Application of Kif13b gene in treatment of abdominal aortic aneurysm

Abstract

The invention discloses application of a Kif13b gene in treatment of abdominal aortic aneurysm, belongs to the technical field of molecular biology and biological medicine, and provides application of the Kif13b gene as a drug target in screening of drugs for preventing, relieving and / or treating abdominal aortic aneurysm. According to the application, macrophage homeostasis and Kif13b gene overexpression can be regulated and controlled through targeted regulation of macrophage Kif13b gene expression, macrophage senescence inflammation can be effectively reversed, then abdominal aortic aneurysm is relieved, and a new target spot and a new strategy are provided for precise treatment of clinical abdominal aortic aneurysm patients.

Description

Technical Field

[0001] The invention belongs to the technical field 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 local permanent dilation of the abdominal aorta exceeding 1.5 times the normal size, which is mainly affected by multiple risk factors such as hypertension, high plasma cholesterol, atherosclerosis, smoking, age and gender. Importantly, once an aneurysm ruptures, the mortality rate will exceed 80%. However, to date, no drug therapy has been approved to inhibit the development of AAA or reduce the risk of aneurysm rupture. Only surgical repair or stent intervention is considered to treat AAA, and the population suitable for surgery is less than 10%, indicating that a large number of AAA patients are still at high risk. The main pathological processes of AAA are inflammatory cell infiltration into the vascular wall, apoptosis of vascular smooth muscle cells, and degradation of extracellular matrix elastin. Increasing new evidence shows that the incidence of AAA is positively correlated with abnormal lipid metabolism, especially triglyceride-rich lipoproteins (TRLs). Recently, a genome-wide association meta-analysis on AAA identified 121 independent risk loci and emphasized the importance of a series of lipoprotein receptor regulators (such as PCSK9) in AAA. In addition, both human and experimental animal studies 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 of abnormal lipoprotein metabolism inducing 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 has a variety of physiological activities, mainly as cargo proteins involved in the transport and localization of intracellular organelles and biomacromolecules. Kinesin-3 family member 13B (Kif13b) is the largest member of the kinesin-3 family. It has recently been found to be a new regulator of lipoprotein receptor function. It is specifically distributed in lipid rafts with high lipid content on the surface of the cell membrane of hepatocytes and mediates the endocytosis of LRP1 to maintain lipid balance. In addition, a series of studies have also reported that under PI3K-dependent conditions, Kif13b binds to microtubules and transports vascular endothelial growth factor receptor 2 (VEGFR2) from the Golgi membrane to the plasma membrane through anterograde transport, participating in the internalization and recycling of VEGFR2. At the same time, Kif13b can also regulate the localization of VEGFR2 on the lipid membrane and achieve receptor phosphorylation. In addition, the serum cholesterol and coagulation factor VIII levels of Kif13b knockout mice are increased; the absorption of low-density lipoprotein by embryonic fibroblasts of Kif13b-deficient mice is reduced. Existing research results suggest that Kif13b may be an important factor involved in maintaining the functional homeostasis of vascular cells and thus treating vascular injury diseases. However, the basic role of Kif13b in other vascular constituent cells has not been widely studied, and whether targeting Kif13b will become a potential therapeutic approach for the treatment of AAA remains to be confirmed. Summary of the invention

[0004] In order 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 patients with abdominal aortic aneurysm in clinical practice.

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

[0006] Preferably, the nucleotide sequence of the Kif13b gene is as shown in 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 Kif13b gene in macrophages through targeted regulation of macrophage homeostasis, thereby preventing, alleviating and / or treating abdominal aortic aneurysm.

[0009] The present invention also provides the use of the drug for promoting the overexpression of the Kif13b gene in the preparation of 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 for promoting 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 promotes the overexpression of Kif13b gene in macrophages through targeted regulation of macrophage homeostasis, thereby 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 found that macrophage homeostasis can be regulated by targeted regulation of the expression of macrophage Kif13b gene. Overexpression of 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 drawings required for use in the embodiments will be briefly introduced below. 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 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 mice macrophages, where A is mRNA expression, WT represents wild-type mice, Kif13b - / -represents Kif13b macrophage conditional knockout mice, "****" represents significant difference, B represents 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 Kif13b conditional knockout mouse AAA model, where A is the gross appearance, Kif13b f / f Representative wild-type mice, Lyz2Kif13b f / f Representative Kif13b macrophage conditional knockout mice, scale bar is 10 mm, 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 difference;

[0024] Figure 7 Transcriptomic enrichment analysis of mouse primary 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] Fig. 9 SA-β-gal staining of primary mouse BMDM, where A is the staining diagram, WT represents wild-type mice, Kif13b - / - represents Kif13b macrophage conditional knockout mice, the scale bar is 50 μm, B is the statistical diagram 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] Fig.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] Fig.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] Fig.12 Primary mouse BMDMγ-H2A.X staining, where A is the staining image, WT represents wild-type mice, Kif13b - / - represents 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] Fig.13 SA-β-gal staining of THP1 cells overexpressing Kif13b, where A is the staining diagram, GFP-HA represents THP1 macrophages infected with LV-GFP-HA, 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 group transfected with control lentivirus LV-GFP-HA, OE represents the group transfected with lentivirus LV-Kif13b-HA, * 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] Fig.14 This is the detection of SASP mRNA in THP1 cells overexpressing Kif13b. GFP+PBS represents THP1 macrophages infected with LV-GFP-HA. Kif13b OE +PBS represents LV-Kif13b-HA lentivirus-infected THP1 macrophages, GFP+LPS represents LPS-treated LV-GFP-HA-infected THP1 macrophages, 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] Fig.15 The gross appearance and diameter statistics of the mouse Kif13b overexpressing AAA model, where A is the gross appearance, the scale is 10 mm, B is the diameter statistics, LV-GFP-HA represents mice injected with LV-GFP-HA by tail vein, LV-Kif13b-HA represents mice injected with LV-Kif13b-HA lentivirus by tail vein, * represents the difference between genotypes, # represents the difference between treatment groups, and "**" represents P < 0.01;

[0033] Fig.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 should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

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

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation 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 a conflict with any incorporated document, the content 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 of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are 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] 1. Construction of Kif13b gene macrophage conditional knockout mice

[0043] Wild type (Kifi3b f / f ) mice were constructed by the Institute of Animal Science, Peking Union Medical College, and Lyz2-cre mice were obtained from the Institute of Model Animals, Wuhan University, and were kindly donated by Professor Li Hongliang of the Department of Cardiology, Renmin Hospital. The two were hybridized to obtain Lyz2Kif13b f / f Mice (eg Figure 1 The experiments used the same littermate Kif13b f / f Mice were used as controls. All animals were bred in the SPF animal room of Peking University Health Science Center (license number: SYXK (Beijing) 2022-0037), with a daily 12-h light cycle, 24±2℃ and 40±5 humidity. All mice were allowed to freely consume food and water unless otherwise specified.

[0044] Eight-week-old wild-type males (Kif13b f / f ) and Kif13b macrophage conditional knockout (Lyz2Kif13b f / f ) mice, primary bone marrow-derived macrophages (BMDM) were extracted, and the relative expression levels of Kif13b mRNA and protein were detected by RT-qPCR and Western Blot: f / f Compared with mice, Lyz2Kif13b f / f Kif13b mRNA and protein were almost undetectable in mouse macrophages, proving that myeloid conditional knockout Kif13b mice were successfully constructed (e.g. Figure 2 Middle A and Figure 2 (shown in B).

[0045] 2. Building Kif13b f / f Mouse and Lyz2Kif13b f / f Mouse AAA Model

[0046] Male mice aged 8-10 weeks, weighing 22-26 g, were selected and incubated in situ with 10 mg / mL porcine pancreatic elastase (PPE, MCE, HY-P2974) for 40 min. The samples were collected two weeks later (e.g. Figure 3 All animal experiments were performed in the SPF animal room of Peking University Health Science Center in accordance with the principles of laboratory animal care (NIH publication No. 85Y23, revised in 1996) and approved by the Peking University Laboratory Animal Ethics Committee (LA2023460).

[0047] 3. Determination of abdominal aorta diameter in mice

[0048] The aorta of PPE-induced abdominal aortic aneurysm mice was collected, fixed with 4% paraformaldehyde for 24 h, dehydrated with 20% sucrose for 24 h, and enfaced to obtain Kif13b 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 Middle 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 dilation, 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 compared with Kif13b f / f The abdominal aorta wall thickness of mice is 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 Media of the Aortic Wall in Mouse Aortic Aneurysms

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

[0055] The results are as follows Figure 6 Middle 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 (BMDM) were extracted from mice. The specific method was as follows: cells were taken from the tibia and femur of mice, then cultured in DMEM supplemented with 10% FBS (FS301-02, TransGen, China) and 1% penicillin-streptomycin, and stimulated with 25ng / mL mouse macrophage colony-stimulating factor 1 (M-CSF; 315-02, Peprotech, USA) for 6 days. Transcriptomics high-throughput sequencing was performed, and pathway enrichment of the differentially expressed genes was performed.

[0058] The results are as follows Figure 7 As shown, Lyz2Kif13b f / f Compared with Kif13b in mice f / f The senescence pathway of 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 mouse primary bone marrow-derived macrophages (BMDM) are as follows: Figure 8 As shown, Lyz2Kif13b f / f Pro-aging genes were upregulated and anti-aging genes were 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 Fig. 9 Middle A and Fig. 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 Fig.10 Middle A and Fig.10As shown in B, Lyz2Kif13b f / f Oxidative stress in mouse BMDM was aggravated.

[0067] 10. Detection of SASPmRNA 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 level of senescence-associated secretory phenotype (SASP) was detected by qPCR.

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

[0070] XI. 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 Fig.12 Middle A and Fig.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 Institute of Cardiovascular Medicine, Peking University) were treated with 100 ng / mL PMA (Solebol, P6741) for 48 h to induce THP1 cells to become macrophages. LV-GFP-HA or LV-Kif13b-HA lentivirus was constructed by: using calcium phosphate (Ca 3 (PO 4 ) 2) method to co-transfect lentiviral vector (15 mg) and pMDLg / pRRE, RSV / Rev and VSV-G (5 mg each) into 293T cells in 10 cm dishes. The culture medium was changed after 16 h of cell culture. Two days after transfection, the viral supernatant was collected and filtered with a 0.45 μm filter (SLHP033RB, Millipore, USA). The filtered supernatant was concentrated to 1e8 TU by ultracentrifugation for in vivo and in vitro experiments. THP1 macrophages were infected with lentivirus at a dose of 1e5 TU, and the cells were treated with 1 μg / mL LPS for 24 h and stained with SA-β-gal.

[0075] The results are as follows Fig.13 Middle A and Fig.13 As shown in 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 to become macrophages. THP1 macrophages were 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 level was detected by qPCR.

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

[0079] XIV. Gross appearance and diameter statistics of the mouse overexpression Kif13b AAA model

[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 “2. Construction of Kif13b f / f Mouse and Lyz2Kif13b f / f 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 to obtain the gross appearance of AAA model of Kif13b-overexpressing mice and control group. ImageJ was used to calculate the ratio of the maximum diameter of abdominal aortic aneurysm to the diameter of adjacent normal abdominal aorta.

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

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

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

[0084] The results are as follows Fig.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 only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of Kif13b gene as a drug target in screening drugs for preventing, alleviating and / or treating abdominal aortic aneurysm.

2. The application according to claim 1, characterized in that: The nucleotide sequence of the Kif13b gene is shown in SEQ ID NO.

1.

3. The application according to claim 1, characterized in that: The drug for preventing, alleviating and / or treating abdominal aortic aneurysm is a drug that promotes overexpression of Kif13b gene.

4. The use according to claim 1, characterized in that: The drug for preventing, alleviating and / or treating abdominal aortic aneurysm promotes the overexpression of Kif13b gene in macrophages through targeted regulation of macrophage homeostasis, thereby preventing, alleviating and / or treating abdominal aortic aneurysm.

5. Use of the drug for promoting overexpression of the Kif13b gene according to claim 1 in the preparation of a drug for preventing, alleviating and / or treating abdominal aortic aneurysm.

6. A drug for preventing, alleviating and / or treating abdominal aortic aneurysm, characterized in that: The invention comprises a drug for promoting overexpression of the Kif13b gene as claimed in claim 1.

7. Use of the Kif13b gene as claimed in claim 1 as a drug target in screening drugs for preventing, alleviating and / or treating macrophage senescent inflammation.

8. The use according to claim 7, characterized in that: The drug for preventing, alleviating and / or treating macrophage senescent inflammation is a drug that promotes overexpression of the Kif13b gene.

9. The use according to claim 7, characterized in that: The drug for preventing, alleviating and / or treating macrophage senescence inflammation promotes the overexpression of Kif13b gene in macrophages through targeted regulation of macrophage homeostasis, thereby preventing, alleviating and / or treating macrophage senescence inflammation.

10. A drug for preventing, alleviating and / or treating macrophage senescent inflammation, characterized in that: The invention comprises a drug for promoting overexpression of the Kif13b gene as claimed in claim 1.

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