A method for constructing an animal model of noncompaction of the ventricular myocardium and its application

Abstract

The present invention provides a method for constructing an animal model of noncompaction of the ventricular myocardium and its application. The method for constructing the animal model of noncompaction of the ventricular myocardium provided by the present invention is simple and efficient in operation; the construction method provided by the present invention has a high modeling rate, making up for the deficiencies of the prior art. Using the animal model provided by the present invention can deeply study the pathogenesis of noncompaction of the ventricular myocardium (NVM), and can provide a theoretical basis for the prevention and / or treatment of this disease.

Description

Technical Field

[0001] The present invention belongs to the field of animal model construction, and relates to a method for constructing an animal model of noncompaction of ventricular myocardium and its application. Background Art

[0002] Non-compaction of ventricular myocardium (NVM) is a special type of hereditary cardiomyopathy first described in 1990, characterized by abnormally thick trabeculae, deep intertrabecular recesses, and a thin compact myocardial layer in the ventricular wall. Since it mainly occurs in the left ventricle, it is also called left ventricular noncompaction (LVNC).

[0003] The clinical manifestations of NVM are non-specific, with large individual differences, and are very easily misdiagnosed or missed. The most common clinical manifestations at the first diagnosis of NVM are heart failure, thromboembolism, and arrhythmia [1] , but patients may also have no clinical symptoms and eventually slowly progress to sudden cardiac death. Due to the high mortality and morbidity of NVM, it has received increasing attention in recent years [2] .

[0004] The etiology and pathogenesis of NVM have not been fully elucidated, mainly involving factors such as embryonic development and genetic variation. In terms of embryonic development, the currently widely accepted is the "theory of noncompaction of ventricular myocardium", that is, abnormal embryonic heart development caused by intrauterine developmental arrest during the normal myocardial compaction process during pregnancy. During the development of the human heart, spongy trabeculae begin to appear at the 4th week of pregnancy. At this time, the coronary vascular circulation has not been established, and oxygen supply mainly depends on the highly developed trabeculae of the endocardium. Subsequently, the subepicardial myocardium forms compact myocardium, the trabeculae gradually thicken, increasing the volume of the compact myocardial layer. At the same time, the intertrabecular recesses develop into capillaries, forming a coronary microcirculation connected to the coronary arteries on the epicardial surface. If any link in the above compaction process is disrupted, it will lead to the occurrence of noncompaction of ventricular myocardium [3] .

[0005] NVM is a polygenic heterogeneous cardiomyopathy, mostly autosomal dominant or X-linked recessive inheritance [4] . Molecular genetic analysis reveals that there are numerous gene mutations in NVM, including NKX2-5, TAZ, LMNA, MYH7, ACTC1, LDB3, TNNT2, and MYBPC3, which mainly encode mitochondrial proteins, sarcomeric proteins, and cytoskeletons [5,6]. In a large retrospective study, researchers detected disease-related DNA variations in 327 cases of LVNC and found that approximately 66 genes were associated with NVM, of which 82% encoded sarcomere proteins [7] . However, these mutated genes are also common in other forms of cardiomyopathy such as hypertrophic cardiomyopathy and dilated cardiomyopathy and are not specific [8] .

[0006] Currently, the diagnosis of NVM mainly relies on ultrasonic cardiogram (UCG) and cardiac magnetic resonance imaging (CMRI) to identify abnormal myocardial structures. Due to the balanced steady-state free precession sequence imaging of CMRI and its high resolution, which can accurately evaluate myocardial anatomical structures and functional information, especially can well distinguish between the compact and non-compact myocardial layers, CMRI has gradually become the preferred method for diagnosing NVM [9] . Currently, there is no drug in clinical practice that can reverse the disease progression of NVM, and the treatment mainly focuses on symptomatic treatment

[0007] Streptozotocin (STZ) was initially an aminoglucose nitrosourea compound isolated from soil microorganisms and has broad-spectrum antibiotic activity

[10] . STZ has a selective destructive effect on pancreatic islet β cells of certain species of animals and can induce diabetes in animals. Since its discovery in 1959, it has been widely used in experimental animals and preclinical studies and is the preferred drug for inducing diabetes in experimental animals

[11] . The preparation of type I and type II diabetic animal models is directly related to the injection dose of STZ. Large-dose injection can widely damage pancreatic islet β cells, resulting in type I diabetes; while a small amount of STZ injection only partially damages pancreatic islet β cells, causing peripheral tissues to be insensitive to insulin. Animal studies have shown that diabetes, especially when combined with a high-fat diet, will severely damage the cardiac function of offspring

[12] . Currently, there is an animal model of type II diabetes established by combining a high-fat diet with STZ injection in male rats

[13] , but there is no relevant research on the heart development of offspring in female rats using the method of high-fat combined with STZ

[0008] Currently, the animal research models of NVM mainly focus on gene knockout mice. However, there are a large number of gene types involved in NVM, and no single gene can be definitely associated with the phenotype. Therefore, it is difficult to establish a comprehensive animal model through gene knockout. Some studies have shown that an animal model of fetal myocardial non-compaction can be prepared by administering 70 mg / kg of all-trans retinoic acid (ATRA) to C57BL / 6 mice on the 8.5th day after pregnancy.

[14] , but the modeling success rate is relatively low. The lack of a reliable animal research model for NVM has led to slow progress in mechanism research.

[0009] Since the etiology and pathogenesis of NVM are still unclear and there is currently no effective treatment method, it is particularly important to establish an effective experimental animal model of NVM to deeply study the specific regulatory mechanisms of its myocardial developmental disorders.

[0010] [References]

[0011] [1]Paluszkiewicz J,Milting H, -Oleksy M,et al.Left VentricularNon-Compaction Cardiomyopathy-Still More Questions than Answers[J].J ClinMed,2022,11(14).DOI:10.3390 / jcm11144135.

[0012] [2]Maron B J,Towbin J A,Thiene G,et al.Contemporary definitions andclassification of the cardiomyopathies:an American Heart AssociationScientific Statement from the Council on Clinical Cardiology,Heart Failureand Transplantation Committee;Quality of Care and Outcomes Research andFunctional Genomics and Translational Biology Interdisciplinary WorkingGroups;and Council on Epidemiology and Prevention[J].Circulation,2006,113(14):1807-1816.

[0013] [3]Towbin J A,Jefferies J L.Cardiomyopathies Due to Left VentricularNoncompaction,Mitochondrial and Storage Diseases,and Inborn Errors ofMetabolism[J].Circulation Research,2017,121(7):838-854.DOI:10.1161 / CIRCRESAHA.117.310987.

[0014] [4]Zhang Z,Xu K,Ji L,et al.A novel loss-of-function mutation in NRAPis associated with left ventricular non-compaction cardiomyopathy[J].FrontCardiovasc Med,2023,10:1097957.DOI:10.3389 / fcvm.2023.1097957.

[0015] [5] Ganame J. Left ventricular non-compaction: from recognition to treatment[J]. Curr Pharm Des, 2015, 21(4): 484-490. DOI: 10.2174 / 138161282104141204143212.

[0016] [6] Probst S, Oechslin E, Schuler P, et al. Sarcomere gene mutations in isolated left ventricular noncompaction cardiomyopathy do not predict clinical phenotype[J]. Circulation Cardiovascular Genetics, 2011, 4(4): 367-374. DOI: 10.1161 / CIRCGENETICS.110.959270.

[0017] [7] Van Waning J I, Caliskan K, Michels M, et al. Cardiac Phenotypes, Genetics, and Risks in Familial Noncompaction Cardiomyopathy[J]. Journal of the American College of Cardiology, 2019, 73(13): 1601-1611. DOI: 10.1016 / j.jacc.2018.12.085.

[0018] [8] Van Waning J I, Caliskan K, Hoedemaekers Y M, et al. Genetics, Clinical Features, and Long-Term Outcome of Noncompaction Cardiomyopathy[J]. Journal of the American College of Cardiology, 2018, 71(7): 711-722. DOI: 10.1016 / j.jacc.2017.12.019.

[0019] [9] Sparrow P, Merchant N, Provost Y, et al. Cardiac MRI and CT features of inheritable and congenital conditions associated with sudden cardiac death[J]. European Radiology, 2009, 19(2): 259 - 270. DOI:10.1007 / s00330-008-1169-5.

[0020]

[10] Reusser F. Mode of action of streptozotocin[J]. Journal of Bacteriology, 1971, 105(2): 580 - 588.

[0021]

[11] Goyal S N, Reddy N M, Patil K R, et al. Challenges and issues with streptozotocin-induced diabetes - A clinically relevant animal model to understand the diabetes pathogenesis and evaluate therapeutics[J]. Chemico-biological Interactions, 2016, 244: 49 - 63. DOI:10.1016 / j.cbi.2015.11.032.

[0022]

[12] Upadhyaya B, Larsen T, Barwari S, et al. Prenatal Exposure to a Maternal High-Fat Diet Affects Histone Modification of Cardiometabolic Genes in Newborn Rats[J]. Nutrients, 2017, 9(4). DOI:10.3390 / nu9040407.

[0023]

[13] D a D E, Kume W T, Correia F S, et al. High-fat diet and streptozotocin in the induction of type 2 diabetes mellitus: a new proposal[J]. Anais Da Academia Brasileira de Ciencias, 2019, 91(1): e20180314. DOI: 10.1590 / 0001-3765201920180314.

[0024]

[14] Cao F, Yang Z, Yin L. A fetal mouse model of ventricular non-compaction using retinoic acid[J]. Pathology, Research and Practice, 2019, 215(8): 152496. DOI: 10.1016 / j.prp.2019.152496. Summary of the Invention

[0025] Problems to be Solved by the Invention

[0026] Aiming at the defects that the etiology and pathogenesis of NVM are still unclear in the prior art and there is a lack of a reliable animal research model for NVM, the present invention provides a method for constructing an animal model of non-compaction of the ventricular myocardium and its application.

[0027] Solutions for Solving the Problems

[0028] In view of the problems existing in the above-mentioned prior art, the inventor of the present invention has conducted in-depth research and repeated experiments, and established an animal model of non-compaction of the ventricular myocardium by using STZ combined with a high-fat diet, thus completing the present invention. That is, the present invention is as described below:

[0029] In a first aspect, the present invention provides a method for constructing an animal model of non-compaction of the ventricular myocardium (NVM), characterized in that the construction method comprises the following steps:

[0030] 1) Select female animals fed with a high-sugar and high-fat diet;

[0031] 2) Select male animals fed with a conventional diet;

[0032] 3) Mate the animals in steps 1) and 2), and detect and confirm that the female animal has a vaginal plug;

[0033] 4) Inject streptozotocin (STZ) into the female animal with a vaginal plug after mating;

[0034] 5) Measure the blood glucose value of the female animal described in step 4). When the blood glucose value of the female animal is greater than 16.7 mmol / L, it indicates that the female animal has gestational hyperglycemia. Wait for the female animal to give birth naturally to obtain an animal model of offspring NVM, where the ratio of the non-compact myocardial thickness to the compact myocardial thickness of the animal model of offspring NVM is greater than 1.4.

[0035] In some specific embodiments, wherein, in step 1), the energy supply ratio of fat in the high-sugar and high-fat diet is 60 kcal%; in some preferred embodiments, the female animal is fed with a high-sugar and high-fat diet for 3 - 5 weeks; in some more preferred embodiments, the female animal is fed with a high-sugar and high-fat diet for 4 weeks.

[0036] In some specific embodiments, wherein, in step 4), the injection dose of STZ is 35 - 50 mg / kg; in some preferred embodiments, the injection amount of STZ is 35 mg / kg; optionally, the female animal with a vaginal plug after mating is fasted for more than 12 hours before injecting STZ.

[0037] In some specific embodiments, wherein, in step 5), the blood glucose value of the female animal is measured on the 3rd day, 7th day, and 13th day after injecting STZ respectively.

[0038] In some specific embodiments, wherein, the animal is a rat; in some preferred embodiments, the animal is an SD rat.

[0039] The present invention in a second aspect provides the use of a combination of streptozotocin (STZ) and a high-sugar and high-fat diet in the preparation of an animal model of non-compaction of the ventricular myocardium (NVM).

[0040] In some specific embodiments, wherein, the injection amount of STZ is 35 - 50 mg / kg; in some preferred embodiments, the injection amount of STZ is 35 mg / kg.

[0041] In some specific embodiments, wherein, the energy supply ratio of fat in the high-sugar and high-fat diet is 60 kcal%.

[0042] In some specific embodiments, wherein, the ratio of the non-compact myocardial thickness to the compact myocardial thickness of the animal model is greater than 1.4.

[0043] In some specific embodiments, wherein, the animal is a rat; in some preferred embodiments, the animal is an SD rat.

[0044] Effects of the Invention

[0045] As can be seen from the technical solution of the present invention, compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0046] 1) The method for constructing an animal model of noncompaction of the ventricular myocardium provided by the present invention is simple and efficient to operate; the construction method provided by the present invention has a high modeling rate, making up for the deficiencies of the prior art.

[0047] 2) The animal model provided by the present invention can be used to deeply study the pathogenesis of NVM and provides a theoretical basis for the prevention and / or treatment of this disease.

[0048] 3) The present invention discovers for the first time that treating rats with streptozotocin and a high-fat diet can cause the offspring of rats to present the phenotype of NVM.

[0049] In order to make the above and other objects, features and advantages of the present invention more obvious and understandable, the following specific preferred embodiments are given and described in detail in conjunction with the accompanying drawings of the specification as follows: BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 Shows a 1:40 microscopic photograph of the left ventricle of the fetal rat heart in the model group.

[0051] Figure 2 Shows a 1:40 microscopic photograph of the left ventricle of the fetal rat heart in the control group. DETAILED DESCRIPTION OF THE INVENTION

[0052] The following will detail various exemplary embodiments, features and aspects of the present invention. The special term "exemplary" used here means "serving as an example, embodiment or illustration". Any embodiment described as "exemplary" here does not have to be construed as superior to or better than other embodiments.

[0053] In addition, in order to better illustrate the present invention, numerous specific details are given in the following detailed description. Those skilled in the art should understand that the present invention can be implemented without some of these specific details. In other instances, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the gist of the present invention.

[0054] Unless otherwise stated, the units used in this specification are all international standard units, and the numerical values and numerical ranges appearing in the present invention should be understood to include the inevitable systematic errors in industrial production.

[0055] In this specification, the meaning expressed by using "can" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0056] In this specification, the "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc. mentioned refer to the specific elements (e.g., features, structures, properties, and / or characteristics) related to the embodiments, which are included in at least one of the embodiments described herein, and may or may not exist in other embodiments. Additionally, it should be understood that the elements can be combined in various embodiments in any suitable manner.

[0057] In this specification, the numerical range represented by "numerical value A to numerical value B" refers to the range including the endpoint numerical values A and B.

[0058] In this specification, when "normal temperature" or "room temperature" is used, the temperature can be 10 to 40 °C.

[0059] Example

[0060] The embodiments of the present invention will be described in detail below in conjunction with examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. For those not specified in the examples, they are carried out under conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained commercially.

[0061] Example 1: Construction method of a rat model of noncompaction of the ventricular myocardium

[0062] 1. Screening of female rats

[0063] Eighteen female Sprague-Dawley (SD) rats (Spebefu (Beijing) Biotechnology Co., Ltd.) at 4 to 5 weeks old and weighing 130 to 160 g were selected. The temperature was controlled at 25 °C, with 12 h of sunlight and 12 h of darkness each day, and free access to water. They were fed with a high-sugar and high-fat diet (Xiaoshu Youtai (Beijing) Biotechnology Co., Ltd., product number: D12492) with 60 kcal% of the energy supply from fat for 4 weeks. After 4 weeks, the body weight of the female SD rats reached 220 g to 270 g. Among them, the formula of the high-sugar and high-fat diet was: protein component 26.2% (w / w), carbohydrate component 26.3% (w / w), and fat component 34.9% (w / w). In 100 kcal% of the energy, the energy supply from fat in the diet accounted for 60 kcal%, the energy supply from protein accounted for 20 kcal%, and the energy supply from carbohydrates accounted for 20 kcal%.

[0064] 2. Mating and checking for plugs

[0065] Select male SD rats with mating ability and fed with regular diet, and conduct 1:1 mating with female SD rats on a high-fat diet for 4 weeks described in step 1 at night. Check the vaginal plugs of female SD rats at 7:30 in the morning the next day. The vaginal plug is a white mass formed by the coagulation of the secretions of the male rat's seminal vesicles and usually fills the female rat's vagina 8-24 hours after night mating. If no obvious vaginal plug is observed, a smear of the female rat's vaginal secretion can be taken and observed under a microscope to confirm successful mating. Female SD rats without a vaginal plug can be taken out separately, caged again at night, and checked for plugs the next day. Repeat this operation until a vaginal plug is seen in the female SD rat.

[0066] 3. Model establishment

[0067] After confirming that the female SD rat has a vaginal plug, fast it overnight on the same day and conduct model establishment the next day. The factors to be considered in model establishment and the specific model establishment methods are as follows:

[0068] (1) Selection of STZ dose: The selection of the STZ dose mainly depends on the body weight and resistance level of the animal. The level of the STZ unit dose is inversely proportional to the body weight.

[0069] For female SD rats weighing 220-270 grams after 4 weeks or more of high-fat feeding and with a vaginal plug after mating, select a STZ dose of 35-50 mg / kg for a single administration.

[0070] (2) Fast the female SD rats with a vaginal plug

[0071] Before injecting STZ, fast the female SD rats overnight but do not restrict water for more than 12 hours. The longer the fasting time, the more obvious the destructive ability of STZ to pancreatic islet β cells. Therefore, when prolonging the fasting time, the dosage of STZ can be appropriately reduced, but it is necessary to consider whether the animal can tolerate long-term fasting.

[0072] (3) Preparation of STZ buffer solution

[0073] Solution A: Add 2.1 g of citric acid (FW: 192.13) to 100 mL of double-distilled water

[0074] Solution B: Add 2.94 g of sodium citrate (FW: 259.069) to 100 mL of double-distilled water

[0075] Mix solution A and solution B in a ratio of 1:1 or 1:1.32, and adjust the pH to 4.2-4.5 (if the pH is too acidic, add double-distilled water to adjust; if the pH is too alkaline, add citric acid to adjust). The resulting mixed solution is the required STZ buffer solution.

[0076] The buffer solution should be prepared and used as soon as possible to prevent oxidation. Therefore, when preparing, it can be scaled down proportionally and prepared in small amounts.

[0077] (4) Injection of STZ solution

[0078] After overnight fasting, measure the fasting blood glucose of female SD rats with visible plugs, weigh them, weigh an appropriate amount of STZ powder at a dose of 35 mg / kg into a dry sterile bottle, and wrap the sterile bottle with tin foil to avoid light. Place the sterile bottle containing STZ powder and the STZ buffer solution prepared in step (3) on ice and take them to the animal house.

[0079] Since STZ is prone to deliquescence and inactivation, it should be quickly weighed using dry aluminum foil / tin foil or dry weighing paper, and strict dryness should be maintained after weighing. The freshly prepared STZ solution should also be used up within 30 minutes.

[0080] Take an appropriate amount of the STZ buffer solution prepared in step (3) to dissolve the STZ powder, prepare a 2% (wt / v) fresh STZ solution, and quickly inject it intraperitoneally into the female SD rats (the specific injection dose is 35 mg / kg). The faster the injection rate, the easier it is to form hyperglycemia. Four hours after the injection of the STZ solution, resume normal diet.

[0081] (5) Record the body weight, dosage, administration time, and fasting blood glucose before administration of female SD rats on the day of administration. Measure the blood glucose of female SD rats on the 3rd, 7th, and 13th days after administration. When the blood glucose of female SD rats > 16.7 mmol / L, it indicates that the female SD rats have gestational hyperglycemia (there will be NVM phenotypes in their offspring embryos), and wait for the female SD rats to give birth naturally to obtain an animal model of offspring NVM.

[0082] The rat model prepared by the above method is called the "STZ group" in Table 1.

[0083] Meanwhile, select 16 female SD (Sprague-Dawley) rats aged 4 - 5 weeks and weighing 130 - 160 g (Specef (Beijing) Biotechnology Co., Ltd.), control the constant temperature at 25 °C, with 12 hours of sunlight and 12 hours of darkness each day, free access to water, and fed with regular feed. Select male SD rats with mating ability and fed with regular feed to mate with them (the same as above), and no other treatment is done to the rest. And take the rats treated as above as the control group (called the "normal group" in Table 1)

[0084] Measure the blood glucose values of female rats in the STZ group before administration, on the 3rd, 7th, and 13th days after administration, and at the same time measure the blood glucose values of female rats in the normal group on the 0th, 3rd, 7th, and 13th days of pregnancy. The specific results are shown in Table 1:

[0085] Table 1 Blood glucose values of female rats in the STZ group and female rats in the normal group

[0086]

[0087] It can be seen from the results in Table 1 that: compared with the rats in the normal group, the blood glucose level of the rats in the STZ group increased significantly after administration.

[0088] Example 2: Heart sections of fetal rats with noncompaction of the myocardium

[0089] In this example, F0 generation positive SD rats were obtained according to the modeling method provided in Example 1. After the F0 generation positive SD rats gave birth naturally, 60 fetal SD rats of the F1 generation (referred to as the "STZ group" in Tables 2 and 3) were obtained. For the fetal SD rats of the F1 generation, the hearts of the fetal rats of the F1 generation were obtained on the 20th day of embryonic development, paraffin sections were cut and stained with hematoxylin-eosin (HE). The thickness of the non-compacted myocardium layer (abbreviated as N) and the compact myocardium layer (abbreviated as C) was measured, and the ratio of the thickness of the non-compacted myocardium layer to the thickness of the compact myocardium layer (Non-compaction / compaction, N / C) was calculated. If N / C > 1.4, it was in line with the diagnosis of noncompaction of the myocardium.

[0090] At the same time, 12 female SD (Sprague-Dawley) rats aged 4-5 weeks and weighing 130-160 g (Spearf Bio (Beijing) Co., Ltd.) were selected. The temperature was controlled at 25 °C, the daily sunlight and darkness time were each 12 h, they drank water freely, and were fed with conventional feed. Male SD rats with mating ability and fed with conventional feed were mated with them (the same as above), and the rest were not treated. The rats treated as above were used as the control group (referred to as the "normal group" in Tables 2 and 3).

[0091] The N, C measurement values and N / C of the fetal rats of the STZ group and the normal group in the left ventricle were measured (see Table 2 for specific results), and the N, C measurement values and N / C of the fetal rats of the STZ group and the normal group in the right ventricle were measured (see Table 3 for specific results).

[0092] Table 2 N, C measurement values and N / C of fetal rats in the left ventricle of the STZ group and the normal group

[0093]

[0094]

[0095] It can be seen from the results in Table 2 that the thickness of the non-compact myocardial layer of the fetal rats in the STZ group in the left ventricle is higher than that of the fetal rats in the normal group; the thickness of the compact myocardial layer of the fetal rats in the STZ group in the left ventricle is lower than that of the fetal rats in the normal group; the ratio of the thickness of the non-compact myocardial layer to the thickness of the compact myocardial layer of the fetal rats in the STZ group in the left ventricle is significantly higher than 1.4, which meets the diagnosis of non-compaction of the myocardium.

[0096] Table 3 Measurement values of N, C and N / C in the right ventricles of fetal rats in the STZ group and fetal rats in the normal group

[0097]

[0098]

[0099] It can be seen from the results in Table 3 that the thickness of the non-compact myocardial layer of the fetal rats in the STZ group in the right ventricle is higher than that of the fetal rats in the normal group; the thickness of the compact myocardial layer of the fetal rats in the STZ group in the right ventricle is lower than that of the fetal rats in the normal group; the ratio of the thickness of the non-compact myocardial layer to the thickness of the compact myocardial layer of the fetal rats in the STZ group in the right ventricle is significantly higher than 1.4, which meets the diagnosis of non-compaction of the myocardium.

[0100] It should be noted that although the technical solutions of the present invention are introduced by specific examples, those skilled in the art can understand that the present invention should not be limited thereto.

[0101] The embodiments of the present invention have been described above. The above description is exemplary and not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art in the technical field without departing from the scope and spirit of the described embodiments. The selection of the terms used herein is intended to best explain the principles of the embodiments, practical applications or improvements to the technology in the market, or to enable other ordinary skill in the art in the technical field to understand the disclosed embodiments.

Claims

1. A method for constructing an animal model of noncompaction of the ventricular myocardium (NVM), characterized in that, The construction method includes the following steps: 1) Select female animals fed with a high-sugar and high-fat diet; 2) Select male animals fed with a conventional diet; 3) Mate the animals in steps 1) and 2), and detect and confirm that the female animals have a vaginal plug; 4) Inject streptozotocin (STZ) into the female animals with a vaginal plug after mating; 5) Measure the blood glucose value of the female animals in step 4). When the blood glucose value of the female animals is greater than 16.7 mmol / L, it indicates that the female animals have gestational hyperglycemia. Wait for the female animals to give natural birth to obtain an animal model of offspring NVM, where the ratio of the non-compact layer myocardial thickness to the compact layer myocardial thickness of the animal model of offspring NVM is greater than 1.

4.

2. The construction method according to claim 1, wherein, In the high-sugar and high-fat diet in step 1), the proportion of energy supply from fat is 60 kcal%.

3. The construction method according to claim 2, wherein, Select female animals fed with a high-sugar and high-fat diet for 3 - 5 weeks.

4. The construction method according to claim 3, wherein, Select female animals fed with a high-sugar and high-fat diet for 4 weeks.

5. The construction method according to any one of claims 1 to 4, wherein, In step 4), the injection dose of STZ is 35 - 50 mg / kg.

6. The construction method according to claim 5, wherein the injection amount of STZ is 35 mg / kg, and the female animals with a vaginal plug after mating are fasted for more than 12 hours before injecting STZ.

7. The construction method according to any one of claims 1 to 4 and 6, wherein, In step 5), measure the blood glucose value of the female animals on the 3rd, 7th, and 13th days after injecting STZ respectively.

8. The construction method according to claim 5, wherein, In step 5), measure the blood glucose value of the female animals on the 3rd, 7th, and 13th days after injecting STZ respectively.

9. The construction method according to any one of claims 1 to 4, 6, and 8, wherein, The animal is a rat.

10. The construction method according to claim 5, wherein the animal is a rat.

11. The construction method according to claim 7, wherein the animal is a rat.

12. The construction method according to claim 9, wherein the animal is an SD rat.

13. The construction method according to claim 10, wherein the animal is an SD rat.

14. The construction method according to claim 11, wherein the animal is an SD rat.

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