OGDHL-mediated atherosclerosis animal model and establishment method thereof

Abstract

The invention discloses an OGDHL-mediated atherosclerosis animal model. The animal model is obtained by inducing systematic high expression of mouse OGDHL through exogenous injection of adenovirus. The invention also discloses an establishment method of the OGDHL-mediated atherosclerosis animal model, the model establishment process is simple and convenient, the success rate is high, the result is stable and reliable, the time for feeding experimental animals is saved, the consequence of model establishment failure is also avoided, and the method is suitable for large-scale popularization and application of the OGDHL-mediated atherosclerosis animal model. The animal model established by the invention has important practical application value in research on etiology, pathology, drug screening, clinical diagnosis and treatment and the like of atherosclerosis, and can be widely popularized.

Description

Technical Field

[0001] The present invention belongs to the field of basic medical technology, and specifically relates to an atherosclerosis animal model mediated by OGDHL, and also relates to a method for establishing the atherosclerosis animal model. Background Art

[0002] Atherosclerosis is a silent chronic vascular pathology and the cause of most cardiovascular ischemic events. The formation of foam cells is one of the very important factors that trigger the formation of atherosclerotic plaques. When the intima of certain parts of the large or medium arteries thickens and a large amount of lipoproteins are deposited, monocytes differentiate into macrophages through the action of the chemokine MCP-1. Macrophages and a small number of smooth muscle cells phagocytize oxidized low-density lipoprotein cholesterol in the blood to form foam cells, which attach to the vascular endothelium to form plaques. As the disease progresses, plaques increase in size and plaque vulnerability increases. Patients may show various serious clinical manifestations, such as myocardial infarction. If plaques rupture to form thrombi, the patient's risk of death increases.

[0003] Oxoglutarate dehydrogenase (OGDH) is the first rate-limiting component of the multienzyme OGDH complex (OGDHC), and its dysfunction is associated with neurodegeneration. The complex plays an important role in the degradation of glucose and glutamate, and the oxoglutarate dehydrogenase-like (OGDHL) gene (a component of OGDHC, expression of which is mainly restricted to the human brain) regulates glutamine metabolism to affect tumor progression and is downregulated in many different cancer types through promoter hypermethylation. These features suggest that OGDHL has a potential growth regulatory role in cancer; however, the molecular mechanism by which OGDHL exerts its growth regulatory function has not been elucidated. OGDHL has been implicated in the pathogenesis of diseases such as atherosclerosis and tissue inflammation, and it also plays an important role in the physiological activities of various cells.

[0004] When conducting experimental research on atherosclerosis, it is necessary to prepare an experimental animal model. In existing technologies, high-fat feeding and balloon injury methods are generally used in the construction of experimental models for small and medium-sized animals. It takes 12 weeks to intervene in experimental animals, which consumes a lot of time and manpower. Summary of the invention

[0005] The first object of the present invention is to provide an animal model of atherosclerosis mediated by OGDHL, provide an animal model that is helpful for studying the growth regulatory function of OGDHL, and shorten the time for establishing the animal model.

[0006] The second object of the present invention is to provide a method for establishing an animal model of atherosclerosis mediated by OGDHL.

[0007] The first technical solution adopted by the present invention is: an animal model of atherosclerosis mediated by OGDHL, wherein the animal model is obtained by exogenous injection of adenovirus to induce systemic high expression of OGDHL in mice.

[0008] The second technical solution adopted by the present invention is: a method for establishing an atherosclerosis animal model mediated by OGDHL, which is specifically carried out according to the following steps: Step 1, constructing an adenovirus expression vector Ad-OGDHL; Step 2: Inject the adenovirus vector Ad-OGDHL obtained in step 1 into the ApoE - / - The mice were fed with a high-fat diet for 6 weeks after injection, and the adenovirus expression vector Ad-OGDHL was injected again in the fourth week. The mice obtained after 6 weeks were the above-mentioned animal model.

[0009] The second technical solution adopted by the present invention is also characterized in that: Furthermore, the specific steps of constructing the adenovirus expression vector in step 1 are as follows: Step 1.1, using Hind Ⅲ and Sal Ⅰ to double-digest mouse OGDHL cDNA plasmid, electroeluting to recover OGDHL cDNA fragment, the nucleotide sequence of cDNA fragment is shown in SEQ ID NO:1, using Hind Ⅲ and Sal Ⅰ to double-digest pCMV-OGDHL plasmid, electroeluting to recover pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment, then ligating OGDHL cDNA fragment and pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment by T4 DNA ligase, and transforming the ligated product into E. coli In DH5α competent cells, the pAdTrack-OGDHL recombinant plasmid was extracted by alkaline lysis after culture; Step 1.2: The pAdTrack-OGDHL recombinant plasmid obtained in step 1.1 was digested with PemⅠ, and the linear fragment was recovered by electroelution. The pAdTrack-OGDHL PemⅠ linear fragment was electroporated into the pAdEasy-1 containing E. coli BJ5183 electrocompetent cells, the transformation products were cultured and the pAdEasy-OGDHL recombinant plasmid was extracted by alkaline lysis method; Step 1.3, using Pac I to digest the pAdEasy-OGDHL obtained in step 1.2 to obtain a Pac I linearized recombinant plasmid pAdEasy-OGDHL fragment, using Pac I enzyme to linearize the recombinant plasmid pAdEasy-OGDHL fragment to transfect HEK293 cells, culturing the cells for 7 to 10 days after transfection, observing cell growth and green fluorescent protein expression every day, and when the green fluorescent protein expression in the cells is "comet-shaped" and cytopathic effect occurs, collecting the HEK293 cells and vortexing to lyse the cells, centrifuging after lysis, collecting the supernatant, which is the Ad-OGDHL virus solution, and storing the Ad-OGDHL virus solution at -80°C for later use; Step 1.4, on the day of infection, the Ad-OGDHL virus solution obtained in step 1.3 was added to 1.5 mL of DEME complete culture medium and cultured for 90 minutes, and then 3.5 mL of DMEM complete culture medium was added for culture. When the cells showed obvious cytopathic effect and were accompanied by shedding, the virus was collected and the infection with Ad-OGDHL virus solution was repeated 4 times. The total amount of the obtained virus was collected to obtain the adenovirus expression vector Ad-OGDHL of the present invention.

[0010] Furthermore, the HEK293 cells were cultured in a culture dish one day before transfection in step 1.3 to ensure that the cell density of the cells during transfection was 30%-50%.

[0011] Furthermore, one day before infection in step 1.4, HEK293 cells were cultured in a culture dish to ensure that the cell density was 75%-85% during infection.

[0012] Furthermore, the cholesterol and fat content of the high-fat feed in step 2 is 1.5% cholesterol + 15% fat.

[0013] Furthermore, in step 2, ApoE is injected - / - The concentration of mouse adenovirus expression vector Ad-OGDHL is 1×10 11 vp / mL.

[0014] The beneficial effect of the present invention is that compared with the existing animal model establishment method, the 12-week experimental animal model construction time is too long and consumes a lot of manpower and material resources. The animal model construction method mentioned in this patent only takes 6 weeks to build the required atherosclerosis model. Compared with the general method of injecting exogenous proteins, the adenovirus transfection method has obvious advantages. It does not require frequent protein injections and only needs to be injected once every 3 weeks. Since the OGDHL gene structure sequence is large, the adenovirus-carrying gene method is the simplest and most convenient. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] FIG1 is a schematic diagram showing a comparison between the animal model of the present invention and the control group when aortic arch plaques are photographed under a microscope after Oil Red O staining; Figure 2 This is a statistical comparison chart obtained by calculating the plaque area of ​​each group in the present invention. DETAILED DESCRIPTION

[0016] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0017] The present invention provides an animal model of atherosclerosis mediated by OGDHL, which can be obtained by exogenous injection of adenovirus to systemically highly express OGDHL. The modeling time of the animal model prepared by the present invention is significantly shortened compared with the existing modeling time.

[0018] The method for establishing an atherosclerosis animal model mediated by OGDHL is specifically carried out in the following steps: Step 1, constructing an adenovirus expression vector Ad-OGDHL; Step 1.1, using Hind Ⅲ and Sal Ⅰ to double-digest mouse OGDHL cDNA plasmid, electroeluting to recover OGDHL cDNA fragment, the nucleotide sequence of cDNA fragment is shown in SEQ ID NO:1, using Hind Ⅲ and Sal Ⅰ to double-digest pCMV-OGDHL plasmid, electroeluting to recover pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment, then ligating OGDHL cDNA fragment and pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment by T4 DNA ligase, and transforming the ligated product into E. coli In DH5α competent cells, the pAdTrack-OGDHL recombinant plasmid was extracted by alkaline lysis after culture; Step 1.2: The pAdTrack-OGDHL recombinant plasmid obtained in step 1.1 was digested with PemⅠ, and the linear fragment was recovered by electroelution. The pAdTrack-OGDHL PemⅠ linear fragment was electroporated into the pAdEasy-1 containing E. coli BJ5183 electrocompetent cells, the transformation products were cultured and the pAdEasy-OGDHL recombinant plasmid was extracted by alkaline lysis method; Step 1.3, using Pac I to digest the pAdEasy-OGDHL obtained in step 1.2 to obtain a Pac I linearized recombinant plasmid pAdEasy-OGDHL fragment, using Pac I enzyme to linearize the recombinant plasmid pAdEasy-OGDHL fragment to transfect HEK293 cells, culturing the cells for 7 to 10 days after transfection, observing cell growth and green fluorescent protein expression every day, and when the green fluorescent protein expression in the cells is "comet-shaped" and cytopathic effect occurs, collecting the HEK293 cells and vortexing to lyse the cells, centrifuging after lysis, collecting the supernatant, which is the Ad-OGDHL virus solution, and storing the Ad-OGDHL virus solution at -80°C for later use; One day before transfection in step 1.3, place HEK293 cells in a culture dish and culture them to ensure that the cell density of the cells is 30%-50% during transfection; Step 1.4: On the day of infection, the Ad-OGDHL virus solution obtained in step 1.3 was added to 1.5 mL of DEME complete culture medium and cultured for 90 min, and then 3.5 mL of DMEM complete culture medium was added for culture. When the cells showed obvious cytopathic effect and were accompanied by shedding, the virus was collected, and the cells were repeatedly infected with the Ad-OGDHL virus solution for 4 times. The total amount of the obtained virus was collected to obtain the adenovirus expression vector Ad-OGDHL of the present invention; One day before infection in step 1.4, place HEK293 cells in a culture dish and culture them to ensure that the cell density is 75%-85% during infection; Step 2: Inject the adenovirus vector Ad-OGDHL obtained in step 1 into the ApoE - / - In mice, high-fat diet was used for 6 weeks after injection, and adenovirus expression vector Ad-OGDHL was injected again in the fourth week. The mice obtained after 6 weeks were the animal model; The cholesterol and fat content of the high-fat diet in step 2 is 1.5% cholesterol + 15% fat; Inject ApoE in step 2 - / - The concentration of mouse adenovirus expression vector Ad-OGDHL is 1×10 11 vp / mL.

[0019] Example 1 The OGDHL-mediated atherosclerosis animal model was obtained by exogenous injection of adenovirus to induce systemic high expression of OGDHL in mice.

[0020] Follow these steps: Step 1, constructing an adenovirus expression vector Ad-OGDHL; Step 2: Inject the adenovirus vector Ad-OGDHL obtained in step 1 into the ApoE- / - The mice were fed with a high-fat diet for 6 weeks after injection, and the adenovirus expression vector Ad-OGDHL was injected again in the fourth week. The mice obtained after 6 weeks were the above-mentioned animal model.

[0021] Example 2 The method for constructing an atherosclerosis animal model mediated by OGDHL specifically includes the following steps: Step 1: Preparation of adenovirus expression vector of Ad-OGDHL gene Step 1.1, the mouse OGDHL cDNA plasmid was double-digested with Hind Ⅲ and Sal Ⅰ, the double-digested products were detected by 1% agarose gel electrophoresis, and the OGDHL cDNA fragment was recovered by electroelution. The pOGDHL-CMV plasmid was double-digested with Hind Ⅲ and Sal Ⅰ, and the pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment was recovered by electroelution. The OGDHL cDNA fragment and the pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment were then ligated with T4 DNA ligase, and the ligated products were transformed into E. coli In DH5α competent cells, Kan ＋ After culturing in LB medium, the pAdTrack-OGDHL recombinant plasmid was extracted by alkaline lysis method, and the recombinant plasmid was identified by double restriction enzyme digestion with Hind Ⅲ and Sal Ⅰ.

[0022] Step 1.2: The pAdTrack-OGDHL recombinant plasmid obtained in step 1.1 was digested with PemⅠ, and the linear fragment was recovered by electroelution. The pAdTrack-OGDHLPemⅠ linear fragment was electrotransformed into the pAdEasy-1 containing E. coli BJ5183 electrocompetent cells, the transformation products were ＋ After culture in LB solid medium, the pAdEasy-OGDHL recombinant plasmid was extracted by alkaline lysis method, and the pAdEasy-OGDHL recombinant plasmid was identified by BamHI and PacⅠ digestion, and the target gene sequence of the pAdEasy-OGDHL recombinant plasmid was determined (Invitrogen).

[0023] Step 1.3, pAdEasy-OGDHL obtained in step 1.2 was digested with Pac I to obtain a Pac I linearized recombinant plasmid pAdEasy-OGDHL fragment. HEK293 cells were cultured in a 60 mm diameter culture dish one day before transfection, and 5 μg of the Pac I linearized recombinant plasmid pAdEasy-OGDHL fragment was used to transfect HEK293 cells. The cell density during transfection was 30%-50%. The cells were cultured for 7-10 days after transfection, and cell growth and green fluorescent protein (GFP) expression in the cells were observed every day. When GFP expression was "comet"-shaped and cytopathic effect (CPE) appeared, the cells were collected in a 15 mL centrifuge tube, and the cells were lysed by vortexing 4 times. After lysis, the cells were centrifuged at 12000 r / min for 3 hours. min, and the supernatant after centrifugation was dispensed into 1.5 mL centrifuge tubes. The supernatant was the Ad-OGDHL virus solution, and the Ad-OGDHL virus solution was stored at -80°C for later use.

[0024] Step 1.4, one day before infection, HEK293 cells were cultured in a culture dish with a diameter of 100 mm. On the day of infection, 200 μL of the Ad-OGDHL virus solution obtained in step 1.3 was added to 1.5 mL of DEME complete culture medium and cultured for 90 min, and then 3.5 mL of DMEM complete culture medium was added for culture. On the third day of infection, when the cells showed obvious cytopathic effect and were accompanied by shedding, the virus was collected and the infection with Ad-OGDHL virus solution was repeated 4 times. The total amount of the virus obtained was collected to obtain the adenovirus expression vector Ad-OGDHL of the present invention.

[0025] Step 2: ApoE - / - After the adenovirus expression vector Ad-OGDHL in step 1.5 was injected into the mouse tail vein, ApoE - / - The mice were fed with a high-fat diet for 6 weeks, and injected with the adenovirus expression vector Ad-OGDHL in step 1.5 again in the fourth week, and the obtained mice were the animal model of the present invention. The cholesterol and fat content of the high-fat diet were both 1.5% cholesterol +15% fat.

[0026] ApoE - / - The concentration of adenovirus expression vector Ad-OGDHL when injected into the mouse tail vein was 1×10 11 vp / mL.

[0027] Example 3 Model verification of Example 2 of the present invention: 1. Experimental subjects (1) The animal model prepared by the present invention was selected as the test group (Ad-OGDHL).

[0028] (2) ApoE - / - Mice were injected with 200 μL of normal saline through the tail vein and then fed with a high-fat diet for 6 weeks to obtain a blank control group (saline).

[0029] (3) ApoE - / - The mice were injected with Ad-GFP adenovirus through the tail vein. The number of adenovirus was 1×10 11 vp / mL, the injected mice were fed with a high-fat diet for 6 weeks, and then injected with Ad-GFP again in the fourth week. The obtained mice were the control group (Ad-GFP). The cholesterol and fat content of the high-fat diet were both 1.5% cholesterol + 15% fat.

[0030] The above-mentioned Ad-GFP adenovirus is prepared according to the following steps: Step a, pRNAT-H1.1 / Adeno was digested with PmeI, recovered by low melting point agarose gel electrophoresis, dephosphorylated with alkaline phosphatase CIAP, extracted with phenol / chloroform, and transformed into supercompetent BJ5183 containing adenovirus backbone vector pAdeasy-1. The positive recombinant plasmid obtained by homologous recombination in bacteria was named pAd-GFP.

[0031] Step b, the positive recombinant plasmid pAd-GFP linearized by PacI was extracted with phenol / chloroform, ethanol / sodium acetate co-precipitated, and then 40 μL of sterilized triple-distilled water was added to dissolve it. After it was completely dissolved, an equal volume of cationic liposome Lipofectamine2000 was added. After incubation at 37°C for 4 hours, HEK293 cells were transfected by cationic liposome Lipofectamine2000. After incubation in an incubator at 37°C and 5% CO2 for 14 to 20 days, the appearance of viral plaques can be observed.

[0032] Step c, after the recombinant adenovirus obtained in step b was centrifuged at 12000r / min for 3 times, 500μL of the recombinant adenovirus supernatant after centrifugation was taken and added to 90% fused HEK293 cells, and cultured in a 37°C, 5% CO2 incubator. After 95% to 100% of the cells showed cytopathic effects under a microscope, the cells were collected by centrifugation, and the cells were repeatedly frozen and thawed 3 times between -80°C and 37°C, and the supernatant was collected by centrifugation at 3000rpm / min for 5min. The recombinant adenovirus supernatant was purified twice by CsCl density gradient centrifugation, and the purified adenovirus was dialyzed twice at 4°C with a dialysate (10mmol / LTris-HCl, 1mmol / LMgCl2, 10% glycerol), each time for 24h, and the dialyzed virus solution was titered by ultraviolet spectrophotometer. Take 100 μL of the purified virus solution, 20 μL of 10% SDS, and 880 μL of PBS, heat-bathe at 65°C for 30 min, vortex three times, and measure the OD260nm and OD280nm of the viral genomic DNA after centrifugation. The number of virus particles and purity are calculated based on this. 1 OD260 = 10 12 vp / mL, if OD260 / OD280>1.3, it indicates high purity, virus titer vp / mL=A260×dilution×10 12 The total amount of viruses obtained is collected to obtain the adenovirus expression vector Ad-GFP.

[0033] 2. Verification steps: After 6 weeks of feeding, the blank control group, control group and experimental group were euthanized by intraperitoneal injection of 1% sodium pentobarbital, and the aortas of each group were separated and frozen. The three groups of frozen aortas were thawed at room temperature for 30 minutes, then fixed in 4% paraformaldehyde for 15 minutes, placed in double distilled water twice (one minute each time), taken out and placed in 60% isopropanol for 5 minutes, and then placed in Oil Red O staining solution for 30 minutes. After the staining was completed, they were placed in 60% isopropanol and double distilled water for 1 minute each, and then sealed with water-soluble sealing agents and placed under a microscope to calculate the plaque area. Figure 1 As shown, Figure 1 The left side shows the aorta of the mouse model with dissection in the control group (saline). Figure 1 The right side shows the aorta dissected from the experimental group (Ad-OGDHL) mouse model. Figure 1 By comparison, the area of ​​plaques on the aorta removed from the experimental group (Ad-OGDHL) mouse model was larger than that of the blank control group (saline) and the control group (Ad-GFP) mouse model. Figure 2As shown, the horizontal axis represents the numbers of the blank control group (saline), the control group (Ad-GFP) and the experimental group (Ad-OGDHL); the vertical axis represents the size of the total injury area of ​​the mice, and the unit length is 10mm 2 , the unit of measurement is square millimeter; it can be seen that the total injury area of ​​the two control groups is about 14mm 2 The total injury area of ​​the animal model constructed by the experimental group was about 31 mm. 2 This indicates that the animal model of accelerated atherosclerosis formation by OGDHL is successfully prepared in the present invention, and the phenotype is obvious.

[0034] Compared with the establishment of a normal animal model, this experiment requires the use of adenovirus for intervention. In order to determine that it is not the adenovirus that causes obvious atherosclerosis in the experimental animals, GFP fluorescent protein is used for comparison with Ad-OGDHL to rule out the impact of adenovirus on the experimental animals. In addition, the successful infection of the experimental animals by adenovirus can be determined by observing the fluorescent reaction.

[0035] The present invention is a method for rapidly preparing an atherosclerosis experimental animal model by injecting adenovirus carrying an OGDHL protein gene in vitro through the circulatory system. The amount of plaque formation on the mouse aorta is observed to detect the role of OGDHL in the construction of an atherosclerosis plaque experimental animal model, and the function of OGDHL is used to rapidly prepare an atherosclerosis experimental animal model. The model establishment process of the present invention is simple, the success rate is high, and the results are stable and reliable, which saves the time of raising experimental animals and avoids the consequences of model construction failure. The animal model established by the present invention has important practical application value in the research of etiology, pathology, drug screening, clinical diagnosis and treatment of atherosclerosis, and can be widely promoted.

Claims

1. An animal model of atherosclerosis mediated by OGDHL, characterized in that: The animal model is obtained by exogenous injection of adenovirus to induce systemic high expression of OGDHL in mice.

2. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 1, characterized in that: Follow these steps: Step 1, constructing an adenovirus expression vector Ad-OGDHL; Step 2: inject the adenovirus expression vector Ad-OGDHL obtained in step 1 into the ApoE - / - The mice were fed with a high-fat diet for 6 weeks after injection, and the adenovirus expression vector Ad-OGDHL was injected again in the fourth week. The mice obtained after 6 weeks were the animal model.

3. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 2, characterized in that: The specific construction steps of the adenovirus expression vector Ad-OGDHL in step 1 are as follows: Step 1.1, using Hind Ⅲ and Sal Ⅰ to double-digest mouse OGDHL cDNA plasmid, electroeluting to recover OGDHL cDNA fragment, the nucleotide sequence of the cDNA fragment is shown in SEQ ID NO: 1, using Hind Ⅲ and Sal Ⅰ to double-digest pCMV-OGDHL plasmid, electroeluting to recover pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment, then ligating the OGDHL cDNA fragment with the pAdTrack-CMV Hind Ⅲ and Sal Ⅰ double-digested linear fragment using T4 DNA ligase, and transforming the ligated product into E. coli In DH5α competent cells, the pAdTrack-OGDHL recombinant plasmid was extracted by alkaline lysis after culture; Step 1.2: The pAdTrack-OGDHL recombinant plasmid obtained in step 1.1 was digested with PemⅠ, and the linear fragment was recovered by electroelution. The pAdTrack-OGDHL PemⅠ digested linear fragment was electrotransformed into the pAdEasy-1 containing E. coli BJ5183 electrocompetent cells, the transformation products were cultured and the pAdEasy-OGDHL recombinant plasmid was extracted by alkaline lysis method; Step 1.3, using PacⅠ to digest the pAdEasy-OGDHL obtained in step 1.2 to obtain a Pac I linearized recombinant plasmid pAdEasy-OGDHL fragment, using PacⅠ enzyme to linearize the recombinant plasmid pAdEasy-OGDHL fragment to transfect HEK293 cells, culturing the cells for 7 to 10 days after transfection, observing cell growth and green fluorescent protein expression every day, and when the green fluorescent protein expression in the cells is "comet"-shaped and cytopathic effect occurs, collecting the HEK293 cells and vortexing to lyse the cells, centrifuging after lysis, collecting the supernatant, which is the Ad-OGDHL virus solution, and storing the Ad-OGDHL virus solution at -80°C for later use; Step 1.4, on the day of infection, the Ad-OGDHL virus solution obtained in step 1.3 was added to 1.5 mL of DEME complete culture medium and cultured for 90 minutes, and then 3.5 mL of DMEM complete culture medium was added for culture. When the cells showed obvious cytopathic effect and were accompanied by shedding, the virus was collected and the infection with Ad-OGDHL virus solution was repeated 4 times. The total amount of the obtained virus was collected to obtain the adenovirus expression vector Ad-OGDHL of the present invention.

4. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 3, characterized in that: In step 1.3, HEK293 cells were cultured in a culture dish one day before transfection to ensure that the cell density of the cells was 30%-50% during transfection.

5. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 3, characterized in that: One day before infection in step 1.4, HEK293 cells were cultured in a culture dish to ensure that the cell density was 75%-85% during infection.

6. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 2, characterized in that: The cholesterol and fat content of the high-fat feed in step 2 is 1.5% cholesterol + 15% fat.

7. The method for establishing an animal model of atherosclerosis mediated by OGDHL according to claim 2, characterized in that: In step 2, ApoE is injected - / - The concentration of mouse adenovirus expression vector Ad-OGDHL is 1×10 11 vp / mL.

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