CN109481428A - A kind of construction method of diabetic cardiomyopathy animal model - Google Patents
A kind of construction method of diabetic cardiomyopathy animal model Download PDFInfo
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- 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
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a kind of construction method of diabetic cardiomyopathy animal model, the SD rat raised through high glucose and high fat is first injected intraperitoneally with STZ more times, after reaching diabetes diagnostic criterion, then is subcutaneously injected with isoprel.After continuing raising 2 weeks, ventricular hemodynamics parameter and electrocardiogram parameters are detected by left ventricular cannulation, three lead electrocardiogram, show that heart function damage occurs in rat model；Heart weight index and left ventricle index show that model group rats ventricle is obviously loose；Rat blood biochemical analysis shows that significant myocardial damage occurs in rat；HE pathological staining meets the pathological change of diabetic cardiomyopathy；Masson collagen staining meets connective tissue proliferation between diabetic cardiomyopathy cardiac muscle.The present invention have it is easy to operate, the modeling time is short, modeling stage mortality of animals is low, the distinguishing features such as modeling success rate height, and can preferably imitate people's type II diabetes myocardial injury pathology pass through.
Description
Technical field
The invention belongs to area of pharmacology, and in particular to a kind of modeling method of NIDDM rats cardiomyopathy model.
Background technique
Diabetes (diabetes mellitus, DM) are a kind of since hypoinsulinism or peripheral tissues are to pancreas islet
Metabolic disease caused by element is insensitive characterized by lasting hyperglycemia state, and may cause various tissues, internal organs (such as
Eye, kidney, heart, blood vessel, nerve etc.) long-term damage, insufficiency or failure, it is in the majority with type II diabetes.According to statistics, 2015
There are about 4.15 hundred million people to suffer from diabetes (illness rate 8.8%) in year 20~79 years old people in the whole world, in addition have 3.18 hundred million people's sugar tolerances by
It damages (early period of illness rate 6.7%).China is global the first big country of diabetic, and sufferer number is up to 1.096 hundred million people within 2015,
1300000 people die of diabetes and its complication.It is predicted simultaneously according to IDF, if intervention is not added, the year two thousand forty whole world diabetic will
Up to 6.42 hundred million, prediabetes crowd 4.81 hundred million, China patient populations will rise to 1.54 hundred million.
Diabetic cardiomyopathy (diabetic cardiomyopathy, DCM), which refers to, betides diabetic, Bu Nengyong
Hypertensive cardiopathy, coronary atherosclerotic heart disease, valvulopathy and other heart diseases are come the cardiac muscle explained
Disease.The disease causes the extensive focal necrosis of cardiac muscle on the basis of metabolic disorder and microangiopathies, subclinical heart function occurs
Can be abnormal, heart failure, arrhythmia cordis and cardiogenic shock are finally progressed to, patient with severe symptoms even dies suddenly.Diabetic cardiomyopathy
Have the characteristics that morbidity is early, progress is fast, the death rate is high in diabetic complication, be mainly shown as in early days cardiac diastolic function by
Damage, it is impaired that stage shows as full impaired cardiac function, especially left ventricular function.
The pathogenesis of diabetic cardiomyopathy is more complex, completely not clear yet so far.It is disorderly to be related to high sugared toxicity, lipid metaboli
Disorderly, cardiac hemodynamic exception and many aspects such as inflammation and oxidative stress.Therefore, mankind's sugar can be imitated by establishing one kind
The animal model for urinating sick dilated cardiomyopathy process can aid in the pathogenesis for understanding diabetic cardiomyopathy in depth, thus more preferably
Prevention and treatment diabetic cardiomyopathy.The current research model in relation to diabetic cardiomyopathy is less, only has been reported that injection streptozotocin
(STZ) animal hyperglycemia state is caused to establish spontaneous model, but its modeling time is long (especially type II diabetes cardiomyopathy),
Mortality of animals height (diabetic cardiomyopathy sheet is as one of reason), model is low at mould rate and pathological change degree difference is big, and
It cannot reflect diabetic cardiomyopathy generation, chronic pathology and Characteristics of Physiological Changes in development process completely.
Isoprel (ISO) is used to construct myocardial hypertrophy and myocardial infarction and ischemia model is long-standing, which makes because of it
The advantages that mould method is simple, stablizes (about 80%) at mould rate, widely uses as pharmacology classical model always.But it is usually
Modeling is carried out for healthy animal.
Summary of the invention
The purpose of the present invention is to provide a kind of construction methods of diabetic cardiomyopathy animal model.
In order to achieve the above objectives, the invention adopts the following technical scheme:
The construction method is the following steps are included: to the obtained diabetes animal model of induction, then with isoprel skin
Under be injected into capable intervention, separated in time is detected referring to diabetic cardiomyopathy test rating after intervention, obtains sugar
Urinate sick cardiomyopathic animals model.
Preferably, the diabetes animal model is the type-2 diabetes mellitus animal model induced by STZ.
Preferably, the abductive approach of the diabetes animal model with high-sugar-fat-diet specifically includes the following steps: fed
It brings up mouse (for example, healthy SD rat), feeds to after the 6th week, increase 15mg/kg STZ intraperitoneal injection processing, wherein STZ is every
Week injection 1 time, continuous injection filter out blood glucose >=16.7mmol/L rat after STZ 4 weeks, obtain diabetes animal model (II
Patients with type Ⅰ DM rat model).
Preferably, the high-sugar-fat-diet includes following components as mass fraction: 15~20% lard, 14~
20% sucrose, 5~11% casein, 2~5% maltodextrin, 1~3% premix and 43~62% breeding
Mouse material.
Preferably, the nursing of the high-sugar-fat-diet uses free foraging pattern.
Preferably, the purity of the STZ is HPLC >=98%, and STZ is using 0.1mol/L citric acid soln and citric acid two
The mixture of sodium buffer is dissolved and (is used while allocating), and the volume fraction of citric acid soln described in mixture is 1%, mixing
The pH of object is 4~4.5.
Preferably, the intervention is specifically includes the following steps: with 2~6mg/kg isoprel continuous subcutaneous injection 5
It~7 days, is injected daily during intervention 1~2 time.The accumulative injection dosage of isoprel is excessively high, then animal is dead during modeling
Rate increase is died, injection dosage is too low, then is unable to reach expected modeling purpose.
Preferably, it for type II diabetes rat, is fed in the intervention using above-mentioned high-sugar-fat-diet.
By keeping metabolic disorder, to finally obtain type II diabetes Dilated Cardiomyopathy Model in Rats.
Preferably, after the final injection of intervention 1~3 week, cardiac determination, cardiac weight and heart weight index is carried out and is surveyed
Fixed, blood parameters measurement and pathological examination (HE dyeing, Masson dyeing).The successful animal of modeling (for example, rat)
It can be used for the research such as pharmacology, the animal pattern without subsequent processing is finally dead due to diabetic cardiomyopathy, i.e., structure of the present invention
The animal model built is stablized.
Preferably, the purity of the isoprel is HPLC >=98%, and isoprel is dissolved using PBS solution
It is subcutaneously injected after (matching while using) in rat neck.
The beneficial effects of the present invention are embodied in:
Compared with existing modeling method, the present invention passes through further isoproterenol on the basis of diabetes animal model
Element is intervened, and diabetic cardiomyopathy model is established, with easy to operate, the modeling time is short, modeling stage mortality of animals is low, modeling
The distinguishing features such as success rate height, and the pathology that can preferably imitate people's diabetes myocardial injury passes through, and has good reality
Application value (for example, building type II diabetes cardiomyopathy model).
Detailed description of the invention
Fig. 1 is blank control group HE stained slice.
Fig. 2 is simple diabetes group HE stained slice.
Fig. 3 is ISO intervention group HE stained slice.
Fig. 4 is blank control group Masson stained slice.
Fig. 5 is simple diabetes group Masson stained slice.
Fig. 6 is ISO intervention group Masson stained slice.
Specific embodiment
Invention is further described in detail with reference to the accompanying drawings and examples.
One, experimental animal
SD rat 70, male, 150~180g of weight is purchased from air force surgeon university Experimental Animal Center.Raising temperature is
22~26 DEG C, relative humidity 35%~70%, lamp, free water.
Two, it is grouped
First group: blank control group (20)
Second group: simple diabetes group (25)
Third group: ISO intervention group (25)
More 5 rats of design of second group and third component are in case experimentation is lost.
Three, modeling method
After adaptive feeding, takes out 20 at random from 70 male rats and be only used as first group (blank control group), remaining
Rat is classified as experimental group.Blank control group is enough daily until experiment terminates to feed normal diet, and on-demand free water, in fact
Test that group is daily until experiment terminates enough to feed high-sugar-fat-diet.
High-sugar-fat-diet formula (as mass fraction): 16.9% lard, 14% sucrose, 10.2% junket egg
(premix commonly uses vitamin enrichment premixed feed for animals, is production for white, 2.2% maltodextrin, 2.1% premix
A kind of additive when feed, usually there is commercialization sale of finished goods) and 54.6% breeding mouse material.
Normal diet is using breeding mouse material.
After feeding 6 weeks, 50 rats of experimental group inject STZ (STZ solution according to the dosage abdominal cavity (ip.) of every 15mg/kg
Preparation method are as follows: 0.1mol/L citric acid is mixed with disodium citrate buffer, is made into the volume point of 0.1mol/L citric acid
The solution that number is 1%, pH4~4.5, tinfoil package are protected from light, and STZ is dissolved under ice bath with the solution, is used while allocating)；Blank
0.1mol/L citric acid and disodium citrate buffer mixed liquor is injected intraperitoneally in control group in equal volume in the same way.Hereafter it presses
This method is injected weekly once, and 4 weeks (4 weeks co-injection STZ 60mg/kg of experimental group) is continuously injected.7d after final injection, Suo You great
Mouse fasting 12h measures fasting blood-glucose (FBG), and experimental group rat FBG >=16.7mmol/L's is considered as type II diabetes modeling success,
Reject wherein fasting blood-glucose person not up to standard.Then experimental group is divided into second group (simple diabetes group) and third group at random
(ISO intervention group).ISO, 5mg/kg, 1 times/d is subcutaneously injected in rear neck in third group (ISO intervention group) every rat, continuous to infuse
It penetrates 5 days；First group (blank control group), second group (simple diabetes group) is not processed.After ISO intervention group continues raising 2 weeks
It carries out index of correlation measurement (the 14th week), after first group (blank control group), second group (simple diabetes group) is continued raising 6 weeks
It carries out index of correlation measurement (the 18th week), referring to table 1.
1. diabetic cardiomyopathy mouse modeling method of table
Four, model is verified
With fasting blood-glucose >=16.7mmol/L and core function abnormality, (especially isovolumic relaxation period left indoor pressure power declines most
Big rate (- dp/dtmax)≤5250mmHg/s), heart weight index increase, blood and Myocardial Biochemical Indexes it is abnormal, and finally with disease
Neo-Confucianism checks that typical myocardial damage symptom (HE dyeing, Masson dyeing) occur models as diabetic cardiomyopathy rat and successfully mark
It is quasi-.
Five, detection method
1. fasting blood-glucose
Each group rat the 8th week (i.e. experimental group inject STZ+7d), the 12nd week, the 14th week, the 18th week (blank control group and
Simple diabetes group), carry out fasting plasma glucose.
2. cardiac determination
The chloraldurate solution 5mL/kg intraperitoneal injection of anesthesia that rat mass fraction is 10%, fixation of lying on the back, neck center
Notch；The free right common carotid artery of operation, being inserted into heart catherization to left ventricle through right common carotid artery (is 1% full of mass fraction
Heparin solution)；Left common carotid artery intubation surveys angiosthenia；Three company of leading of rat limbs records ECG signal.After stablizing 20min,
Left indoor pressure curve is recorded with System of organism signal, measures arterial pressure (systolic pressure, diastolic pressure), left ventricular contraction
The maximum rate (- dp/ for pressing (LVSP), left ventricular diastolic pressure terminal pressure (LVEDP), isovolumic relaxation period left indoor pressure power to decline
dtmax), isovolumic contraction period left indoor pressure power rise maximum rate (dp/dtmax), electrocardiogram parameters etc..Wherein isovolumic relaxation period
The maximum rate (- dp/dt of left indoor pressure power declinemax)≤5250mmHg/s shows that apparent myocardial function occurs in rat
Lesion.
3. cardiac weight and heart weight index
Rat is put to death, heart is quickly removed, filter paper blots after normal saline flushing.Measurement cardiac weight, and according to
Weight calculates heart weight index.
Heart weight index=heart weight/weight
4. Biochemical Indices In Serum
Rat eye socket metaplexus venous blood sampling, measures blood glucose value immediately.Residual blood 2000r/15min low-speed centrifugal, takes blood
Clear measurement rat biochemical indicator:
Myocarditis: lactic dehydrogenase (LDH), α-hydroxybutyrate dehydrogenase (α-HBDH), creatine kinase (CK), creatine kinase are same
Work enzyme (CK-MB)
Liver function class: alanine aminotransferase (ALT), aspartate amino transferase (AST), alkaline phosphatase
(ALP), gamma glutamyltransferase (γ-GT), total bilirubin (Tbil), bilirubin direct (Dbil)
Kidney function class: creatinine (CREA), urea (UREA)
Blood-lipoids: total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL-C), low-density lipoprotein
(LDL-C)
5. pathological examination
HE dyeing: the fixed cardiac muscular tissue of 12% formalin, gradient alcohol dehydration, paraffin embedding, 5 μm of cardiac muscular tissue are continuous
Slice, row Hematoxylin-eosin dyeing production wax disk(-sc) are main to observe (myocardial ultramicrostructure arranging situation, cardiac muscle cell gap fiber
Change and cicatricial tissue formational situation, cell infiltration situation).
Masson dyeing: after heart sections dewaxing, Bouin ' s liquid is put into acting on 2h in 37 DEG C of incubators, flowing water flushing is cut
Piece to yellow disappears；Weiger ' s Garapa uniformly dyeing 10min, 1% hydrochloride alcohol break up the several seconds, Ponceaux acid fuchsin liquid dye
10min, 1% phosphomolybdic acid aqueous solution handle about 5min, and brilliant green redyes 5min, and 1% glacial acetic acid handles 1min, gradient alcohol dehydration,
Dimethylbenzene is transparent, neutral gum sealing；Micro- sem observation.Image analysis system calculates area of collagen and accounts for gross area ratio, mainly
It observes (collagen tissue proliferative conditions after cardiac muscle cell apoptosis).
Six, statistical method
It is for statistical analysis using EXCEL software, measurement data withIt indicates, is relatively examined with t between group, in group.
Seven, testing result and analysis
The model construction is repeated 3 times altogether, and as a result height is consistent, and following result is subject to first time experimental result.
1) basic condition is tested
1. rat entirety survival condition
This experiment uses SD rat 70, rats death situation such as table 2 during experiment:
2. diabetic cardiomyopathy rat model somatic death rate of table
2. Glycemia Decline success rate:
Using fasting blood-glucose >=16.7mmol/L as standard, simple diabetes group is 100%, ISO intervention group is 100%.
3. diabetic cardiomyopathy modeling success rate:
Meet following whole indexs: fasting blood-glucose >=16.7mmol/L, core function abnormality (the especially left room of isovolumic relaxation period
The maximum rate (- dp/dt of interior pressure declinemax)≤5250mmHg/s), heart weight index increases, blood parameters are abnormal and disease
Neo-Confucianism checks typical myocardial damage symptom occur, the above index with blank control group there are statistical difference between group (P≤
0.05) subject to.
Simple diabetes group is 60% (12/20), ISO intervention group is 100% (20/20).And ISO intervention group and simple sugar
It urinates sick group to compare, myocardial damage becomes apparent, and main indicator changes more close with document report.
Experiment shows modeling method of the present invention compared with the spontaneous cardiomyopathy modeling method of traditional simple diabetes, in model
It verifies index, modeling success rate and shows significant superiority on the modeling time.
2) fasting blood-glucose (table 3)
3. diabetic cardiomyopathy rat model blood glucose statistical form of table
3) rat left ventricle hemodynamic index and electrocardiogram parameters
Each group rat is cooked left ventricular cannulation through right common carotid artery, left common carotid artery intubation detects its hemodynamic index；
Three company of leading of rat limbs records its ECG signal.It the results are shown in Table 4-1, table 4-2, table 5-1 and table 5-2.
Table 4-1. diabetic cardiomyopathy rat model hemodynamic index statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group, ▲ indicate
P≤0.05, ▲ ▲ indicate P≤0.01
Table 4-2. diabetic cardiomyopathy rat model hemodynamic index statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group, ▲ indicate
P≤0.05, ▲ ▲ indicate P≤0.01
Table 5-1. diabetic cardiomyopathy rat model electrocardiogram statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
Table 5-2. diabetic cardiomyopathy rat model electrocardiogram statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
The result shows that simple diabetes group is compared with blank control group, left ventricle in terms of haemodynamics main indicator
Internal pressure maximum fall off rate (- dp/dtmax) be remarkably decreased (P≤0.01), the actual measurement of myocardium of left ventricle contractile element is maximum to shorten speed
Shortening speed (Vmax) when degree (Vpm) and 0 load of myocardium of left ventricle contractile element also decreases (P≤0.05), shows to grow
Phase hyperglycemia state can lead to Left Ventricular Diastolic Function in Rats attenuating, and there are a degree of myocardial damages.
There are significant differences with blank control group for the multiple indexs of ISO intervention group: artery mean pressure AP, left ventricular pressure are most
Big climbing speed dp/dtmax, maximal descending rate of internal-dp/dtmax, the maximum contracting of myocardium of left ventricle contractile element actual measurement
Shortening speed Vmax (P≤0.01) when short velocities Vp m, myocardium of left ventricle 0 load of contractile element declines, and left ventricle starts to receive
It is reduced to dp/dtmaxInterval time t-dp/dtmaxExtend, and auterial diastole presses DP, artery mean pressure AP, left ventricular systolic pressure
LVSP, left ventricular diastolic pressure LVDP, left ventricle mean pressure LVAP, myocardial contraction ingredient when pressure development is 40mmHg in left ventricle
Shorten speed V40 to be also substantially reduced, and there are significant difference (P≤0.05) with blank control group.Show that ISO intervention group is big
Mouse Myocardial damage is significant, left ventricular function reduction occurs, and there are pathologic left ventricular hypertrophy and the possibility of myocardial fibrosis.
In terms of electrocardiogram parameters, simple diabetes group and blank control group are compared, T wave amplitude significantly reduce (P≤
0.01), R wave amplitude increases (P≤0.05), shows that simple diabetes development can lead to the change of Rat Ecg T wave, amplitude subtracts
It is few, and R wave amplitude increases, and shows possibility of the rat there are left ventricular hypertrophy.Simple diabetes group and ISO intervention group compare, ISO
Intervention group P wave amplitude further increases (P≤0.05), show ISO intervention group rat there are pulmonary hypertension may, and it is serious
Left ventricle lesion can lead to the generation of pulmonary hypertension；ISO intervention group T wave amplitude is further reduced, and shows ISO intervention group rat
The change of T wave is more obvious, amplitude reduction, inverted possible (P≤0.05) occurs；ISO intervention group R wave amplitude dramatically increase (P≤
0.01), show that rat left ventricle hypertrophy has obvious serious trend compared with simple diabetes group；ISO intervention group QT interval prolongation (P
≤ 0.05), QT interval prolongation is often caused by ventricular bipolar postpones, and sees that the hearts such as left ventricular hypertrophy and Myocardial damage are organic to be changed
Become.
ISO intervention group and blank control group compare, and P wave amplitude significantly increases (P≤0.01), T wave amplitude is substantially reduced (P
≤ 0.01), the T wave time extends (P≤0.01), R wave amplitude increases (P≤0.01), QRS interval prolongation (P≤0.05), PR interphase
Extend (P≤0.05), QT interphase significantly extends (P≤0.01).
The above parameter change shows ISO intervention group compared with blank control group, and rat left ventricle and cardiac muscle have significant disease
Become, and rat myocardium from injury severity caused by ISO intervention group is greater than simple diabetic model group.
4) cardiac weight and heart weight index (table 6)
The 6. diabetic cardiomyopathy rat model heart of table weight index statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
The result shows that model group (simple diabetes group, ISO intervention group) compared with blank control group, heart weight index is all
It significantly increases (P≤0.01), and also there is significant difference (P≤0.01) between ISO intervention group and simple diabetes group, react
There are such as ventricular hypertrophy, the pathologic such as myocardial fibrosis in model group (simple diabetes group, ISO intervention group) rat heart
Change, and ISO intervention group severity is more than simple diabetes group.
5) Biochemical Indices In Serum
Eye socket metaplexus venous collection rat vein blood, 2000r/15min low-speed centrifugal take serum measurement rat biochemistry to refer to
Mark.It the results are shown in Table 7-1 and table 7-2.
Table 7-1. diabetic cardiomyopathy rat model Biochemical Indices In Serum statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
Table 7-2. diabetic cardiomyopathy rat model Biochemical Indices In Serum statistical form
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
The result shows that simple diabetes group compared with blank control group, ALT, ALP, UREA, TC, TG, LDL-C, CK-
MB, LDH generally increase, and there are statistical differences.ISO intervention group compared with blank control group, ALT, AST, ALP, UREA,
TC, TG, LDL-C, CK-MB, α-HBDH, LDH generally increase, and there are statistical difference, show two model group animals with just
Normal animal is compared, and there are different degrees of hepatic injury, myocardial damage, renal function (urea metabolism reduced capability) damage and lipid generations
Thank to disorder.For myocardium situation, ISO intervention group in LDH, α-HBDH, CK, CK-MB cardiac muscle index of correlation than simple glycosuria
Sick group shows even more serious myocardial damage, meets model foundation imagination.
6) pathological examination
6.1 HE dyeing
As it can be seen that simple diabetes group (Fig. 2), ISO intervention group (Fig. 3) rat myocardial cell sarcoplasm are loose under mirror, myogen is fine
Dimension dissolution, cardiac muscle cell's vacuolar degeneration, myocardium interstitial collagen hyperplasia, interstitial cell infiltration.And blank control group (Fig. 1) rat
Cardiac muscle cell's marshalling rule, even dyeing, karyon is clear in structure, and size is uniform, and cardiac muscle fibre band understands, part sample
Only a small amount of bleeding and cell infiltration.
6.2 Masson dyeing
The result shows that simple diabetes group (Fig. 5), ISO intervention group (Fig. 6) have collagen in addition to blank control group (Fig. 4)
Dyeing, blue area of collagen percentage statistics are shown in Table 8.
Table 8.Masson blue area of collagen percentage
Note: compared with ISO intervention group, * indicates that P≤0.05, * * indicate P≤0.01；Compared with blank control group,▲Indicate P
≤ 0.05,▲▲Indicate P≤0.01
According to table 8, ISO intervention group is compared with blank control group, and blue area of collagen percentage increases (P≤0.05), table
There is significant necrosis in bright ISO intervention group cardiac muscle, fibrosis changes.ISO intervention group rat model compared with simple diabetes group,
There are significantly more Myocardial damages.
In short, the present invention mainly passes through the high glucose and high fat raising of 6 weeks early periods of SD rat, it is injected intraperitoneally with a small amount of STZ more times,
After detection blood glucose reaches the rat diabetes diagnostic criteria of 16.7mmol/L, then with isoprel 5mg/kg, once a day
Continuous subcutaneous injection 5d.After continuing raising 2 weeks, ventricular hemodynamics ginseng is detected by left ventricular cannulation, three lead electrocardiogram
Several and electrocardiogram parameters, show that heart function damage occurs in rat model；Heart weight index and left ventricle index show model group rats
Ventricle is obviously loose；Rat blood biochemical analysis shows that significant myocardial damage occurs in the modeling method rat；HE pathological staining
Meet the pathological change of diabetic cardiomyopathy；Masson collagen staining meets connective tissue proliferation between diabetic cardiomyopathy cardiac muscle.
Experiment shows to carry out multiple injection under high sugared feeding environment high in fat to rat with low dose of STZ, induce NIDDM rats
Afterwards, then with 5mg/kg/d isoprel 5d is subcutaneously injected, efficient, easy duplicate diabetic cardiomyopathy rat can be established out
Model is shown to be intervened using isoprel (ISO), can promote the myocardial damage on Diabetes Foundation.With it is existing
Diabetic cardiomyopathy modeling method is compared, the present invention and simple diabetes spontaneity myocardial damage (for example, simple diabetes group)
Compared to the technical advantage obvious with myocardial damage, modeling success rate is high, repeatability is strong, the modeling time is short, can preferably imitate
The pathology of people's type II diabetes myocardial injury passes through.
Claims (8)
1. a kind of construction method of diabetic cardiomyopathy animal model, it is characterised in that: the construction method is the following steps are included: right
Obtained diabetes animal model is induced, then is intervened with isoprel subcutaneous injection, is spaced after intervention certain
Time is detected referring to diabetic cardiomyopathy test rating, obtains diabetic cardiomyopathy animal model.
2. a kind of construction method of diabetic cardiomyopathy animal model according to claim 1, it is characterised in that: the glycosuria
Sick animal model is the type-2 diabetes mellitus animal model induced by STZ.
3. a kind of construction method of diabetic cardiomyopathy animal model according to claim 2, it is characterised in that: the glycosuria
The abductive approach of sick animal model is fed to after the 6th week, is increased specifically includes the following steps: with high-sugar-fat-diet nursing rat
Add 15mg/kg STZ intraperitoneal injection to handle, wherein STZ is injected weekly 1 time, continuous injection filtered out after STZ 4 weeks blood glucose >=
The rat of 16.7mmol/L, obtains diabetes animal model.
4. a kind of construction method of diabetic cardiomyopathy animal model according to claim 3, it is characterised in that: described high in fat
High sugar feed includes following components as mass fraction: 15~20% lard, 14~20% sucrose, 5~11% junket egg
White, 2~5% maltodextrin and 43~62% breeding mouse material.
5. a kind of construction method of diabetic cardiomyopathy animal model according to claim 3, it is characterised in that: described high in fat
The nursing of high sugar feed uses free foraging pattern.
6. a kind of according to claim 1,2 or 3 construction method of diabetic cardiomyopathy animal model, it is characterised in that: institute
Intervention is stated specifically includes the following steps: injecting 1~2 daily with 2~6mg/kg isoprel continuous subcutaneous injection 5~7 days
It is secondary.
7. a kind of construction method of diabetic cardiomyopathy animal model according to claim 6, it is characterised in that: the intervention
It is fed in the process using high-sugar-fat-diet.
8. a kind of construction method of diabetic cardiomyopathy animal model according to claim 6, it is characterised in that: in intervention
After final injection 1~3 week, cardiac determination, cardiac weight and heart weight assessment of indices, blood parameters measurement and pathology are carried out
It learns and checks.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111657227A (en) * | 2020-06-10 | 2020-09-15 | 新疆医科大学第一附属医院 | β1Preparation method of cardiomyopathy model of adrenergic receptor |
| CN113207799A (en) * | 2021-03-19 | 2021-08-06 | 中山大学 | Construction method of type II diabetes mouse rapid heart failure model |
| CN115024279A (en) * | 2022-07-26 | 2022-09-09 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
-
2018
- 2018-11-20 CN CN201811384775.8A patent/CN109481428A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111657227A (en) * | 2020-06-10 | 2020-09-15 | 新疆医科大学第一附属医院 | β1Preparation method of cardiomyopathy model of adrenergic receptor |
| CN113207799A (en) * | 2021-03-19 | 2021-08-06 | 中山大学 | Construction method of type II diabetes mouse rapid heart failure model |
| CN113207799B (en) * | 2021-03-19 | 2022-03-15 | 中山大学 | Construction method of type II diabetes mouse rapid heart failure model |
| CN115024279A (en) * | 2022-07-26 | 2022-09-09 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
| CN115024279B (en) * | 2022-07-26 | 2024-04-12 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
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Application publication date: 20190319 |