Application of sulforaphane in the preparation of products for treating aplastic anemia

Abstract

The present invention discloses the application of sulforaphane in the preparation of a product for treating aplastic anemia, and belongs to the field of biotechnology. The present invention finds that sulforaphane (SPN) can not only reverse the disease degree of aplastic anemia, but also achieve the purpose of treating AA by inhibiting the occurrence of ferroptosis. Therefore, the use of sulforaphane in the preparation of a product for preventing and treating aplastic anemia must have a good application prospect, and the present invention provides a new solution for the treatment of aplastic anemia.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to the application of sulforaphane in preparing a product for treating aplastic anemia. Background Art

[0002] Sulforaphane (Sulforaphane, Synonyms, SFN) is an orally active Keap1 / Nrf2 / ARE inducer. SPN can promote the transcription of tumor suppressor proteins and effectively inhibit the activity of HDACs. Studies have shown that SPN can inhibit ferroptosis by activating the Keap1 / Nrf2 / ARE pathway and further inducing the expression of HO-1. In addition, SPN also has anti-cancer and anti-inflammatory activities.

[0003] Aplastic anemia (AA) is a serious bone marrow failure disease, the main pathological feature of which is impaired bone marrow hematopoietic function, resulting in reduced production of all blood cells. Currently, for patients with severe aplastic anemia (SAA), HLA-matched sibling donor hematopoietic stem cell transplantation (MSD-HSCT) is still the first choice for patients under 40 years old; for SAA patients who are not suitable for first-line MSD-HSCT, other hematopoietic therapies can be considered on the basis of immunosuppressive therapy combined with TPO receptor agonists (TPO-RA).

[0004] Ferroptosis is a new form of programmed cell death, characterized by the lethal accumulation of lipid reactive oxygen species (ROS), which is iron-dependent and different from other traditional forms of cell death such as apoptosis, necrosis and autophagy in terms of morphology, biochemistry and genetics. With the continuous deepening of ferroptosis research, it has been found in recent years that ferroptosis is closely related to blood system diseases such as leukemia, lymphoma, multiple myeloma (MM), myelodysplastic syndromes (MDS) and anemia, but the relationship between ferroptosis and AA is still rare.

[0005] Currently, the cost of treating aplastic anemia (AA) with Western medicine is usually high. Although sulforaphane has been widely used in tumor prevention and treatment, there are no reports on the use of sulforaphane in the prevention and treatment of aplastic anemia. Ferroptosis is an important form of programmed cell death and is closely related to cell activity. Studying the ferroptosis regulation mechanism of sulforaphane (SFN) on aplastic anemia is crucial for a deeper understanding of sulforaphane as a drug for aplastic anemia. Summary of the invention

[0006] The present invention provides an application of sulforaphane in preparing a product for treating aplastic anemia, wherein the CAS registration number of the sulforaphane is 4478-93-7.

[0007] Preferably, sulforaphane is the only active ingredient in the product.

[0008] Preferably, the product is a medicine.

[0009] More preferably, the medicine further contains medically acceptable excipients.

[0010] More preferably, the dosage form of the drug is capsule, tablet, granule, powder, solution or lozenge.

[0011] The present invention also provides a traditional Chinese medicine composition for treating aplastic anemia, which is composed of the following traditional Chinese medicine components in parts by weight: 20 to 30 parts of sulforaphane; 1 to 5 parts of lycopene; 1 to 5 parts of cryptoxanthin; 1 to 5 parts of hesperidin; 1 to 5 parts of dioscorea; 1 to 5 parts of limonin; 1 to 5 parts of chrysanthemum ketone; and 1 to 5 parts of kaempferol.

[0012] Preferably, the Chinese medicine composition is composed of the following Chinese medicine components in parts by weight: 25 parts of sulforaphane; 1 part of lycopene; 1 part of cryptoxanthin; 1 part of hesperidin; 1 part of dioscorea; 1 part of limonin; 1 part of chalcanthone; and 1 part of kaempferol.

[0013] The present invention also provides a traditional Chinese medicine preparation for treating aplastic anemia, wherein the traditional Chinese medicine preparation is prepared from the traditional Chinese medicine composition.

[0014] Preferably, the preparation method of the traditional Chinese medicine preparation comprises the following steps:

[0015] (1) adding water to the Chinese medicine composition and soaking it at 50-60° C. for 30-45 minutes;

[0016] (2) filtering and collecting the filtrate;

[0017] (3) Spray drying to form granules or vacuum concentration to form spray preparations.

[0018] More preferably, the step (1) further comprises grinding the traditional Chinese medicine composition.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] (1) The present invention effectively alleviates the severity of aplastic anemia through the effect of sulforaphane on aplastic anemia cells;

[0021] (2) The present invention uses sulforaphane to treat aplastic anemia mouse model and finds that sulforaphane has a significant preventive and therapeutic effect on aplastic anemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the blood routine level test of SAA model mice before and after SPN treatment in Example 3.

[0023] Figure 2 This is the sternal HE detection of SAA model mice before and after SPN treatment in Example 3.

[0024] Figure 3 Flow cytometric detection of ROS levels in bone marrow cells of SAA model mice before and after SPN treatment in Example 3.

[0025] Figure 4 This is a bar graph analysis of ROS levels in bone marrow cells of SAA model mice before and after SPN treatment in Example 3.

[0026] Figure 5 This is the ELISA detection of ferroptosis-related markers in SAA model mice before and after SPN treatment in Example 3.

[0027] Figure 6 This is the WB detection of the levels of ferroptosis-related proteins in SAA model mice before and after SPN treatment in Example 3.

[0028] Figure 7 This is a bar graph analysis of the levels of ferroptosis-related proteins in SAA model mice before and after SPN treatment in Example 3.

[0029] Figure 8 The ROS levels of 32D cells before and after IFNγ treatment were detected by flow cytometry in Example 4.

[0030] Fig. 9 This is a bar graph analysis of ROS levels in 32D cells before and after IFNγ treatment in Example 4.

[0031] Fig.10 This is the CCK8 experiment in Example 4 to detect the cell activity of 32D cells before and after IFNγ treatment.

[0032] Fig.11 The WB method in Example 4 was used to detect the levels of ferroptosis-related proteins in 32D cells before and after IFNγ treatment.

[0033] Fig.12 This is a bar graph analysis of the levels of ferroptosis-related proteins in 32D cells before and after IFNγ treatment in Example 4.

[0034] Fig.13The CCK8 experiment in Example 5 detected the cell activity of aplastic anemia 32D cells before and after SPN treatment.

[0035] Fig.14 The flow cytometer in Example 5 detected the apoptosis level of aplastic anemia 32D cells before and after SPN treatment.

[0036] Fig.15 The flow cytometer in Example 5 was used to detect the intracellular ROS levels in aplastic anemia 32D cells before and after SPN treatment.

[0037] Fig.16 This is a bar graph analysis of intracellular ROS levels in aplastic anemia 32D cells before and after SPN treatment in Example 5.

[0038] Fig.17 The flow cytometer in Example 5 detected the intracellular lipid peroxidation level of aplastic anemia 32D cells before and after SPN treatment.

[0039] Fig.18 This is a bar graph analysis of intracellular lipid peroxidation levels in aplastic anemia 32D cells before and after SPN treatment in Example 5.

[0040] Fig.19 The WB method in Example 5 was used to detect the levels of ferroptosis-related proteins in aplastic anemia 32D cells before and after SPN treatment.

[0041] Fig. 20 This is a bar graph analysis of the levels of ferroptosis-related proteins in aplastic anemia 32D cells before and after SPN treatment in Example 5.

[0042] Fig.21 The RT-PCR method in Example 5 was used to detect the expression levels of ferroptosis-related genes in aplastic anemia 32D cells before and after SPN treatment.

[0043] Fig. 22 This is a routine blood test of SAA model mice before and after the treatment of the traditional Chinese medicine complex mainly composed of SPN in Example 9.

[0044] Fig.23 This is the detection of ROS levels in bone marrow cells of SAA model mice before and after the treatment of the SPN-based traditional Chinese medicine complex in Example 9.

[0045] Fig.24 HE detection of the anterior and posterior sternum of SAA model mice treated with the SPN-based traditional Chinese medicine compound in Example 9. DETAILED DESCRIPTION

[0046] Example 1 Establishment of SAA model mice

[0047] Establishment of SAA mice: B6D2F1 (F1) female mice were prepared by mating B6 female mice with DBA / 2 male mice, and B6 female mice (donor mice) of the same age were selected as lymph node mononuclear cell donor mice; F1 female mice received TBI with a dose of 5 Gy in the irradiation equipment of the linear accelerator, and the B6 donor lymph node mononuclear cell suspension was prepared at the same time; 8×10 donor mice were injected intraperitoneally into the F1 female mice within 4-6 hours after irradiation. 8 Lymph node mononuclear cells were injected into the B6D2F1 female mice; the B6D2F1 female mice injected with mononuclear cells were returned to their original place for feeding, the day of injection was regarded as day 0, and after waiting for 12 days, the model was constructed.

[0048] Example 2 Drug administration to SAA model mice

[0049] Twelve SAA model mice constructed by the above method were randomly divided into two groups, with 6 mice in each group. One group was given SFN by gavage, and the dose of 20 mg / kg SFN was continuously gavaged for 10 days, which was set as the SFN+AA group; the other group of model mice was given an equal volume of PBS by gavage for 10 days, which was set as the AA group. Six normal F1 female mice were set as the control group, and PBS was continuously gavaged for 10 days, which was set as the control group. The gavage volume in each group was 100 μL / mouse.

[0050] Example 3 Collection of Mouse Specimens

[0051] After the above experimental mice in each group were treated for 10 consecutive days, 1.0-2.0 mL of retro-orbital venous blood was obtained from the mice on the 11th day by enucleation of the eyeballs after painless anesthesia, and the blood routine of each mouse was analyzed by an automatic hematology analyzer after anticoagulation.

[0052] After the experiment, the mice were euthanized and the sternums were removed under sterile conditions. The sternums were fixed with 4% paraformaldehyde for 15 min and then stained with hematoxylin-eosin (HE).

[0053] After the experiment, the mice were euthanized, and the spleen of the mice was removed under sterile conditions. About 0.1 g of spleen was weighed and ground by ultrasound, and total protein was extracted using RIPA lysis buffer and protease inhibitor mixture (mixing ratio 100:1). Finally, western blot was used to test ferroptosis-related proteins. The proteins detected in this part mainly include: Gpx4, Slc7a11, Nrf-2 and HO-1.

[0054] After the experiment, the mice were euthanized, and the femurs on both sides were removed under sterile conditions. RPMI 1640 medium was quickly extracted using a 1 mL syringe to flush out the nucleated cells in the bone marrow. The flushing was repeated 3 to 4 times, and the femoral bone marrow mononuclear cell suspension was obtained after red lysis. The level of ROS in the bone marrow cells of each experimental group was detected by flow cytometry, and the method was operated according to the instructions of the reagent H2DCFDA detection probe (MCE, HY-D0940).

[0055] Example 4 Construction of a 32D cell aplastic anemia model

[0056] 4.1 Cell culture and plating

[0057] 32D cells frozen in liquid nitrogen were selected and quickly revived in a 37°C water bath. The complete culture medium used was DMEM containing 10% FBS and 1% double antibody. The culture conditions were 37°C and 5% CO2.

[0058] Cell passaging: When the cell density reaches 80%-90%, the cells should be passaged in time. The passaged cells should be placed back in a 37°C, 5% CO2 cell culture incubator for continued culture.

[0059] Cell plating: 32D cells in the logarithmic phase of the above culture were plated at 5×10 5 The cells were plated in 6-well plates and cultured for 24 h before subsequent treatment.

[0060] 4.2 Detection of 32D cells induced by different concentrations of INF-γ and related indicators

[0061] (1) Inoculate 32D cells in good condition until the density reaches about 80% and starve them (in DMEM medium without FBS) for 12 h;

[0062] (2) After starvation, cells were seeded in a 96-well plate at 1000 cells / well;

[0063] (3) INF-γ (0, 400, 800, 1000 U / mL) was added to the designated wells and the cells were intervened for 12 h, 24 h, and 48 h, respectively.

[0064] (4) CCK-8 reagent was used to detect the changes in cell activity before and after treatment of 32D cells with different concentrations of INF-γ.

[0065] 4. Detection of ROS levels in 332D cells

[0066] After normal 32D cells and 32D cells with in vitro AA models were collected, the ROS levels in cells of each experimental group were detected by flow cytometry using the H2DCFDA probe.

[0067] 4.432D cell protein extraction and detection of related protein indicators

[0068] The cells of the protein samples to be extracted were collected and dissolved with RIPA buffer and protease inhibitor mixture (mixing ratio 100:1). The protein concentration was measured using the BCA protein detection kit. The expression levels of ferroptosis-related proteins, including Gpx4, Slc7a11, Nrf2 and Ho-1, were detected by western blot.

[0069] Example 5 Treatment of INF-γ-induced 32D aplastic cells with different concentrations of SFN

[0070] 5.1 Cell culture and plating

[0071] 32D cells frozen in liquid nitrogen were selected and quickly revived in a 37°C water bath. The complete culture medium used was DMEM containing 10% FBS and 1% double antibody. The culture conditions were 37°C and 5% CO2.

[0072] Cell passaging: When the cell density reaches 80%-90%, the cells should be passaged in time. The passaged cells should be placed back in a 37°C, 5% CO2 cell culture incubator for continued culture.

[0073] INF-γ induced AA cell model: The 32D cells in the logarithmic phase were starved for 12 hours. 800 U / mL INF-γ was added to the culture dish and cultured for another 24 hours.

[0074] 5.2 Treatment of IFN-γ-induced 32D cells with different concentrations of SFN

[0075] (1) Inoculate cells in good condition (32D cells treated with IFN-γ) in a 6-well plate and pre-culture for 24 h;

[0076] (2) When the cell density reached about 80%, SFN at the specified concentration (0, 1, 2, 5, 10 and 15 μM, respectively) was added to each well and culture was continued for 12 h, 24 h and 48 h.

[0077] 5.3 Detection of related indicators after treatment of IFN-γ-induced 32D cells with different concentrations of SFN

[0078] (1) CCK-8 reagent was used to detect the changes in cell activity over time in IFN-γ-induced 32D cells before and after treatment with different concentrations of SFN. The detection time was 0h, 12h, 24h, and 48h, respectively.

[0079] (2) IFN-γ-induced 32D cells were treated with different concentrations of SFN. The cells were collected after 24 h and the levels of ROS and lipid peroxidation in the cells were detected using H2DCFDA probe (MCE, HY-D0940) and BODIPY 581 / 591C11 (MCE, HY-D1301) probes, respectively.

[0080] (4) Proteins of 32D cells treated with INF-γ for 24 h were collected and the expression levels of ferroptosis-related proteins were detected by western blot, including Gpx4, Slc7a11, Nrf2 and Ho-1.

[0081] (5) After 24 h of treatment with INF-γ, 32D cells were collected and total RNA was extracted using the trizol method. RT-PCR was used to detect the expression levels of ferroptosis-related genes, including Gpx4, Slc7a11, Nrf2 and Ho-1.

[0082] Example 6 Preparation of a Chinese medicinal compound for treating aplastic anemia

[0083] 1. Take the following Chinese medicinal components by weight: 25 parts of sulforaphane; 1 part of lycopene; 1 part of cryptoxanthin; 1 part of hesperidin; 1 part of diosgenin; 1 part of limonin; 1 part of calcineurin; 1 part of calcineurin; 1 part of calcineurin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0084] 2. Add 2 times the weight of water to the above powder, dissolve it at 60°C under ultrasound for 45 minutes, collect the filtrate after filtering, spray dry to obtain solid powder, make it into granular preparation, and finally prepare Chinese medicine combination drug C1 for treating aplastic anemia.

[0085] Example 7 Preparation of a Chinese medicinal compound for treating aplastic anemia

[0086] 1. Take the following Chinese medicinal components by weight: 20 parts of sulforaphane; 1 part of lycopene; 1 part of cryptoxanthin; 1 part of hesperidin; 1 part of diosgenin; 1 part of limonin; 1 part of calcineurin; 1 part of calcineurin; 1 part of calcineurin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0087] 2. Add 2 times the weight of water to the above powder, dissolve it at 60°C under ultrasound for 45 minutes, collect the filtrate after filtering, spray dry to obtain solid powder, make it into granular preparation, and finally prepare Chinese medicine combination drug C2 for the treatment of aplastic anemia.

[0088] Example 8 Preparation of a Chinese medicinal compound for treating aplastic anemia

[0089] 1. Take the following Chinese medicinal components by weight: 25 parts of sulforaphane; 1 part of lycopene; 1 part of cryptoxanthin; 1 part of hesperidin; 1 part of diosgenin; 1 part of limonin; 1 part of calcineurin; 1 part of calcineurin; 1 part of calcineurin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0090] 2. Add 2 times the weight of water of the mixed raw materials to the above powder, dissolve it at 50°C under ultrasound for 30 minutes, collect the filtrate after filtration, and vacuum concentrate it to the same weight volume as the mixed raw materials to prepare a spray preparation, and finally prepare the Chinese medicine combination drug C3 for the treatment of aplastic anemia.

[0091] Example 9 Treatment of SAA model mice with composite Chinese medicine mainly based on SFN

[0092] 9.1 Treatment of SAA model mice with traditional Chinese medicine

[0093] Twenty-four SAA model mice constructed by the above method were randomly divided into 4 groups, with 6 mice in each group. The groups were set as follows: C1 was given by gavage at a dose of 20 mg / kg for 10 consecutive days, which was set as the C1+AA group; C2 was given by gavage at a dose of 20 mg / kg for 10 consecutive days, which was set as the C2+AA group; C3 was given by gavage at a dose of 20 mg / kg for 10 consecutive days, which was set as the C3+AA group; the fourth group was the model group, which was given an equal volume of PBS for 10 consecutive days, which was set as the AA group, and the gavage volume in each group was 100 μL / mouse.

[0094] 9.2 Collection of mouse specimens and detection of related indicators

[0095] After the above experimental mice in each group were treated for 10 consecutive days, 1.0-2.0 mL of retro-orbital venous blood was obtained from the mice on the 11th day by enucleation of the eyeballs after painless anesthesia, and the blood routine of each mouse was analyzed by an automatic hematology analyzer after anticoagulation.

[0096] After the experiment, the mice were euthanized and the sternums were removed under sterile conditions. The sternums were fixed with 4% paraformaldehyde for 15 min and then stained with hematoxylin-eosin (HE).

[0097] From the experimental results of the mouse model, we can see that in the constructed severe aplastic anemia (SAA) mouse model, the blood routine of each mouse (including white blood cell count, hemoglobin level and platelet count) was analyzed using a fully automatic blood cell analyzer. The results showed that the white blood cell count, hemoglobin level and platelet count in the SAA model mice were significantly decreased. However, after treatment with sulforaphane (SPN, 20mg / kg), it was found that the blood routine indicators of SAA were restored, indicating that SPN can alleviate the symptoms of AA ( Figure 1). Subsequently, we further verified the pathological state of the mouse sternum tissue by HE staining. The results showed that the SAA model group mice had obvious bone marrow damage, reduced hematopoietic tissue in the medullary cavity, large or lipid droplet vacuoles, and obvious fatification. These results showed that the SAA model mice were successfully constructed. After treatment with SPN, HE staining showed that the mouse bone marrow had partially recovered, the hematopoietic tissue in the medullary cavity had increased, and the lipid droplet vacuoles had decreased, indicating that the mouse bone marrow hematopoietic function had been significantly improved ( Figure 2 ). To further verify that SPN can improve the symptoms of aplastic anemia, we detected the level of ROS in mouse bone marrow cells by flow cytometry. The results showed that compared with normal mice, the level of ROS in bone marrow cells of SAA mice was significantly increased, and the level of ROS was reduced after SPN treatment, indicating that the activity of mouse bone marrow cells was restored after SPN treatment ( Figure 3 and Figure 4 ). Ferroptosis is an important type of programmed cell death that affects cell activity. Therefore, we then used ELISA to detect related indicators of ferroptosis in SAA model mice, mainly including MDA, Fe and 4-HNE. The results of the ELISA method showed that the levels of MDA, Fe and 4-HNE were significantly increased in SAA model mice, indicating that a high level of cellular ferroptosis occurred in SAA model mice. However, after treatment with SPN, these indicators were significantly restored, further indicating that SPN can alleviate the symptoms of aplastic anemia by inhibiting the occurrence of ferroptosis ( Figure 5 ). The levels of ferroptosis-related proteins (including HO-1, Nfr2, Slc7a11 and Gpx4) were detected by western blot. The results showed that in SAA model mice, the protein levels of HO-1, Nfr2, Slc7a11 and Gpx4 were significantly downregulated, indicating that the system against ferroptosis in SAA model mice was damaged, thereby promoting the occurrence of ferroptosis and further aggravating the symptoms of SAA. Interestingly, after treatment with SPN, the levels of the above proteins were significantly restored, which was able to inhibit the occurrence of ferroptosis ( Figure 6 and Figure 7 ).

[0098] Subsequently, the therapeutic effect of SPN was further tested in an in vitro cell model. First, a model of aplastic anemia was constructed using 32D cells. IFNγ treatment of 32D cells was able to construct an in vitro cell model of AA. Therefore, flow cytometry was used to detect the level of intracellular ROS after 32D cells were treated with different concentrations of IFNγ. The results showed that IFNγ treatment significantly increased the level of ROS in 32D cells in a dose-dependent manner ( Fig. 9). Subsequently, we used CCK8 experiments to verify the changes in cell activity after IFNγ treatment of 32D cells. The results showed that with the treatment of IFNγ, the activity level of 32D cells was significantly inhibited in a dose-dependent manner ( Fig.10 In addition, the levels of ferroptosis-related proteins (including HO-1, Nfr2, Slc7a11 and Gpx4) were detected by western blot in IFNγ-treated 32D cells. The results showed that the protein levels of HO-1, Nfr2, Slc7a11 and Gpx4 in 32D cells decreased significantly with IFNγ treatment ( Fig.11 and Fig.12 ). These results indicate that IFNγ can significantly induce ferroptosis of 32D cells, thereby inhibiting cell activity. In the above-constructed aplastic anemia cell model (plastic anemia 32D cells), the CCK8 experiment was used to detect the effect of SPN on the activity of aplastic anemia 32D cells. The results showed that with the treatment of SPN, the cell activity can be significantly restored ( Fig.13 ). The level of cell apoptosis was detected by flow cytometry, and the results showed that the level of cell apoptosis in 32D cells of aplastic anemia was significantly reduced with the treatment of SPN ( Fig.14 ). Subsequently, flow cytometry was used to detect the levels of ROS and lipid peroxidation in aplastic anemia 32D cells after SPN treatment. The results showed that with the treatment of SPN, the level of ROS in the cells could be significantly inhibited ( Fig.15 and Fig.16 ). Similar experimental results were also confirmed in the detection of lipid peroxidation ( Fig.17 and Fig.18 ). These experimental results show that SPN can significantly inhibit the occurrence of ferroptosis in aplastic anemia. Subsequently, we detected the levels of ferroptosis-related proteins (including HO-1, Nfr2, Slc7a11 and Gpx4) by western blot. The results showed that with the treatment of SPN, the above proteins in aplastic anemia 32D cells increased significantly ( Fig.19 and Fig. 20 ). RT-PCR experiments showed that SPN treatment could significantly increase the mRNA levels of HO-1, Nfr2, Slc7a11 and Gpx4 genes ( Fig.21 ). The above experiments showed that SPN can significantly inhibit ferroptosis, thereby enhancing cell activity and alleviating the symptoms of aplastic anemia.

[0099] Finally, we verified the therapeutic effect of multiple Chinese herbal combination drugs (C1, C2 and C3) based on SPN on aplastic anemia in vivo experiments. First, after the model mice were treated, we tested the blood routine of each group of mice. The results showed that compared with the AA model mice, the blood routine indicators of the mice treated with Chinese herbal combination drugs were restored, that is, the symptoms of aplastic anemia were alleviated ( Fig. 22 ). The level of ROS in mouse bone marrow cells was further detected by flow cytometry. The results showed that compared with AA model mice, the level of ROS in small bone marrow cells treated with the Chinese herbal combination drug was significantly downregulated ( Fig.23 ). Finally, the pathological state of the sternum of each mouse was detected by HE staining. The results of HE staining showed that after treatment with the Chinese medicine combination, the bone marrow of the mouse was significantly restored by HE staining, which indicated that the hematopoietic function of the mouse bone marrow was improved ( Fig.24 ).

[0100] In summary, sulforaphane (SPN) can not only reverse the severity of aplastic anemia, but also achieve the purpose of treating AA by inhibiting the occurrence of ferroptosis. Therefore, the use of sulforaphane to prepare drugs for the prevention and treatment of aplastic anemia is expected to have good application prospects.

[0101] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. The use of sulforaphane in the preparation of a product for treating aplastic anemia, characterized in that: The CAS registration number of the sulforaphane is 4478-93-7; and the product is a drug.

2. The use according to claim 1, characterized in that: Sulforaphane is the only active ingredient in the product.

3. The use according to claim 1, characterized in that: The medicine also contains medically acceptable auxiliary materials.

4. The use according to claim 3, characterized in that: The dosage form of the drug is capsule, tablet, granule, powder, solution or lozenge.

5. Use of a Chinese medicine composition in preparing a drug for treating aplastic anemia, characterized in that: The Chinese medicine composition consists of the following Chinese medicine components in parts by weight: 20-30 parts of sulforaphane; 1-5 parts of lycopene; 1-5 parts of cryptoxanthin; 1-5 parts of hesperidin; 1-5 parts of dioscorea; 1-5 parts of limonin; 1-5 parts of chalcanthone; and 1-5 parts of kaempferol.

6. The use according to claim 5, characterized in that: The Chinese medicine composition consists of the following Chinese medicine components in parts by weight: 25 parts of sulforaphane; 1 part of lycopene; 1 part of cryptoxanthin; 1 part of hesperidin; 1 part of diosmetin; 1 part of limonin; 1 part of chrysoprotone; and 1 part of kaempferol.

7. Use of a traditional Chinese medicine preparation in the preparation of a drug for treating aplastic anemia, characterized in that: The Chinese medicine preparation is prepared from the Chinese medicine composition described in claim 5 or 6.

8. The use according to claim 7, characterized in that: The preparation method of the Chinese medicine preparation comprises the following steps: (1) Add water to the Chinese medicine composition and soak at 50-60°C for 30-45 minutes; (2) Filter and collect the filtrate; (3) Spray drying to form granules or vacuum concentration to form spray preparations.

9. The use according to claim 8, characterized in that: Prior to step (1), the step also includes grinding the traditional Chinese medicine composition.

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