Application of platelet factor 4 in preparation of medicine for treating and / or preventing senile deafness

Abstract

The invention relates to the field of biological medicines, in particular to application of a platelet factor 4 in preparation of a medicine for treating and / or preventing senile deafness. Experimental results show that PF4 not only does not aggravate hearing loss of a subject and structural damage of cochlea tissues, but also can reduce an auditory induction threshold of the subject, so that the hearing loss is obviously improved, and the effect of improving senile deafness is achieved. The PF4 can be used as a medicine for treating and / or preventing the senile deafness, and the occurrence and development of the senile deafness can be effectively regulated.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the use of platelet factor 4 in the preparation of medicaments for the treatment and / or prevention of age-related hearing loss. Background Technology

[0002] Hearing loss is a common clinical auditory dysfunction that causes varying degrees of hearing impairment. Specifically, it refers to lesions in the auditory system, including the sound-conducting, sound-sensing, and auditory pathways, particularly the auditory nerve and its various centers, leading to varying degrees of decreased auditory sensitivity, elevated hearing thresholds, auditory dysfunction, and even complete hearing loss. Depending on the degree of hearing loss, mild cases are termed hearing impairment, hearing loss, or hearing decline, while severe cases are termed deafness or total deafness. Common causes of hearing loss in humans include exposure to noise or ototoxic drugs, congenital genetic factors, aging, and trauma. However, due to the complexity of the inner ear's anatomy and the uncertainty of its pathological mechanisms, there are currently few drugs available for the prevention and treatment of hearing loss, and their efficacy is often unsatisfactory. To date, there are no effective and highly targeted preventative and therapeutic measures.

[0003] Age-related hearing loss, also known as presbycusis, is a progressive, irreversible, bilateral, symmetrical sensorineural hearing loss. It is the second most common disease among the elderly worldwide and the third most prevalent disease globally. The pathogenesis of presbycusis is complex, and there is currently no specific treatment; the primary approach is to use hearing aids to compensate for the hearing loss. Additionally, there are reports that improving blood circulation in the inner ear can restore impaired function in patients with presbycusis. However, due to the complex pathological mechanisms of presbycusis, there are few effective interventions specifically targeting it in current technologies. Summary of the Invention

[0004] In a first aspect, the present invention provides the use of platelet factor 4 in the preparation of a medicament for treating and / or preventing age-related hearing loss in a subject.

[0005] In some embodiments, the subject has or is at risk of developing age-related hearing loss.

[0006] As used herein, “subject” means a subject who needs treatment and / or prevention of a disease (such as age-related hearing loss), and more specifically, means mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle, but the invention is not limited thereto.

[0007] In some embodiments, the platelet factor 4 is human platelet factor 4 or its functional fragments, derivatives or analogs.

[0008] As used herein, a “functional fragment” is a substance that substantially retains the biological activity of platelet factor 4. In some embodiments, a “functional fragment” substantially retains the biological activity of unmodified platelet factor 4, for example, retaining at least about 20%, 30%, 40%, 50%, 60%, 75%, 85%, 90%, 95%, 97%, 99% or more of the biological activity of platelet factor 4 (e.g., human platelet factor 4) (and may even have a higher level of activity than platelet factor 4).

[0009] As used herein, a “derivative” is a variant of platelet factor 4 obtained through chemical or biological modification. In some embodiments, a “derivative” substantially retains the biological activity of unmodified platelet factor 4 (e.g., human platelet factor 4).

[0010] As used herein, an “analyte” is a substance that has a similar structure or biological activity to platelet factor 4. In some embodiments, an analogue may be synthesized by protein engineering or rational design and is generally structurally similar to platelet factor 4 (e.g., human platelet factor 4) and substantially retains the biological activity of platelet factor 4.

[0011] In some embodiments, the drug uses platelet factor 4 as the main active ingredient.

[0012] In some embodiments, the drug uses platelet factor 4 as the sole active ingredient.

[0013] In some embodiments, the medicament further includes a pharmaceutically acceptable carrier.

[0014] In some embodiments, the carrier includes liposomes, nanoparticles, or hydrogels.

[0015] In some embodiments, the drug is an injectable formulation, such as administered via intraauricular injection or intravenous injection.

[0016] In some embodiments, the concentration of platelet factor 4 in the drug comprises 0.001-0.1 mg / mL.

[0017] In some embodiments, the concentration of platelet factor 4 in the drug is 0.005 mg / mL.

[0018] In some embodiments, the drug is prepared in dose units and formulated as a single-use, independent dose containing an effective amount of platelet factor 4.

[0019] In some embodiments, the effective amount can be calculated based on ≥15 μg platelet factor 4 / kg body weight, for example, 20 μg platelet factor 4 / kg body weight.

[0020] In some embodiments, the expression levels of CCL5 and / or IL-6 in the subjects are elevated.

[0021] In some embodiments, the expression level of CCL5 and / or IL-6 refers to the absolute (quantitative) or relative (qualitative) value or concentration of CCL5 and / or IL-6 in the sample. In some embodiments, suitable methods may be used to analyze the collected samples to detect the level of CCL5 and / or IL-6 in the samples. After determining the level of CCL5 and / or IL-6 in the samples, the level of CCL5 and / or IL-6 can be compared with reference values. In some embodiments, the reference values ​​may be derived from a normal population. If the subject's CCL5 and / or IL-6 level is elevated compared to the reference value (i.e., not lower than, for example, the same as the reference value, substantially the same as the reference value, not statistically different from the reference value, or within the established range of the reference value), then the subject is assessed as having elevated CCL5 and / or IL-6 levels.

[0022] In some embodiments, the sample may contain any biological material suitable for detecting the desired indicator and may contain cellular and / or non-cellular material. In some embodiments, the sample may be isolated from any suitable biological tissue or fluid. In some embodiments, the sample may be biological material isolated from a subject. In some embodiments, the sample may be whole blood, serum, or plasma, i.e., the subject's whole blood, serum, or plasma has elevated levels of CCL5 and / or IL-6.

[0023] In some embodiments, the drug is used to lower the hearing threshold of the subject.

[0024] In some embodiments, the drug is used to reduce the hearing threshold of the subject by 5-10 dB HL.

[0025] In some embodiments, the drug is used to promote M2 polarization of the inner ear macrophages of the subject.

[0026] In some embodiments, the drug is used to improve the survival rate of the cochlear hair cells of the subject.

[0027] Secondly, the present invention provides the use of platelet factor 4 in the preparation of CC chemokine ligand 5 (CCL5) inhibitors.

[0028] In some embodiments, the CCL5 inhibitor is used to inhibit CC motif chemokine receptor 5 (CCR5) in inner ear macrophages.

[0029] In some embodiments, the CCL5 inhibitor is used to treat and / or prevent age-related hearing loss in the subject.

[0030] In some embodiments, the subject has or is at risk of developing age-related hearing loss.

[0031] In some embodiments, the platelet factor 4 is human platelet factor 4 or its functional fragments, derivatives or analogs.

[0032] In some embodiments, the platelet factor 4 is configured to be 0.001-0.1 mg / mL.

[0033] In some embodiments, the platelet factor 4 is configured at 0.005 mg / mL.

[0034] In some embodiments, the expression levels of CCL5 and / or IL-6 in the subjects are elevated.

[0035] In some embodiments, the CCL5 inhibitor is used to lower the hearing threshold of the subject.

[0036] In some embodiments, the CCL5 inhibitor is used to reduce the hearing threshold of the subject by 5-10 dBHL.

[0037] In some embodiments, the CCL5 inhibitor is used to promote M2 polarization of the subject's inner ear macrophages.

[0038] In some embodiments, the CCL5 inhibitor is used to improve the survival rate of the cochlear hair cells of the subject.

[0039] Thirdly, the present invention provides the use of detection reagents for CCL5 (CC class chemokine ligand 5) and / or IL-6 (interleukin-6) in the preparation of kits for evaluating the therapeutic and / or preventive effects of platelet factor 4 on age-related hearing loss.

[0040] In some embodiments, when the subject's CCL5 and / or IL-6 levels are elevated, platelet factor 4 is assessed as having a therapeutic and / or preventive effect on the subject's age-related hearing loss.

[0041] In some embodiments, the elevation of CCL5 and / or IL-6 levels refers to an elevation of CCL5 and / or IL-6 levels in the subject's whole blood, serum, or plasma.

[0042] In some embodiments, the sample is a whole blood, serum, or plasma sample.

[0043] Compared with the prior art, the beneficial effects of the present invention include at least the following aspects: Platelet factor 4 (PF4), also known as CXCL4, is a cytokine released by platelets. It consists of 70 amino acids and has a molecular weight of 7.8 kDa. PF4 is mainly found in platelet particles and is released during platelet activation and thrombus formation. PF4 has been reported to have biological functions such as inhibiting vascular endothelial cell growth and promoting platelet aggregation.

[0044] Studies show that vascular diseases can directly lead to spasms and embolisms in the blood vessels of the inner ear, eventually forming thrombi and causing microcirculatory disturbances in the ear. It was previously believed that platelet factors are released when blood vessels are damaged, participating in the coagulation process and resulting in poor blood flow, which may further lead to circulatory disorders. A review article, "Advances in the Pathogenesis and Intervention Research of Age-Related Hearing Loss," reported that vascular lesions in the inner ear may be related to age-related hearing loss, and that hemodynamic changes caused by vascular damage in the inner ear are one of the causes of age-related hearing loss. The article also suggested that vasodilators can intervene in age-related hearing loss by improving microcirculation in the inner ear (e.g., reducing platelet factor levels in subjects). Currently, there are no reports of treating age-related hearing loss using platelet factors (e.g., PF4).

[0045] Hearing depends on specialized cells called hair cells located in the basilar membrane of the cochlea. These cells are named for the hair-like fiber bundles found on their surface. When sound waves enter the ear, these fibers move, activating the hair cells. The hair cells then send signals to nearby neurons through synaptic contact points. Hair cells must transmit information rapidly and continuously for humans to recognize complex sound streams and understand language. However, as a highly specialized sensory organ with a significant blood supply barrier, the cochlea faces considerable challenges in drug delivery; conventional drugs often fail to reach the cochlear tissue and achieve effective local concentrations.

[0046] Surprisingly, the experimental results of this invention show that PF4 not only did not worsen the hearing loss and structural damage to the cochlear tissue in the subjects, but it also reduced the auditory evoked threshold (ABR threshold), resulting in a significant improvement in hearing loss, for example, reducing the pure-tone hearing threshold by an average of 10-20 dB HL. In other words, PF4 can be used as a drug for the treatment and / or prevention of age-related hearing loss, effectively regulating the occurrence and development of age-related hearing loss. This invention also found that after PF4 treatment, the survival rate of cochlear hair cells in subjects with age-related hearing loss was significantly improved, especially the outer hair cells related to high-frequency hearing, whose loss rate was reduced by 30%-50%.

[0047] The experimental results of this invention also show that PF4 can reduce the expression level of CCL5 to inhibit the specific expression of CCR5 in inner ear macrophages, leading to M2 polarization of inner ear macrophages, and ultimately improving age-related hearing loss.

[0048] Furthermore, this invention also found that CCL5 and / or IL-6 levels differ between young and elderly subjects, and that PF4 intervention can downregulate CCL5 and / or IL-6 expression levels in subjects with elevated CCL5 and / or IL-6 expression levels, thereby improving age-related hearing loss. Therefore, the potential therapeutic and / or preventative effects of PF4 on age-related hearing loss can be assessed by examining whether and to what extent CCL5 and / or IL-6 levels are elevated before PF4 intervention, thus enabling precise intervention and individualized treatment. Additionally, the therapeutic and / or preventative effects of PF4 on age-related hearing loss can be further evaluated by examining whether and to what extent CCL5 and / or IL-6 levels are reduced after PF4 intervention.

[0049] In summary, this invention provides a new approach to the treatment of hearing loss and is expected to be applied to patients with different types of hearing loss. Attached Figure Description

[0050] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0051] Figure 1 The figure shows the results of PF4 alleviating hearing loss in age-related deafness in mice; Figure 2 The results of CCL5 expression levels in each experimental group are shown in the figure. Figure 3 This is a graph showing the results of single-cell sequencing. Figure 4 The graph shows the results of the expression levels of inflammatory factors in each experimental group. Detailed Implementation

[0052] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0053] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0054] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0055] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0056] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, and even more typically + / -0.5% of the value.

[0057] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within that range. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0058] Example 1: Preparation of PF4-based pharmaceutical compositions Active ingredient: Recombinant human PF4 (rhPF4) or its functional fragments, derivatives or analogs are used as the main active ingredient.

[0059] Drug carriers: Proteins or pharmaceutically acceptable carriers with good biocompatibility can be used to deliver PF4 via local administration (e.g., intraauricular injection) or systemic administration (e.g., intravenous injection), such as liposomes, nanoparticles, or hydrogels, to improve the stability and targeting of PF4.

[0060] The concentration of PF4 (dissolved in solvents such as physiological saline or buffer) is 0.001-0.1 mg / mL, and the dosage is 20 μg / kg. The pharmaceutical composition may also include antioxidants, such as vitamin E.

[0061] Example 2 PF4 can delay age-related hearing loss in mice and improve cochlear-related pathological changes. 100 μl of PF4 (5 μg / ml in this example) was injected intraperitoneally into middle-aged mice (12-week-old male C57BL / 6J mice). The injections were repeated 8 times over 24 days for 4 consecutive months. The control group received an equal volume of saline intraperitoneally according to the same injection protocol. The effects of PF4 on auditory function (e.g., auditory brainstem response (ABR)) and inner ear morphology were evaluated.

[0062] Auditory function testing: Mice were anesthetized with xylazine hydrochloride (10 mg / kg) and salbutamol (55 mg / kg), fixed in a soundproof box, and subjected to ABR and DPOAE (distortion product otoacoustic emission) tests.

[0063] Inner ear tissue preparation, immunofluorescence staining, and confocal scanning: The inner ears of mice in the experimental and control groups, after fixation and decalcification, were dissected under a stereomicroscope to expose the stria vascularis and basement membrane. The specimens were fixed and sealed at room temperature, incubated with primary antibody at 4°C overnight, and then incubated with secondary antibody at room temperature. Finally, the specimens were mounted and scanned using a laser confocal microscope.

[0064] Basement membrane galactosidase staining: The mouse inner ear basement membrane was dissected, fixed at room temperature, stained with SA-β-gal, incubated overnight at 37 ℃, mounted with neutral resin, and observed under a regular optical microscope.

[0065] The results showed that with increasing age, the hearing threshold of middle-aged mice increased, leading to hearing loss (i.e., symptoms of age-related hearing loss). Intraperitoneal injection of PF4 reduced the ABR threshold in the experimental group (PF4 group) at seven stimulation frequencies (4kHz, 5.6kHz, 8kHz, 11.3kHz, 16kHz, 22.6kHz, and 32kHz), significantly improving hearing loss and reducing the pure-tone hearing threshold of subjects by an average of 5-10 dB HL (e.g., ...). Figure 1 (As shown). In addition, intraperitoneal injection of PF4 can reduce aging indicators of the inner ear basement membrane and improve the morphology of inner ear tissues.

[0066] Example 3 Spleen sample preparation: The spleen was dissected from the mouse abdominal cavity, cut into small pieces, ground thoroughly, and the cell suspension was collected. The filtered cell suspension was added to red blood cell lysis buffer at a volume ratio of 1:5 and mixed well. After standing for 5-10 minutes until the liquid became clear, it was centrifuged. The resulting precipitate was resuspended, washed, centrifuged again, and the supernatant was discarded. The precipitate was then resuspended in 100 µL PBS for staining.

[0067] Flow cytometry sorting of mouse spleen T cells: First, the prepared spleen cell suspension was surface stained: Surface fluorescent antibodies (CD45, CD3, and APC) were added according to the recommended dosage and incubated in the dark. After incubation, 1 mL of PBS was added to terminate the reaction, and the supernatant was discarded by centrifugation. This washing process was repeated twice. 1 mL of Foxp3 fixation / permeabilization working solution was added to each sample tube and gently pipetted repeatedly. Fixation was performed at 2-8 °C in the dark for 30-60 min. 2 mL of 1X permeabilization buffer was added to each sample tube, and centrifugation (500 g, 5 min) was performed, followed by discarding the supernatant. The precipitate was resuspended in 300 µL of PBS (gently pipetting repeatedly) and analyzed by flow cytometry. Finally, T cells were extracted from the samples using Flowjo software according to the standard procedure, and the data were integrated. Then, dimensionality reduction analysis was performed using the FltSNE plugin (max iterations = 1000, perplexity = 20 and learning rate = 200, https: / / doi.org / 10.48550 / arXiv.1712.09005) based on the following metrics: CD45 percp, CD3 PE, and APCPE / Cyanine7.

[0068] In vitro culture of mouse T cells: Immunomagnetic beads were mixed with biotinylated antibodies CD3 and CD28 at a ratio of 1:1:1 and incubated at 4 degrees Celsius for 2 hours. Subsequently, the flow-cytometry-sorted mouse spleen T cells were resuspended in 1640 medium containing IL-2 (1:1000), and then added to appropriate culture dishes according to the cell count, ensuring that the total cell count of the control and experimental groups was approximately the same. The incubated magnetic bead antibody solution was then added to the culture dishes at a volume ratio (1:2000), and finally incubated in an incubator.

[0069] Luminex liquid-phase chip technology for detecting inflammatory factor levels: The protein expression level of CCL5 in blood or T cell culture medium was detected using Luminex liquid-phase chip technology.

[0070] Inner ear single-cell sequencing analysis: scRNA-seq samples included 1-month-old, 2-month-old, 5-month-old, 12-month-old, and 15-month-old mice (5 males and 5 females per group). Cochleas from mice of the same age and sex were pooled for scRNA-seq. Single cells were captured using droplet-based microfluidics, and single-cell transcriptomic libraries were constructed using a Chromium 10× single-cell instrument (10×Genomics), a 10×Genomics Chromium single-cell 3' GEM library, and a Gel Bead Kit v3. The datasets were aligned and quantified using the Cell Ranger single-cell software suite (10×Genomics) with default parameters. Quantification of sample-specific FASTQ files was assessed using the cellranger counting function, which aligned with the mouse reference genome (mm10) and generated a gene expression matrix. The filtered gene expression matrix was used for downstream analysis. Downstream analysis of the scRNA-seq data was performed using the single-cell toolkit Seurat (version 3.2.0) in R (version 3.6.3).

[0071] Splenic T cells from young (8-week-old) and senescent (52-week-old) mice were cultured in vitro in a culture medium (specifically, ImmunoCult™-XF T Cell Expansion Medium was used in this example, with a T cell concentration of 1×10⁻⁶). 6 PF4 (1 µg / ml in this example) was added to the T cells (cells / ml). The experimental group in this example was divided into the following 4 groups: Old group (senescent group), Young group (young group), Old-PF4 group (senescent-PF4 group) and Young-PF4 group (young-PF4 group), and the protein expression level of CCL5 in the culture medium of T cells isolated from the above 4 groups was detected.

[0072] The single-cell sequencing results of this embodiment ( Figure 3 The results showed that CCR5 (CC motif chemokine receptor 5) is specifically expressed in inner ear macrophages.

[0073] The detection results of this embodiment are as follows: Figure 2 As shown, the expression level of CCL5 (CC chemokine ligand 5) in the aging group was significantly higher than that in the young group, indicating that there is a difference in the expression level of CCL5 between young and old mice. However, intraperitoneal injection of PF4 can significantly reduce the expression level of CCL5 in old mice.

[0074] In addition, such as Figure 2As shown, the expression levels of CCL5 in the young group and the young-PF4 group were similar, indicating that intraperitoneal injection of PF4 did not further reduce the expression level of CCL5 in young mice. These results demonstrate that intraperitoneal injection of PF4 can specifically reduce the expression level of CCL5 in aged mice, thereby improving hearing loss in age-related deafness.

[0075] In summary, PF4 can downregulate CCL5 to inhibit CCR5 specifically expressed on inner ear macrophages, leading to M2 polarization of inner ear macrophages and ultimately improving age-related hearing loss.

[0076] Example 4 The experimental groups in this embodiment were divided into the following six groups: Old group (senescent group), Young group (young group), Old-PF4 group (senescent-PF4 group), Young-PF4 group (young-PF4 group), HEI-OC1 group, and HEI-OC1+PF4 group. Among them, the Old group (senescent group), Young group (young group), Old-PF4 group (senescent-PF4 group), and Young-PF4 group (young-PF4 group) used the supernatant of the culture medium corresponding to the experimental group in Example 3 to culture inner ear hair cell-like cells (HEI-OC1). The HEI-OC1 group used ordinary culture medium (ImmunoCult™-XF T Cell Expansion Medium) to culture inner ear hair cell-like cells. The HEI-OC1+PF4 group used ordinary culture medium with the addition of PF4. The concentration of inner ear hair cell-like cells in each experimental group in this embodiment was 1×10⁻⁶. 6 The concentration of PF4 was 1 µg / ml. This example further examined the expression levels of P21, P16, TNF-α, iNos, IL-6, and IL-1 in the culture media of the above six groups.

[0077] This embodiment focuses on whether PF4 can simultaneously reduce the expression levels of various inflammatory factors in the Old-PF4 group (which reflects the comprehensive effect of PF4 on T cells and thus on elderly subjects) and the HEI-OC1+PF4 group (which reflects the effect of PF4 on inner ear hair cell-like cells) to comprehensively evaluate the ameliorative effect of PF4 on age-related hearing loss.

[0078] like Figure 4As shown, although PF4 can comprehensively treat and / or prevent age-related hearing loss, its mechanism of improvement is complex. Only the Old-PF4 group and the HEI-OC1+ PF4 group showed a simultaneous decrease in IL-6 expression levels. While PF4 can reduce P21 expression levels in the Old-PF4 group, it may lead to an increase in P21 expression levels in the HEI-OC1+ PF4 group. Similarly, while PF4 can reduce P16 expression levels in the HEI-OC1+ PF4 group, it may lead to an increase in P21 expression levels in the Old-PF4 group. Regarding IL-1, PF4 may lead to an increase in IL-1 expression levels in both the Old-PF4 group and the HEI-OC1+ PF4 group. Although PF4 can reduce TNF-α expression levels in the HEI-OC1+ PF4 group, there was no significant difference in TNF-α expression levels between the Old and Young groups. Although PF4 can reduce the expression level of iNos in the Old-PF4 group, there is no significant difference in the expression level of iNos between the HEI-OC1 group and the HEI-OC1+PF4 group.

[0079] The above results suggest that PF4 may be more suitable for improving age-related hearing loss in subjects with higher IL-6 expression levels. Furthermore, the effectiveness of PF4 in improving age-related hearing loss in subjects can be assessed by measuring (and changes in) their IL-6 expression levels.

[0080] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. Use of platelet factor 4 in the preparation of a medicament for the treatment and / or prevention of age-related hearing loss in a subject.

2. The use as described in claim 1, characterized in that, The platelet factor 4 is human platelet factor 4 or its functional fragments, derivatives or analogs.

3. The use as described in claim 1, characterized in that, The drug uses platelet factor 4 as its main active ingredient.

4. The use as described in claim 1, characterized in that, The concentration of platelet factor 4 in the drug includes 0.001-0.1 mg / mL.

5. The use as described in claim 1, characterized in that, The subjects showed elevated levels of CCL5 and / or IL-6 expression.

6. The use as described in claim 1, characterized in that, The drug is used to lower the hearing threshold of the subject.

7. The use as described in claim 1, characterized in that, The drug is used to promote M2 polarization of the inner ear macrophages of the subject.

8. The use of platelet factor 4 in the preparation of CC-type chemokine ligand 5 inhibitors, characterized in that, The CC chemokine ligand 5 inhibitor is used to inhibit CC motif chemokine receptor 5 in inner ear macrophages.

9. Use of CCL5 and / or IL-6 detection reagents in the preparation of kits for evaluating the therapeutic and / or preventive effects of platelet factor 4 on age-related hearing loss.

10. The use as described in claim 9, characterized in that, When the subjects' CCL5 and / or IL-6 levels were elevated, the platelet factor 4 was assessed as having a therapeutic and / or preventive effect on the subjects' age-related hearing loss.

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