the daily stomach-filling of remaining each group；Model control group separately sets the i.e. non-inoculated tumour animal of intact animal to equal volume distilled water, and inoculation same amount of normal saline gavages equal volume distilled water as Normal group as false inoculation group 10 daily, once a day, continuous 7 days.Animal experimental model of the invention grows tumour in animal body, verification process is shorter, calculates inhibiting rate by weighing knurl weight, verification the verifying results have convincingness by aseptically injecting tumour sterile diluent directly into experimental animal body.
Description
Technical field
The invention belongs to cultivate vertebrate technical field more particularly to a kind of experimental animal model for anticancer agents and its building
Method.
Background technique
Currently, the prior art commonly used in the trade is such that cancer is the common disease and frequently-occurring disease in the current mankind, due to
The death rate is high, and becomes dead synonym.Cancer seriously threatens the health and life of the mankind, and countries in the world are put into largely
Human and material resources expand attack to cancer in all directions.Research has shown that traditional Chinese medicine not only has preferable treatment to the treatment of cancer
Effect, and can be reduced side effect and recurrence rate, can effectively extend the life cycle of patient and improve the existence matter of patient
Amount, and stable to a certain extent or diminution tumour, therefore Chinese medicine and therapy of combing traditional Chinese and Western medicine cancer have become and clinically generally adopt
One of method.Tumour is multifactor, multi-step a complex biological process.Traditional Chinese medicine thinks swollen
Why tumor generates, and is because healthy energy is insufficient, and perverse trend is illegally occupied, and causes " product at "." heresy " is most main in cancer development process
The pathological manifestations and pathological factor wanted specific to show as the stagnation of the circulation of vital energy, blood stasis, heat toxin, phlegm wet etc. more.So should also pay attention in the treatment
Using activating microcirculation and removing stasis medicinal, strengthening vital QI to eliminate pathogenic factors, detoxicating and resolving a mass etc. " eliminating evil " treatment means and be aided with the method for " righting ", play good effect more.It passes
The antitumor animal model of system needs to cultivate kinds of tumor cells, is then transplanted in animal body, is diffused it,
Then by inject anti-tumor drug be grouped verifying, whole process takes a long time, in verification process experimental animal understand because
A variety of causes and die, cause verification the verifying results without convincingness.
In conclusion problem of the existing technology is: traditional antitumor animal model is needed to kinds of tumor cells
It is cultivated, is then transplanted in animal body, be diffused it；Verifying, process are grouped by injecting anti-tumor drug
It takes a long time, experimental animal can die because of a variety of causes in verification process, cause verification the verifying results without convincingness.
Solve the difficulty and meaning of above-mentioned technical problem:
Tumor-cell antigen is not strong or critical antigen is hidden, for tumor surface or tumour product, to tumour
The existence of itself does not play a crucial role；The appearance of tumour, which generates the immune function of body, to be inhibited or closes, and tumour is escaped
The immune surveillance function of body keeps body weak to the immune response of tumour or nothing；Diversity, the complexity of tumor invasion, even if
Same tumour different stage, different differentiation degrees, the complexity of gene mutation, antigenicity are multifarious.
Summary of the invention
In view of the problems of the existing technology, the present invention provides a kind of experimental animal model for anticancer agents and its building sides
Method.
The invention is realized in this way a kind of experimental animal model for anticancer agents, the experimental animal model for anticancer agents is to take
Laboratory mice or rat press sterile working in its right axillary and inoculate mouse or rat tumor sterile diluent, claim after 24 hours
It is grouped again；It weighs after 24 hours, it is random to be grouped, every group 10, the daily 20mg/kgi.p of CTX, the daily stomach-filling of remaining each group；Model
Control group separately sets the i.e. non-inoculated tumour animal of intact animal, inoculation same amount of normal saline is as false inoculation to equal volume distilled water
Group 10, gavages equal volume distilled water as Normal group daily, once a day, continuous 7 days.
It is described anti-swollen another object of the present invention is to provide a kind of construction method of experimental animal model for anticancer agents
The construction method of tumor experimental animal model the following steps are included:
Step 1, takes different types of mouse and rat several are cultivated, weight 17g~110g, male and female dual-purpose；
Step 2 is aseptically extracted tumor liquid, is diluted by a certain percentage with physiological saline, takes laboratory mice
Or rat inoculates mouse or rat tumor sterile diluent by sterile working in its right axillary, grouping of weighing after 24 hours；
Step 3 is weighed after 24 hours, and random to be grouped, every group 10, the daily 20mg/kgi.p of CTX, remaining each group is daily
Stomach-filling；
Step 4, model control group separately set the i.e. non-inoculated tumour animal of intact animal to equal volume distilled water, are inoculated with equivalent
Physiological saline gavages equal volume distilled water as Normal group as false inoculation group 10 daily, once a day, continuous 7 days；
Step 5, next day of being discontinued put to death mouse, weigh and remove subcutaneous tumor mass, claim knurl weight, count the death rate, weight, tumor
Tumour inhibiting rate is weighed and calculates, t is examined between as a result organizing, and ATK is compared with KL middle dose group.
In conclusion advantages of the present invention and good effect are as follows: animal experimental model of the invention passes through in aseptic condition
Tumour sterile diluent to be injected directly into experimental animal body down, grows tumour in animal body, verification process is shorter,
Inhibiting rate is calculated by weighing knurl weight, verification the verifying results have convincingness.
Detailed description of the invention
Fig. 1 is the construction method flow chart of experimental animal model for anticancer agents provided in an embodiment of the present invention.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
It needs to carry out incubation to kinds of tumor cells for traditional antitumor animal model to take a long time, verifying effect
Fruit does not have the problem of convincingness；Animal experimental model of the invention into experimental animal body by aseptically directly infusing
Tumour sterile diluent is penetrated, grows tumour in animal body, verification process is shorter, calculates inhibition by weighing knurl weight
Rate, verification the verifying results have convincingness.
Application principle of the invention is explained in detail with reference to the accompanying drawing.
Experimental animal model for anticancer agents provided in an embodiment of the present invention is to take laboratory mice or rat in its right axillary by nothing
Bacterium operation inoculates mouse or rat tumor sterile diluent, grouping of weighing after 24 hours；It weighs after 24 hours, random point
Group, every group 10, the daily 20mg/kgi.p of CTX, the daily stomach-filling of remaining each group；Model control group is separately set to equal volume distilled water
Intact animal is non-inoculated tumour animal, and inoculation same amount of normal saline gavages isometric(al) distillation as false inoculation group 10 daily
Water is as Normal group, once a day, continuous 7 days.
As shown in Figure 1, the construction method of experimental animal model for anticancer agents provided in an embodiment of the present invention the following steps are included:
S101: taking different types of mouse and rat several are cultivated, weight 17g~110g, male and female dual-purpose；
S102: aseptically extract tumor liquid, be diluted by a certain percentage with physiological saline, take laboratory mice or
Rat inoculates mouse or rat tumor sterile diluent by sterile working in its right axillary, grouping of weighing after 24 hours；
It weighs after S103:24 hours, random to be grouped, every group 10, the daily 20mg/kgi.p of CTX, remaining each group fills daily
Stomach；
S104: model control group separately sets the i.e. non-inoculated tumour animal of intact animal to equal volume distilled water, and inoculation equivalent is raw
Salt water is managed as false inoculation group 10, gavages equal volume distilled water daily as Normal group, once a day, continuous 7 days；
S105: next day of being discontinued puts to death mouse, weighs and removes subcutaneous tumor mass, claims knurl weight, count the death rate, weight, knurl weight
And tumour inhibiting rate is calculated, t is examined between as a result organizing, and ATK is compared with KL middle dose group.
Experiment mice provided in an embodiment of the present invention is divided into lotus knurl control group, the large, medium and small dosage group of test medicine, and peace is replaced
Can group, cyclophosphamide group, wherein animal 20 in lotus knurl control group.
Application effect of the invention is described in detail below with reference to experiment.
1, to rat liver cancer H22The Inhibition test of solid tumor
Aseptically extract H22Tumor liquid takes BALB/c mouse with physiological saline 1:3 dilution, presses sterile behaviour in right axillary
Make to inoculate mouse tumor sterile diluent 0.2ml/ only, it is (dynamic to be grouped into lotus knurl control group at random for grouping of weighing after 24 hours
More one times of object number), the large, medium and small dosage group of test medicine, peace for can (ATK) group, cyclophosphamide group (CTX), every group 10.CTX
Daily 20mg/kgi.p, the daily stomach-filling of remaining each group, model control group is to equal volume distilled water.Dosage is shown in aforementioned table.Separately
If intact animal (non-inoculated tumour, inoculation same amount of normal saline is as false inoculation group) 10, gavages equal volume distilled water daily
As Normal group.Once a day, continuous 7 days, next day of being discontinued put to death mouse, weighs and removes subcutaneous tumor mass, claims knurl weight, system
The meter death rate, weight, knurl weight simultaneously calculate tumour inhibiting rate.As a result t is examined between organizing, and ATK is compared with KL middle dose group.The result shows that health
Imperial capsule is to rat liver cancer H22Solid tumor has significant inhibiting effect, is shown in Table 1.
1 Kang Long capsule of table is to rat liver cancer H22Solid tumor inhibiting effect
Note: compared with lotus knurl model group, * *: P < 0.01；Compared with suitable multiple ATK, P > 0.05.
2, to the inhibiting tumor assay of Mice Bearing Lewis Lung Cancer
The lung cancer kind mouse with well-grown solid tumor is taken, removes tumor mass under aseptic condition, physiological saline 1:4 is made even
Slurry, takes C57/6J black rat, male and female dual-purpose, weight 17-22g, sterile dilute in right axillary subcutaneous vaccination mouse tumor by sterile working
After releasing liquid 0.2ml/ only, grouping of weighing after 24 hours, grouping and administration and result treatment are tested with Murine Hepatoma22.The result shows that
Kang Long capsule has significant inhibiting effect to Mice Bearing Lewis Lung Cancer, is shown in Table 2.
Inhibiting effect of the 2 Kang Long capsule of table to Mice Bearing Lewis Lung Cancer
Note: compared with lotus knurl model group, * *: P < 0.01；Compared with suitable multiple ATK, △: P < 0.05.
3, the resistance of rat sarcoma W256 is tested
Well-grown W256 tumor liquid is extracted under aseptic condition, with physiological saline 1:3 dilution, is taken SD rat, is pressed in right axillary
After sterile working inoculates mouse tumour sterile diluent 0.2ml/ only, grouping of weighing after 24 hours, grouping is the same, administration class amount
It see the table below.The daily 15mg/kgi.p of CTX, the daily gastric infusion of remaining each group, once a day, continuous 7 days, next day of being discontinued put to death small
Mouse weighs and removes subcutaneous tumor mass, claims knurl weight, and calculate tumour inhibiting rate.T is examined between each batch of result carries out group.The result shows that Kang Long
Capsule in Rats sarcoma W256 has significant inhibiting effect, is shown in Table 3.
The inhibiting effect of 3 Kang Long Capsule in Rats sarcoma W256 of table
The present invention is to Kang Long capsule to rat liver cancer H22, Mice Bearing Lewis Lung Cancer and rat ehrlich ascites tumor inhibiting effect
It is studied.As a result show that normal blank group animal is showed no false lump and is formed, weight and the death rate meet the requirements, and show to swell
Tumor well-grown.Compared with normal blank control group, tumor animal weight has certain downward trend；Tumor animal has on a small quantity in
Way is dead, and each comparison among groups of death toll, weight are not statistically significant (P > 0.05).Kang Long capsule is in the inhibiting effect of solid tumor
Certain dose dependent, clinical equimultiple dosage show preferable activity.In test Kang Longteng is also shown with one
The effect of fixed raising immunity of organisms, the characteristics of having fully demonstrated Chinese medicine compound prescription multipath tumor suppression.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (2)
1. a kind of experimental animal model for anticancer agents, which is characterized in that the experimental animal model for anticancer agents is to take laboratory mice
Or rat inoculates mouse or rat tumor sterile diluent by sterile working in its right axillary, grouping of weighing after 24 hours；24
It weighs after hour, it is random to be grouped, every group 10, the daily 20mg/kgi.p of CTX, the daily stomach-filling of remaining each group；Model control group is given
Equal volume distilled water separately sets the i.e. non-inoculated tumour animal of intact animal, and inoculation same amount of normal saline is as false inoculation group 10, often
Day gavage equal volume distilled water as Normal group, once a day, continuous 7 days.
2. a kind of construction method of experimental animal model for anticancer agents as described in claim 1, which is characterized in that the antitumor reality
Test the construction method of animal model the following steps are included:
Step 1, takes different types of mouse and rat several are cultivated, weight 17g~110g, male and female dual-purpose；
Step 2 is aseptically extracted tumor liquid, is diluted by a certain percentage with physiological saline, and laboratory mice or big is taken
Mouse inoculates mouse or rat tumor sterile diluent by sterile working in its right axillary, grouping of weighing after 24 hours；
Step 3 is weighed after 24 hours, random to be grouped, and every group 10, the daily 20mg/kgi.p of CTX, the daily stomach-filling of remaining each group；
Step 4, model control group separately set the i.e. non-inoculated tumour animal of intact animal to equal volume distilled water, are inoculated with normal
Salt water gavages equal volume distilled water as Normal group as false inoculation group 10 daily, once a day, continuous 7 days；
Step 5, next day of being discontinued put to death mouse, weigh and remove subcutaneous tumor mass, claim knurl weight, the statistics death rate, weight, knurl weight are simultaneously
Tumour inhibiting rate is calculated, t is examined between as a result organizing, and ATK is compared with KL middle dose group.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110250095A (en) * | 2019-06-18 | 2019-09-20 | 浙江省肿瘤医院 | A kind of animal model of middle liver mixture to lotus liver cancer and sarcoma mouse tumor-inhibiting action |
| CN110432225A (en) * | 2019-07-23 | 2019-11-12 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | The construction method of blood stasis phlegm solidifying card and laryngocarcinoma Disease Syndrome integrated animal model |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110250095A (en) * | 2019-06-18 | 2019-09-20 | 浙江省肿瘤医院 | A kind of animal model of middle liver mixture to lotus liver cancer and sarcoma mouse tumor-inhibiting action |
| CN110432225A (en) * | 2019-07-23 | 2019-11-12 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | The construction method of blood stasis phlegm solidifying card and laryngocarcinoma Disease Syndrome integrated animal model |
| CN110432225B (en) * | 2019-07-23 | 2021-10-22 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Construction method of animal model combining blood stasis phlegm coagulation and laryngeal cancer |
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