CN110036951A - Tilapia mossambica fatty liver damage model and its construction method and application - Google Patents
Tilapia mossambica fatty liver damage model and its construction method and application Download PDFInfo
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- CN110036951A CN110036951A CN201910327015.1A CN201910327015A CN110036951A CN 110036951 A CN110036951 A CN 110036951A CN 201910327015 A CN201910327015 A CN 201910327015A CN 110036951 A CN110036951 A CN 110036951A
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- 208000010706 fatty liver disease Diseases 0.000 title claims abstract description 28
- 208000004930 Fatty Liver Diseases 0.000 title claims abstract description 24
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- 231100000240 steatosis hepatitis Toxicity 0.000 title claims abstract description 24
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- 210000002966 serum Anatomy 0.000 claims description 11
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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
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/168—Steroids
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
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Abstract
The invention discloses Tilapia mossambica fatty liver damage model and its construction method and application, the construction method of the model includes domestication, the preparation of high lipid food, induction and tests and analyzes step.Compared with prior art, the invention has the following advantages that the high lipid food that (1) the method for the invention uses is formulated, simple, easily operated, ingredient is easy to control, modeling success rate is high and stable, the modeling time is shorter, there is good application prospect；(2) animal model constructed by the present invention can be used for fatty,fiss hepatic injury pathogenesis caused by studying many factors and the correlation of fat metabolic disturbance and hepatic injury, it can be used for screening the drug of prevention and treatment fatty,fiss liver, and the pharmacological mechanism of assessment fish hepatic simultaneously.
Description
Technical field
The invention belongs to technical field of aquaculture, are related to a kind of construction method of fish damage model, specially Rofe
Fish fats liver injury model and its construction method and application.
Background technique
Fatty liver damage is the wide hair venereal disease of one of main liver diseases of fish and long-standing problem culture fishery
Evil, risk factor is more, such as nutritional imbalance, environmental factor stresses, physiological function can lead to fish liver rouge extremely
Fat excessive buildup and disorders of lipid metabolism, and then cause hepatic disease.Fatty liver disease, which involves the nearly all master in China, breeds fish class,
It mainly includes reducing growth, efficiency of feed utilization, immunity, stress tolerance, product sensory etc. that it, which is endangered, is made to culture fishery
At great negative effect.Though being studied for many years and prevention practice, cannot still be contained at all so far, occurrence and development mechanism
Also it does not illustrate completely.
In view of the limitation of investigative technique, the progress of fatty,fiss hepatic injury is slow.Animal model is research fat
Property hepatic injury pathogenesis, screening effectively prevent the important means of drug.In mouse, fatty liver is constructed using high lipid food
Injured animal model is presently most used method, has the advantages that easy to operate, reproducible, but there is also the modeling times
The defects of length, experimental cost is higher.In fish, although existing research is shown, researching fish rouge can be induced by high lipid food
The pathogenesis of fat liver, but the report at present about building fatty,fiss liver injury model is less.
A kind of grass carp fatty liver disease experimental model method for building up of Chinese patent CN201110001159.1, the side disclosed
Method disposably injects the normal saline solution of thioacetamide to grass carp the following steps are included: 1) by the way of intraperitoneal injection,
It is cultivated after injection；In the normal saline solution of the thioacetamide concentration of thioacetamide be 100~
200mg/ml；2) in breeding process, the mixed feed that grease total amount is 4~6% is fed, is cultivated 2~6 weeks；Since the present invention is right
Grass carp injection thioacetamide causes grasscarp liver tissue, cellular damage, interferes lipid-metabolism and the energetic supersession of grass carp, together
When feed the mixed feed of appropriate fat content, can rapidly obtain the test grass carp of fatty liver disease.The deficiency of this method
Be: 1) this method be injection method, easily cause fish mechanicalness hepatic injury, at the same to fish stress be larger；2) this method
Easily cause fish dead (highest survival rate 85%)；3) model is mainly the liver as caused by chemical substance (thioacetamide)
Hepatic injury caused by metabolic disorder, and do not pay close attention to the generation of the main fatty liver of fish (nutritional fatty liver), development
Mechanism is combined not close with production reality.
Summary of the invention
The technical issues of solution: for overcome the deficiencies in the prior art, fatty liver disease incidence machine is preferably studied
System and screening associated treatment drug and a kind of ideal animals experimental model for constructing, the present invention provides Tilapia mossambica fatty livers
Damage model and its construction method and application.
Technical solution: the construction method of Tilapia mossambica fatty liver damage model the described method comprises the following steps:
Step 1, domestication
The healthy Tilapia mossambica without wound is chosen, 142.15 ± 7.55g of initial weight is supported and tamed in circulation, is tamed
Condition are as follows: 30 ± 2 DEG C of temperature；Dissolved oxygen > 6mg/L；PH 7.4-8.1, the domestication time is 2 weeks, daily according to the 2- of fish total weight
3% feeds basal feed feed, periodically feeds 2 times, respectively 9:00 and 16:00；
Step 2 prepares high lipid food
The high lipid food includes following components by weight: 85-92 parts of Tilapia mossambica basal feeds, 7-13 parts of lards,
0.75-1.5 portions of soybean oils and 0.25-0.5 parts of cholesterol；
Step 3, induction
Tilapia mossambica after choosing step 1 domestication, feeds high lipid food, twice daily, 9:00 and 16:00, daily feeding volume
It for the 2-3% of fish total weight, continuously feeds 8-12 weeks, obtains Tilapia mossambica fatty liver damage model；
Step 4, evaluation analysis
Serum biochemical markers object, liver lipids level, Histopathology is carried out for the Tilapia mossambica after step 3 induction to examine
It is disconnected, the hepatic injury degree of judgment models.
Preferably, Tilapia mossambica basal feed described in step 2 are as follows: 28 parts of crude protein, 11.6 parts of crude fibre, coarse ash 15
Part, 6 parts of crude fat, 0.7 part of total phosphorus, 1.3 parts of total amino acid.
Preferably, high lipid food described in step 2 includes following components by weight: 85 parts of Tilapia mossambica basal feeds, 13
Part lard, 1.5 portions of soybean oils and 0.5 part of cholesterol.
The Tilapia mossambica fatty liver damage model of any description above method building.
Application of the Tilapia mossambica fatty liver damage model described above in fatty,fiss liver study of incident mechanism model.
Application of the Tilapia mossambica fatty liver damage model described above in fatty,fiss liver drug screening.
The principle of Tilapia mossambica fatty liver damage model construction method of the present invention: since intake fat is excessive, cause
Fat deposition in Tilapia mossambica hepatic tissue, and then cause adipose metabolism disorder, oxidative stress and inflammatory reaction, finally cause liver
Meronecrosis and liver tissue injury.In fish, nutritional fatty liver be most important liver damage disease, principal causative because
Element is overnutrition, therefore this experimental model agrees with production practices completely, facilitates the generation and the hair that disclose fatty,fiss hepatopathy
Exhibition mechanism and demand effectively preventing measure.
The utility model has the advantages that the high lipid food that (1) the method for the invention uses is formulated, simple, easily operated, ingredient is easy to control,
It is high to model success rate, and stablize, the modeling time it is shorter, have good application prospect；(2) animal model constructed by the present invention can
For studying the mutual of fatty,fiss hepatic injury pathogenesis and fat metabolic disturbance caused by many factors and hepatic injury
Relationship, while can be used for screening the drug of prevention and treatment fatty,fiss liver, and the pharmacological mechanism of assessment fish hepatic.
Detailed description of the invention
Fig. 1 is Tilapia mossambica hepatic tissue pathology slice HE colored graph, and wherein A is Normal group, and B is model group；
Fig. 2 is Tilapia mossambica hepatic tissue pathology slice oil red O stain figure, and wherein A is Normal group, and B is model group.
Specific embodiment
Following embodiment further illustrates the contents of the present invention, but should not be construed as limiting the invention.Without departing substantially from
In the case where spirit of that invention and essence, to modification made by the method for the present invention, step or condition and replaces, belong to the present invention
Range.Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art.
Embodiment 1
(1) experimental design
The healthy Tilapia mossambica (142.15 ± 7.55g of initial weight) without wound is chosen, is randomly divided into 2 groups: model group and blank control
Group, every group of 30 fishes.All Tilapia mossambicas start formally to feed after adaptive feeding 2 weeks.Model group feeds high lipid food:
85% Tilapia mossambica basal feed, 13% lard, 1.5% soybean oil and 0.5% cholesterol.Blank control group feeds basal feed.
Two groups of feeding volumes are identical, twice daily (9:00 and 16:00), and daily feeding volume is the 2~3% of fish total weight, continuously feed 8
Week.After modeling 4 weeks, there is fatty liver symptom in Tilapia mossambica, and after 8 weeks, Tilapia mossambica shows apparent fat deposition and hepatic injury.
(2) Tilapia mossambica hepatic injury evaluation method and interpretation of result
Evaluation method includes serum biochemical markers object, liver lipids level, Tissue pathological diagnosis.After feeding 8 weeks, fasting
Then 12h acquires blood and hepatic tissue.
1, serum biochemical markers object changes: blood serum designated object is mainly glutamic-oxalacetic transaminease (GOT), glutamic-pyruvic transaminase (GPT)
With malonaldehyde (MDA), the results are shown in Table 1.
1 high lipid food of table feeds the variation of serum GPT, GOT and MDA after 8 weeks
| GPT(IU/L) | GOT(IU/L) | MDA(μM) | |
| Blank control group | 8.23±0.91 | 6.47±0.99 | 4.74±0.33 |
| Model group | 23.90±1.54** | 13.39±1.76** | 6.22±0.32* |
Table 1 shows mould the results show that the serum biochemical markers object level of model group Tilapia mossambica is significantly higher than blank control group
There is obvious damage in the liver of type group Tilapia mossambica.
2, liver lipids are horizontal: free fatty acid (FFA), triglycerides (TG), cholesterol (TC), the results are shown in Table 2.
The variation of hepatic tissue FFA, TG and TC content after 2 high lipid food of table is fed 8 weeks
Table 2 is as the result is shown: compared with blank control group, the liver lipids horizontal (FFA, TG, TC) of model group Tilapia mossambica are aobvious
It writes and increases, show to deposited a large amount of lipid in model group liver of hybrid tilapia.
3, histopathological analysis: HE stained slice and oil red O stain slice, as depicted in figs. 1 and 2.
Fig. 1 is as the result is shown: there is apparent vacuole, lipid degeneration, meronecrosis and inflammatory in model group Tilapia mossambica hepatic tissue
Cellular infiltration.Fig. 2 result further demonstrates that, a large amount of lipidosis in model group Tilapia mossambica hepatic tissue.
Embodiment 2
(1) experimental design
Based on this animal model, the protective effect that jamaicin and polygonin damage Tilapia mossambica fatty liver is had rated.If
Set blank control group, model group and drug-treated group (jamaicin and polygonin processing group).Blank control group feeds basal feed；
Model group feeds high lipid food (85% Tilapia mossambica basal feed, 13% lard, 1.5% soybean oil and 0.5% cholesterol)；Barberry
Alkali group feeds the high lipid food of the jamaicin containing 50mg/kg；Polygonin group feeds the high lipid food of the polygonin containing 1g/kg.Every group 30
Fish is three and parallel repeats.This test culture-cycle is 8 weeks, carries out sampling analysis at the 8th week.
(2) jamaicin and polygonin hepatoprotective effect evaluation method and interpretation of result
Evaluation method includes serum biochemical markers object, liver lipids level, hepatic tissue antioxidant levels.After feeding 8 weeks, prohibit
12h is eaten, blood and hepatic tissue are then acquired.
1, serum biochemical markers object changes: GOT and GPT, the results are shown in Table 3.
The influence of 3 jamaicin of table and polygonin to tilapia serum GPT and GOT
| GPT(IU/L) | GOT(IU/L) | |
| Blank control group | 10.72±1.13** | 6.23±1.42** |
| Model group | 23.29±1.87 | 19.05±1.93 |
| Jamaicin group | 11.93±1.29** | 7.25±1.01** |
| Polygonin group | 11.72±1.67** | 9.66±0.47** |
The results show that compared with model group, jamaicin and polygonin drug-treated group significantly inhibit in tilapia serum table 3
GPT and GOT vigor significantly alleviates Tilapia mossambica hepatic injury after showing jamaicin and polygonin drug-treated.
2, liver lipids are horizontal: FFA, TG, TC, the results are shown in Table 4.
The influence of 4 jamaicin of table and polygonin to hepatic tissue FFA, TG and TC content
| FFA(μmol/gliver) | TG(μmol/gliver) | TC(μmol/gliver) | |
| Blank control group | 0.97±0.08** | 17.41±1.83** | 12.22±1.21** |
| Model group | 2.87±0.16 | 31.43±3.77 | 28.17±2.98 |
| Jamaicin group | 1.43±0.13* | 20.16±1.91* | 17.95±2.31* |
| Polygonin group | 1.52±0.11* | 19.33±2.18* | 16.38±2.34* |
Table 4 is as the result is shown: compared with model group, jamaicin and polygonin processing cause fat content in Tilapia mossambica hepatic tissue
It is remarkably decreased, shows that drug-treated significantly suppresses liver of hybrid tilapia fat metabolic disturbance, and alleviate the rouge in hepatic tissue
Fat deposition.3, liver antioxidant levels: superoxide dismutase (SOD) and malonaldehyde (MDA), the results are shown in Table 5.
The influence of 5 jamaicin of table and polygonin to Tilapia mossambica oxidation resistance
| SOD(U/mgprot) | MDA(μmol/gprot) | |
| Blank control group | 183.55±18.23* | 1.04±0.09* |
| Model group | 149.42±10.52 | 1.87±0.12 |
| Jamaicin group | 187.15±7.44* | 1.09±0.11* |
| Polygonin group | 179.47±16.21* | 0.93±0.07** |
Table 5 is as the result is shown: compared with model group, it is living that jamaicin and polygonin processing significantly improve Tilapia mossambica hepatic tissue SOD
Power shows the oxidative stress that two kinds of drugs can inhibit high lipid food to induce；Jamaicin and polygonin significantly reduce liver simultaneously
MDA content in tissue shows that drug-treated significantly suppresses lipid peroxidation in liver of hybrid tilapia, alleviates oxidative stress to liver
The damage of cell.
Claims (6)
1. the construction method of Tilapia mossambica fatty liver damage model, which is characterized in that the described method comprises the following steps:
Step 1, domestication
The healthy Tilapia mossambica without wound is chosen, 142.15 ± 7.55g of initial weight is tamed, acclimation conditions through experimental situation are as follows: temperature 30 ±
2℃；Dissolved oxygen > 6mg/L；PH 7.4-8.1, domestication time are 2 weeks, feed basal feed according to the 2-3% of fish total weight daily and raise
Material, periodically feeds 2 times, respectively 9:00 and 16:00；
Step 2 prepares high lipid food
The high lipid food includes following components by weight: 85-92 parts of Tilapia mossambica basal feeds, 7-13 parts of lards, 0.75-
1.5 portions of soybean oils and 0.25-0.5 parts of cholesterol；
Step 3, induction
Tilapia mossambica after choosing step 1 domestication, feeds high lipid food, and twice daily, 9:00 and 16:00, daily feeding volume are fish
The 2-3% of total weight, continuously feeds 8-12 weeks, obtains Tilapia mossambica fatty liver damage model；
Step 4, evaluation analysis
Serum biochemical markers object, liver lipids level, Tissue pathological diagnosis are carried out for the Tilapia mossambica after step 3 induction, is sentenced
The hepatic injury degree of disconnected model.
2. the construction method of Tilapia mossambica fatty liver damage model according to claim 1, which is characterized in that by weight
Meter, Tilapia mossambica basal feed described in step 2 are as follows: 28 parts of crude protein, 11.6 parts of crude fibre, 15 parts of coarse ash, 6 parts of crude fat,
0.7 part of total phosphorus, 1.3 parts of total amino acid.
3. the construction method of Tilapia mossambica fatty liver damage model according to claim 1, which is characterized in that step 2 institute
State high lipid food includes following components by weight: 85 parts of Tilapia mossambica basal feeds, 13 parts of lards, 1.5 portions of soybean oils and 0.5
Part cholesterol.
4. the Tilapia mossambica fatty liver damage model of any the method building of claim 1-3.
5. the answering in fatty,fiss liver study of incident mechanism model of Tilapia mossambica fatty liver damage model described in claim 4
With.
6. application of the Tilapia mossambica fatty liver damage model described in claim 4 in fatty,fiss liver drug screening.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113080338A (en) * | 2021-04-29 | 2021-07-09 | 中国水产科学研究院南海水产研究所 | Micropterus salmoides fatty liver model and construction method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101810866A (en) * | 2010-05-13 | 2010-08-25 | 江苏省中医药研究院 | New method for screening anti-liver injury medicament by using model organism zebra fish |
| CN102027886A (en) * | 2011-01-05 | 2011-04-27 | 苏州大学 | Method for establishing grass carp fatty liver disease experimental model |
| CN104542389A (en) * | 2014-12-23 | 2015-04-29 | 中国科学院苏州生物医学工程技术研究所 | Preparation method for non-alcoholic fatty liver disease zebra fish |
| CN105400732A (en) * | 2015-11-19 | 2016-03-16 | 南京理工大学 | Establishment method for alcoholic fatty liver zebra fish model |
-
2019
- 2019-04-22 CN CN201910327015.1A patent/CN110036951A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101810866A (en) * | 2010-05-13 | 2010-08-25 | 江苏省中医药研究院 | New method for screening anti-liver injury medicament by using model organism zebra fish |
| CN102027886A (en) * | 2011-01-05 | 2011-04-27 | 苏州大学 | Method for establishing grass carp fatty liver disease experimental model |
| CN104542389A (en) * | 2014-12-23 | 2015-04-29 | 中国科学院苏州生物医学工程技术研究所 | Preparation method for non-alcoholic fatty liver disease zebra fish |
| CN105400732A (en) * | 2015-11-19 | 2016-03-16 | 南京理工大学 | Establishment method for alcoholic fatty liver zebra fish model |
Non-Patent Citations (1)
| Title |
|---|
| 李钰等: "斑马鱼营养性脂肪肝模型构建", 《水产学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113080338A (en) * | 2021-04-29 | 2021-07-09 | 中国水产科学研究院南海水产研究所 | Micropterus salmoides fatty liver model and construction method and application thereof |
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