Application of artemether, artesunate or dihydroartemisinin in preparation of medicine for preventing or treating aflatoxin B1 poisoning

Abstract

The invention discloses an application of artemether, artesunate or dihydroartemisinin in preparation of a medicine for preventing or treating aflatoxin B1 poisoning. Belongs to the technical field of medicine. The technical problem of detoxification of aflatoxin B1 is solved. The invention provides an application of artemether, artesunate or dihydroartemisinin in preparation of a medicine for treating aflatoxin B1 poisoning diseases. The body can be promoted to detoxify the AFB1.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to the application of artemether, artesunate, or dihydroartemisinin in the preparation of drugs for the prevention or treatment of aflatoxin B1 poisoning. Background Technology

[0002] Aflatoxins are a class of toxic secondary metabolites mainly produced by Aspergillus parasiticus and Aspergillus flavus. To date, more than 20 aflatoxins have been discovered, among which aflatoxin B1 (AFB1) is currently the most toxic and harmful mycotoxin known and has been classified as a Group 1 carcinogen by the International Agency for Research on Cancer. AFB1 is widely found in plant-based agricultural products and feed. Ingestion of AFB1-contaminated feed by livestock and poultry can lead to acute or chronic poisoning, causing hepatotoxicity, enterotoxicity, nephrotoxicity, immunotoxicity, neurotoxicity, and reproductive toxicity, resulting in significant economic losses to the livestock industry. Simultaneously, the biotransformation and metabolites of AFB1 in livestock and poultry, such as aflatoxin M1 (AFM1), remain and accumulate in animal-derived agricultural products such as meat, eggs, and dairy products, directly threatening food safety and human health through the food chain. However, currently, there is a lack of efficient methods to remove AFB1 or reduce its toxicity. Therefore, identifying and screening products that can promote AFB1 detoxification is of great significance.

[0003] The primary measure to reduce the harm of AFB1 is mold prevention and detoxification. However, due to the highly stable physicochemical properties of AFB1, and considering the safety, stability, economy, and loss of nutrients in food and feed during the detoxification process, currently used methods, including physical methods, chemical methods, adsorbent methods, and biodegradation methods, cannot completely eliminate AFB1 in agricultural production or prevent human and livestock from ingesting it. Therefore, there is an urgent need to screen products that can promote the body's detoxification of AFB1. Summary of the Invention

[0004] The present invention aims to propose the application of artemether, artesunate, or dihydroartemisinin in the preparation of drugs for the prevention or treatment of aflatoxin B1 poisoning, so as to solve the problem of detoxification of aflatoxin B1.

[0005] This invention provides the use of artemether, artesunate, or dihydroartemisinin in the preparation of medicaments for treating aflatoxin B1 poisoning.

[0006] This invention provides the use of artemether, artesunate, or dihydroartemisinin in the preparation of medicaments for the prevention of aflatoxin B1 poisoning.

[0007] This invention provides the application of artemether, artesunate, or dihydroartemisinin in the preparation of health products for the adjuvant prevention / treatment of aflatoxin B1 poisoning.

[0008] This invention provides the application of artemether, artesunate, or dihydroartemisinin in the preparation of feed additives for the adjuvant prevention / treatment of aflatoxin B1 poisoning.

[0009] Further, it was found to inhibit the reduction in hepatocyte viability and increased DNA damage in animals caused by aflatoxin B1.

[0010] Further specifying, the dosage form of the drug is any one of tablets, capsules, granules, powders, or liquid preparations.

[0011] This invention provides a method for detoxifying aflatoxin B1 for non-therapeutic and non-diagnostic purposes, characterized in that artemether, artesunate, or dihydroartemisinin is added to human liver cell culture medium and reacted for 12 hours.

[0012] Further specified, the sample was HL-7702 cells.

[0013] Further specifying, the concentration of artemether, artesunate, or dihydroartemisinin is 1-10 μM.

[0014] Further limiting, the concentration of AFB1 is 10-80 μM.

[0015] Beneficial effects: Artemether, artesunate, or dihydroartemisinin can promote the detoxification of AFB1, significantly reduce the level of AFB1-induced γ-H2AX in AFB1-infected cells, and significantly enhance cell viability. Attached Figure Description

[0016] Figure 1 The effect of aflatoxin B1 (AFB1) alone on the cell viability of normal human hepatocytes (HL-7702).

[0017] Figure 2 The effect of artemether (ATM) alone on the cell viability of normal human hepatocytes (HL-7702).

[0018] Figure 3 Pretreatment with artemether (ATM) for 12 h was used to alleviate the cytotoxicity of aflatoxin B1 (AFB1) induced in HL-7702 cells.

[0019] Figure 4 The effect of supplementing with artemether (ATM) at different time points (0, 3, 6, 12 h) after exposure to aflatoxin B1 (AFB1) on alleviating the cytotoxicity of HL-7702 cells.

[0020] Figure 5This study investigated the inhibitory effect of artemether (ATM) on aflatoxin B1 (AFB1)-induced DNA damage in HL-7702 cells. Changes in γ-H2AX levels in cells were analyzed using immunofluorescence staining (10 μm scale).

[0021] Figure 6 This study investigated the inhibitory effect of artemether (ATM) on aflatoxin B1 (AFB1)-induced DNA damage in HL-7702 cells. Changes in γ-H2AX levels in cells were detected using Western blot.

[0022] Figure 7 The effect of ATM on GST enzyme activity in normal human hepatocytes (HL-7702).

[0023] Figure 8 The effects of ATM on the serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) activities in mice after AFB1 exposure.

[0024] Figure 9 The effect of ATM on the residual amount of AFB1-albumin adduct in the serum of mice after AFB1 exposure.

[0025] Figure 10 The effect of ATM on the residual amount of AFB1 in the liver tissue of mice after AFB1 exposure.

[0026] Figure 11 The effects of ATM on the morphology of mouse liver tissue after AFB1 exposure were investigated. The NC group served as the control group, ATM-L as the low-dose artemether treatment group, and ATM-H as the high-dose artemether treatment group. NC+AFB1 was the AFB1-inducing group alone, and ATM-L+AFB1 was the combined low-dose artemether and AFB1 treatment group. ATM-H+AFB1 was the combined high-dose artemether and AFB1 treatment group (magnification 400X, scale 50μm).

[0027] Figure 12 To investigate the effect of ATM on AFB1-induced DNA damage in mouse hepatocytes, the levels of γ-H2AX in the liver tissues of mice in each group were detected by immunohistochemistry. The NC group served as the control group; ATM-L was the low-dose artemether treatment group; ATM-H was the high-dose artemether treatment group; NC+AFB1 was the AFB1-inducing group alone; ATM-L+AFB1 was the low-dose artemether and AFB1 combined treatment group; and ATM-H+AFB1 was the high-dose artemether and AFB1 combined treatment group (magnification 400X, scale 50μm).

[0028] Note: AFB1 represents aflatoxin B1; ATM represents artemether; *P<0.05 compared with the control group; **P<0.01 compared with the control group; ***P<0.001 compared with the control group.

[0029] Figure 13 The figure shows the effect of artemether, artesunate, and dihydroartemisinin pretreatment for 12 h on alleviating aflatoxin B1 (AFB1, 40 μM)-induced cytotoxicity in HL-7702 cells. Detailed Implementation

[0030] HL-7702 cellsWang W, Li D, Xu Q, Cheng J, Yu Z, Li G, Qiao S, Pan J,Wang H, Shi J, Zheng T, Sui G. G-quadruplexes promote the motility in MAZphase-separated condensates to activate CCND1 expression and contribute to hepatocarcinogenesis. Nat Commun. 2024 Feb 5;15(1):1045. Artemether was purchased from MCE, catalog number HY-N0402; artesunate was purchased from MCE, catalog number HY-N0193; dihydroartemisinin was purchased from MCE, catalog number HY-N0176.

[0031] Example 1: In vitro experiment I. Effects of AFB1 on the viability of HL-7702 cells 3.0×10 3 HL-7702 cells per well were seeded into 96-well plates and cultured for 24 h. Then, AFB1 concentration gradients of 10 μM, 20 μM, 40 μM, and 80 μM were added. After incubation for 24 h, CCK8 solution (10 μL per well) was added. After 1-2 h of culture, the absorbance at 450 nm was measured.

[0032] Result: As Figure 1 As shown, cell viability gradually decreased as the AFB1 concentration gradually increased.

[0033] II. Effects of Artemether on HL-7702 Cell Viability 3.0×10 3HL-7702 cells per well were seeded into 96-well plates and cultured for 24 h. Then, artemether was added at different concentrations: 1 μM, 5 μM, 10 μM, 25 μM, 50 μM, 100 μM, and 150 μM. After incubation for 24 h, CCK8 solution (10 μL per well) was added. After 1-2 h of culture, the absorbance at 450 nm was measured.

[0034] Result: As Figure 2 As shown, cell viability did not decrease when the artemether concentration was below 25 μM, but cell viability decreased significantly when the artemether concentration exceeded 50 μM.

[0035] III. Artemether can prevent AFB1-induced decrease in HL-7702 cell viability. 3.0×10 3 HL-7702 cells per well were seeded into 96-well plates and cultured for 24 h. The cells were then divided into 20 groups: 5 groups treated with artemether (0 μM), 5 groups treated with artemether (1 μM), 5 groups treated with artemether (5 μM), and 5 groups treated with artemether (10 μM). After 12 h, each artemether-treated group was further divided into 5 subgroups, treated with AFB1 at 0 μM, 10 μM, 20 μM, 40 μM, and 80 μM respectively. After 24 h of incubation, CCK8 solution (10 μL per well) was added, and the absorbance at 450 nm was measured after 1-2 h of culture.

[0036] Result: As Figure 3 As shown, compared with the artemether (0 μM) treatment group, pretreatment with 1 μM, 5 μM, and 10 μM artemether significantly enhanced cell viability after AFB1 exposure in a dose-dependent manner. This indicates that artemether can prevent AFB1 poisoning, and 10 μM artemether has the best detoxification effect on 80 μM AFB1.

[0037] 3.0×10 3 HL-7702 cells per well were seeded into 96-well plates and cultured for 24 h. The cells were then divided into 20 groups. The control group was the solvent of the drug (DMSO) and the control group was the solvent of AFB1 (DMSO). The cells were pretreated with artemether (1 μM), artesunate (1 μM), and dihydroartemisinin (1 μM) for 12 h. Then, 40 μM of MAFB1 was added to each cell and the cells were incubated for 24 h. CCK8 solution (10 μL per well) was then added and the absorbance at 450 nm was measured after 1-2 h of culture.

[0038] Figure 13 As shown, pretreatment with artemether (1 μM), artesunate (1 μM), and dihydroartemisinin (1 μM) for 12 h significantly improved cell viability after AFB1 (40 μM) exposure.

[0039] IV. Artemether can promote the detoxification of AFB1. 3.0×10 3 HL-7702 cells per well were seeded into 96-well plates and cultured for 24 h. Then, AFB1 (40 μM) was added for treatment. At different time points (0, 3, 6, 12 h) after adding AFB1, artemether (10 μM) was added for treatment. After incubation with AFB1 for 36 h, CCK8 solution (10 μL per well) was added. After culturing for 1-2 h, the absorbance at 450 nm was measured.

[0040] Result: As Figure 4 As shown, compared with the AFB1-only treatment group, artemether supplementation at 0, 3, and 6 h after AFB1 exposure significantly improved cell viability, indicating that artemether can promote the detoxification effect of AFB1.

[0041] V. Artemether alleviates AFB1-induced DNA damage in HL-7702 cells 4.0×10 4 HL-7702 cells per well were seeded into confocal culture dishes and cultured for 24 h. Afterward, the cells were treated with 40 μM AFB1 for 24 h with and without artemether (1 μM, 5 μM, 10 μM). The phosphorylation level of the DNA damage marker H2AX (γ-H2AX, Ser139) was analyzed by immunofluorescence staining. The culture medium was discarded, and the cells were washed with PBS and incubated at room temperature for 15 min with 4% paraformaldehyde (Beyotime Biotechnology). After washing with PBS, the cells were incubated at 37°C for 15 min with PBS containing Triton X-100. After washing with PBS again, the cells were incubated at room temperature for 5 min with antigen retrieval solution (Saiwell Biotechnology). The cells were blocked with PBS containing 10% donkey serum for 1 h. γ-H2AX antibody (Cell Signaling Technology) was then added. Incubate overnight at 4°C with 1:1000 diluted DAPI; add fluorescent secondary antibody (1:1000) and incubate at 37°C for 1 h in the dark, then at room temperature for 2 h; wash cells 3 times with PBS; add DAPI solution for nuclear staining and incubate at room temperature for 20 min; take pictures using a fluorescence microscope (GEDelta Vision Elite).

[0042] After treating HL-7702 cells as described above, the phosphorylation level of the DNA damage marker H2AX was detected by Western blotting: RIPA lysis buffer (Beyotime Biotechnology) was added to cell culture dishes and lysed on ice for 5 min; after collecting the lysis buffer, 5×SDS buffer was added and the cells were heated in a metal bath for 8 min; proteins were separated by SDS-PAGE electrophoresis, and the proteins were transferred to a PVDF membrane by wet transfer (200 mA, 30 min), and then blocked with 5% BSA at 37℃ for 1 h; primary antibodies (γ-H2AX and α-Tubulin) were added and incubated overnight at 4℃; secondary antibody (1:3000) was added and incubated at 37℃ for 1 h, and then ultrasensitive ECL working solution (Tanon) was added, and the cells were photographed using a chemiluminescence imaging system (Tanon).

[0043] Result: As Figure 5 As shown, compared with the artemether (0 μM) treatment group, co-treatment with 1 μM, 5 μM, and 10 μM artemether significantly reduced the number of γ-H2AX foci. Western blotting also revealed that artemether (1 μM, 5 μM, and 10 μM) significantly reduced AFB1-induced γ-H2AX levels. Figure 6 ).

[0044] VI. Artemether promotes AFB1 detoxification by activating the activity of phase II metabolic enzymes GSTs. 1.0×10 5 HL-7702 cells were seeded per well into 6-well plates and cultured for 24 h. Then, artemether (1 μM, 5 μM, 10 μM) was added for 24 h each. Intracellular GSTs enzyme activity was detected using 1-chloro-2,4-dinitrobenzene (CDNB) as a substrate: the culture medium was discarded, cells were digested with trypsin and counted, and homogenized on ice to lyse the cells. The cells were centrifuged at 8000g for 10 min at 4 °C, and the supernatant was collected for testing. The test sample and reagent II were added according to the kit instructions, and the cells were incubated at 37 °C for 15 min. Reagent III was added, and the cells were incubated at 37 °C for 5 min. The absorbance at 340 nm was measured using a multi-mode microplate reader.

[0045] Result: As Figure 7 As shown, compared with the artemether (0 μM) treatment group, the addition of 1 μM, 5 μM, and 10 μM artemether significantly enhanced the activity of GSTs enzymes in a concentration-dependent manner.

[0046] Example 2: In vivo experiment All animal experimental procedures were approved by the Animal Protection and Use Ethics Committee of Northeast Forestry University (No. 2023009). Male ICR mice aged 6-8 weeks (weighing 25-30g) were purchased from Liaoning Changsheng Biotechnology Co., Ltd. Thirty-six mice were randomly divided into six groups: control group (8 mL / kg corn oil by gavage, once daily for 17 days), low-dose artemether group (30 mg / kg artemether by gavage, once daily for 17 days), high-dose artemether group (60 mg / kg artemether by gavage, once daily for 17 days), AFB1 group (8 mL / kg corn oil by gavage, once daily for 17 days, followed by intraperitoneal injection of 2 mg / kg AFB1 every other day starting on day 14), low-dose artemether + AFB1 group (30 mg / kg artemether by gavage, once daily for 17 days, followed by intraperitoneal injection of 2 mg / kg AFB1 every other day starting on day 14), and high-dose artemether + AFB1 group (60 mg / kg artemether by gavage, once daily for 17 days, followed by intraperitoneal injection of 2 mg / kg AFB1 every other day starting on day 14). On the evening of day 17, all mice were fasted. On the morning of day 18, the weight of mice in each group was measured. After anesthetizing the mice with sodium pentobarbital via intraperitoneal injection, blood was collected by enucleation. The blood was then incubated overnight at 4°C. The next day, the blood was centrifuged at 3000g at 4°C, and the serum was collected and frozen after 20 minutes. Mice were euthanized by cervical dislocation, and liver tissue was quickly collected and weighed. The liver tissue was separated and placed in Bouin's solution (approximately 7 mL), and fixed overnight at 4°C.

[0047] result: 1. Effects of artemether on serum ALP, ALT, and AST enzyme activities induced by AFB1 in mice Frozen serum samples were thawed on ice and then processed according to the instructions of the alkaline phosphatase assay kit (Nanjing Jiancheng, A059-2-2), alanine aminotransferase assay kit (Nanjing Jiancheng, C009-2-1), and aspartate aminotransferase assay kit (Nanjing Jiancheng, C010-2-1).

[0048] like Figure 7 As shown, compared with the AFB1-only treatment group, both low-dose and high-dose artemether gavage significantly reduced serum ALP, ALT, and AST enzyme activities after AFB1 exposure. Figure 8 ).

[0049] 2. Effect of artemether on serum AFB1-albumin adduct levels in mice after AFB1 exposure Thaw the frozen serum samples on ice and follow the instructions of the aflatoxin B1 ELISA kit (Jianglai Biotechnology, JL-49096).

[0050] like Figure 8 As shown, compared with the AFB1-only treatment group, both low-dose and high-dose artemether gavage significantly reduced the serum AFB1-albumin adduct levels after AFB1 exposure. Figure 9 ).

[0051] 3. Effect of artemether on AFB1 content in liver tissue of mice after AFB1 exposure Take an appropriate volume of liver tissue, weigh it, grind it under liquid nitrogen, add pre-cooled PBS, and centrifuge at 5000g for 10 min; take 100 μL of supernatant and place it in a new EP tube, add 20 μL of 5% BSA solution, and incubate at room temperature for 5 min; then add 10 μL of methanol, vortex for 10 s, and incubate at room temperature for 2 min; filter the above mixture using a 0.22 μm filter to remove impurities (the filter needs to be rinsed with pre-cooled PBS beforehand); follow the instructions of the aflatoxin B1 ELISA kit (Jianglai Biotechnology, JL-49096).

[0052] like Figure 10 As shown, compared with the AFB1-only treatment group, both low-dose and high-dose artemether gavage significantly reduced the AFB1 content in the liver tissue of mice after AFB1 exposure. Figure 10 ).

[0053] 4. Tissue sections Liver tissue was fixed for no more than 24 hours (longer fixation time would lead to excessive tissue permeability). The liver tissue was then transferred to 70% anhydrous ethanol (approximately 7 mL). The next day, the liver tissue samples from each group of mice were dehydrated: samples were placed in 70% ethanol for 30 min, 80% ethanol for 30 min, 90% ethanol for 30 min, 95% ethanol for 30 min, anhydrous ethanol I for 30 min, and anhydrous ethanol II for 30 min in sequence; Clearing: Samples were transferred to anhydrous ethanol:xylene (volume ratio 1:1) mixed solution for 30 min, and then placed in xylene for 10 min; Paraffin infiltration: Samples were transferred to a xylene:paraffin (volume ratio 1:1) mixed solution preheated to 60℃ and placed for 30 min, and then treated overnight in liquid paraffin; Embedding: Each group of samples was labeled and embedded in paraffin using an embedding cassette. The embedded paraffin blocks were then trimmed. When trimming, it is best to remove as much paraffin as possible near the tissue to reduce the size of the sections and ensure maximum utilization of the sample. Sectioning: Fix the trimmed tissue block in the slot of the paraffin microtome (LEICA-RM2235), adjust the sample position to be parallel to the blade, manually rotate the handle to ensure uniform sectioning speed, select a section thickness of 5μm, place the intact tissue section on the adhesion slide, and add 40℃ water to the slide beforehand, and mark the sample name on the slide with a pencil; Spreading: Place the above sections on a 40℃ spreader until the sections are fully spread, then absorb the water on the slide and mount it; Drying: Place the mounted slides in a 40℃ oven and bake overnight.

[0054] 5. H&E staining: 1) Dewaxing: Place the prepared sections in xylene I for 20 min and xylene II for 5 min to remove excess paraffin around the samples.

[0055] 2) Rehydration: Place the dewaxed sections in anhydrous ethanol for 5 min, 95% ethanol for 5 min, 90% ethanol for 5 min, 80% ethanol for 5 min, and 70% ethanol for 5 min respectively.

[0056] 3) Hematoxylin staining: Place the rehydrated sections in hematoxylin staining solution for 3-5 min to stain the cell nuclei. After staining, rinse the sections three times under running water, 5 min each time.

[0057] 5) Eosin staining: Place the differentiated sections in eosin staining solution for 20-30 seconds to stain the cytoplasm. After staining, rinse the sections three times under running water, 5 minutes each time.

[0058] 6) Mounting: Place the stained liver tissue sections in 70% ethanol for 1 min, 80% ethanol for 1 min, 90% ethanol for 1 min, 95% ethanol for 1 min, anhydrous ethanol for 1 min, and xylene for 1 min, respectively. Finally, mount with neutral resin mounting medium, and then observe and photograph under a microscope.

[0059] 6. Immunohistochemical staining Drying: Place the slices in a 60℃ oven for 1 hour.

[0060] Gradient dewaxing: Xylene I and Xylene II for 30 min each, rinsing every 5 min during this period; gradient alcohol for 5 min each at 100%, 95%, 90%, and 80% in sequence; then rinse three times with running water, switching between different directions for 5 min each time.

[0061] Antigen retrieval: Prepare 200 mL of EDTA antigen retrieval solution (pH=9.0) (EDTA antigen retrieval solution can effectively remove cross-links, fully expose antigen epitopes, and improve immunostaining effects). Microwave the solution until boiling, then place the slides in the solution and start timing. Microwave retrieval for a total of 30 min. After retrieval, transfer the slides to cold tap water to cool to room temperature (approximately 20 min).

[0062] Soak and clean the tissue with distilled water, wipe the fluid off the edges of the tissue, and mark the tissue with an immunohistochemical pen.

[0063] Wash three times with PBS, 5 min each time.

[0064] Inactivation of endogenous peroxidase: Cover the tissue with 3% H2O2 and incubate at room temperature in the dark for 25 min to block endogenous peroxidase, thereby reducing non-specific staining. Wash three times with PBS-T, 5 min each time.

[0065] Wash three times with 1xPBS, each time for 5 minutes.

[0066] Punching: Prepare a 3‰ Triton X-100 solution using preheated PBS, incubate at 37°C for 20 min, wash 3 times with PBS-T for 5 min each time, and wash 3 times with PBS for 5 min each time.

[0067] Blocking: Dilute the primary antibody to an appropriate concentration with 10% donkey serum. Add the primary antibody to cover the tissue (100-200 μL per section), and incubate the sections overnight at 4°C in a humidified chamber.

[0068] The next day, wash three times with PBS, each time for 5 minutes.

[0069] Incubation with HRP-labeled secondary antibody: Dilute the secondary antibody to an appropriate concentration with 10% BSA, add the secondary antibody to the tissue, incubate at room temperature for 1 h, and then wash three times with PBS for 5 min each time.

[0070] AEC staining: Prepare the staining working solution according to the AEC staining kit (Solepro) instructions, with a ratio of 50:50:50:850 (solution A:solution B:solution C:ddH2O). Add 100-200 μL of the staining working solution to each sample and incubate at room temperature in the dark for 30 min. Terminate the reaction by adding ddH2O.

[0071] Stain with hematoxylin for 2 minutes, then rinse the sections three times under running water for 5 minutes each time.

[0072] After staining, the slides were mounted at 60°C using glycerin-gelatin mounting medium.

[0073] Depend on Figure 11 It was found that AFB1-treated mice exhibited disordered hepatocyte cord arrangement, granular degeneration, fatty degeneration, and inflammatory infiltration. Low- and high-dose groups significantly improved the pathological morphology of mouse livers, with a significant reduction in fatty degeneration and inflammatory infiltration. γ-H2AX is a marker of DNA damage, derived from... Figure 12 It can be seen that AFB1-treated mice showed significant DNA damage in their hepatocytes, while the DNA damage level in the low-dose and high-dose groups was significantly reduced.

[0074] The above embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art will recognize that various variations or modifications can be made based on the above description. Therefore, any obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. Use of artemether, artesunate or dihydroartemisinin in the preparation of a drug for treating diseases caused by aflatoxin B1 poisoning.

2. Use of artemether, artesunate or dihydroartemisinin in the preparation of a drug for preventing diseases caused by aflatoxin B1 poisoning.

3. Use of artemether, artesunate or dihydroartemisinin in the preparation of a health product for assisting in the prevention / treatment of diseases caused by aflatoxin B1 poisoning.

4. Use of artemether, artesunate or dihydroartemisinin in the preparation of a feed additive for assisting in the prevention / treatment of diseases caused by aflatoxin B1 poisoning.

5. Use according to any one of claims 1 to 4, characterized in that, Inhibition of the decrease in the activity of animal liver cells and the increase in DNA damage caused by aflatoxin B1.

6. Use according to any one of claims 1 to 4, characterized in that, The dosage form of the drug is any one of tablets, capsules, granules, powders, and liquid preparations.

7. A method for detoxification of aflatoxin B1 for non-therapeutic and diagnostic purposes, characterized by, Artemether, artesunate or dihydroartemisinin is added to human liver cell culture solution and reacted for 12 hours.

8. The method of detoxification according to claim 7, wherein, The sample is HL-7702 cells.

9. The method of detoxification according to claim 7, wherein, The concentration of artemether, artesunate or dihydroartemisinin is 1-10 μM.

10. The method of detoxification according to claim 7, wherein, The concentration of AFB1 is 10-80 μM.

Images