Application of Kif13b gene in the treatment of abdominal aortic aneurysm

Abstract

The present invention discloses the use of the Kif13b gene in the treatment of abdominal aortic aneurysms, belonging to the fields of molecular biology and biomedicine technology. It provides the use of the Kif13b gene as a drug target in screening drugs for the prevention, alleviation, and / or treatment of abdominal aortic aneurysms. The present invention finds that macrophage homeostasis can be regulated by targeted regulation of macrophage Kif13b gene expression. Overexpression of the Kif13b gene can effectively reverse senescent inflammation in macrophages, thereby alleviating abdominal aortic aneurysms. This provides a new target and new strategy for the precise treatment of patients with abdominal aortic aneurysms in clinical practice.

Description

Technical Field

[0001] The present invention belongs to the technical fields of molecular biology and biomedicine, and in particular relates to application of Kif13b gene in treating abdominal aortic aneurysm. Background Art

[0002] Abdominal aortic aneurysm (AAA) is a fatal vascular disease characterized by a localized, permanent dilation of the abdominal aorta exceeding 1.5 times its normal size. It is primarily influenced by multiple risk factors, including hypertension, high plasma cholesterol, atherosclerosis, smoking, age, and sex. Importantly, once an aneurysm ruptures, the mortality rate exceeds 80%. However, to date, no pharmacological therapies have been approved to inhibit AAA progression or reduce the risk of rupture. Surgical repair or stenting are the only options for AAA, and less than 10% of the population is suitable for surgery, indicating that a significant number of AAA patients remain at high risk. The primary pathological processes underlying AAA development are inflammatory cell infiltration into the vessel wall, apoptosis of vascular smooth muscle cells, and degradation of extracellular matrix elastin. Accumulating evidence indicates that the incidence of AAA is positively correlated with abnormal lipid metabolism, particularly triglyceride-rich lipoproteins (TRLs). A recent genome-wide association meta-analysis of AAA identified 121 independent risk loci and highlighted the importance of a series of lipoprotein receptor regulators, such as PCSK9, in AAA. In addition, studies in humans and experimental animals have confirmed that lipoprotein receptors such as low-density lipoprotein receptor-related protein 1 (LRP1) are associated with the occurrence and development of AAA, indicating that lipoproteins and their receptors play an indispensable role in AAA. However, it is worth noting that the detailed molecular mechanism by which abnormal lipoprotein metabolism induces AAA and how lipoprotein receptors participate in the pathogenesis of AAA remain largely unclear, thus limiting the further application of targeting lipoproteins and their receptors to treat AAA.

[0003] The kinesin-3 family of proteins possesses diverse physiological activities, primarily acting as cargo proteins involved in the transport and localization of intracellular organelles and biomacromolecules. Kinesin-3 family member 13B (Kif13b), the largest member of the kinesin-3 family, has recently been identified as a novel regulator of lipoprotein receptor function. It is specifically localized in lipid rafts, the lipid-rich surface of hepatocytes, and mediates the internalization of LRP1 to maintain lipid homeostasis. Furthermore, a series of studies have reported that Kif13b binds to microtubules and transports vascular endothelial growth factor receptor 2 (VEGFR2) from the Golgi membrane to the plasma membrane via anterograde transport, contributing to VEGFR2 internalization and recycling. Kif13b also regulates VEGFR2 localization on lipid membranes, enabling receptor phosphorylation. Furthermore, Kif13b knockout mice exhibit elevated serum cholesterol and factor VIII levels, and embryonic fibroblasts of Kif13b-deficient mice exhibit reduced low-density lipoprotein uptake. Existing research results suggest that Kif13b may be an important factor involved in maintaining the functional homeostasis of vascular cells and thus treating vascular damage diseases. However, the fundamental role of Kif13b in other vascular components has not been extensively studied, and whether targeting Kif13b will become a potential therapeutic approach for AAA remains to be confirmed. Summary of the Invention

[0004] To solve the above technical problems, the present invention proposes the application of Kif13b gene in the treatment of abdominal aortic aneurysm. Kif13b gene therapy can effectively reverse macrophage senescent inflammation and provide new targets and new strategies for the precise treatment of abdominal aortic aneurysm patients in clinical practice.

[0005] To achieve the above objectives, the present invention provides the use of Kif13b gene as a drug target in screening drugs for preventing, alleviating and / or treating abdominal aortic aneurysms.

[0006] Preferably, the nucleotide sequence of the Kif13b gene is shown as SEQ ID NO.1.

[0007] Preferably, the drug for preventing, alleviating and / or treating abdominal aortic aneurysm is a drug that promotes overexpression of the Kif13b gene.

[0008] Preferably, the drug for preventing, alleviating and / or treating abdominal aortic aneurysm promotes overexpression of the Kif13b gene in macrophages through targeting, regulating macrophage homeostasis, and thereby preventing, alleviating and / or treating abdominal aortic aneurysm.

[0009] The present invention also provides use of the drug for promoting overexpression of the Kif13b gene in preparing a drug for preventing, alleviating and / or treating abdominal aortic aneurysm.

[0010] The present invention also provides a drug for preventing, alleviating and / or treating abdominal aortic aneurysm, including a drug that promotes overexpression of the Kif13b gene.

[0011] The present invention also provides the use of the Kif13b gene as a drug target in screening drugs for preventing, alleviating and / or treating macrophage senescent inflammation.

[0012] Preferably, the drug for preventing, alleviating and / or treating macrophage senescent inflammation is a drug that promotes overexpression of the Kif13b gene.

[0013] Preferably, the drug for preventing, alleviating and / or treating macrophage senescence inflammation targets and promotes overexpression of the Kif13b gene in macrophages, thereby regulating macrophage homeostasis, and preventing, alleviating and / or treating macrophage senescence inflammation.

[0014] The present invention also provides a drug for preventing, alleviating and / or treating macrophage aging inflammation, including a drug for promoting overexpression of the Kif13b gene.

[0015] Compared with the prior art, the present invention has the following advantages and technical effects:

[0016] The present invention discovered that macrophage homeostasis can be regulated by targeted regulation of the expression of the macrophage Kif13b gene. Overexpression of the Kif13b gene can effectively reverse macrophage aging inflammation, thereby alleviating abdominal aortic aneurysm, providing new targets and new strategies for the precise treatment of patients with abdominal aortic aneurysm in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0018] Figure 1 The construction process of Kif13b conditional knockout mice. In the figure, WT represents wild-type mice, Floxed represents conditional gene knockout mice, and Myeloid Specific deletion represents myeloid conditional Kif13b gene knockout mice.

[0019] Figure 2 The mRNA and protein expression of Kif13b conditional knockout mouse macrophages were determined. A represents mRNA expression, WT represents wild-type mice, and Kif13b - / -represents Kif13b macrophage conditional knockout mice, "****" represents significant difference, B is protein expression determination, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice;

[0020] Figure 3 Schematic diagram for the construction of Kif13b conditional knockout mouse AAA model, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice;

[0021] Figure 4 The gross appearance and diameter statistics of the Kif13b conditional knockout mouse AAA model, where A represents the gross appearance, Kif13b represents the f / f Representative wild-type mice, Lyz2Kif13b f / f Representative Kif13b macrophage conditional knockout mice, scale bar is 10mm, B is diameter statistics, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice, and “***” represents significant difference;

[0022] Figure 5 HE staining of Kif13b conditional knockout mouse AAA model, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice;

[0023] Figure 6 The EVG staining images of the AAA model of Kif13b conditional knockout mice, where A is the EVG staining image, B is the area ratio of the elastic fiber staining positive area in the vascular wall tunica media to the tunica media area, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice, and “*” represents significant differences;

[0024] Figure 7 For transcriptomic enrichment analysis of primary mouse BMDM, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice;

[0025] Figure 8Heat map of differentially expressed genes in the aging pathway of primary mouse BMDM, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f represents Kif13b macrophage conditional knockout mice;

[0026] Figure 9 SA-β-gal staining of primary mouse BMDM, where A is the staining image, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, the scale bar is 50 μm, B is the statistical graph of the number of stained cells, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, WT+LPS represents lipopolysaccharide-treated wild-type mice, Kif13b - / - +LPS represents lipopolysaccharide-treated Kif13b macrophage conditional knockout mice, * represents the difference between genotypes, # represents the difference between treatment groups, “**” represents P < 0.01, “***” represents P < 0.001, “#” represents P < 0.05, and “##” represents P < 0.01;

[0027] Figure 10 ROS staining of primary mouse BMDM, A is the staining diagram, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, the scale bar is 100 μm, B is the statistics of reactive oxygen species positive cells, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, WT+LPS represents lipopolysaccharide-treated wild-type mice, Kif13b - / - +LPS represents lipopolysaccharide-treated Kif13b macrophage conditional knockout mice, * represents the difference between genotypes, # represents the difference between treatment groups, “*” represents P < 0.05, “**” represents P < 0.01, “##” represents P < 0.01, and “###” represents P < 0.001;

[0028] Figure 11 For the detection of SASP mRNA in primary mouse BMDM, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, WT+LPS represents lipopolysaccharide-treated wild-type mice, Kif13b - / -+LPS represents lipopolysaccharide-treated Kif13b macrophage conditional knockout mice, * represents the difference between genotypes, # represents the difference between treatment groups, “ns” represents no statistically significant difference, “*” represents P < 0.05, “**” represents P < 0.01, “***” represents P < 0.001, “##” represents P < 0.01, and “###” represents P < 0.001;

[0029] Figure 12 Primary mouse BMDMγ-H2A.X staining, where A is the staining image, WT represents wild-type mice, Kif13b - / - Representative Kif13b macrophage conditional knockout mice, scale bar is 10 μm, B is the average γ-H2A.X density of each cell nucleus, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, WT+LPS represents lipopolysaccharide-treated wild-type mice, Kif13b - / - +LPS represents lipopolysaccharide-treated Kif13b macrophage conditional knockout mice, * represents the difference between genotypes, # represents the difference between treatment groups, “ns” represents no statistically significant difference, “**” represents P < 0.01, “****” represents P < 0.0001, and “##” represents P < 0.01;

[0030] Figure 13 SA-β-gal staining of THP1 cells overexpressing Kif13b, where A is the staining image, GFP-HA represents THP1 macrophages infected with LV-GFP-HA, and Kif13b-HA represents THP1 macrophages infected with LV-Kif13b-HA lentivirus, the scale bar is 200 μm, B is the statistics of SA-β-gal positive cells, NC represents the control lentivirus LV-GFP-HA transfection group, OE represents the lentivirus LV-Kif13b-HA transfection group, * represents the difference between genotypes, # represents the difference between treatment groups, "ns" represents no statistically significant difference, "****" represents P < 0.0001, and "####" represents P < 0.0001;

[0031] Figure 14 For the detection of Kif13b SASP mRNA overexpression in THP1 cells, GFP+PBS represents LV-GFP-HA infected THP1 macrophages, Kif13b OE +PBS represents THP1 macrophages infected with LV-Kif13b-HA lentivirus, GFP+LPS represents THP1 macrophages infected with LPS-treated LV-GFP-HA, Kif13b OE+LPS represents LPS-treated LV-Kif13b-HA lentivirus-infected THP1 macrophages, * represents the difference between genotypes, # represents the difference between treatment groups, “**” represents P < 0.01, “****” represents P < 0.0001, and “####” represents P < 0.0001;

[0032] Figure 15 The gross appearance and diameter statistics of the Kif13b overexpressing AAA model in mice, where A is the gross appearance, the scale is 10 mm, and B is the diameter statistics. LV-GFP-HA represents mice injected with LV-GFP-HA in the tail vein, and LV-Kif13b-HA represents mice injected with LV-Kif13b-HA lentivirus in the tail vein. * represents the difference between genotypes, # represents the difference between treatment groups, and "**" represents P < 0.01.

[0033] Figure 16 HE and EVG staining of the mouse AAA model overexpressing Kif13b. LV-GFP-HA represents mice injected with LV-GFP-HA through the tail vein, and LV-Kif13b-HA represents mice injected with LV-Kif13b-HA lentivirus through the tail vein. DETAILED DESCRIPTION

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0037] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.

[0038] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0039] Experimental Example 1

[0040] Kif13b gene synthesis sequence (CDS sequence: abbreviation of coding sequence. DNA is transcribed into mRNA, and mRNA is translated into protein after splicing and other processing. CDS is a DNA sequence that corresponds one-to-one with the protein sequence, and the sequence does not contain other sequences that do not correspond to the protein. Sequence changes during mRNA processing are not considered. In short, the CDS sequence completely corresponds to the protein codon. CDS is a sequence that encodes a protein product and is a term in structural genomics).

[0041]

[0042] I. Construction of conditional knockout mice of Kif13b gene in macrophages

[0043] Wild-type (Kifi3b f / f ) mice were constructed by the Institute of Laboratory Animals, Peking Union Medical College. Lyz2-cre mice were from the Institute of Model Animals, Wuhan University, and were kindly donated by Professor Li Hongliang of the Department of Cardiovascular Medicine, Renmin Hospital. The two were crossed to obtain Lyz2Kif13b f / f mice (as Figure 1 shown). Littermate Kif13b f / f mice were used as controls in the experiment. All animals were bred in the SPF-class animal house of Peking University Health Science Center (License number: SYXK (Beijing) 2022-0037), ensuring 12-hour cyclic light exposure per day, maintaining a temperature of 24±2°C and a humidity of 40±5%. Without special instructions, all mice had free access to food and water.

[0044] Eight-week-old male wild-type (Kif13b f / f ) and conditional knockout of Kif13b in macrophages (Lyz2Kif13b f / f ) mice were selected, and primary bone marrow-derived macrophages (BMDM) were extracted. The relative expression levels of Kif13b mRNA and protein were detected by RT-qpcr and Western Blot: Compared with Kif13b f / f mice, almost no Kif13b mRNA and protein were detected in the macrophages of Lyz2Kif13b f / f mice, demonstrating the successful construction of myeloid conditional knockout Kif13b mice (as shown in A in Figure 2 and B in Figure 2 ).

[0045] II. Construction of AAA models in Kif13b f / f mice and Lyz2Kif13b f / f mice

[0046] Eight- to ten-week-old male mice weighing 22-26 g were selected, and the infrarenal abdominal aortic segments were incubated in situ with 10 mg / mL porcine pancreatic elastase (PPE, MCE, HY-P2974) for 40 min. Samples were taken two weeks later (as shown in Figure 3 ). All animal experiments were conducted in the SPF-class animal house of Peking University Health Science Center, carried out in accordance with the principles of laboratory animal care (revised in NIH publication No. 85Y231996), and approved by the Laboratory Animal Ethics Committee of Peking University (LA2023460).

[0047] III. Measurement of abdominal aortic diameter in mice

[0048] The aorta of PPE-induced abdominal aortic aneurysm mice was collected, fixed with 4% paraformaldehyde for 24 h, and dehydrated with 20% sucrose for 24 h. The aorta was enfaced and Kif13b was obtained. f / f and Lyz2Kif13b f / f Gross view of the mouse AAA model, and the ratio of the maximum diameter of the abdominal aortic aneurysm to the diameter of the adjacent normal abdominal aorta was calculated using ImageJ.

[0049] The results are as follows Figure 4 China A and Figure 4 As shown in B, Lyz2Kif13b f / f The diameter of the mouse abdominal aorta compared with Kif13b f / f The dilation of the mouse aorta was significantly increased.

[0050] IV. Observation of the Aorta Characterization of Abdominal Aortic Aneurysm in Mice

[0051] The abdominal aortic aneurysm was cut at the site of obvious expansion, embedded in paraffin and sliced ​​(5 μm thickness), and then Kif13b f / f and Lyz2Kif13b f / f HE staining of mouse AAA model.

[0052] The results are as follows Figure 5 As shown, Lyz2Kif13b was observed f / f The abdominal aorta wall of mice is compared with Kif13b f / f The abdominal aorta wall thickness of mice increased, and Lyz2Kif13b f / f The number of inflammatory cells with large nuclei and dark staining increased in the abdominal aorta wall of mice.

[0053] 5. Characterization and Observation of the Medial Tumor of the Aortic Wall in Mouse Aortic Aneurysms

[0054] Prepare paraffin sections in the same manner as in "IV. Observation of aorta characterization of abdominal aortic aneurysm in mice" and perform Kif13b f / f and Lyz2Kif13b f / f EVG staining of mouse AAA model was performed, and the proportion of the area of ​​the tunica media in which elastic fibers were positively stained was calculated using ImageJ.

[0055] The results are as follows Figure 6 China A and Figure 6 As shown in B, Lyz2Kif13b f / f The degree of elastic fiber disintegration in the abdominal aorta of mice was aggravated.

[0056] VI. Transcriptomics High-throughput Sequencing

[0057] From 8-week-old male Kif13b f / f and Lyz2Kif13bf / f Primary bone marrow-derived macrophages (BMDMs) were isolated from mice by harvesting from the tibia and femur. BMDMs were cultured in DMEM supplemented with 10% FBS (FS301-02, TransGen, China) and 1% penicillin-streptomycin and stimulated with 25 ng / mL mouse macrophage colony-stimulating factor 1 (M-CSF; 315-02, Peprotech, USA) for 6 days. High-throughput transcriptomics sequencing was performed, and pathway enrichment was performed on the differentially expressed genes identified.

[0058] The results are as follows Figure 7 As shown, Lyz2Kif13b f / f Compared with Kif13b in mice f / f The senescence pathway in mouse BMDM was significantly upregulated.

[0059] 7. Analyze transcriptome sequencing results

[0060] Analysis of Kif13b f / f and Lyz2Kif13b f / f The transcriptome sequencing results of primary mouse bone marrow-derived macrophages (BMDM) are as follows: Figure 8 As shown, Lyz2Kif13b f / f Pro-aging genes are upregulated and anti-aging genes are downregulated in mouse BMDM.

[0061] 8. Determination of Senescent Cell Content in Primary Mouse Bone Marrow-derived Macrophages

[0062] Kif13b f / f and Lyz2Kif13b f / f Mouse primary bone marrow-derived macrophages (BMDM) were treated with 10 ng / mL LPS (Sigma, L2630) for 24 h and then stained with SA-β-gal.

[0063] The results are as follows Figure 9 China A and Figure 9 As shown in B, Lyz2Kif13b f / f The number of mouse BMDM senescent cells (SA-β-gal positive staining) increased.

[0064] IX. Oxidative Stress Assay in Primary Mouse Bone Marrow-derived Macrophages

[0065] Kif13b f / f and Lyz2Kif13b f / f Mouse primary bone marrow-derived macrophages (BMDM) were treated with 10 ng / mL LPS for 24 h, and ROS staining was performed.

[0066] The results are as follows Figure 10 China A and Figure 10As shown in B, Lyz2Kif13b f / f Oxidative stress in mouse BMDM was aggravated.

[0067] 10. Detection of SASP mRNA in Primary Mouse BMDM

[0068] Kif13b f / f and Lyz2Kif13b f / f Mouse primary bone marrow-derived macrophages (BMDM) were treated with 10 ng / mL LPS for 24 h, and the mRNA levels of the senescence-associated secretory phenotype (SASP) were detected by qPCR.

[0069] The results are as follows Figure 11 As shown, Lyz2Kif13b f / f Mouse BMDM activated the SASP.

[0070] 11. γ-H2A.X staining of primary mouse BMDM

[0071] Kif13b f / f and Lyz2Kif13b f / f Mouse primary bone marrow-derived macrophages (BMDM) were treated with 10 ng / mL LPS for 24 h and stained with γ-H2A.X.

[0072] The results are as follows Figure 12 China A and Figure 12 As shown in B, Lyz2Kif13b f / f The amount of γ-H2A.X in the nucleus of mouse BMDM increased, and DNA damage was aggravated.

[0073] 12. SA-β-gal staining of THP1 cells overexpressing Kif13b

[0074] THP1 cells (from the Cardiovascular Institute of Peking University) were treated with 100 ng / mL PMA (Solebol, P6741) for 48 hours to induce THP1 cells into macrophages. LV-GFP-HA or LV-Kif13b-HA lentiviruses were constructed by co-transfecting 293T cells plated in 10 cm dishes with the lentiviral vector (15 mg) and 5 mg each of pMDLg / pRRE, RSV / Rev, and VSV-G using the calcium phosphate (Ca3(PO4)2) method. The medium was changed after 16 hours of cell culture. Two days after transfection, the viral supernatant was collected and filtered through a 0.45 μm filter (SLHP033RB, Millipore, USA). The filtered supernatant was concentrated by ultracentrifugation to 1e8 TU for use in in vitro and in vivo experiments. THP1 macrophages were infected with 1e5 TU of lentivirus and treated with 1 μg / mL LPS for 24 hours before staining with SA-β-gal.

[0075] The results are as follows Figure 13 China A and Figure 13 As shown in middle B, the number of senescent cells (SA-β-gal positive staining) overexpressing Kif13b was reduced.

[0076] 13. Detection of SASP mRNA in THP1 cells overexpressing Kif13b

[0077] THP1 cells were treated with 100 ng / mL PMA (Solebol, P6741) for 48 h to induce THP1 cells into macrophages. THP1 macrophages were then infected with LV-GFP-HA or LV-Kif13b-HA lentivirus (1e5 TU) and treated with 1 μg / mL LPS for 24 h. The senescence-associated secretory phenotype (SASP) mRNA levels were detected by qPCR.

[0078] The results are as follows Figure 14 As shown, SASP can be significantly inhibited by expressing Kif13b.

[0079] 14. Gross appearance and diameter statistics of the Kif13b AAA model in mice

[0080] Eight-week-old WT mice were selected and injected with LV-GFP-HA and LV-Kif13b-HA lentiviruses (1e7 TU) through the tail vein. One week later, a PPE-induced AAA model was established (the method was the same as in “II. Construction of Kif13b f / f Mouse and Lyz2Kif13b f / f Two weeks later, the aortas of mice with PPE-induced abdominal aortic aneurysms were collected, fixed with 4% paraformaldehyde for 24 hours, and dehydrated with 20% sucrose for 24 hours. Aortic enfaces were then performed to obtain gross images of the AAA model in Kif13b-overexpressing mice and the control group. The ratio of the maximum diameter of the abdominal aortic aneurysm to the diameter of the adjacent normal abdominal aorta was calculated using ImageJ.

[0081] The results are as follows Figure 15 China A and Figure 15 As shown in middle B, the expansion of the abdominal aorta diameter was significantly reduced in mice overexpressing Kif13b.

[0082] 15. HE and EVG staining of the Kif13b overexpressing AAA model in mice

[0083] The area where the abdominal aortic aneurysm was obviously dilated was cut, embedded in paraffin and sectioned (5 μm thickness). Then, HE and EVG staining of the WT mouse overexpressing Kif13b AAA model were performed. ImageJ was used to calculate the proportion of the area of ​​the tunica media where elastic fibers were positively stained.

[0084] The results are as follows Figure 16 As shown, the thickness of the abdominal aorta wall of mice overexpressing Kif13b was reduced, the number of inflammatory cells with large nuclei and dark staining was reduced, and the degree of elastic fiber disintegration was alleviated.

[0085] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. Use of a drug that promotes overexpression of the Kif13b gene in the preparation of a drug for alleviating and / or treating abdominal aortic aneurysm.

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