Application of miRNA-148a-3p in preparation of medicine for relieving inflammation

Abstract

The invention provides application of miRNA-148a-3p in preparation of a medicine for relieving inflammation, and belongs to the technical field of biological medicine. In the invention, the miRNA-148a-3p can reduce the expression of p65 and up-regulate the expression level of SIRT1 by targeting Tp65 in a 3'untranslated region of the miRNA-148a-3p so as to relieve inflammatory response and pathological injury degree. Through verification, the miRNA-148a-3p can reduce the expression of p65, up-regulate the expression of SIRT1 gene and inhibit the expression of proinflammatory factors in intestinal tracts of dextran sodium sulfate induced colitis model mice so as to relieve inflammation. Therefore, the camel milk exosome has an important protective effect on prevention and treatment of enteritis by regulating and controlling the miR-148a-3p / p65 / SIRT1 signal channel.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of miRNA-148a-3p in the preparation of drugs for alleviating inflammation. Background Art

[0002] Intestinal inflammation is a disease involving a complex immune response, and its occurrence may be related to a variety of factors, including infection, genetic susceptibility, environmental factors, and unhealthy lifestyles. Studies have shown that macrophage polarization is an important link in the intestinal inflammatory response. Although the normal inflammatory response is an important part of the body's defense mechanism, in some cases, the disorder of the inflammatory response can lead to chronic inflammation, which in turn causes colitis and other intestinal diseases. The manifestations of colitis usually include abdominal pain, diarrhea, and weight loss, and in severe cases can lead to intestinal damage and complications. Therefore, controlling and alleviating intestinal inflammatory responses is crucial to maintaining intestinal health.

[0003] Common drugs currently used to treat colitis include nonsteroidal anti-inflammatory drugs, steroids, and immunosuppressants. Although these drugs have certain efficacy in relieving inflammation and controlling symptoms, their side effects cannot be ignored. For example, long-term use of nonsteroidal anti-inflammatory drugs may cause gastrointestinal discomfort, peptic ulcers or bleeding; long-term use of steroids may cause a variety of side effects, such as immunosuppression, osteoporosis, and metabolic disorders. Therefore, seeking new natural anti-inflammatory substances has become a clinical need. Summary of the invention

[0004] The purpose of the present invention is to provide the use of miRNA-148a-3p in the preparation of drugs for alleviating inflammation. miRNA-148a-3p can reduce the expression of pro-inflammatory cytokines and increase the expression of anti-inflammatory cytokines, thereby being used to alleviate intestinal inflammation, providing new targets and ideas for the development of new, effective and safe drugs for the treatment of intestinal inflammation.

[0005] The present invention provides use of miRNA-148a-3p or a miRNA-148a-3p agonist in preparing a drug for alleviating inflammation, wherein the nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1.

[0006] Preferably, the nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.12.

[0007] Preferably, the inflammation comprises intestinal inflammation.

[0008] Preferably, the intestinal inflammation includes colitis.

[0009] Preferably, the colitis comprises dextran sulfate sodium induced colitis.

[0010] Preferably, the dosage form of the drug includes injection or oral preparation.

[0011] The present invention also provides the use of miRNA-148a-3p or a miRNA-148a-3p agonist in the preparation of at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1, and the nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.12.

[0012] Preferably, the pro-inflammatory cytokines include TNF-α and / or iNOS; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

[0013] The present invention also provides the use of camel milk exosomes in preparing a medicine for alleviating intestinal inflammation.

[0014] The present invention also provides the use of camel milk exosomes in preparing at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The pro-inflammatory cytokines include TNF-α; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

[0015] The present invention provides the use of miRNA-148a-3p or miRNA-148a-3p agonist in the preparation of a drug for relieving inflammation, wherein the nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1. In the present invention, miRNA-148a-3p can significantly reduce the expression of p65 and upregulate the expression level of Sirtuin 1 (SIRT1) by targeting Tp65 (p65) in its 3' untranslated region (3' UTR) to relieve colonic inflammatory response and pathological damage. It has been verified that miRNA-148a-3p can reduce the expression of p65, upregulate the expression of SIRT1 gene, inhibit the expression of pro-inflammatory factors (TNF-α, iNOS) in the intestine of dextran sulfate sodium (DSS)-induced colitis model mice, and thus relieve inflammation. This indicates that camel milk exosomes play an important protective role in the prevention and treatment of enteritis by regulating the miR-148a-3p / p65 / SIRT1 signaling pathway, providing a new therapeutic target for the treatment of colitis and further laying the foundation for the development of new, effective and safe therapeutic drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] 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 labor.

[0017] Figure 1 This is the morphology of camel milk exosomes; Figure 2 Schematic diagram of the average particle size of camel milk exosomes; Figure 3 This is the result diagram of the mRNA expression level of the pro-inflammatory factor iNOS; Figure 4 This is the result diagram of the mRNA expression level of the anti-inflammatory factor Arg-1; Figure 5 This is the result diagram of the mRNA expression level of the pro-inflammatory factor TNF-α; Figure 6 This is the result diagram of the mRNA expression level of the anti-inflammatory factor IL-10; Figure 7 This is the result diagram of the dual luciferase experiment to verify the miRNA target gene; Figure 8 Target gene SIRT1 Expression level result diagram of Fig. 9 Downstream genes of target genes in inflammatory pathways ( p65Gene) expression level results. DETAILED DESCRIPTION

[0018] The present invention provides use of miRNA-148a-3p or a miRNA-148a-3p agonist in preparing a drug for alleviating inflammation. The nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1, specifically: 5'-UAAAGUGCUGUCUCUGCUGCUG-3'.

[0019] In the present invention, miRNA-148a-3p is a functional miRNA in camel milk exosomes, and the targeted signaling pathway is the p65 / SIRT1 pathway. miRNA-148a-3p can act as an inflammatory inhibitor to inhibit p65 Expression, up-regulated SIRT1 The expression level of miRNA-148a-3p was reduced, thereby reducing the expression of pro-inflammatory cytokines TNF-α and iNOS, and increasing the expression of anti-inflammatory cytokines Arg-1 and IL-10. This provides a theoretical basis for regulating inflammatory pathways based on miRNA-148a-3p to alleviate inflammation in the future, and can be used to alleviate inflammation.

[0020] In the present invention, miRNA-148a-3p can be artificially synthesized with low cost and can achieve large-scale production.

[0021] In the specific implementation of the present invention, the nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.12, specifically: 5'-UCAGUGCACUACAGAACUUUGU-3'.

[0022] In the specific implementation of the present invention, the inflammation includes intestinal inflammation; the intestinal inflammation includes colitis; the colitis includes colitis induced by sodium dextran sulfate.

[0023] In the specific implementation of the present invention, the dosage form of the drug includes injection or oral preparation.

[0024] In the specific implementation of the present invention, the relief of intestinal inflammation is achieved through at least one of the following aspects: 1) Inhibition p65 Gene expression; 2) Increase SIRT1 Gene expression; 3) Reduce the expression of pro-inflammatory cytokines; 4) Increase the expression of anti-inflammatory cytokines.

[0025] In the specific implementation of the present invention, the pro-inflammatory cytokines include TNF-α and / or iNOS; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

[0026] The present invention also provides the use of miRNA-148a-3p or a miRNA-148a-3p agonist in the preparation of at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1, and the nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.12.

[0027] In the specific implementation of the present invention, the pro-inflammatory cytokines include TNF-α and / or iNOS; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

[0028] The present invention also provides the use of camel milk exosomes in preparing a medicine for alleviating intestinal inflammation.

[0029] The present invention also provides the use of camel milk exosomes in preparing at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The pro-inflammatory cytokines include TNF-α; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

[0030] In the present invention, the camel milk exosomes are extracted from camel milk by differential centrifugation; the camel milk exosomes have a typical exosome structure and an average particle size of 123.6 nm.

[0031] In the present invention, camel milk exosomes can be stored at low temperature.

[0032] In the present invention, the pro-inflammatory cytokines are pro-inflammatory cytokines in serum; the anti-inflammatory cytokines are anti-inflammatory cytokines in serum.

[0033] To further illustrate the present invention, the application of miRNA-148a-3p provided by the present invention in preparing drugs for alleviating inflammation is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0034] Example 1 Isolation, purification and characterization of camel milk exosomes An appropriate amount of camel milk was placed in a 50 mL centrifuge tube, and camel milk exosomes were extracted by differential centrifugation, see [Badawy, Abdelnaser A.; El-Magd, Mohammed A.; AlSadrah, Sana A..Therapeutic Effect of Camel Milk and Its Exosomes on MCF7 Cells In Vitro and In Vivo.[J].Integrative Cancer Therapies (INTEGR CANCER THER), 2018, Vol. 17(4): 1235-1246]; the exosomes obtained in Example 1 were characterized by transmission electron microscopy (TEM) and nanoparticle size tracking analysis (NTA). Figure 1 As shown, TEM results showed that the isolated exosomes had a typical exosome structure; Figure 2 As shown, NTA results showed that the average particle size of exosomes was 123.6 nm.

[0035] Example 2 Using bioinformatics methods to predict and screen candidate miRNAs in exosomes for alleviating enteritis According to the exosomes prepared in Example 1, the miRNA expression profile was constructed. New precursor miRNAs were predicted by miREvo and mirdeep2 software. Then, according to the predicted results of miRNA, its expression level was counted, and DESeq2 based on negative binomial distribution was used for analysis to identify the upregulated miRNAs in exosomes. Cluster analysis was performed on the screened upregulated miRNAs, and target gene prediction was performed using miRanda and RNAhybrid software, and the final result was the intersection of the two softwares.

[0036] By performing gene ontology (GO) analysis on miRNA target genes, the GO classification terms of upregulated miRNA target genes can be identified, and then the relationship between differential miRNAs and gene function changes in different samples can be explored.

[0037] In addition, the KEGG (Kyoto Encyclopedia of Genes and Genomes) database is a major public database that focuses on the study of signaling pathways. Using the resources of this database, enrichment analysis can be performed on dysregulated genes involved in the KEGG pathway. In the present invention, the goal is to conduct pathway enrichment studies on the target genes of those upregulated miRNAs.

[0038] Based on the GO database and KEGG public database, the up-regulated miRNAs were enriched in functions and pathways. Based on the enrichment results, the signaling pathway most closely related to inflammatory response was screened out: NF-κB signaling pathway. The key genes related to enteritis in this pathway were selected. SIRT1 , further explore (or verify) the potential role of miRNA (miRNA-148a-3p) in alleviating enteritis.

[0039] Example 3 Effects of exosomal miRNA on macrophage polarization and expression levels of inflammatory factors in DSS-induced colitis According to the candidate key miRNA-148a-3p in Example 2 (nucleotide sequence as shown in SEQ ID NO.1), the corresponding miRNA-148a-3p agonist (miRNA group, nucleotide sequence as shown in SEQ ID NO.12) and miRNA-148a-3p inhibitor (NC group, nucleotide sequence as shown in SEQ ID NO.13, specifically: 5'-CAGUACUUUUUGUGUAGUACG-3') were synthesized in vitro by Genomatix according to its mature sequence.

[0040] 1. Establishment of mouse colitis (UC) model Three-week-old male mice were selected and randomly divided into a blank group (n=10) and an experimental group (n=50). The experimental group was induced with dextran sulfate sodium (DSS) to establish a mouse UC model. The experimental group was further divided into a model group (DSS group), a miRNA group, a control group (NC group), an exosome group, and a camel milk group. The steps for constructing a mouse UC model are as follows: 1) 48 male mice aged 8 weeks and weighing 22±2 g were selected and randomly divided into 6 groups, with 8 mice in each group. From the beginning of modeling (day 0), mice in the blank group were allowed to drink distilled water freely, and mice in the experimental group were allowed to drink distilled water containing 3% DSS freely. Drinking water and DSS water were changed every day for 9 consecutive days.

[0041] 2) The method to determine whether the colitis model was successfully established is through the DAI (Disease Activity Index) score. The DAI score comprehensively evaluates weight loss, stool characteristics, and the degree of occult blood. The scoring criteria are as follows: weight loss score (scored according to the percentage of weight loss), stool viscosity score (scored from normal to severe diarrhea), and stool bleeding score (scored according to occult blood and the degree of bleeding). The DAI score is the average of the three scores. In the experiment, the scores of the blank group mice were basically stable, while the scores of the experimental group mice increased significantly. Starting from the 3rd day, there were obvious changes, especially the weight decreased significantly after the 4th day, and the DAI score was significantly higher than the blank group on the 7th day. This shows that the mouse UC model was successfully established.

[0042] The treatments for each group are as follows: Blank group: mice were injected with 0.2 ml PBS via tail vein once a day for 9 consecutive days; DSS group: 0.2 ml PBS was injected into the tail vein of UC mice once a day for 9 consecutive days; miRNA group: 4OD miRNA-148a-3p agonist was injected into the tail vein of UC mice once a day for 9 consecutive days; NC group: UC mice were injected with 4OD miRNA-148a-3p inhibitor via tail vein once a day for 9 consecutive days; Exosome group: 0.025 ml of exosomes isolated and purified in Example 1 were injected into the tail vein of mice with UC model once a day for 9 consecutive days; Camel milk group: UC model mice were gavaged with 0.2 ml camel milk once a day for 9 consecutive days.

[0043] After the experiment, peritoneal macrophages and intestinal tissues of each group of mice were collected for subsequent experiments.

[0044] Real-time fluorescence quantitative PCR and ELISA were used to detect and analyze the relative expression levels of related pro-inflammatory factors and anti-inflammatory factors in the above groups. The specific methods are as follows: 2. Real-time fluorescence quantitative PCR (RT-qPCR) detection RT-qPCR was used to detect and analyze the relative expression levels of related pro-inflammatory factors and anti-inflammatory factors in the above groups. The specific steps are as follows: (a) Total RNA extraction Trizol reagent was used to extract total RNA from peritoneal macrophages and intestinal tissues of each group of mice.

[0045] (b) Reverse transcription system Primer-ScriptTM RT-PCR kit was used, and the specific reverse transcription system and conditions are shown in Table 1.

[0046] Table 1 Reverse transcription reaction system and conditions

[0047] Reaction conditions: 37℃ for 15min; 85℃ for 5s; storage at 4℃.

[0048] (c) RT-qPCR reaction system The primers for detecting pro-inflammatory factors and anti-inflammatory factors include: The primers used to detect iNOS are shown in SEQ ID NO.2 and SEQ ID NO.3 respectively; The primers used to detect Arg-1 are shown in SEQ ID NO.4 and SEQ ID NO.5 respectively; The primers used to detect the internal reference gene GAPDH are shown in SEQ ID NO.6 and SEQ ID NO.7 respectively; All the above primer sequences are shown in Table 2.

[0049] Table 2 Primer sequence information

[0050] SYBR Premix Ex TaqTM kit was used. The reaction system and conditions are shown in Table 3: Table 3 RT-PCR reaction system and conditions

[0051] Reaction conditions: pre-denaturation at 95°C for 30 s; denaturation at 95°C for 5 s; extension at 60°C for 30 s, for a total of 40 cycles.

[0052] (d) Experimental results All data were analyzed in GraphPad Prism 8 software using one-way ANOVA and Tukey post hoc test for multiple comparisons. P <0.05,** P <0.01,*** P The difference was considered significant when the difference was less than 0.001. The relative quantitative formula 2-ΔΔCt was used to calculate the fold change of relative expression according to the Ct value of the internal reference GADPH.

[0053] like Figure 3 and Figure 4 As shown, the results showed that the expression level of pro-inflammatory factors in the DSS group was increased compared with the blank group. After treatment with miRNA (miRNA-148a-3p agonist), the pro-inflammatory factor iNOS was significantly reduced, and the anti-inflammatory factor Arg-1 was significantly increased, indicating that miRNA-148a-3p can reduce the expression of pro-inflammatory factors in the intestine caused by DSS and increase the expression of anti-inflammatory factors in the intestine caused by DSS. Similarly, after treatment with camel milk and exosomes, they can effectively improve the symptoms of DSS-induced enteritis by regulating the inflammatory environment of the mouse intestine and increasing the expression of anti-inflammatory factors (Arg-1).

[0054] 3. ELISA detection: The treatment was carried out according to Example 3, and the serum of each group of mice was collected. The levels of TNF-α and IL-10 were determined according to the operating instructions of the mouse interleukin 10 (IL-10) ELISA research kit and the mouse tumor necrosis factor (TNF-α) research kit purchased from Jiangsu Enzyme Biotechnology Co., Ltd.

[0055] The experimental results are as follows: All data were analyzed in GraphPad Prism 8 software using one-way ANOVA and Tukey post hoc test for multiple comparisons. P <0.05,** P <0.01,*** P The difference was considered significant when the difference was less than 0.001. The relative quantitative formula 2-ΔΔCt was used to calculate the fold change of relative expression according to the Ct value of the internal reference GADPH.

[0056] like Figure 5 and Figure 6 As shown, the results showed that compared with the control group, the protein expression level of proinflammatory factors in the model group was increased. After treatment with miRNA (miRNA-148a-3p agonist), the protein expression level of the proinflammatory factor TNF-α was significantly reduced, and the protein expression level of the anti-inflammatory factor IL-10 was significantly increased, indicating that miRNA-148a-3p can reduce the expression of proinflammatory factor proteins in the serum of colitis caused by DSS and increase the expression of anti-inflammatory factor proteins in the serum of colitis caused by DSS. Similarly, after treatment with camel milk and exosomes, the expression of the proinflammatory factor TNF-α can be inhibited, further alleviating the symptoms of enteritis caused by DSS.

[0057] Example 4 Dual luciferase validation experiment of exosomal miRNA and target genes of inflammatory pathway and changes in expression of key genes downstream of target genes.

[0058] Based on the miRNA-148a-3p screened in Example 2, RNAhybrid was used to predict the 3'UTR binding sites of the candidate miRNA and the target gene mRNA online. Based on the prediction results, Heyuan Biotechnology (Shanghai) Co., Ltd. was commissioned to construct the corresponding vector pMIR-REPORT Luciferase-3′-UTR, and then verified by dual luciferase reporter experiments. The experimental groups are as follows: A: wild-type miR-148a-3p-NC group; B: wild-type miR-148a-3p-mimics group; C: mutant miR-148a-3p-NC group; D: Mutant miR-148a-3p-mimics group.

[0059] The treatments for each group are as follows: The wild-type miR-148a-3p-NC group was prepared by inserting the 3'UTR sequence of miR-148a-3p (nucleotide sequence as shown in SEQ ID NO. 14, specifically: 5'-UAAUACGUGAAUUGCGGUAUCA-3') into the vector pMIR-REPORT; The mutant miR-148a-3p-NC group was prepared by mutating the 3'UTR sequence of miR-148a-3p (the nucleotide sequence is shown in SEQ ID NO.15, specifically: 5'-AAAACGUGAAUUGCGGUAUCA-3') and then inserting it into the vector pMIR-REPORT.

[0060] Dual luciferase assay results like Figure 7 As shown, the fluorescence expression of wild-type miR-148a-3p-mimics was significantly reduced compared with other groups, indicating that miRNA-148a-3p can silence the expression of P65, a key gene in the inflammatory pathway NF-κB, thereby exerting an anti-inflammatory effect.

[0061] Example 5 Referring to step 1 in Example 3, a mouse colitis (UC) model was established, and the treatment of each group was also the same as step 1 in Example 3.

[0062] RT-qPCR detection and analysis of target genes SIRT1 【 SIRT1 The relative expression level of the gene regulates the activity of NF-κB through deacetylation, inhibiting its transcription of inflammatory factors (such as TNF-α and IL-1β), thus playing an important role in the regulation of inflammation (inhibiting inflammation) Figure 8 ) and key genes in the inflammatory pathway NF-κB P65 The changes in expression levels of Fig. 9 ).

[0063] The intestinal tissue total RNA extraction, reverse transcription system, RT-qPCR reaction system, and data analysis were the same as those in Example 3.

[0064] The primers for detecting related genes include: The primers used to detect P65 are shown in SEQ ID NO.8 and SEQ ID NO.9 respectively; The primers used to detect SIRT1 are shown in SEQ ID NO.10 and SEQ ID NO.11 respectively; All the above primer sequences are shown in Table 4.

[0065] Table 4 Primer sequence information

[0066] The experimental results are as follows: like Figure 8 and Fig. 9 As shown in the figure, compared with the blank group, the expression level of p65 gene in the DSS group was increased. After treatment with miRNA (miRNA-148a-3p agonist) or camel milk exosomes, the expression level of p65 was significantly reduced, and the expression level of SIRT1 gene was significantly increased, further indicating that camel milk exosomes or miRNA-148a-3p can silence the expression of p65, a key gene in the inflammatory pathway NF-κB, thereby exerting an anti-inflammatory effect.

[0067] In summary, camel milk exosomes and miRNA-148a-3p derived from camel milk exosomes can significantly inhibit the expression of proinflammatory factors TNF-α and iNOS in the NF-κB pathway, thereby achieving its anti-inflammatory effect.

[0068] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Use of miRNA-148a-3p or a miRNA-148a-3p agonist in the preparation of a drug for alleviating inflammation, wherein the nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that: The nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.

12.

3. The use according to claim 1, characterized in that: The inflammation includes intestinal inflammation.

4. The use according to claim 3, characterized in that: The intestinal inflammation includes colitis.

5. The use according to claim 4, characterized in that: The colitis includes dextran sulfate sodium induced colitis.

6. The use according to claim 1, characterized in that: The dosage form of the drug includes injection or oral preparation.

7. Use of miRNA-148a-3p or a miRNA-148a-3p agonist in the preparation of at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The nucleotide sequence of the miRNA-148a-3p is shown in SEQ ID NO.1, and the nucleotide sequence of the miRNA-148a-3p agonist is shown in SEQ ID NO.

12.

8. The use according to claim 7, characterized in that: The pro-inflammatory cytokines include TNF-α and / or iNOS; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

9. Application of camel milk exosomes in the preparation of drugs for alleviating intestinal inflammation.

10. Use of camel milk exosomes in the preparation of at least one of the following preparations: 1) p65 Gene inhibitors; 2) SIRT1 Gene agonists; 3) pro-inflammatory cytokine inhibitors; 4) Anti-inflammatory cytokine agonists; The pro-inflammatory cytokines include TNF-α; the anti-inflammatory cytokines include Arg-1 and / or IL-10.

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