CN105850868A - Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice - Google Patents
Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice Download PDFInfo
- Publication number
- CN105850868A CN105850868A CN201610189192.4A CN201610189192A CN105850868A CN 105850868 A CN105850868 A CN 105850868A CN 201610189192 A CN201610189192 A CN 201610189192A CN 105850868 A CN105850868 A CN 105850868A
- Authority
- CN
- China
- Prior art keywords
- mice
- model
- group
- fatty liver
- fat
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 13
- 238000013232 NAFLD rodent model Methods 0.000 title description 2
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 claims abstract description 33
- 210000004185 liver Anatomy 0.000 claims abstract description 25
- 238000011740 C57BL/6 mouse Methods 0.000 claims abstract description 23
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 claims abstract description 18
- 241000699670 Mus sp. Species 0.000 claims abstract description 17
- 210000004369 blood Anatomy 0.000 claims abstract description 12
- 239000008280 blood Substances 0.000 claims abstract description 12
- 208000032928 Dyslipidaemia Diseases 0.000 claims abstract description 8
- 208000017170 Lipid metabolism disease Diseases 0.000 claims abstract description 8
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 claims abstract description 8
- 210000005228 liver tissue Anatomy 0.000 claims abstract description 5
- 239000012188 paraffin wax Substances 0.000 claims abstract description 4
- 150000002632 lipids Chemical class 0.000 claims description 22
- 238000010171 animal model Methods 0.000 claims description 16
- 235000013305 food Nutrition 0.000 claims description 16
- 238000001514 detection method Methods 0.000 claims description 7
- 210000002966 serum Anatomy 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 230000037396 body weight Effects 0.000 claims description 4
- 230000006698 induction Effects 0.000 claims description 4
- 230000007170 pathology Effects 0.000 claims description 4
- 239000011547 Bouin solution Substances 0.000 claims description 2
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 2
- 229930040373 Paraformaldehyde Natural products 0.000 claims description 2
- 230000004888 barrier function Effects 0.000 claims description 2
- 210000005252 bulbus oculi Anatomy 0.000 claims description 2
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 claims description 2
- 229960002327 chloral hydrate Drugs 0.000 claims description 2
- 238000004043 dyeing Methods 0.000 claims description 2
- 239000000834 fixative Substances 0.000 claims description 2
- 239000006166 lysate Substances 0.000 claims description 2
- 229920002866 paraformaldehyde Polymers 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 206010002091 Anaesthesia Diseases 0.000 claims 1
- 230000037005 anaesthesia Effects 0.000 claims 1
- 210000000232 gallbladder Anatomy 0.000 claims 1
- 239000007928 intraperitoneal injection Substances 0.000 claims 1
- 210000001519 tissue Anatomy 0.000 abstract description 8
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 238000011713 fatty liver animal model Methods 0.000 abstract description 4
- 235000005911 diet Nutrition 0.000 abstract description 3
- 230000002440 hepatic effect Effects 0.000 abstract description 3
- 235000012000 cholesterol Nutrition 0.000 abstract description 2
- 230000037213 diet Effects 0.000 abstract description 2
- 230000003908 liver function Effects 0.000 abstract description 2
- 238000012423 maintenance Methods 0.000 abstract description 2
- 230000003044 adaptive effect Effects 0.000 abstract 1
- 238000010186 staining Methods 0.000 abstract 1
- 208000010706 fatty liver disease Diseases 0.000 description 11
- 208000004930 Fatty Liver Diseases 0.000 description 10
- 206010019708 Hepatic steatosis Diseases 0.000 description 10
- 231100000240 steatosis hepatitis Toxicity 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 9
- 230000001575 pathological effect Effects 0.000 description 9
- 238000011161 development Methods 0.000 description 6
- 230000018109 developmental process Effects 0.000 description 6
- 108010028554 LDL Cholesterol Chemical group 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000009825 accumulation Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 230000037356 lipid metabolism Effects 0.000 description 3
- 230000008506 pathogenesis Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
- 102000013918 Apolipoproteins E Human genes 0.000 description 2
- 108010025628 Apolipoproteins E Proteins 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 208000029767 Congenital, Hereditary, and Neonatal Diseases and Abnormalities Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 208000024335 physical disease Diseases 0.000 description 2
- 208000007082 Alcoholic Fatty Liver Diseases 0.000 description 1
- 206010016262 Fatty liver alcoholic Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 108010023302 HDL Cholesterol Chemical group 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241001142635 Lema Species 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 208000026594 alcoholic fatty liver disease Diseases 0.000 description 1
- 230000003444 anaesthetic effect Effects 0.000 description 1
- 238000013096 assay test Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 235000009200 high fat diet Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000031142 liver development Effects 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 201000001862 viral hepatitis Diseases 0.000 description 1
Classifications
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Fodder In General (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to establishment of a non-alcoholic fatty liver disease animal model by utilizing ApoE-/-mice, specifically to the establishment of a high-fat induced non-alcoholic fatty liver disease animal model with severe dyslipidemia. The establishment is realized through the following steps: selecting SPF-grade male AopE-/-mice and C57BL/6 mice, and according to the variety of the mice, randomly dividing the mice into model groups and contrast groups, i.e., 4 groups in total and 8 mice in each group; subjecting all mice to adaptive feeding with an ordinary maintenance feed for a week, then supplying a high-fat feed to the model groups, and supplying a contrast feed to the contrast groups; after ten weeks, killing the mice and collecting blood, and detecting four routine blood indexes and liver function; taking fresh liver tissue homogenates, and determining the contents of cholesterol and triglyceride in tissues; and taking hepatic tissues, preparing the hepatic tissues into paraffin and frozen slices, respectively carrying out H.E. and oil red O staining, and determining the effect and the quality of the non-alcoholic fatty liver disease (NAFLD) model. According to the invention, simple high-fat and high-calorie diet is used to establish a model, and a severe non-alcoholic fatty liver disease (NAFLD) model is established within a short period of 10 weeks; and indexes of blood fat, liver fat and the like of the model have excellent stability and high repeatability; thus, a reliable method is provided for the establishment of the non-alcoholic fatty liver disease (NAFLD) model with high quality.
Description
Technical field
The present invention relates to the foundation of non-alcoholic fatty liver disease (NAFLD) animal model, be specially with serious dyslipidemia
The foundation of the non-alcoholic fatty liver disease animal model of one class height fat induction.
Background technology
Non-alcohol fatty liver (NAFLD) refers in addition to ethanol and other clear and definite damage liver factors, in causing hepatocyte
Fat over-deposit is the clinical pathology syndrome of principal character.NAFLD is made up of the disease of a series of continuous development, the most only
There is simple fatty liver (SFL) and non-alcoholic stellato-hepatitis (NASH).Along with the development of pathological changes, finally cause liver
The fibrosis of tissue, and cause the formation of liver cirrhosis.Obesity and dyslipidemia caused by NAFLD are also to cause other simultaneously
Chief culprit is returned in the crime of dysbolismus disease, such as atherosclerosis (AS), hypertension and type-II diabetes etc..Along with society
Progressive, people's dietary structure there occurs change, large quantities of higher fatty acid, hypercholesterolemia, and the food of high heat enters the dining table of people,
Incident is that the continuous of China's pathogenesis of fatty liver rate promotes, it has also become be only second to the second largest hepatic disease of viral hepatitis.
People commonly use various means to study various disease, and one of most important of which method builds animal model exactly.In order to more
Fully understand the generating process of fatty liver, have been carried out many clinical researches.But, NAFLD Disease evolution is with development time very
Long, this largely limits the data that we obtain having reasonable dismissal.Then, attention is gone to development conjunction by researcher
Fatty liver forming process is verified in suitable animal model direction.Set up the NAFLD that reliable and stable, Pathologic process meets
Animal model, pathogenesis and Therapeutic Method to research NAFLD are significant.
Existing non-alcoholic fatty liver disease animal model classification is numerous and diverse, and animal selects also to be respectively arranged with length, and rodent mostly is SD
Rat, wistar rat, C57BL/6 mice, Holland rabbit etc., non-rodent includes pig, dog etc..These animal models
Though non-alcoholic fatty liver disease Development process can be reacted to a certain extent, but mostly existence and stability is poor, become the feature of mould cycle length.
Use herein especially the ApoE-/-clpp gene deratization using C57BL/6 as background replace wild type C57BL/6 as object of study,
ApoE-/-mice, because the shortage of apo E, causes congenital disorder of lipid metabolism, the high fat food of the daily ingestion of abundance of mice
Thing can not metabolism in time go out external, causes lipid in the accumulation of liver.The more important thing is, ApoE-/-mouse model occurs serious
Dyslipidemia, this is an important indicator in fatty liver development process, mutual for discussion NAFLD and cardiovascular disease further
Pathogenesis for effect, it is provided that a kind of method reliably.
Summary of the invention
The present invention provides a kind of ApoE-of utilization/-mice to set up the simple fatty liver with serious dyslipidemia induced through high fat
Pathological changes animal model.
The present invention selects the male AopE-of SPF level/-mice and C57BL/6 mice, according to Mus kind random sub-model group and matched group,
Totally 4 groups, often group 8.After all mices feed 1 week with common maintenance feedstuff adaptability, model group gives high lipid food, right
Give control feed according to group, sooner or later respectively once quantitatively surely throw feedstuff, it is ensured that 40g/ cage, 4 mices of every cage.Take at blood after 10 weeks
Extremely, the conventional four items of blood lipid tests (TC, TG, LDL-c, HDL-c) of detection and liver function (ALT and AST)；Take fresh and alive liver group
Knit homogenate, measure tissue inner cholesterol and content of triglyceride；Separately take hepatic tissue and do paraffin, frozen section, carry out H.E. respectively
And oil red O stain, it is determined that NAFLD modelling effect and quality.
Feature of the present invention uses the ApoE-/-clpp gene deratization with C57BL/6 as background, is formed non-through high fat diet induction development
Alcoholic fatty liver model.ApoE-/-mice, because the shortage of apo E, causes congenital disorder of lipid metabolism so that every
Day the lipid ingested can not normal transport in vivo, cause its accumulation in liver, simultaneously with serious dyslipidemia, with
NAFLD clinical pathology similarity is high.C57BL/6 mice uses as comparison in the present invention.Animal in the inventive method
Feedstuff is control feed (ND) and high lipid food (HFD) is purchased from Jiangsu Mei Disen biological medicine company limited.Side of the present invention
The male AopE-of SPF level/-mice in method is purchased from this laboratory animal company limited of Changzhou Cavan, Jiangsu Province；SPF level is male
C57BL/6 mice is purchased from Beijing Vital River Experimental Animals Technology Co., Ltd..
The present invention uses pure height fat high caloric diet modeling, can also produce severe NAFLD within 10 weeks the shortest cycles
Model.The more important thing is, the index excellent stability such as model blood fat, liver fat, repeatable high.High-quality for NAFLD
The foundation of amount model, it is provided that a kind of method reliably.
Accompanying drawing explanation
Fig. 1 is the C57BL/6 mouse liver pathological section figure (H.E. dye X400) that control feed is fed；
Fig. 2 is AopE-/-mouse liver pathological section figure (H.E. dye X400) that control feed is fed；
Fig. 3 is the C57BL/6 mouse liver pathological section figure (H.E. dye X400) that high lipid food is fed；
Fig. 4 is AopE-/-mouse liver pathological section figure (H.E. dye X400) that high lipid food is fed；
Fig. 5 is C57BL/6 mouse liver pathological section figure (oil red O stain X400) that control feed is fed；
Fig. 6 is AopE-/-mouse liver pathological section figure (oil red O stain X400) that control feed is fed；
Fig. 7 is C57BL/6 mouse liver pathological section figure (oil red O stain X400) that high lipid food is fed；
Fig. 8 is AopE-/-mouse liver pathological section figure (oil red O stain X400) that high lipid food is fed.
Detailed description of the invention
The preparation of embodiment 1 animal model of the present invention
1, laboratory animal: the male AopE-of SPF level/-mice, 6～8 week old, body weight (22 ± 2) g, purchased from Jiangsu Province's Changzhou
This laboratory animal company limited of Cavan [SCXK (Soviet Union) 2011-003].SPF level male C57BL/6 mice, 6～8 week old, body
Weight (22 ± 2) g, purchased from Beijing Vital River Experimental Animals Technology Co., Ltd. [SCXK (capital) 2011-0011].
2, laboratory animal is all raised in our department's barrier environment, temperature 22～25 DEG C, and humidity 50%, within 12 hours, day alternates with night.
3, laboratory animal is after adaptability feeds 1 week, random point four groups, often group 8, in detail packet situation such as table 1 below.Make
Mould 10 weeks, respectively once quantitatively throws feedstuff, it is ensured that 40g/ cage, 4 mices of every cage, period free diversion, water source is sooner or later surely
The aquesterilisa of SPF level, weighs in once every two weeks.Animal feed is control feed (MD12031) and high lipid food
(MD12032) Jiangsu Mei Disen biological medicine company limited it is purchased from.
Animal is grouped table 1 in detail
4, after modeling 10 weeks, each group mice same time all puts to death, fasting 12 hours before putting to death.4% chloral hydrate abdominal cavity
Injecting anesthetic, plucks after eyeball takes blood, and whole blood 37 DEG C stands 1 hour, places clots for 4 DEG C and separates out, and 3000rmp is centrifuged 10min
Take upper serum.TC, TG, LDL-c, HDL-c, AST and ALT etc. in automatic clinical chemistry analyzer detection serum refer to
Mark, result is as shown in table 2.
Table 2 respectively organize Biochemical Indices In Serum change table (N=6)
Note: TC, TG, LDL-C and HDL-C unit be mmol/L, AST and ALT unit be U/L.A: compare with A group,
P<0.01；B: compare with B group, P < 0.01；C: compare with C group, P < 0.01.
AopE-/-mice group that high lipid food is fed all has compared to other three groups of TC, TG and LDL-C and significantly rises
(P<0.01)；High lipid food feed C57BL/6 mice group feed compared to control feed C57BL/6 mice group TC, TG
All having with LDL-C rising (P < 0.01) is significantly dropped but numerical value to compare gap little；What is interesting is and be both what control feed was fed
C57BL/6 mice group TC that AopE-/-mice group is fed compared to control feed, TG and LDL-C all have and significantly drop rising
(P < 0.01) and numerical difference are away from bigger.Between each group, in serum, AST and ALT index does not has significant difference, demonstrates that high fat lures
Lead 10 weeks, each group liver not relatively macrolesion, judge according to NAFLD PD, in the present invention, model should be pure fat
Fat liver.Data above shows that ApoE-/-mice odds ratio on serum lipids promotes is more apparent.
5, the detection of liver lipids, concrete grammar, as a example by a mice, respectively takes 100mg fresh liver tissue according to reagent
Box carries lysate and respectively adds 1ml and be homogenized, and a respective homogenate part is used for detecting triglyceride (TG) and T-CHOL (TC)
Content, another part measures its protein content respectively, finally must with every milligram of protein concentration correct triglyceride (TG) and
T-CHOL.Triglyceride (tissue) enzymic measuring reagent box (E1013) and T-CHOL (tissue) assay test kit
(E1015) purchased from Beijing Puli's lema gene Technology Co., Ltd..Result is as shown in table 3.
Table 3 respectively organize liver lipids change table (N=6)
The unit of note: TC and TG is mmol/kg.A: compare with A group, P < 0.01；B: compare with B group, P < 0.01；C: with
C group compares, P < 0.01.
Corresponding with blood fat is that ApoE-/-mouse liver accumulation of lipid ratio is more serious, AopE-/-mice that wherein high lipid food is fed
Group is compared to other three groups, and liver TG and TC rises substantially (P < 0.01), the C57BL/6 mice group phase that high lipid food is fed
C57BL/6 mice group liver TG and the TC rising fed compared with control feed is also that obvious (P < 0.01) illustrates C57BL/6
Mice height fat is induced successfully, but effect is inferior to ApoE-/-mice.
6, the detection of pathology of hepar, concrete grammar, as a example by a mice, takes the same position of liver, divides two parts.
6a, a part of Bouin ' s fixative fixes, and carries out H.E. dyeing as paraffin section.
6b, another part are fixed with 4% paraformaldehyde fixative, carry out oil red O stain as frozen section.
By pathological section it can be seen that control feed feed C57BL/6 murine liver tissue compact conformation, radioactivity liver lock construction
Clearly, cell caryoplasm full (Fig. 1 and 5)；AopE-/-murine liver tissue structure that control feed is fed is the most clear, but carefully
Born of the same parents periphery has occurred that slight fat invades profit, should be developed to mild fatty liver (Fig. 2 and 6)；The C57BL/6 that high lipid food is fed
There is fat lesion in mice group hepatocyte, occurred that cavity area has reached more than 1/2, moderate fatty liver (Fig. 3 should have been developed into
With 7)；AopE-/-mice group that high lipid food is fed, has occurred that serious hepatocyte fat fills the air and has gathered, dripped sample in big fat,
Have developed into severe fatty liver (Fig. 4 and 8).Further illustrate the success of gene knockout ApoE-/-mice NAFLD model.
In sum, ApoE-/-mice is utilized to be successfully established the non-alcoholic fatty liver disease with serious dyslipidemia through the induction of high fat
NAFLD animal model.For research NAFLD pathogeny especially NAFLD and cardiovascular disease abnormalities of sugar/lipid metabolism institute shape
The interaction become provides a kind of Novel movable object model.
Claims (1)
1. utilizing the method that ApoE-/-mice sets up nonalcoholic fatty liver model, the phase is characterised by, sets up the step of model
Suddenly it is:
1) Mus kind, is chosen: choosing the male AopE-of SPF level/-mice respectively, 6～8 week old, body weight 22 ± 2g, SPF level is male
C57BL/6 mice, 6～8 week old, body weight 22 ± 2g, raise in our department's barrier environment, temperature 22～25 DEG C, humidity 50%,
Within 12 hours, day alternates with night, and adaptability is fed 1 week；
2), packet and modeling: after adaptability feeds 1 week, random point four groups, wherein AopE-/-mice two groups, named right
According to group B and high fat group D, C57BL/6 mice two groups, named control group A and high fat group C, often group 8；Modeling 10
Week, period matched group feeding control feed, high fat group feeding high lipid food, sooner or later respectively once quantitatively surely throw feedstuff, it is ensured that 40g/
Cage, 4 mices of every cage, free diversion, water source is the aquesterilisa of SPF level, weighs in every two weeks once；
3), taking a blood sample and detect: after modeling 10 weeks, each group mice same time all puts to death, fasting 12 hours before putting to death, and 4%
Chloral hydrate intraperitoneal injection of anesthesia, plucks after eyeball takes blood, and whole blood 37 DEG C stands 1 hour, places clots for 4 DEG C and separates out, 3000rmp
Centrifugal 10min takes upper serum, TC, TG, LDL-c, HDL-c, the AST in automatic clinical chemistry analyzer detection serum
With ALT index；
4), the detection of liver lipids: each group, as a example by a mice, respectively takes 100mg fresh liver tissue according to test kit certainly
Band lysate respectively adds 1ml and is homogenized, and a respective homogenate part is used for detecting containing of triglyceride TG and T-CHOL TC
Amount, another part measures its protein content respectively, finally must correct triglyceride TG and total gallbladder with every milligram of protein concentration
Sterin TC；
5), the detection of pathology of hepar: each group, as a example by a mice, takes the same position of liver, divides two parts, a part
Fixing with Bouin ' s fixative, carry out H.E. dyeing as paraffin section, another part is fixed with 4% paraformaldehyde fixative,
Oil red O stain is carried out as frozen section；
6), to step 3) step 5) be analyzed, determine with ApoE-/-mice through high fat induction be successfully established with
The non-alcoholic fatty liver disease NAFLD animal model of serious dyslipidemia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610189192.4A CN105850868A (en) | 2016-03-28 | 2016-03-28 | Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610189192.4A CN105850868A (en) | 2016-03-28 | 2016-03-28 | Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105850868A true CN105850868A (en) | 2016-08-17 |
Family
ID=56627036
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610189192.4A Pending CN105850868A (en) | 2016-03-28 | 2016-03-28 | Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105850868A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107197823A (en) * | 2017-06-20 | 2017-09-26 | 遵义医学院 | A kind of Establishment of Rat Model method of NASH |
| CN109105333A (en) * | 2017-06-22 | 2019-01-01 | 中美冠科生物技术（太仓）有限公司 | Animal model for nonalcoholic fatty liver disease |
| CN109730028A (en) * | 2019-01-31 | 2019-05-10 | 青岛市市立医院 | A method of Models of Nonalcoholic Fatty Liver Disease is established using mouse |
| CN110151787A (en) * | 2018-02-12 | 2019-08-23 | 玛旺干细胞医学生物科技股份有限公司 | Liver-protecting combination and application thereof |
| CN111727935A (en) * | 2020-07-27 | 2020-10-02 | 中美冠科生物技术（太仓）有限公司 | Method for preparing fatty liver disease animal model by using MS-NASH mice |
| CN111789078A (en) * | 2020-07-20 | 2020-10-20 | 北京航空航天大学 | Method for establishing rat non-obese non-alcoholic fatty liver disease model |
| CN114365716A (en) * | 2021-12-06 | 2022-04-19 | 中粮集团有限公司 | Preparation method of non-alcoholic simple fatty liver golden hamster model |
| CN114503955A (en) * | 2022-03-15 | 2022-05-17 | 北京基谱生物科技有限公司 | Non-alcoholic fatty liver animal model and establishment method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101011585A (en) * | 2007-01-22 | 2007-08-08 | 浙江大学 | Process for preparing non-alcohol fatty liver mouse model |
| CN102972345A (en) * | 2012-12-12 | 2013-03-20 | 贵州大学 | Modeling processing method of non-alcoholic fatty liver mouse model |
| CN105052830A (en) * | 2015-08-04 | 2015-11-18 | 施军平 | Fatty-liver-related liver cancer model building method based on knockout mice |
| US20150327578A1 (en) * | 2014-05-15 | 2015-11-19 | Purina Animal Nutrition Llc | Methods of feeding high fat poured feed blocks |
| CN105168303A (en) * | 2015-09-30 | 2015-12-23 | 泰山医学院 | Application of Viburnum sargentii alcohol extract in preparation of NAFLD (non-alcoholic fatty liver disease) treatment drug |
-
2016
- 2016-03-28 CN CN201610189192.4A patent/CN105850868A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101011585A (en) * | 2007-01-22 | 2007-08-08 | 浙江大学 | Process for preparing non-alcohol fatty liver mouse model |
| CN102972345A (en) * | 2012-12-12 | 2013-03-20 | 贵州大学 | Modeling processing method of non-alcoholic fatty liver mouse model |
| US20150327578A1 (en) * | 2014-05-15 | 2015-11-19 | Purina Animal Nutrition Llc | Methods of feeding high fat poured feed blocks |
| CN105052830A (en) * | 2015-08-04 | 2015-11-18 | 施军平 | Fatty-liver-related liver cancer model building method based on knockout mice |
| CN105168303A (en) * | 2015-09-30 | 2015-12-23 | 泰山医学院 | Application of Viburnum sargentii alcohol extract in preparation of NAFLD (non-alcoholic fatty liver disease) treatment drug |
Non-Patent Citations (3)
| Title |
|---|
| 曹玉珍 等: "高脂饲料诱发C57BL/6J小鼠非酒精性脂肪肝相关指标的动态观察", 《东北农业大学学报》 * |
| 程勤 等: "高脂饲料诱发非酒精性脂肪肝实验研究", 《淮海医药》 * |
| 赵昕怡 等: "利拉鲁肽改善非酒精性脂肪性肝病小鼠肝脂肪沉积的机制探讨", 《中华肝脏病杂志》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107197823A (en) * | 2017-06-20 | 2017-09-26 | 遵义医学院 | A kind of Establishment of Rat Model method of NASH |
| CN109105333A (en) * | 2017-06-22 | 2019-01-01 | 中美冠科生物技术（太仓）有限公司 | Animal model for nonalcoholic fatty liver disease |
| CN109105333B (en) * | 2017-06-22 | 2022-08-05 | 浙江药源新地生物科技有限公司 | Animal model for non-alcoholic fatty liver disease |
| CN110151787A (en) * | 2018-02-12 | 2019-08-23 | 玛旺干细胞医学生物科技股份有限公司 | Liver-protecting combination and application thereof |
| CN109730028A (en) * | 2019-01-31 | 2019-05-10 | 青岛市市立医院 | A method of Models of Nonalcoholic Fatty Liver Disease is established using mouse |
| CN111789078A (en) * | 2020-07-20 | 2020-10-20 | 北京航空航天大学 | Method for establishing rat non-obese non-alcoholic fatty liver disease model |
| CN111789078B (en) * | 2020-07-20 | 2022-02-25 | 北京航空航天大学 | Method for establishing rat non-obese non-alcoholic fatty liver disease model |
| CN111727935A (en) * | 2020-07-27 | 2020-10-02 | 中美冠科生物技术（太仓）有限公司 | Method for preparing fatty liver disease animal model by using MS-NASH mice |
| CN114365716A (en) * | 2021-12-06 | 2022-04-19 | 中粮集团有限公司 | Preparation method of non-alcoholic simple fatty liver golden hamster model |
| CN114503955A (en) * | 2022-03-15 | 2022-05-17 | 北京基谱生物科技有限公司 | Non-alcoholic fatty liver animal model and establishment method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105850868A (en) | Method for establishment of non-alcoholic fatty liver disease model by utilizing ApoE-/-mice | |
| Sandnes et al. | Hyperferritinemia—a clinical overview | |
| Welsh et al. | Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches | |
| Lin et al. | NMR-based fecal metabolomics fingerprinting as predictors of earlier diagnosis in patients with colorectal cancer | |
| CN105850867A (en) | Method for modeling mouse models with non-alcoholic fatty liver disease (NAFLD) | |
| Primrose et al. | Metabolomics and human nutrition | |
| CN107773566B (en) | A kind of construction method of tire source property metabolic syndrome animal model and its application | |
| Stupp et al. | Isotopic ratio outlier analysis global metabolomics of Caenorhabditis elegans | |
| CN106526156B (en) | A method of detection, screening and identification syndrome of deficiency of kidney yang metabolism biological marker | |
| CN108588210B (en) | Hepatic injury biomarker, method and application comprising biological micromolecule and gene | |
| CN101285834A (en) | Method for accurately and rapidly checking aspergillus flavus toxin | |
| Bronowicka-Szydełko et al. | Association of novel advanced glycation end-product (AGE10) with complications of diabetes as measured by enzyme-linked immunosorbent assay | |
| Tee et al. | Ketohexokinase-mediated fructose metabolism is lost in hepatocellular carcinoma and can be leveraged for metabolic imaging | |
| CN103478071A (en) | Method for establishing diabetes-inducing model | |
| Wang et al. | Metabolomics reveals discrimination of Chinese propolis from different climatic regions | |
| Zulkifli et al. | Chemometric-empowered spectroscopic techniques in pharmaceutical fields: A bibliometric analysis and updated review | |
| CN104237504A (en) | Immunofluorescence method and kit for pathological autopsy of anaphylactic shock | |
| CN110278914A (en) | A kind of alcoholic fibrosis and inflammatory animal model and its construction method and purposes | |
| CN101846685A (en) | Enzyme-linked immunoassay kit for chlamys ferreri blood cells and preparation method thereof | |
| CN104721238B (en) | The hepatitis b virus infected construction method for merging mice alcoholic fatty liver model | |
| Shi et al. | Compound pollen protein nutrient increases serum albumin in cirrhotic rats | |
| Wen et al. | The influence of corn silk polysaccharide on signal pathway of TGF-β1 in type 2 diabetic mellitus rat | |
| Cao et al. | Analysis of five active ingredients of Er‐Zhi‐Wan, a traditional Chinese medicine water‐honeyed pill, using the biopharmaceutics classification system | |
| CN106645748A (en) | Mammalian differential serum protein and differentially expressed protein system of penile smooth muscle tissue of abnormal and phlegmatic type of impotence syndrome and screening method and application | |
| Zhou et al. | Five‐in‐One: Simultaneous isolation of multiple major liver cell types from livers of normal and NASH mice |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160817 |