Application of hemoglobin oxygen carrier in preparation of antiviral drugs

Abstract

The invention relates to the technical field of biological medicine, in particular to application of hemoglobin oxygen carriers (HBOCs) in preparation of antiviral drugs. The invention provides a brand new action mechanism of the HBOCs, and reveals that the HBOCs changes immune cell subset distribution in mouse bone marrow and up-regulates the expression level of an antiviral gene Isg15, so that the resistance of a body to viruses is enhanced, and a new thought is provided for development and preparation of antiviral drugs.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the application of hemoglobin oxygen carrier in the preparation of antiviral drugs. Background Art

[0002] Oxygen is an indispensable necessity for maintaining physiological processes such as cell growth, differentiation, metabolism and tissue homeostasis. As a natural oxygen carrier, red blood cells can bind oxygen in the lungs and transport it to the whole body. Hemoglobin (Hb) is the main oxygen carrier in red blood cells, facilitating oxygen transfer through its well-known oxygen-carrying function. Many malignant factors, such as trauma, blood loss and pernicious anemia, may lead to a decrease in the level of red blood cells and hemoglobin in the blood. Timely transfusion of red blood cells, restoration of blood volume and ensuring adequate oxygen supply are the key to saving lives. Hemoglobin oxygen carriers (HBOCs) are a class of blood substitutes based on natural hemoglobin obtained by polymerization, cross-linking and polymer modification. Due to its excellent oxygen binding and release properties, it is widely used in the clinic for patients with tissue ischemia or hypoxia. At present, glutaraldehyde cross-linked bovine hemoglobin (HBOC-201) has been approved for the treatment of acute pernicious anemia in South Africa and Russia in 2001 and 2010, respectively.

[0003] In view of this, the present invention is proposed. Summary of the invention

[0004] The purpose of the present invention is to provide an application of hemoglobin oxygen carrier in the preparation of antiviral drugs. The present invention mainly uses scRNA-seq technology to explore the new mechanism of action of HBOCs and provide an effective strategy for preparing drugs that enhance the body's resistance to viruses. It reveals that HBOCs have the ability to significantly enhance the antiviral ability of immune cells in mouse bone marrow, providing a new idea for the development and preparation of new antiviral drugs.

[0005] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted: One aspect of the present invention relates to the use of hemoglobin oxygen carrier in the preparation of antiviral drugs.

[0006] The present invention uses single-cell RNA sequencing (scRNA-seq) technology to reveal that HBOCs change the distribution of immune cell subsets in the bone marrow of mice and upregulate the expression level of the antiviral gene Isg15, thereby enhancing the body's ability to resist viruses. The present invention provides a new idea for the development and preparation of antiviral drugs.

[0007] Hemoglobin oxygen carrier is a red blood cell substitute with oxygen carrying capacity, which is made from hemoglobin by chemical modification such as cross-linking, polymerization, coupling or encapsulation in a synthetic membrane. The present invention does not specifically limit the type or preparation method of hemoglobin oxygen carrier, and all existing hemoglobin oxygen carriers in the art can be used to implement the technical solution of the present invention. In some specific embodiments, the hemoglobin oxygen carrier includes but is not limited to: glutaraldehyde cross-linked bovine hemoglobin.

[0008] Furthermore, the hemoglobin oxygen carrier exerts an antiviral effect by activating signaling pathways related to immune cell subpopulations and the body's viral defense and immune response.

[0009] Furthermore, mouse bone marrow cells were grouped using classic immune cell markers to analyze the distribution changes of immune cell subpopulations.

[0010] Furthermore, differential gene expression (DEGs) analysis and signal pathway enrichment analysis were used to screen the differential genes among the groups of mice, and to identify the important signal pathways in which they were enriched.

[0011] Furthermore, the immune cell subpopulation includes, but is not limited to, at least one of neutrophils, dendritic cells, monocytes, T cells and B cells.

[0012] Furthermore, the hemoglobin oxygen carrier regulates gene Isg15 The expression of cytokines plays an antiviral role.

[0013] Another aspect of the present invention also relates to a composition comprising a hemoglobin oxygen carrier and auxiliary materials.

[0014] The composition obtained by combining the hemoglobin oxygen carrier and the auxiliary materials required for antiviral drugs can be used to enhance the body's antiviral ability.

[0015] The present invention does not specifically limit the type of excipients, and the excipients include, but are not limited to, conventional excipients in the field of antiviral drugs.

[0016] Furthermore, the composition can also be used in combination with other conventional antiviral drugs in the art to synergistically exert antiviral capabilities.

[0017] Another aspect of the present invention also relates to the use of the composition in the preparation of antiviral drugs.

[0018] The present invention verifies through relevant experiments that hemoglobin oxygen carrier has the ability to enhance the body's antiviral ability, and reveals the antiviral mechanism of hemoglobin oxygen carrier. The composition using hemoglobin oxygen carrier as the main active ingredient is used for antiviral and has a good antiviral effect.

[0019] Another aspect of the present invention also relates to a method for analyzing the antiviral mechanism of hemoglobin oxygen carrier, comprising the following steps: (a) Hemoglobin oxygen carriers were injected intravenously into mice to analyze the distribution changes of immune cell subsets in the mouse bone marrow and screen for significantly differentially expressed genes and signal enrichment pathways; (b) analyzing the expression levels of the significantly differentially expressed genes in different virus models to verify the effects of the significantly differentially expressed genes; (c) Pathological analysis and RT-qPCR experiments were performed on the spleen tissues of mice injected with hemoglobin oxygen carriers to verify the application potential of hemoglobin oxygen carriers in the preparation of antiviral drugs.

[0020] The method for analyzing the antiviral mechanism of hemoglobin oxygen carriers has the advantages of being simple, easy to operate, and not requiring reliance on complex process methods. It reveals and verifies the antiviral mechanism of hemoglobin oxygen carriers from multiple perspectives, providing a solid theoretical basis for the antiviral effect of hemoglobin oxygen carriers.

[0021] Furthermore, single-cell RNA sequencing (scRNA-seq) was used to analyze the distribution changes of immune cell subsets in mouse bone marrow.

[0022] scRNA-seq is a technology that studies gene expression differences at the single-cell level. This technology reveals the unique gene expression pattern of each cell by sequencing RNA transcripts in individual cells, providing a powerful means for studying cell heterogeneity. Its basic principle is to convert mRNA transcripts in individual cells into cDNA and then perform high-throughput sequencing.

[0023] The present invention does not specifically limit the type of virus model, and any virus model with typical characteristics in the field can be used to implement the technical solution of the present invention. In some specific embodiments, the virus model includes but is not limited to: at least one of severe acute respiratory syndrome coronavirus 2, Sendai virus, hepatitis C virus or hepatitis B virus.

[0024] The present invention analyzes the antiviral mechanism of hemoglobin oxygen carrier from three aspects, which are as follows: In the first aspect, the present invention performs scRNA-seq analysis on mouse bone marrow cells, analyzes the distribution changes of immune cell subsets in the bone marrow of mice injected with hemoglobin oxygen carriers and the control group, and screens significantly differentially expressed genes and signal enrichment pathways to reveal the antiviral mechanism of HBOCs; The bone marrow cells of mice in different treatment groups were grouped and subgrouped according to the classic markers of immune cells, and the composition ratio changes of important immune cell groups (such as neutrophils, dendritic cells, monocytes, T cells and B cells) were analyzed. The results showed that the number of neutrophils and monocytes decreased, and the number of dendritic cells (DC), B cells and T cells increased, indicating the influence of HBOCs on the immune function of mice. DEGs analysis and GO analysis of these immune cell populations showed that HBOCs could significantly activate the signaling pathways of immune cells' defense response to viruses. Among them, the expression level of Isg15 mRNA was significantly upregulated in all five immune cells, suggesting that Isg15 Genes play a role in antiviral responses; In the second aspect, the present invention analyzes bulk RNA-seq data in the public GEO database to detect the expression level of Isg15mRNA in different models of antiviral response, thereby verifying Isg15 The key role of genes in the body's defense against viruses; By analyzing the bulk RNA-seq data of mouse models or humans infected with different viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Sendai virus (SeV), hepatitis C virus (HCV) and hepatitis B virus (HBV), the results showed that the expression level of Isg15 mRNA in the virus-infected group was significantly increased compared with the normal group, indicating that Isg15 Plays an important role in the body's defense against viruses; In a third aspect, the present invention verifies the potential of HBOCs as antiviral drugs through hematoxylin-eosin (H&E) staining of tissue sections and RT-qPCR experiments; Pathological analysis of H&E stained sections of spleen tissues of mice in different treatment groups showed that neutrophil infiltration was massive in the HBOC group, and the boundary between red pulp and white pulp was blurred, indicating the antiviral effect of spleen tissue. RT-qPCR experiments were used to detect the expression of Isg15 mRNA in spleen tissues of mice. The results showed that compared with the normal group, the Isg15 mRNA level in the HBOC group was significantly increased, indicating that HBOCs can increase Isg15 The expression of mitochondria can enhance the body's antiviral ability, suggesting its development potential in the preparation of antiviral drugs.

[0025] Isg15 mRNA is the messenger RNA encoding Isg15 protein. Isg15 is a ubiquitin-like protein induced by interferon and widely exists in mammals, birds, insects and other organisms. It has multiple biological functions, including antiviral, anti-tumor, immune regulation, and participation in cell autophagy and metabolic processes.

[0026] Compared with the prior art, the present invention has the following beneficial effects: The present invention uses single-cell RNA sequencing technology to reveal the mechanism by which HBOCs change the distribution of immune cell subsets in the mouse bone marrow and upregulate the expression level of the antiviral gene Isg15, thereby enhancing the body's resistance to viruses, providing a new research idea for the development and preparation of antiviral drugs.

[0027] The composition obtained by combining the hemoglobin oxygen carrier and the auxiliary materials required for antiviral drugs in the present invention is used to enhance the body's antiviral ability. The composition can also be used in combination with other conventional antiviral drugs in the field to synergistically exert antiviral capabilities and broaden the use of antiviral drugs.

[0028] The present invention verifies through relevant experiments that hemoglobin oxygen carrier has the ability to enhance the body's antiviral ability, and reveals the antiviral mechanism of hemoglobin oxygen carrier. The composition using hemoglobin oxygen carrier as the main active ingredient is used for antiviral and has a good antiviral effect.

[0029] The method for analyzing the antiviral mechanism of hemoglobin oxygen carriers provided by the present invention has the advantages of being simple, easy to operate, and not requiring reliance on complex process methods. It reveals and verifies the antiviral mechanism of hemoglobin oxygen carriers from multiple perspectives, and provides a solid theoretical basis for the antiviral effect of hemoglobin oxygen carriers. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0031] Figure 1 Figure 1 is an exploration of the antiviral mechanism of HBOCs. (A) UMAP diagram of bone marrow cell clusters in mice in the control group / HBOC group (n=42414); (B) Bubble diagram showing the characteristic markers of different bone marrow cell clusters; (C) Composition ratio diagram of different cell clusters; (D) GO analysis diagram of significantly upregulated genes among 5 immune cell groups, showing 10 representative signal enrichment pathways, and the p value was corrected by the Benjamini-Hochberg method, and p<0.05 after correction; (E) Venn diagram showing the corresponding genes involved in the viral defense response in 5 immune cells, among which the expression of gene Isg15 was significantly upregulated in all 5 immune cells.

[0032] Figure 2Figure 1 is a validation diagram of the role of Isg15 in the body's antiviral response. (AE) Box plots showing the expression levels of Isg15 mRNA in different virus infection models in the GEO database (n=3). (A) Bulk RNA-seq data of normal or SARS-CoV-2 infected mouse lung tissue (GSE264189). (B) Bulk RNA-seq data of normal or SeV infected mouse lung tissue (GSE267631). (C) Bulk RNA-seq data of normal or HCV infected mouse liver tissue (GSE200811). (D) Bulk RNA-seq data of normal or HBV infected mouse liver tissue (GSE163285). (E) Bulk RNA-seq data of liver tissue of healthy or SARS-CoV-2 infected patients (GSE206635). Two-tail Student's t-test was used to test statistical differences, *p<0.05, ***p<0.001, ****p<0.0001.

[0033] Figure 3 The following is a validation diagram of the antiviral mechanism of HBOCs. (A) H&E staining of spleen tissue of mice in the control group / HBOC group (the scale bar on the left is 50 μm, and the scale bar on the right is 5 μm). The white arrow indicates the boundary between red pulp and white pulp, and the black arrow indicates neutrophils. (B) Isg15 mRNA expression in mouse spleen tissue, where the error bar indicates the Mean±SD of three repeated experiments (n=3), and the two-tail Student's t-test was used to analyze the statistical difference, * p<0.05. DETAILED DESCRIPTION

[0034] The technical scheme of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and specific embodiments, but it will be understood by those skilled in the art that the following described embodiments are part of embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If specific conditions are not specified in the embodiments, they are carried out according to normal conditions or conditions recommended by the manufacturer. If the manufacturer is not specified in the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0035] Example 1 Study on the antiviral mechanism of HBOCs In this example, scRNA-seq analysis was performed on the bone marrow cells of mice in the control group and HBOC group, respectively, to explore the distribution changes of important immune cell groups in the mouse bone marrow, and to screen the significantly different genes and signal enrichment pathways between the groups, revealing the new antiviral mechanism of HBOCs. Figure 1 .

[0036] The scRNA-seq analysis experiment was performed as follows: (1) Sample processing: Healthy and suitable male C57BL / 6J mice were randomly divided into two groups (n=3): control group and HBOC group. The mice in the HBOC group were intravenously injected with 600 mg / kg of Orfodine once a day for 3 consecutive days; the control group was given an equal volume of PBS in the same way. On the fourth day, the femur and tibia of the mice were taken for subsequent bone marrow cell isolation, and the spleen tissue was used for subsequent analysis.

[0037] (2) Bone marrow cell isolation: Remove the femur and tibia with the trimmed ends in a sterile environment, remove the surrounding muscle and fibrous tissue, and rinse the bone marrow cells thoroughly with PBS. Filter the cell suspension through a cell strainer and centrifuge. Remove the supernatant, wash the cell pellet twice with PBS and centrifuge. After removing the red blood cells with red blood cell lysis buffer, resuspend the cells in PBS containing 0.04% BSA and use trypan blue reagent to stain to evaluate the viability of the single cell suspension.

[0038] (3) scRNA-seq: Mouse bone marrow scRNA-seq was performed on the 10× Genomics Chromium Platform, and all experimental steps were performed according to the manufacturer's instructions. Single-cell suspensions were encapsulated into single-cell gel-bead emulsions (GEMs) using the Chromium Next GEM Chip G Single Cell Kit (10× Genomics) on the Chromium Controller, barcoded cDNA was synthesized on a C1000 Touch Thermal cycler (Bio-Rad), and amplified using the Amplification Master Mix Kit (10× Genomics). The final library was constructed using the Chromium Next GEMSingle Cell 3' Reagent Kit v3.1 (10× Genomics) and quantified by the Agilent BioAnalyzer 2100. Sequencing was performed on an Illumina NovaSeq X Plus with paired-end reads of 150 bp.

[0039] (4) Data preprocessing: The original base call files (BCL) of the sequencing were converted into FASTQ files using Cell Ranger software and aligned with the mouse reference genome (mm10 / GRCm38) to generate a count matrix for each sample.

[0040] (5) Data analysis: R software was used to normalize the matrix, screen highly variable genes, perform principal component analysis (PCA) analysis, perform batch processing, and perform unbiased clustering to obtain a two-dimensional visualization map (UMAP) of different cell subsets, and annotate the cell subsets based on classic immune cell markers. The genes with significant differences between the five important immune cell groups (neutrophils, dendritic cells, monocytes, T cells, and B cells) were analyzed, and GO analysis was performed on these genes. Signal pathways with a p-value less than 0.05 after adjustment by the Benjamini-Hochberg method were selected. The jveen tool was used to identify genes co-regulated by HBOCs in related signal pathways.

[0041] In this example, scRNA-seq was performed on bone marrow cells of C57BL / 6J mice with different treatments. Based on the characteristic markers of bone marrow cells reported previously, 10 cell subsets were identified by unbiased clustering: neutrophils, B cells, monocytes, T cells, dendritic cells, erythrocytes, macrophages, hematopoietic stem cells, mast cells, and mesenchymal stem cells (MSCs) ( Figure 1 AB). Compared with the control group, neutrophils in the HBOC group accounted for the largest proportion of all immune cells, followed by B cells ( Figure 1 C). Notably, in the HBOC group, the numbers of B cells, T cells, and dendritic cells increased, while the numbers of neutrophils and monocytes decreased ( Figure 1 A, C). These results indicate that HBOCs have a certain impact on the composition and functional status of various immune cells in the mouse bone marrow.

[0042] This example focuses on neutrophils, dendritic cells, monocytes, T cells, and B cells, and performs GO analysis on DEGs among the five immune cell subgroups, and finds that signaling pathways related to the body's viral defense and immune response are significantly activated ( Figure 1 D), suggesting that HBOCs may enhance the antiviral activity of these immune cells.

[0043] In addition, among genes related to viral defense, the Isg15 gene was found to be upregulated by HBOCs in all five immune cells ( Figure 1 E), indicating the correlation between the antiviral mechanism of HBOCs and Isg15. Therefore, this example reveals a new antiviral mechanism of HBOCs through scRNA-seq technology.

[0044] Example 2 Verification of the role of gene Isg15 in the body's antiviral response This example analyzes multiple related bulk RNA-seq data in the GEO database, detects the expression level of Isg15 mRNA in different virus infection models, and verifies the key role of gene Isg15 in the body's antiviral response process. Figure 2 .

[0045] Bulk RNA-seq data analysis was performed as follows: (1) Data acquisition: Bulk RNA-seq data of mouse or human tissues infected with different viruses were obtained from the GEO database, and the relevant count matrices were downloaded, including normal or SARS-CoV-2 infected mouse lung tissue data (GSE264189), normal or SeV infected mouse lung tissue data (GSE267631), normal and HCV infected mouse liver tissue data (GSE200811), normal or HBV infected mouse liver tissue data (GSE163285), and data of healthy or SARS-CoV-2 infected patients (GSE206635).

[0046] (2) Data analysis: R software was used to normalize the matrix, perform numerical conversion, and visualize the expression of Isg15 mRNA in the normal group / virus infection group. Two-tail Student's t-test was used to test statistical differences, and the p value should be less than 0.05.

[0047] In this example, bulk RNA-seq data of mice or human models infected with different viruses in public databases were selected to analyze the expression level of Isg15 mRNA. Figure 2 AE shows the expression levels of Isg15 mRNA in these virus infection models. The results in the figure show that compared with the normal group, the expression levels of Isg15 mRNA in the virus infection group increased significantly, suggesting that the gene Isg15 Therefore, this example verifies Isg15 The key role of HBOCs in antiviral response Isg15 expression to exert antiviral effects.

[0048] Example 3 Verification of the antiviral mechanism of HBOCs In this example, H&E staining and RT-qPCR experiments were performed on the spleen tissues of mice in the control group and HBOC group to explore the pathological changes in spleen tissue and the differences in Isg15 mRNA expression levels, and to verify the antiviral mechanism of HBOCs. Figure 3 .

[0049] 1. HE staining experiment is carried out as follows: Mouse spleen tissue was fixed in 10% neutral formalin buffer solution, dehydrated and embedded in paraffin blocks. The paraffin blocks were evenly cut into 5 μm thick sections, dewaxed, rehydrated, and stained with hematoxylin and eosin, respectively. The sections were observed under a microscope, and histological images were obtained for subsequent pathological analysis.

[0050] 2. RT-qPCR experiment was carried out as follows: (1) RNA extraction: After ultrasonic homogenization of the tissue, lyse it with TRNzol (Invitrogen) reagent and collect the supernatant by centrifugation. Collect the RNA precipitate with chloroform / isopropanol, wash twice with 75% ethanol and centrifuge. Aspirate the ethanol and dissolve the extracted RNA with RNase-free ddH2O.

[0051] (2) Reverse transcription and RT-qPCR detection: Genomic DNA removal and reverse transcription reaction were performed using PrimeScript™RT reagentKit (Takara Bio) according to the manufacturer's instructions to synthesize cDNA. The relative RNA expression level of the samples was detected using Universal BlueqPCR SYBR Green Master Mix (Yeasen). The primer sequences are as follows: Isg15 forward 5'-GGAACGAAAGGGGCCACAGCA-3' (SEQ ID No. 1), reverse 5'-CCTCCATGGGCCTTCCCTCGA-3 (SEQ ID No. 2); GAPDH forward: 5'-CTCAGGAGAGTGTTTCCTCGTC-3' (SEQ ID No. 3), reverse 5'-ATGGGCTTCCCGTTGATGAC-3' (SEQ ID No. 4). The experiment was repeated three times, and GAPDH was used as a quantitative internal reference. The -ΔΔCt standard method was used to calculate the mRNA expression level.

[0052] In this example, pathological analysis of HE-stained spleen tissue sections of mice in the control group and HBOC group was performed. The results showed that compared with the normal group, the spleen of mice in the HBOC group was infiltrated with a large number of neutrophils, and the boundary between the red pulp and the white pulp was blurred, suggesting that HBOCs enhanced the antiviral effect of the mouse spleen ( Figure 3 A).

[0053] In this example, the expression of Isg15 mRNA in the spleen of mice was detected by RT-qPCR experiment. The results showed that the Isg15 mRNA level in the HBOC group mice was significantly higher than that in the normal group ( Figure 3 B), indicating that HBOCs can be regulated by Isg15 expression to enhance the body's antiviral ability.

[0054] In summary, this example shows that HBOCs have the ability to enhance the antiviral effect of the mouse spleen, suggesting that HBOCs have certain prospects and development potential in the preparation of new antiviral drugs.

[0055] Although the present invention has been illustrated and described with specific embodiments, it should be appreciated that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Those skilled in the art should understand that the technical solutions described in the above embodiments may be modified, or some or all of the technical features thereof may be replaced by equivalents without departing from the spirit and scope of the present invention. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention. Therefore, this means that all such replacements and modifications within the scope of the present invention are included in the appended claims.

Claims

1. Application of hemoglobin oxygen carrier in the preparation of antiviral drugs.

2. The use according to claim 1, characterized in that: The hemoglobin oxygen carrier includes: glutaraldehyde cross-linked bovine hemoglobin.

3. The use according to claim 1, characterized in that: The hemoglobin oxygen carrier exerts an antiviral effect by activating signaling pathways related to immune cell subpopulations and the body's viral defense and immune response.

4. The use according to claim 3, characterized in that: The immune cell subpopulation includes at least one of neutrophils, dendritic cells, monocytes, T cells and B cells.

5. The use according to claim 1, characterized in that: The hemoglobin oxygen carrier regulates genes Isg15 The expression of cytokines plays an antiviral role.

6. A composition, characterized in that It comprises a hemoglobin oxygen carrier and auxiliary materials.

7. Use of the composition as claimed in claim 6 in the preparation of antiviral drugs.

8. A method for analyzing the antiviral mechanism of hemoglobin oxygen carriers, characterized in that: The following steps are involved: (a) Hemoglobin oxygen carriers were injected intravenously into mice to analyze the distribution changes of immune cell subsets in the mouse bone marrow and screen for significantly differentially expressed genes and signal enrichment pathways; (b) analyzing the expression levels of the significantly differentially expressed genes in different virus models to verify the effects of the significantly differentially expressed genes; (c) Pathological analysis and RT-qPCR experiments were performed on the spleen tissues of mice injected with hemoglobin oxygen carriers to verify the application potential of hemoglobin oxygen carriers in the preparation of antiviral drugs.

9. The method for analyzing the antiviral mechanism of hemoglobin oxygen carrier according to claim 8, characterized in that: Single-cell RNA sequencing was used to analyze the distribution changes of immune cell subsets in mouse bone marrow.

10. The method for analyzing the antiviral mechanism of hemoglobin oxygen carrier according to claim 8, characterized in that: The virus model includes at least one of severe acute respiratory syndrome coronavirus 2, Sendai virus, hepatitis C virus or hepatitis B virus.

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