Application of exosome derived from olfactory mucosa mesenchymal stem cells pretreated based on active peptide in improvement of neuroinflammation

Abstract

The invention relates to application of an exosome derived from olfactory mucosa mesenchymal stem cells pretreated based on active peptide in improvement of neuroinflammation, in particular to a treatment strategy for neuroinflammation of Alzheimer's disease (AD). An IFN-FGF fusion protein (SEQ ID NO: 3) and a coding gene (SEQ ID NO: 4) of the IFN-FGF fusion protein are constructed, and the IFN-FGF fusion protein and the coding gene (SEQ ID NO: 4) are used for in-vitro pretreatment of OM-MSCs, so that the A-MS Cs-Exo exosome with anti-inflammatory and neuroprotective functions is obtained. In an A beta oligomer-induced AD neuroinflammation model, the A-MSCs-Exo significantly alleviates neuroinflammation and cognitive impairment by inhibiting excessive activation of hippocampal Iba1 + microglia cells, reducing serum IL-6 level, improving SOD activity and improving Morris water maze behavioral performance, the effect is better than that of traditional FGF-1 pretreatment (B-MSCs-Exo) and untreated exosome (C-MSCs-Exo), and the A-MSCs-Exo can be used for preparing the A beta oligomer-induced AD neuroinflammation model. The invention provides a novel exosome intervention strategy for AD treatment.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and particularly to the application of exosomes derived from olfactory mucosa mesenchymal stem cells (OM-MSCs) pretreated with bioactive peptides in the regulation of neuroinflammation. Background Art

[0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized by neuroinflammation, β-amyloid (Aβ) deposition, and cognitive decline. Abnormal activation of microglia is the core mechanism of AD neuroinflammation, which exacerbates neuronal damage by releasing pro-inflammatory factors such as IL-6. Currently, most clinical drugs are aimed at symptom relief, lacking effective intervention means for the neuroinflammatory pathway.

[0003] As a carrier for intercellular communication, exosomes can regulate the activity of target cells by delivering functional proteins or miRNAs. Exosomes derived from mesenchymal stem cells (MSCs) have attracted much attention due to their anti-inflammatory and neuroprotective potential, but their efficacy is limited by the deficiency of donor cell pretreatment strategies. Pretreatment with FGF-1 can partially enhance the function of exosomes, but there are problems such as a single action target and poor stability.

[0004] The present invention constructs an IFN-FGF fusion protein, combines the immunomodulatory function of interferon γ (IFN-γ) with the nerve repair activity of fibroblast growth factor (FGF), optimizes the pretreatment protocol of OM-MSCs, and significantly improves the anti-inflammatory and antioxidant stress capabilities of exosomes. Experiments show that A-MSCs-Exo has significant advantages over exosomes pretreated with traditional FGF-1 (B-MSCs-Exo) and untreated exosomes (C-MSCs-Exo) in inhibiting microglial activation and improving cognitive impairment in AD model mice, providing an innovative solution for the treatment of AD. Summary of the Invention

[0005] In order to solve the problems of insufficient targeting and single function of existing mesenchymal stem cell exosomes in the regulation of neuroinflammation, the present invention first provides an exosome for treating Alzheimer's disease (AD) neuroinflammation, which is characterized in that: the exosome is derived from olfactory mucosa mesenchymal stem cells (OM-MSCs) pretreated with an IFN-FGF fusion protein;

[0006] In some embodiments, the IFN-FGF fusion protein is composed of an active fragment of human interferon γ (IFN-γ) (SEQ ID NO: 1) and an active fragment of fibroblast growth factor receptor 1 (FGFR1) (SEQ ID NO: 2) connected by a flexible linker peptide (GGGGSGGGGS), and its amino acid sequence is as shown in SEQ ID NO: 3;

[0007] In some embodiments, the exosomes significantly alleviate AD-related neuroinflammation and cognitive impairment by inhibiting microglial overactivation, reducing the level of pro-inflammatory factor IL-6, increasing the activity of antioxidant enzyme SOD, and improving neuronal function.

[0008] The present invention further provides a method for preparing the exosomes, comprising the following steps:

[0009] Cell pretreatment: Seed OM-MSCs in a serum-free medium, add IFN-FGF fusion protein at a final concentration of 150 ng / mL, and pretreat at 37 °C for 48 hours;

[0010] Exosome induction and collection: Replace with exosome-free serum medium and continue culturing for 24 hours, and collect the cell supernatant;

[0011] Exosome purification and verification: Extract exosomes by gradient centrifugation combined with the ExoQuick-TC kit, and verify the positivity of exosome markers CD63 and TSG101 and negativity of Calnexin by Western Blot.

[0012] Finally, the present invention provides the use of the exosomes and their pharmaceutical compositions in the preparation of drugs for treating neuroinflammatory diseases, including:

[0013] Treating neuroinflammation related to Alzheimer's disease (AD);

[0014] Inhibiting abnormal activation of microglia and release of pro-inflammatory factors;

[0015] Improving oxidative stress injury and neuronal synaptic dysfunction;

[0016] The pharmaceutical composition contains a therapeutically effective amount of A-MSCs-Exo and a pharmaceutically acceptable carrier.

[0017] Compared with the prior art, the present invention has at least the following beneficial effects:

[0018] Innovative fusion protein design: For the first time, the immunomodulatory function of IFN-γ and the nerve repair activity of FGFR1 are integrated through a flexible linker peptide, breaking through the limitations of single growth factor pretreatment and significantly enhancing the anti-inflammatory and neuroprotective synergistic effects of exosomes.

[0019] Exosome function optimization: By pretreating OM-MSCs with IFN-FGF fusion protein, the obtained exosomes (A-MSCs-Exo) have better inflammatory regulation ability than traditional FGF-1 pretreatment (B-MSCs-Exo) and untreated exosomes (C-MSCs-Exo), specifically manifested as: Iba1 in the hippocampal region +The inhibitory rate of microglial activation increased by 40%-50%; the serum IL-6 level decreased to below that of the model group (p<0.01); the SOD activity recovered to more than 85% of the normal level (p<0.05); the escape latency of Morris water maze was shortened to about 25% of that of the model group (p<0.01).

[0020] The treatment mechanism is clear: it is revealed for the first time that A-MSCs-Exo improves the pathological process of AD through multi-pathway synergy (inhibiting microglial activation / regulating oxidative stress / protecting neurons), providing a new strategy for exosome intervention in neurodegenerative diseases.

[0021] Clinical application potential: The exosomes have low immunogenicity, high targeting and stability, and can be developed into non-invasive nasal administration or intravenous injection preparations, providing a safe and effective new biological agent for the treatment of AD. Description of the Drawings

[0022] Figure 1 Detection effect of His-IFN-FGF protein by SDS-PAGE.

[0023] Figure 2 Verification of the marker protein of exosomes by Western Blot.

[0024] Figure 3 Effects of exosomes from each group on the inflammatory level and oxidative stress response of AD neuroinflammatory model mice.

[0025] Figure 4 Effects of exosomes from each group on the activation of hippocampal microglia in AD neuroinflammatory model mice. Detailed Description of the Invention

[0026] To make the technical problems, technical solutions and advantages to be solved by the present invention clearer, the following will be described in detail with reference to the drawings and specific embodiments.

[0027] Example 1 Preparation of IFN-FGF Fusion Protein

[0028] On NCBI, according to interferon gamma precursor [Homo sapiens], NCBI Reference Sequence: NP_000610.2, select the active fragment:

[0029] QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNYSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRG (SEQ ID NO:1);

[0030] Select the active fragment on NCBI according to Chain B, FGF RECEPTOR 1, PDB: 1FGK_B:

[0031] QDGPLYVIVEYASKGNLREYLQARRPPGLEYSYNPSHNPEEQLSSKDLVSCAYQVARGMEYLASKKCIHRDLAARNVLVTEDNVMKIADFGLARDIHHIDYYKKTTNGRLPVKWMAPEALFDRIYTHQSDVWSFGVLLWEIFTLGGSPYPGVPVEELFKLLKEGHRMDKPSNCTNELYMMMRDCWHAVPSQRPTFKQLVEDLDRIVALTSNQE (SEQ ID NO:2);

[0032] Construct an IFN-FGF fusion protein, where GGGGSGGGGS is the linker to enhance the protein folding stability. The amino acid sequence of the fusion protein is as follows:

[0033] QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQS

[0034] QIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNY

[0035] SVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRGGGGGSGGGGS

[0036] QDGPLYVIVEYASKGNLREYLQARRPPGLEYSYNPSHNPEEQLSSKDLVSCA

[0037] YQVARGMEYLASKKCIHRDLAARNVLVTEDNVMKIADFGLARDIHHIDYY

[0038] KKTTNGRLPVKWMAPEALFDRIYTHQSDVWSFGVLLWEIFTLGGSPYPGV

[0039] PVEELFKLLKEGHRMDKPSNCTNELYMMMRDCWHAVPSQRPTFKQLVED

[0040] LDRIVALTSNQE (SEQ ID NO:3);

[0041] Design the nucleotide sequence encoding the IFN-FGF fusion protein according to the codon preference of Escherichia coli:

[0042] caggatccgtatgtgaaagaagcggaaaacctgaaaaaatattttaacgcgggccatagcgatgtggcggataacggcaccctgtttctgggcattctgaaaaactggaaagaagaaagcgatcgcaaaattatgcagagccagattgtgagcttttattttaaactgtttaaaaactttaaagatgatcagagcattcagaaaagcgtggaaaccattaaagaagatatgaacgtgaaattttttaacagcaacaaaaaaaaacgcgatgattttgaaaaactgaccaactatagcgtgaccgatctgaacgtgcagcgcaaagcgattcatgaactgattcaggtgatggcggaactgagcccggcggcgaaaaccggcaaacgcaaacgcagccagatgctgtttcgcggcggcggcggcggcagcggcggcggcggcagccaggatggcccgctgtatgtgattgtggaatatgcgagcaaaggcaacctgcgcgaatatctgcaggcgcgccgcccgccgggcctggaatatagctataacccgagccataacccggaagaacagctgagcagcaaagatctggtgagctgcgcgtatcaggtggcgcgcggcatggaatatctggcgagcaaaaaatgcattcatcgcgatctggcggcgcgcaacgtgctggtgaccgaagataacgtgatgaaaattgcggattttggcctggcgcgcgatattcatcatattgattattataaaaaaaccaccaacggccgcctgccggtgaaatggatggcgccggaagcgctgtttgatcgcatttatacccatcagagcgatgtgtggagctttggcgtgctgctgtgggaaatttttaccctgggcggcagcccgtatccgggcgtgccggtggaagaactgtttaaactgctgaaagaaggccatcgcatggataaaccgagcaactgcaccaacgaactgtatatgatgatgcgcgattgctggcatgcggtgccgagccagcgc ccgacctttaaacagctggtggaagatctggatcgcattgtggcgctgaccagcaaccaggaa(SEQ ID NO:4).

[0043] Design primers by referring to the nucleotide sequence of the IFN-FGF fusion protein, and perform PCR amplification of the target gene using upstream and downstream primers; after verifying the PCR amplification product by gel electrophoresis, cut the gel and recover it, digest it at 37°C for 3 h, and then insert it into the pET28a empty vector digested under the same conditions. After resistance screening, perform sequencing verification, and name the successfully constructed vector pET28a-IFN-FGF. Transform the recombinant plasmid into BL21(DE3) competent cells, plate it and incubate it upside down at 37°C for 12 h. Pick monoclonal colonies from the plate into LB liquid medium (K+), and when the OD600 value of the bacterial solution reaches 0.6, add IPTG with a final concentration of 1 mol / L, and induce it at 37°C for 6 h, then collect the bacteria. Ultrasonically disrupt the collected bacteria, fully dissolve the collected bacterial precipitate with 8 mol / L urea, filter it through a filter membrane, and then use a His-tag protein purification kit (Beyotime, China) to purify the target protein by affinity chromatography column. Collect the elution fraction with higher purity, place it at 4°C overnight, and then renature it in 2 mol·L-1 urea buffer solution. Centrifuge at 4°C and 10,000 g for 15 min, and the collected supernatant is the purified His-IFN-FGF protein. Detect the purification effect by SDS-PAGE, as shown in Figure 1 。

[0044] Figure 1 The results showed that the molecular weight of His-IFN-FGF was 41.37 kDa, which was in line with expectations.

[0045] Preparation of olfactory mucosa mesenchymal stem cell-derived exosomes in each group after pretreatment in Example 2

[0046] Isolation and culture of olfactory mucosa mesenchymal stem cells (OM-MSCs): Anesthetize and decapitate wild-type C57BL / 6 mice (4 weeks old). In a laminar flow hood, cut open the nasal cavity of the mice and remove the olfactory mucosa, transfer it to PBS solution containing 3% penicillin-streptomycin, soak it for 5 min, and then pick out the mucosa with forceps and cut it into pieces. The cut olfactory mucosa was resuspended in DMEM / F-12 complete medium (containing 10% mesenchymal stem cell-specific serum), transferred to a culture flask with a dropper, and cultured in a cell culture incubator for one week. Use DMEM / F12 medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin, culture it at 37°C and 5% CO2, change the medium every 3 days, and when passaged to the 3rd generation, detect the CD90+ / CD44+ / CD34- / CD45- phenotype by flow cytometry to confirm that the purity of OM-MSCs > 95%.

[0047] Group A: The 4th generation of OM-MSCs was used at 1×10 6Inoculate at a density of 1×10 cells / mL into serum-free medium supplemented with recombinant His-IFN-FGF protein (final concentration 150 ng / mL), and pre-treat at 37°C for 48 hours.

[0048] Group B: Seed passage 4 OM-MSCs at a density of 1×10 6 cells / mL into serum-free medium supplemented with FGF-1 (R&D Systems (Bio-Techne), catalog number: 232-FA) (final concentration 150 ng / mL), and pre-treat at 37°C for 48 hours.

[0049] Group C: Seed passage 4 OM-MSCs at a density of 1×10 6 cells / mL into serum-free medium supplemented with an equal volume of PBS, and pre-treat at 37°C for 48 hours.

[0050] After pre-treating the OM-MSCs in groups A-C, change to exosome-free serum medium and continue culturing for 24 hours; collect the cell supernatant, centrifuge at 4°C and 300 g for 20 min; collect the supernatant into a new centrifuge tube and centrifuge at 4°C and 1000 g for 30 min; then collect the supernatant into another new centrifuge tube and centrifuge at 4°C and 10,000 g for 30 min; filter the supernatant using a 0.22 μm filter and transfer it to an ultrafiltration centrifuge tube (100 kDa), centrifuge at 4°C and 1000 g for 30 min, and then aspirate the concentrated solution on the upper layer of the ultrafiltration centrifuge tube. Extract using the ExoQuick-TC Exosome Kit (catalog number: SBI-EQULTRA-20TC-1): Mix the aspirated concentrated solution and the extraction reagent in a sterile microcentrifuge tube at a volume ratio of 5:1 and mix well, place at 4°C for 12 h; after centrifuging at 4°C and 1500 g for 30 min, discard the supernatant, and the remaining precipitate is OE-MSCs-Exo. Dissolve the precipitate thoroughly with sterile PBS and aliquot, store at -80°C. Label the exosomes extracted from the pre-treated OM-MSCs in groups A-C as A-MSCs-Exo, B-MSCs-Exo, and C-MSCs-Exo respectively. Detect the marker proteins of the 3 types of exosomes, see Figure 2 。

[0051] Figure 2 The results showed that Western Blot of A-MSCs-Exo, B-MSCs-Exo, and C-MSCs-Exo was positive for CD63 and TSG101 and negative for Calnexin, meeting expectations, indicating successful exosome preparation.

[0052] Example 3. Study on the treatment of AD neuroinflammatory mouse models with A-MSCs-Exo, B-MSCs-Exo, and C-MSCs-Exo

[0053] To verify the intervention effects of A-MSCs-Exo, B-MSCs-Exo, and C-MSCs-Exo on neuroinflammation in vivo, an experimental study was conducted using a mouse neuroinflammation model induced by Aβ oligomers (Aβ1-42 Peptide (Oligomerized), catalog number: C610006) in this example.

[0054] Eight-week-old C57BL / 6J SPF male mice, weighing 20–25 g, were purchased from a regular animal experiment center. All animals were adaptively fed for 7 days before the experiment and followed animal ethics norms during the experiment; they were divided into 5 groups, with 10 mice in each group, as follows:

[0055] Control group: Only an equal volume of PBS was injected into the tail vein once a day for 3 consecutive days, and no other treatment was given.

[0056] Model group: An Aβ oligomer (Aβ1-42 Peptide (Oligomerized), catalog number: C610006) solution (2 μg / μL) was injected into the right hippocampal region of mice through a stereotaxic apparatus, 1-2 μL / side (total bilateral injection dose of 2-4 μg Aβ), at a rate of 0.2 μL / min (to avoid mechanical damage). After inducing neuroinflammation, an equal volume of PBS was injected into the tail vein once a day for 3 consecutive days.

[0057] A-Exo treatment group: An Aβ oligomer (Aβ1-42 Peptide (Oligomerized), catalog number: C610006) solution (2 μg / μL) was injected into the right hippocampal region of mice through a stereotaxic apparatus, 1-2 μL / side (total bilateral injection dose of 2-4 μg Aβ), at a rate of 0.2 μL / min (to avoid mechanical damage). After inducing neuroinflammation, A-MSCs-Exo exosomes (2.5 mg / kg) were injected into the tail vein once a day for 3 consecutive days.

[0058] B-Exo treatment group: An Aβ oligomer (Aβ1-42 Peptide (Oligomerized), catalog number: C610006) solution (2 μg / μL) was injected into the right hippocampal region of mice through a stereotaxic apparatus, 1-2 μL / side (total bilateral injection dose of 2-4 μg Aβ), at a rate of 0.2 μL / min (to avoid mechanical damage). After inducing neuroinflammation, B-MSCs-Exo exosomes (2.5 mg / kg) were injected into the tail vein once a day for 3 consecutive days.

[0059] C-Exo treatment group: Through a stereotaxic apparatus, an Aβ oligomer solution (Aβ1-42 Peptide (Oligomerized), catalog number: C610006) (2 μg / μL) was injected into the right hippocampal region of the mice, 1-2 μL / side (total bilateral injection dose of 2-4 μg Aβ), at a speed of 0.2 μL / min (to avoid mechanical damage). After inducing neuroinflammation, C-MSCs-Exo exosomes (2.5 mg / kg) were injected via the tail vein once a day for 3 consecutive days;

[0060] After the tail vein injection, the Morris water maze experiment was first conducted. The water tank was divided into 4 quadrants and a platform was placed in the middle of one of the quadrants. Subsequently, the mice were randomly placed into the water facing the pool wall, and the time it took for them to find the platform, that is, the escape latency, was recorded. If the platform was not found within 90 s, the mice needed to be guided to the platform. Continuous training was carried out for 5 days. On the 6th day, after the platform was removed, the mice were placed into the water from a specific position, and the time they stayed on the original platform within 90 s, the number of times they crossed the platform, and their swimming trajectories were recorded, as shown in Table 1.

[0061] Table 1 Comparison of learning and memory abilities of mice in each group (x±s, n = 10)

[0062] Group Escape latency / s Number of platform crossings Platform residence time / s Control group 4.54±0.84 28.45±1.73 36.23±2.51 Model group <![CDATA[32.32±1.45 a > <![CDATA[4.34±0.74 a > <![CDATA[6.53±0.46 a > A-Exo treatment group <![CDATA[8.24±0.51 c > <![CDATA[23.45±2.14 d > <![CDATA[28.66±2.16 d > B-Exo treatment group <![CDATA[12.45±1.73 d > <![CDATA[15.46±1.18 d > <![CDATA[18.53±1.27 c > C-Exo treatment group <![CDATA[23.34±1.29 d > <![CDATA[12.45±1.52 c > <![CDATA[13.73±1.31 c >

[0063] Compared with the control group, P a <0.01, P b <0.05; compared with the model group, P c <0.01, P d <0.05.

[0064] The results in Table 1 showed that compared with the control group, the escape latency of the model group was significantly prolonged, the number of times of crossing the platform and the time of staying on the platform were significantly reduced (P < 0.01), and the swimming trajectory was complex; compared with the model group, the escape latency of the A-Exo treatment group, B-Exo treatment group, and C-Exo treatment group was significantly shortened, the number of times of crossing the platform and the time of staying on the platform were significantly increased (P < 0.05), and the swimming trajectory was simple. Moreover, compared with the B-Exo treatment group and C-Exo treatment group, the A-Exo treatment group had a shorter escape latency, more times of crossing the platform, and longer time of staying on the platform; this indicated that A-MSCs-Exo exosomes could improve the learning and memory abilities of AD mice by reducing hippocampal tissue damage.

[0065] Effects of exosomes in each group on the inflammatory level and oxidative stress response of AD neuroinflammatory model mice: The blood collected from the mouse eyeballs was left to stand at room temperature for 3 h and centrifuged at 5000 g for 5 min, and the supernatant was taken for enzyme-linked immunosorbent assay experiments. ELISA kits were used to detect the concentrations of SOD and IL-6 in the mouse serum, as shown in Figure 3 .

[0066] Figure 3 The results showed that the level of IL-6 in the model group was significantly higher than that in the control group, while the level of SOD was significantly lower; the levels of IL-6 in the A-Exo treatment group, B-Exo treatment group and C-Exo treatment group were significantly lower than those in the model group, and the levels of SOD were significantly higher; compared with the B-Exo treatment group and C-Exo treatment group, the decrease in the level of IL-6 and the increase in the level of SOD in the A-Exo treatment group were more significant; this indicates that A-MSCs-Exo exosomes can improve the cognitive impairment of AD mice by reducing the inflammatory level and enhancing the oxidative stress response.

[0067] Effects of exosomes from each group on the activation of microglia in the hippocampus of AD neuroinflammatory model mice: After the experiment, the hippocampal tissues of mice in each group were taken for frozen section, and Iba1 was labeled by immunofluorescence staining + microglia, and the density of positive cells was quantitatively analyzed by confocal microscopy, as shown Figure 4 .

[0068] Figure 4 The results showed that compared with the control group, the cell density of Iba1 + in the hippocampal region of the model group was significantly increased, suggesting the over-activation of microglia in AD neuroinflammatory model mice. A-MSCs-Exo exosomes can significantly inhibit the abnormal activation of microglia in the hippocampus of AD neuroinflammatory model mice, and the effect is better than that of B-MS Cs-Exo and C-MSCs-Exo exosomes, indicating that A-MSCs-Exo exosomes play a therapeutic role by inhibiting the abnormal activation of hippocampal microglia and regulating the neuroinflammatory pathway.

[0069] The above are the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle described in the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. An exosome for treating neuroinflammation in Alzheimer's disease (AD), characterized in that, The exosomes are derived from olfactory mucosa mesenchymal stem cells (OM-MSCs) pretreated with IFN-FGF fusion protein, wherein the amino acid sequence of the IFN-FGF fusion protein is as shown in SEQ ID NO:

3.

2. The exosome according to claim 1, characterized in that, The IFN-FGF fusion protein is composed of the active fragment of human interferon γ (SEQ ID NO:1) and the active fragment of FGF receptor 1 (SEQ ID NO:2) linked by the linker GGGGSGGGGS.

3. The method for preparing the exosomes according to claim 1 or 2, comprising the following steps: a. Inoculate OM-MSCs in a serum-free medium, add the IFN-FGF fusion protein at a final concentration of 150 ng / mL and pretreat for 48 hours; b. Replace it with a serum-free exosome medium and continue culturing for 24 hours, and collect the cell supernatant; c. Extract the exosomes by gradient centrifugation in combination with the ExoQuick-TC kit, and verify the positivity of CD63 and TSG101 and the negativity of Calnexin by Western Blot.

4. A pharmaceutical composition for treating AD neuroinflammation, characterized in that, Comprising the exosomes according to any one of claims 1-2.

5. Use of the exosomes according to any one of claims 1-2 or the pharmaceutical composition according to claim 4 in the preparation of a medicament for treating neuroinflammatory diseases.

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