Polypeptide for inhibiting formation of P-body in cell and application thereof

Abstract

The invention discloses a polypeptide for inhibiting formation of P-bodies in cells and application of the polypeptide. Wherein the amino acid sequence of the polypeptide is as shown in SEQ ID NO. 4. According to the invention, structural analysis is carried out on the key protein of the P-body, so that a key structural domain required for forming the P-body is determined; the method comprises the following steps: designing an inhibitory peptide of a P-small body key protein through AlphaFold 3 and a bioinformatics means; the polypeptide with the amino acid sequence shown as SEQ ID NO.4 is obtained by combining the cell-penetrating peptide with the designed inhibitory peptide, and the bioavailability of the polypeptide in cells is enhanced. The polypeptide can effectively inhibit and block formation of P-bodies in cells, can significantly reduce expression of endothelial cell inflammatory factors at a concentration of 30 [mu] M, has an important effect on protection of a blood-brain barrier, and has important significance in treatment of cardiovascular and cerebrovascular diseases.

Description

Technical Field

[0001] The present invention relates to the fields of biomedicine and biotechnology, and in particular to a polypeptide for inhibiting the formation of P-bodies in cells and an application thereof. Background Art

[0002] The P-body (Processing body, P-body) is a membrane-less organelle located in the cytoplasm. It is mainly involved in the metabolic process of mRNA, including degradation, storage and translation regulation. P-bodies are granular structures with the following key features: (1) Composition: P-bodies are composed of a variety of RNA-binding proteins and RNA, which play an important role in the function of P-bodies; (2) Degradation site: P-bodies are the main site of mRNA degradation and can effectively regulate gene expression. (3) Translation inhibition: mRNA can be sequestered in P-bodies, thereby inhibiting its translation; (4) Environmental response: P-bodies can respond to environmental changes in the cell, such as stress or nutrient deficiency, thereby regulating the fate of RNA and participating in stress response; (5) Dynamic characteristics: P-bodies are not fixed structures. They will change dynamically according to the physiological state of the cell and changes in the external environment.

[0003] The main protein components that make up the P-body include DDX6, EDC4, 4E-T, LSM14a and PATL1. These protein molecules bind to each other through key domains and together form the P-body. Each of these proteins is necessary for the assembly of the P-body. At present, there are literatures that prove the domain information of the binding between protein molecules in the P-body, and explore genetic modification methods to inhibit the formation of P-body. However, there is currently no research and product on the inhibition of P-body formation in cells by peptides.

[0004] Therefore, the existing technology still needs to be improved and developed. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a polypeptide for inhibiting the formation of P-bodies in cells and its application, aiming to solve the problem of the lack of polypeptide drugs that inhibit the formation of P-bodies in cells in the prior art.

[0006] The technical solution of the present invention is as follows:

[0007] In a first aspect, a polypeptide for inhibiting the formation of P-bodies in cells is provided, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO.4.

[0008] In a second aspect, a nucleic acid is provided, wherein the nucleic acid encodes the polypeptide as described in the first aspect.

[0009] In a third aspect, a recombinant vector is provided, wherein the recombinant vector expresses the polypeptide as described in the first aspect.

[0010] In a fourth aspect, a composition is provided, comprising the polypeptide as described in the first aspect.

[0011] In a preferred technical solution, the composition further comprises a pharmaceutically acceptable solvent.

[0012] In a further preferred technical solution, the solvent is selected from one or more of water, physiological saline, aqueous glucose solution, and dimethyl sulfoxide.

[0013] In a preferred technical solution, the composition further comprises a pharmaceutically acceptable carrier.

[0014] In a further preferred technical solution, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a filler, a binder, a lubricant, a disintegrant, and a stabilizer.

[0015] In the fifth aspect, a method is provided for preparing an agent for inhibiting the formation of P-bodies in cells by using the polypeptide as described in the first aspect, the nucleic acid as described in the second aspect, the recombinant vector as described in the third aspect, or the composition as described in the fourth aspect.

[0016] In a sixth aspect, a method is provided for preparing a drug for treating cardiovascular and cerebrovascular diseases by using the polypeptide described in the first aspect, the nucleic acid described in the second aspect, the recombinant vector described in the third aspect, or the composition described in the fourth aspect.

[0017] Beneficial effects: The present invention determines the key domains required for the formation of P-bodies by structural analysis of the key proteins of P-bodies; designs inhibitory peptides of the key proteins of P-bodies by means of AlphaFold 3 and bioinformatics; obtains a polypeptide with an amino acid sequence as shown in SEQ ID NO.4 by combining the cell-penetrating peptide with the designed inhibitory peptide, and enhances its bioavailability in cells. The polypeptide can effectively inhibit and block the formation of P-bodies in cells, and can significantly reduce the expression of inflammatory factors in endothelial cells at a concentration of 30 μM, which plays an important role in protecting the blood-brain barrier and is of great significance in the treatment of cardiovascular and cerebrovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a binding analysis diagram of polypeptide-1 and DDX6 protein.

[0019] Figure 2 This is a binding analysis diagram of polypeptide-2 and DDX6 protein.

[0020] Figure 3 This is a binding analysis diagram of polypeptide-3 and DDX6 protein.

[0021] Figure 4 This is a binding analysis diagram of polypeptide-4 and 4E-T protein.

[0022] Figure 5 This is a graph showing the effects of peptides-1 to -4 on the cell activity of beend.3 cells.

[0023] Figure 6 This is a graph showing the intensity change of peptide-1-Cy5 entering beend.3 cells as the incubation time increases.

[0024] Figure 7 This is a graph showing the effect of polypeptide-1 on the number of P-bodies in beend.3 cells.

[0025] Figure 8 This is a graph showing the effect of polypeptide-2 on the number of P-bodies in beend.3 cells.

[0026] Fig. 9 This is a graph showing the effect of peptide-3 on the number of P-bodies in beend.3 cells.

[0027] Fig.10 This is a graph showing the effect of peptide-4 on the number of P-bodies in beend.3 cells.

[0028] Fig.11 This is a graph showing the effect of peptide-5 on the number of P-bodies in beend.3 cells.

[0029] Fig.12 This is a graph showing the results of peptide-4 alleviating BMEC damage caused by OGD / R.

[0030] Fig.13 This is a graph showing the results of peptide-4 reducing the mRNA levels of inflammatory factors in bEnd.3 cells induced by OGD / R. DETAILED DESCRIPTION

[0031] The present invention provides a polypeptide for inhibiting the formation of P-bodies in cells and its application. In order to make the purpose, technical scheme and effect of the present invention clearer and more specific, the present invention is further described in detail below.

[0032] P-bodies play a crucial role in the fine regulation of RNA regulation and gene expression in cells. Dysregulation of P-body function and RNA regulation is associated with a variety of diseases, including cancer and neurodegenerative diseases. Understanding P-body RNA regulation is crucial to elucidating the complexity of gene regulation and the response of cells to environmental changes. However, there are currently no reagents and drugs that regulate P-body structure and function.

[0033] Based on this, an embodiment of the present invention provides a polypeptide for inhibiting the formation of P-bodies in cells, and the amino acid sequence of the polypeptide is shown in SEQ ID NO.4.

[0034] The present invention conducts structural analysis on the key proteins of P-bodies and determines the key domains required for the formation of P-bodies. By means of AlphaFold 3 and bioinformatics, corresponding inhibitory peptides (inhibitory peptides-1 to 4) are designed for each key protein molecule, and a non-specific peptide is also designed as an invalid control (invalid control peptide-5). In order to improve the cell membrane permeability of the inhibitory peptide, the present invention connects a cell-penetrating peptide (TAT) with transmembrane ability to the N-terminus of the above-mentioned P-body inhibitory peptide, and its amino acid sequence is YGRKKRRQRRR. By combining the cell-penetrating peptide with the designed inhibitory peptide, a composite polypeptide-1 to 4 (SEQ ID NO.1 to 4) is formed to enhance its bioavailability in cells. The results of the study using polypeptides-1 to 4 show that the polypeptides with the cell-penetrating peptide sequence added reach the highest peak at 12 hours after aggregation in the cells. At the same time, polypeptide-4, whose amino acid sequence is shown in SEQ ID NO.4, effectively inhibits the formation of P-bodies at a concentration of 30 μM and significantly reduces the expression of inflammatory factors in endothelial cells, which plays an important role in protecting the blood-brain barrier. Other polypeptides, including polypeptide-1, polypeptide-2 and polypeptide-3, failed to effectively inhibit the formation of P-bodies. Therefore, polypeptide-4 is of great significance in the treatment of cardiovascular and cerebrovascular diseases.

[0035] An embodiment of the present invention provides a nucleic acid encoding the polypeptide as described above.

[0036] An embodiment of the present invention provides a recombinant vector, wherein the recombinant vector expresses the polypeptide as described above.

[0037] An embodiment of the present invention provides a composition, which includes the polypeptide as described above.

[0038] In one embodiment, the composition further comprises a pharmaceutically acceptable solvent.

[0039] In a more specific embodiment, the solvent is selected from one or more of water, physiological saline, aqueous glucose solution, and dimethyl sulfoxide (DMSO).

[0040] In one embodiment, the composition further comprises a pharmaceutically acceptable carrier.

[0041] In a more specific embodiment, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a filler, a binder, a lubricant, a disintegrant, and a stabilizer.

[0042] The embodiments of the present invention provide a use of the above-mentioned polypeptide, the above-mentioned nucleic acid, the above-mentioned recombinant vector, and the above-mentioned composition in the preparation of an agent for inhibiting the formation of P-bodies in cells.

[0043] The embodiments of the present invention provide a use of the above-mentioned polypeptide, the above-mentioned nucleic acid, the above-mentioned recombinant vector, and the above-mentioned composition in the preparation of a drug for treating cardiovascular and cerebrovascular diseases.

[0044] The present invention will be further described below by means of specific examples.

[0045] Example 1

[0046] 1. Experimental plan

[0047] 1. Design of peptides that inhibit P-body formation

[0048] Protein sequences of key P-body binding proteins collected at NCBI, with information from GeneBank, include:

[0049] (1)DDX6 (Mus musculus): P54823;

[0050] (2)4E-T(Mus musculus): Q9EST3;

[0051] (3)LSM14a (Mus musculus): Q8K2F8;

[0052] (4)PATL1 (Mus musculus): Q3TC46.

[0053] Based on the sites of their interaction, five inhibitory peptides were designed in this example, as follows:

[0054] (1) Inhibitory peptide-1 that occupies the DFDF and TGF structural pockets in the RECA2 region of DDX6 (sequence from positions 219-238 of mouse 4E-T): EWFSAGPTSQSETIELTGFD;

[0055] (2) Inhibitory peptide-2 occupying the TGF structural pocket (sequence from position 27-40 of mouse PATL-1): EEIDQFNDDTFGSG;

[0056] (3) Inhibitory peptide-3 occupying the DFDF structural pocket (sequence from positions 290-304 of mouse LSM14-A): KFEKDFDFESANAQFNK;

[0057] (4) Inhibitory peptide-4 occupying the structural pocket of DDX6 (sequence from positions 317-334 of mouse DDX6): TERQKVHCLNTLFSRLQI;

[0058] (5) Nonspecific ineffective control peptide-5: DKENQDFEFEAKEFRKEHINFSAD.

[0059] On this basis, a cell-penetrating peptide sequence (YGRKKRRQRRR) was added to the N-terminus of the initial polypeptide to construct a polypeptide for inhibiting the formation of P-bodies in cells. The sequences of the five polypeptides are as follows:

[0060] (1) 4E-T based polypeptide-1: YGRKKRRQRRREWFSAGPTSQSETIELTGFD (SEQ ID NO. 1);

[0061] (2) PATL-1-based polypeptide-2: YGRKKRRQRRREEIDQFNDDTFGSG (SEQ ID NO. 2);

[0062] (3) LSM14-A-based polypeptide-3: YGRKKRRQRRRKFEKDFDFESANAQFNK (SEQ ID NO. 3);

[0063] (4) DDX6-based polypeptide-4: YGRKKRRQRRRTERQKVHCLNTLFSRLQI (SEQ ID NO. 4);

[0064] (5) Ineffective control polypeptide-5: YGRKKRRQRRRDKENQDFEFEAKEFRKEHINFSAD (SEQ ID NO. 5).

[0065] Through NCBI protein sequence comparison, we ensured that these designed peptides had no homologous sequences in the mouse proteome. At the same time, AlphaFold 3 was used to simulate these peptides and the corresponding P-body domains to determine the spatial position of the peptides on the relevant domains.

[0066] 2. Dissolution and preservation of peptides

[0067] Custom-synthesized peptides 1 to 5 and peptide 1-Cy5 with fluorescent labels. Peptide 1 and peptide 1-Cy5 were dissolved in DMSO, prepared into storage solution, and then aliquoted and frozen in a -80℃ refrigerator. Peptide 2 to 5 were dissolved in ultrapure water, prepared into storage solution, filtered, and aliquoted and frozen in a -80℃ refrigerator.

[0068] 3. Construction of plasmid and viral vector

[0069] The DNA sequence of Ddx6 (NM_001357703.1) was cloned into the pLentiV-EGFP vector to construct the Ddx6-egfp recombinant plasmid (using BamHI / AfeI restriction sites). The recombinant plasmid was co-transfected into HEK293T cells with pMD2.G and psPAX2 using Lipo-3000 reagent. Complete medium was replaced 4 hours after transfection and cultured for 48 hours. Subsequently, the cell supernatant was collected, filtered through a 0.22 μm filter, and centrifuged at 3000 rpm for 10 minutes to collect the virus particles. The collected viruses were stored at -80 °C.

[0070] 4. Cell line construction

[0071] The collected virus was added to 10% (v / v) FBS-DMEM complete medium containing 1% (v / v) polybrene and incubated with bEnd.3 cells. After 48 hours, the cells were observed under a microscope (Carl Zeiss, Axio Observer7, Germany). After confirming the presence of EGFP signal in the cells under 488nm excitation, the cells were digested with trypsin and made into a single cell suspension. Monoclonal cell sorting was performed using a 100μm nozzle using a BD FACSAria SORP flow cytometer, and the sorted cells were inoculated in a 96-well plate. After 14 days, the successfully surviving monoclonal cells were selected for further culture.

[0072] 5. Cytotoxicity studies

[0073] bEnd.3 vascular endothelial cell line was cultured at 1×10 4 / mL of cell density was planted in a 96-well plate and cultured in DMEM complete medium containing 10% (v / v) FBS. When the cells grew to 70% to 80% density, they were replaced with a medium containing the polypeptide designed in this embodiment, and the concentrations of the polypeptides were 0.003μM, 0.03μM, 0.3μM, 3μM, 15μM and 30μM, respectively, and the control group contained an equal amount of solvent. After incubation for 24 hours, 10μL CCK-8 solution was added to each well, and after incubation for 1 hour, the absorbance value was detected using an enzyme marker to calculate the cell viability.

[0074] 6. Study on the efficacy of blocking P-body formation

[0075] The bEnd.3 vascular endothelial cell line expressing DDX-6-EGFP constructed in step 4 was cultured in DMEM complete medium containing 10% (v / v) FBS at a rate of 1×10 4 / mL of cell density was inoculated in a glass-bottomed culture dish. When the cells grew to a density of 70% to 80%, a microscope was used to take pictures, and then the culture medium containing the polypeptide designed in this embodiment was replaced, and the concentrations of the polypeptide were 0.3μM, 3μM and 30μM respectively; the control group added an equal amount of polypeptide-5. The cells were incubated in the new culture medium for 5 hours, and then photographed again after 24 hours and the oxygen glucose deprivation (OGD) experiment was started. After removing the culture medium, the cells were washed twice with DPBS, and then an EBSS solution without glucose treated with 95% (v / v) nitrogen and 5% (v / v) carbon dioxide was added. The cells were placed in a hypoxic chamber, and a mixed gas of 95% (v / v) nitrogen and 5% (v / v) carbon dioxide was introduced for 15 minutes to reduce the oxygen concentration in the chamber to below 0.5% (v / v). After stopping the gas input, the chamber containing the cells was placed in an incubator at 37°C for 4 hours. After the treatment, the chamber was opened and the cells were fixed with 4% (w / v) paraformaldehyde, and then photographed under a microscope. After oxygen-glucose deprivation, another group of cells were replaced with complete medium containing the polypeptide designed in this example, and incubated for another 4 hours, and then fixed with 4% (w / v) paraformaldehyde and photographed. Finally, ImageJ software was used to count and statistically analyze P-bodies.

[0076] 7. Study on the protective effect of polypeptide-4 on primary mouse brain microvascular endothelial cells (BMEC)

[0077] Extraction of primary vascular endothelial cells: 10 adult mice (3-4 months old) were taken, anesthetized and killed, brain tissue was removed, meninges were removed, cortical tissue was separated and chopped. Type II collagenase (1 mg / mL) was added and digested at 37°C for 40 minutes by shaking. Then centrifuged at 1000g for 5 minutes at 4°C, the supernatant was removed, 25 times of 20% (w / v) BSA solution was added, mixed and centrifuged again at 1000g for 20 minutes at 4°C, and the supernatant was removed. Then, type II collagenase-dispase (1 mg / mL) was added, digested at 37°C for 40 minutes by shaking, centrifuged again at 1000g for 5 minutes at 4°C, and the supernatant was removed. After adding a small amount of DMEM and mixing, it was placed in a 30% (w / v) Percoll solution and centrifuged at 1000g for 10 minutes at 4°C. Take the red precipitate near the bottom and the yellow-white part above it, add DMEM to dilute, and then centrifuge at 1000g for 10 minutes at 4°C. The obtained brain microvessel segment was inoculated into a fibronectin-coated culture plate. During the 48-hour culture, puromycin (4 μg / mL) was added to the culture medium to remove non-endothelial cells, and then replaced with normal culture medium (containing high-glucose DMEM, 20% (v / v) FBS, 100 μg / mL heparin sodium, 2 ng / mL bFGF, 2 mM glutamine and double antibody (Pen-Strep)).

[0078] Cell culture and treatment: After 14 days of culture of primary vascular endothelial cells, the culture medium containing 30 μM polypeptide-4 in this embodiment was replaced and incubated for 24 hours. After that, the culture medium was removed, washed twice with DPBS, and a glucose-free DMEM solution treated with 95% (v / v) nitrogen and 5% (v / v) carbon dioxide was added. The cells were placed in a hypoxic chamber and a mixed gas of 95% (v / v) nitrogen and 5% (v / v) carbon dioxide was introduced for 15 minutes to make the oxygen concentration in the chamber lower than 0.5% (v / v). After stopping the gas input, the cell-containing chamber was placed in a 37°C incubator for 4 hours. After the treatment was completed, it was replaced with complete culture medium and continued to incubate for 24 hours. 10 μL CCK-8 solution was added to each well, and after incubation for 1 hour, the absorbance value was detected using an enzyme reader to calculate the cell viability.

[0079] 8. Effect of polypeptide-4 on the mRNA expression of inflammatory factors in bEnd.3 cells induced by OGD / R

[0080] The dose of polypeptide-4 was 30 μM, and the control group added an equal amount of solvent. After incubation with bEnd.3 cells for 24 hours, OGD treatment was performed for 6 hours. During OGD treatment, 30 μM polypeptide-4 or solvent was added to the sugar-free DMEM medium used. After OGD treatment, the medium was restored to DMEM medium containing glucose and oxygen and 30 μM polypeptide-4 or solvent was added, and incubated for 2 hours or 4 hours. Cells were collected at three time points, OGD-6h, OGD-6h / R-2h, and OGD-6h / R-4h, and total cell RNA was extracted using a PCR extraction kit. The mRNA levels of TNF-α, G-CSF, GM-CSF, CXCL1, CXCL2, ICAM1, and VCAM1 in the cells were determined using a Real-Time PCR detection kit and instrument.

[0081] 2. Experimental Results

[0082] 1. Peptide structure

[0083] Alpha Fold 3 was used to analyze the binding of peptide-1, peptide-2 and peptide-3 to DDX6 protein, and the binding of peptide-4 to 4E-T protein. The results are as follows: Figures 1 to 4 shown. Figures 1 to 4 In the figure, orange represents DDX6, silvery white represents 4E-T, bright purple represents the binding site identified in the literature, and green / cyan / blue / orange represent peptides-1 to 4, respectively. Figures 1 to 4 It can be seen that polypeptides-1 to 4 can bind to their related proteins.

[0084] 2. The peptide has no cytotoxicity to beend.3 cells

[0085] Cytotoxicity study results Figure 5 As shown. Figure 5 It can be seen that the five synthesized polypeptides had no significant effect on the survival rate of beend.3 cells in the concentration range from 0.003 to 30 μM. This result shows that these five polypeptides have good safety.

[0086] 3. Determination of the time window for fluorescently labeled peptides to enter beend.3 cells

[0087] In this example, polypeptide-1 was labeled with a Cy5 fluorescent label and co-incubated with beEend.3 cells at different time points. Fluorescence images of beEend.3 cells were obtained using a fluorescence microscope under the same exposure parameters. The results are as follows: Figure 6 As shown. Figure 6 It can be seen that the fluorescence intensity reaches the maximum value after 12 hours of incubation.

[0088] 4. Peptide-4 significantly inhibits P-body formation

[0089] The experimental results of the five peptides at different doses and during oxygen glucose deprivation and recovery (OGD / R) are as follows: Figures 7 to 11 As shown. Figure 7 It can be seen that peptide-1 cannot prevent the formation of P-bodies under non-OGD / R conditions, but it can significantly inhibit the formation of P-bodies in cells after OGD at concentrations of 3μM and 30μM. Fig.10 It can be seen that peptide-4 also failed to prevent the formation of P-bodies under non-OGD / R conditions, but at a concentration of 30 μM, it significantly inhibited the formation of P-bodies in cells during OGD and recovery. Figure 8 , 9 As shown in Figures 1 and 11, peptide-2, peptide-3 and peptide-5 could not prevent the formation of P-bodies in either non-OGD / R or OGD / R conditions. Therefore, peptide-4 significantly inhibited the formation of P-bodies during the OGD / R process, showing a good blocking effect.

[0090] 5. Peptide-4 alleviates BMEC damage caused by OGD / R

[0091] The results of the study on the protective effect of peptide-4 on BMEC are as follows Fig.12 As shown. Fig.12 It can be seen that BMEC cells are damaged during OGD / R, and cell viability decreases by 30%. Peptide-4 can significantly reduce BMEC cell damage, and cell viability doubles, only decreasing by 15%. Therefore, peptide-4 can reduce BMEC damage caused by OGD / R.

[0092] 6. Peptide-4 reduces the level of inflammatory factors expressed by beend.3 cells induced by OGD / R

[0093] The results of the effect of polypeptide-4 on the mRNA expression of inflammatory factors in bEnd.3 cells induced by OGD / R are as follows Fig.13 As shown. Fig.13 It can be seen that during the OGD / R process, the mRNA levels of all inflammatory factors increased significantly. The use of 30μM peptide-4 significantly inhibited the mRNA levels of multiple inflammatory factors expressed by bEnd.3 cells, including: G-CSF, VCAM1 and ICAM1. Therefore, peptide-4 has a significant inhibitory effect on the expression of inflammatory factors in vascular endothelial cells caused by OGD / R.

[0094] It should be understood that the application of the present invention is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. A polypeptide for inhibiting the formation of P-bodies in cells, characterized in that The amino acid sequence of the polypeptide is shown in SEQ ID NO.

4.

2. A nucleic acid, characterized in that The nucleic acid encodes the polypeptide of claim 1.

3. A recombinant vector, characterized in that: The recombinant vector expresses the polypeptide according to claim 1.

4. A composition, characterized in that The composition comprises the polypeptide of claim 1.

5. The composition according to claim 4, characterized in that The composition also includes a pharmaceutically acceptable solvent.

6. The composition according to claim 5, characterized in that The solvent is selected from one or more of water, physiological saline, glucose aqueous solution, and dimethyl sulfoxide.

7. The composition according to claim 4, characterized in that The composition also includes a pharmaceutically acceptable carrier.

8. The composition according to claim 7, characterized in that The pharmaceutically acceptable carrier is selected from one or more of an excipient, a filler, a binder, a lubricant, a disintegrant, and a stabilizer.

9. Use of the polypeptide according to claim 1, the nucleic acid according to claim 2, the recombinant vector according to claim 3, or the combination according to any one of claims 4 to 8 in the preparation of an agent for inhibiting the formation of P-bodies in cells.

10. Use of the polypeptide according to claim 1, the nucleic acid according to claim 2, the recombinant vector according to claim 3, or the combination according to any one of claims 4 to 8 in the preparation of a drug for treating cardiovascular and cerebrovascular diseases.

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