Nano-selenium preparation for targeted regulation of GPx3 expression as well as preparation method and application of nano-selenium preparation
By developing nanoselenium preparations targeted to regulate GPx3 expression, using the mixed reaction of shiitake polysaccharide and vitamin C with sodium selenite, the problem of insufficient GPx3 expression in the prior art was solved, higher anti-tumor activity and lower toxicity were achieved, and the therapeutic effect of prostate cancer was significantly improved.
Patent Information
- Application Number
- CN202510374624.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-27
- Publication Date
- 2025-06-27
AI Technical Summary
The prior art is difficult to effectively improve GPx3 expression in prostate cancer tissues, resulting in poor anti-tumor effect.
A nanoselenium preparation targeted to regulate GPx3 expression was developed. Nanoselenium preparation targeted to regulate GPx3 expression was prepared by mixing sodium selenite solution with shiitake polysaccharide (LET) and vitamin C (VC) solutions.
This nanoselenium preparation can significantly improve GPx3 expression, induce DNA damage in tumor cells, inhibit the proliferation, migration and invasion of tumor cells, enhance the immune response, improve the efficacy of anti-tumor drugs, and reduce toxicity.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of synthetic pharmaceutical chemistry, and particularly relates to a nano-selenium preparation for targeted regulation of GPx3 expression, a preparation method of the nano-selenium preparation for targeted regulation of GPx3 expression, and its application in anti-tumor drugs. Background Art
[0002] Prostate cancer is a major health problem, with approximately 13 million new cases diagnosed globally each year. Surgical resection, radiotherapy, hormone therapy, and chemotherapy are the main methods for treating prostate cancer. However, patients may experience serious complications, iatrogenic injuries, drug toxicity, and drug resistance. For example, in the past few decades, androgen deprivation therapy (ADT; also known as medical castration) has been the main treatment option for advanced prostate cancer. However, the survival benefit of ADT therapy is questionable, and many patients experience a rapid increase in prostate-specific antigen (PSA) or a short PSA doubling time after treatment. Although adjuvant therapies such as taxane chemotherapy and novel anti-androgen drugs have been shown to improve overall survival, not all patients can receive adjuvant therapy due to biological, economic, or geographical limitations. Therefore, in the current clinical treatment of prostate cancer, a new type of easily available clinical treatment drug is needed.
[0003] A large number of clinical studies have found that GPx3 is lowly expressed in prostate cancer tissues, and as the malignancy of the tumor increases, the abundance of GPx3 expression decreases. At the same time, the immune infiltration of tumor tissues decreases, and the immunotherapy effect of tumor cells is poor. This suggests that GPx3 may be a potential target for the treatment of prostate cancer. Currently, there is still no study pointing out how to increase the expression of GPx3 in prostate cancer tissues or drugs. Therefore, how to increase the expression of GPx3 in prostate cancer tissues to improve the anti-tumor effect has important scientific significance.
[0004] Selenium is an essential trace element in the human body, and the anti-cancer and anti-tumor effects of selenium have been confirmed by a large number of clinical and pre-clinical research results. Selenium mainly exerts its biological functions by regulating the expression of selenoproteins. It is still unclear whether selenium can or which form of selenium can target and regulate the expression of GPx3 in prostate cancer. Therefore, developing drugs that can target and regulate the expression of GPx3 in prostate cancer is expected to improve the cancer treatment effect. Summary of the Invention
[0005] In view of the above-mentioned disadvantages of the prior art, the purpose of the present invention is to provide a new nano-selenium preparation with higher anti-tumor activity and lower toxicity against prostate cancer.
[0006] The present invention also aims to provide a preparation method of the above-mentioned new nano-selenium preparation with higher anti-tumor activity and lower toxicity against prostate cancer.
[0007] The present invention also aims to provide the application of the above novel nano-selenium preparation with higher anti-tumor activity and lower toxicity against prostate cancer in anti-tumor drugs.
[0008] The present application provides a preparation method of a nano-selenium preparation for targeted regulation of GPx3 expression, including the following steps: mixing and reacting a sodium selenite solution with an LET (lentinan) solution, water, and a VC solution (vitamin C solution) respectively, and then separating, purifying, and filtering the reaction solution to obtain the nano-selenium preparation for targeted regulation of GPx3 expression.
[0009] Specifically, the concentrations of the sodium selenite solution, the LET solution, and the VC solution are 7.5 - 10 mg / ml, 10.5 - 14.5 mg / ml, and 33 - 37 mg / ml respectively.
[0010] Specifically, the volume parts added during the mixing reaction of the sodium selenite solution, the LET solution, water, and the VC solution are 0.9 - 1.1 parts, 1.8 - 2.2 parts, 5 - 7 parts, and 0.9 - 1.1 parts respectively.
[0011] Specifically, it includes the following steps: slowly dropwise adding the LET solution to the sodium selenite solution, stirring after adding until completely mixed; slowly dropwise adding water and stirring until completely mixed; uniformly and slowly dropwise adding the VC solution, stirring after adding until completely mixed; standing and then stirring and reacting for a certain period of time; loading into a dialysis bag for dialysis, recovering into a centrifuge tube, centrifuging, and then filtering with a filter head to obtain the nano-selenium preparation for targeted regulation of GPx3 expression.
[0012] Specifically, slowly dropwise add the LET solution to the sodium selenite solution, stir for 4 - 6 minutes after adding until completely mixed; slowly dropwise add water and stir for 4 - 6 minutes until completely mixed; uniformly and slowly dropwise add the VC solution, stir for 4 - 6 minutes after adding until completely mixed; stand at 3°C - 5°C for 25 - 40 minutes, then stir and react for 8 - 12 hours; load into a dialysis bag with a molecular weight of 9000 - 11000 for dialysis, recover into a centrifuge tube, centrifuge at 2500 - 3500 rpm for 8 - 12 minutes, and then filter with a 0.15 - 0.25 μm filter head to obtain the nano-selenium preparation for targeted regulation of GPx3 expression
[0013] Specifically, it includes the following steps: slowly dropwise add the LET solution into the sodium selenite solution, stir for 5 minutes after adding until completely mixed; slowly dropwise add water and stir for 5 minutes until completely mixed; uniformly and slowly dropwise add the VC solution, stir for 5 minutes after adding until completely mixed; stand at 4°C for 30 minutes, and then stir and react for 14 hours; load into a dialysis bag with a molecular weight cut-off of 10,000, change ultrapure water every 2 hours for the first 12 hours; then change ultrapure water every 12 hours, and dialyze for a total of 3 days; recover into a centrifuge tube, centrifuge at 3000 rpm for 10 minutes, and then filter with a 0.20 μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
[0014] Specifically, it includes the following steps: slowly dropwise add 2 ml of a 12.5 mg / ml LET solution into 1 ml of an 8.64 mg / ml sodium selenite solution, stir for 5 minutes after adding until completely mixed; slowly dropwise add 6 ml of water and stir for 5 minutes until completely mixed; uniformly and slowly dropwise add 1 ml of a 35.2 mg / ml VC solution at a rate of one drop every 20 s, stir for 5 minutes after adding until completely mixed; stand at 4°C for 30 minutes, and then stir and react for 10 hours; load into a dialysis bag with a molecular weight cut-off of 7000, change ultrapure water every 2 hours for the first 12 hours; then change ultrapure water every 12 hours, and dialyze for a total of 3 days; recover into a centrifuge tube, centrifuge at 3000 rpm for 10 minutes, and then filter with a 0.20 μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
[0015] A nano-selenium preparation for targeted regulation of GPx3 expression, which is prepared by using the preparation method described in any one of the above schemes.
[0016] An application of a nano-selenium preparation for targeted regulation of GPx3 expression in anti-tumor drugs.
[0017] Specifically, the anti-tumor drug is an anti-prostate cancer drug targeting GPx3.
[0018] The improvements of this application bring the following advantages:
[0019] Nanometerization treatment: Nanometerize single-atom selenium, aiming to improve the stability and anti-tumor activity of the drug.
[0020] GPx3 targeted regulation: Utilize lentinan nano-selenium to target and regulate the expression of prostate cancer GPx3 protein, induce DNA damage in tumor cells, and thus inhibit the proliferation, migration and invasion abilities of tumor cells.
[0021] Immune enhancement and antioxidant: Combine the immune enhancement effect of lentinan and the antioxidant, anti-inflammatory, anti-apoptosis and other biological activities of selenium. Load nano-selenium with lentinan to control the metabolism and toxicity of nano-selenium in the body, and at the same time increase its anti-tumor effect. Description of the Drawings
[0022] Figure 1 A is the scanning result of transmission electron microscopy (TEM); Figure 1 B is the detection result of dynamic light scattering (DLS) of lentinan nano-selenium and nano-selenium; Figure 1 C is the detection result of dynamic light scattering (DLS) of lentinan nano-selenium for 5 consecutive days in different environments.
[0023] Figure 2 A is the protein thermal migration detection of GPx3 and lentinan nano-selenium; Figure 2 B is the best binding model of lentinan and GPx3 in 50 simulations of intermolecular interactions. Figure 2 C is the top 5 binding sites and binding energies with the highest binding frequency of lentinan and GPx3 in 50 simulations of intermolecular interactions. Figure 2 D is the detection kit for glutathione peroxidase activity reflecting the change of GPx3 activity.
[0024] Figure 3 For the detection of cell DNA damage and cell cycle. Figure 3 A shows that DNA damage occurs in PC3 cells after treatment with lentinan nano-selenium.
[0025] Figure 4 For the cell colony formation assay.
[0026] Figure 5 For the cell cycle detection.
[0027] Figure 6 For the results of cell immunofluorescence staining and co-culture. Detailed implementation manners
[0028] The following specific examples illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.
[0029] Lentinan has an immunopotentiating effect and can play its antitumor role by enhancing the phagocytosis of macrophages, activating T lymphocytes to release activation factors, and increasing the activity of NK cells. Taking lentinan during chemotherapy can effectively improve the sensitivity of tumors to chemotherapeutic drugs, enhance the efficacy of chemotherapy, reduce the toxic effects of chemotherapy, improve the physical condition of patients, and prolong their lifespan. Currently, it has been used in clinical treatment as an adjuvant therapeutic drug in combination with radiotherapy and chemotherapy drugs for various malignant tumors such as lung cancer, liver cancer, gastric cancer, and breast cancer. Selenium can participate in antioxidant reactions and protect cells from oxidative damage. In addition, selenium also has various biological activities such as anti-inflammatory and anti-apoptotic effects. These properties make selenium-containing drugs have broad application prospects in the field of tumor clinical research. Compared with general selenium compounds, nano-selenium exhibits stronger antitumor activity and is regarded as a new type of antitumor drug. By loading nano-selenium with lentinan, it is expected to control the metabolism and toxicity of nano-selenium in the body while increasing the antitumor effect of nano-selenium.
[0030] Example 1 Preparation method of a nano-selenium preparation for targeted regulation of GPx3 expression
[0031] Slowly drop 2 ml of 12.5 mg / ml LET solution into 1 ml of 8.64 mg / ml sodium selenite solution. After adding, stir for 5 minutes until completely mixed; slowly add 6 ml of water and stir for 5 minutes until completely mixed; slowly and uniformly drop 1 ml of 35.2 mg / ml VC solution at a rate of one drop every 20 s. After adding, stir for 5 minutes until completely mixed; let it stand at 4°C for 30 minutes, then stir and react for 10 hours; load it into a dialysis bag with a molecular weight cut-off of 10,000. In the first 12 hours, change ultrapure water every two hours; then change ultrapure water every 12 hours, and dialyze for a total of 3 days; recover it into a centrifuge tube, centrifuge at 3000 rpm for 10 minutes, and then filter with a 0.20 μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
[0032] Example 2 Preparation method of a nano-selenium preparation for targeted regulation of GPx3 expression
[0033] Slowly add 2 ml of 10.5 mg / ml LET solution dropwise to 1 ml of 7.5 mg / ml sodium selenite solution. After adding, stir for 4 minutes until completely mixed. Slowly add 6 ml of water and stir for 4 minutes until completely mixed. Slowly add 1 ml of 33 mg / ml VC solution dropwise at a rate of one drop every 15 s. After adding, stir for 4 minutes until completely mixed. Let it stand at 4 °C for 25 minutes, then stir and react for 8 hours. Load it into a dialysis bag with a molecular weight cut-off of 9000. In the first 10 hours, change ultrapure water every two hours. Then change ultrapure water every 10 hours, and dialyze for a total of 4 days. Recover it into a centrifuge tube, centrifuge at 2500 rpm for 12 minutes, and then filter with a 0.15 μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
[0034] Example 3 Preparation method of a nano-selenium preparation for targeted regulation of GPx3 expression
[0035] Slowly add 2 ml of 14.5 mg / ml LET solution dropwise to 1 ml of 10 mg / ml sodium selenite solution. After adding, stir for 6 minutes until completely mixed. Slowly add 6 ml of water and stir for 6 minutes until completely mixed. Slowly add 1 ml of 37 mg / ml VC solution dropwise at a rate of one drop every 25 s. After adding, stir for 6 minutes until completely mixed. Let it stand at 4 °C for 35 minutes, then stir and react for 12 hours. Load it into a dialysis bag with a molecular weight cut-off of 11000. In the first 14 hours, change ultrapure water every two hours. Then change ultrapure water every 14 hours, and dialyze for a total of 3 days. Recover it into a centrifuge tube, centrifuge at 3500 rpm for 8 minutes, and then filter with a 0.25 μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
[0036] Example 4 Characterization of a nano-selenium preparation (lentinan nano-selenium) for targeted regulation of GPx3 expression
[0037] Drop 4 - 5 drops of the drug solution onto a copper grid. After it is completely dried, it can be used for electron microscopy scanning and shooting. Under the electron microscope, it can be observed that the synthesized lentinan nano-selenium is round, with clear boundaries and uniform size, and the diameter is about 100 nm. Figure 1 B is the ZATA particle size analysis structure. Place the liquid in a special sample tube and detect the particle size and charge of the solute in the solution, and repeat the detection more than three times. Compared with the nano-selenium particles not bound to Let, the particle size of the synthesized lentinan nano-selenium is significantly lower than that of the pure nano-selenium particles, and the particle size of the synthesized lentinan nano-selenium is 100 nm. Figure 1C was also for ZATA particle size analysis. Specifically, the synthesized lentinan nano-selenium was resuspended in ultrapure water (DDH2O), PBS buffer and cell lysate, and stored at 4 °C. After sample preparation, the particle size of lentinan nano-selenium was detected at fixed time points for 5 consecutive days, with three replicates for each detection. Summarizing the particle size detection results for 5 days, it was found that lentinan nano-selenium could maintain a stable particle size in different solutes within 5 days, and there was no significant change in the particle size within 5 days.
[0038] Figure 1 The results showed that the synthesized lentinan nano-selenium was evenly distributed in the solution, with a uniform size, between 80 - 100 nm ( Figure 1 A,B), and had a stable morphology ( Figure 1 C). These results indicated that stable lentinan nano-selenium was successfully prepared in this example.
[0039] Example 5
[0040] Approximately four million cells were lysed on ice with RIPA lysis buffer, centrifuged at 10,000 rpm for 15 minutes at 4 °C, and the supernatant protein sample was aspirated. Further, the protein sample was mixed with the drug at 4 °C for 24 h, and then the sample was incubated at different temperatures for 30 min. After centrifugation at 10,000 rpm for 10 min at 4 °C, the supernatant was taken for Western blot detection. The gray value of the protein band was detected using Image J to represent the abundance of the target protein. If the protein could bind to the drug, the protein content after treatment at different temperatures would decrease. It was observed that the abundance of GPx3 decreased with the increase in the pretreatment temperature, indicating that GPx3 could bind to lentinan nano-selenium. Figure 2 B and C were for using Autodock docking software to simulate the docking of GPx3 (PDB id: 2R37) with lentinan (C 42 H 70 O 35 ). Specifically, Figure B showed the binding model with the lowest binding energy among 50 simulated dockings. It was observed that the GPx3 protein could form a binding pocket at the positions of D191, H199, and Y214 and bind to lentinan. Figure C was the summary of the binding sites of GPx3 and lentinan in 50 docking models among 50 docking simulations. It was found that the binding frequencies of 5 sites (GLY61, ASP191, TYR214, ILE195, PHE92) were the highest. It was considered that under natural conditions, lentinan would bind to these 5 amino acid residues of GPx3. Figure 2D shows the change in the enzyme activity of GPx3 protein after treatment with lentinan nano-selenium using a GPx3 protease activity detection kit (CAS: JN24284). Specifically, the culture medium of PC3 cells was taken for detection. Lentinan nano-selenium was added to the aliquoted culture medium in reverse chronological order (0, 3, 6, 12, 24 h) at a ratio of 1:100. According to the kit instructions, the O.D. value (absorbance) of different samples was measured at a wavelength of 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the sample concentration was calculated. The results showed that the enzyme activity of GPx3 gradually increased after treatment with lentinan nano-selenium.
[0041] Figure 2 The results showed that in PC3 cells, lentinan nano-selenium could bind to and increase the expression of GPx3 protein and improve the enzyme activity of the protein ( Figure 2 A, B, C), and the binding energy between nano-selenium and GPx3 was strong ( Figure 2 D).
[0042] Example 6
[0043] After electrophoresis and staining of cells treated with 15 μM lentinan nano-selenium for 24 h and cells without drug treatment according to the comet assay protocol, the cell morphology was observed under a fluorescence microscope. It was observed that the control group cells were round and had no tail, indicating that the control group cells had no DNA damage, but the drug-treated group cells were spindle-shaped and had an obvious tail, indicating that the cells had DNA damage. Figure 3 B Cell cycle detection. Specifically, 50,000 cells from different groups were fixed, hydrated, and stained with DCF probe, and then the DNA content and cell cycle of each cell were detected by flow cytometry. It was observed that as the concentration of the treatment drug increased, the proportion of cells in the G2 / M phase per 10,000 cells increased, and the increase was significant, indicating that lentinan nano-selenium could cause DNA damage to cells ( Figure 3 A) and lead to cell cycle arrest in the G2 / M phase.
[0044] Example 7
[0045] Figure 4 A Specifically, a fixed number (2000) of cells were cultured in a culture dish, and different concentrations of drugs were added to different dishes. After 14 days, the dishes were photographed and the number of cell clusters in the dishes was calculated to quantitatively analyze the effect of the drug on cell proliferation ability. It was observed that as the concentration of the treatment drug increased, the number of cell clusters in the dishes gradually decreased, indicating that lentinan nano-selenium could weaken the cell proliferation ability. Figure 4 B shows the summary results of three repeated cloning experiments. It was observed that the cell proliferation ability of cells treated with 15 μM was significantly lower than that of the control group.
[0046] Figure 4It is shown that lentinan nano-selenium can affect the proliferation viability of tumor cells.
[0047] Example 8
[0048] Figure 5 Specifically for group A, 50,000 cells were taken from different groups respectively. After the processes of fixation, hydration and DCF probe staining, the DNA content and the cell cycle stage of each cell were detected by flow cytometry. It was observed that for the cells pretreated with GCK733 (an ATM inhibitor), after treatment with lentinan nano-selenium, the cells in G2 / M phase did not increase with the increase of drug concentration, and the content of cells in G2 / M phase was lower than that of the control group, indicating that GCK733 can reverse the therapeutic effect of lentinan nano-selenium. Figure 5 Specifically for B, it is a summary of 3-cycle detections. It was observed that GCK733 can weaken the effect of lentinan nano-selenium on the tumor cell cycle, and the proportion of cells in G2 / M phase in the drug treatment group was lower than that of the control group.
[0049] Figure 5 It is shown that the ATM inhibitor can weaken the effect of nano-selenium in treating prostate cancer, indicating that after lentinan nano-targetedly regulates GPx3, it affects DNA repair through the ATM protein.
[0050] Example 9
[0051] Figure 6 Specifically for A, clinical sections of prostate cancer tissues and normal prostate tissues were stained. The cell nuclei were blue, PD-L1 was green, and GPx3 was red. It was observed that the expression of PD-L1 was low and the expression of GPx3 was high in normal tissues, and this trend was opposite in prostate cancer tissues. It is considered that the expression of GPx3 is negatively correlated with PD-L1. Figure 6 Specifically for B, 400,000 PC3 cells and 1.2 million Jurkat cells were co-cultured in a 48-well plate, and different concentrations of drugs were added during the process. It was observed that the number of dead tumor cells in the group of immune cells Jurkat treated with lentinan nano-selenium was more, and with the increase of the concentration of lentinan nano-selenium, the killing ability of immune cells Jurkat against tumor cells increased. It is shown that lentinan nano-selenium can enhance the effect of tumor immunotherapy.
[0052] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.
Claims
1. Application of a nano-selenium preparation for targeted regulation of GPx3 expression in anti-tumor drugs.
2. According to the use of claim 1, the nano-selenium preparation achieves the anti-cancer effect by targeted upregulation of GPx3 protein expression.
3. The use according to claim 1, wherein the anti-tumor drug is an anti-prostate cancer drug.
4. A method for preparing a nano-selenium preparation for targeted regulation of GPx3 expression, characterized in that: The following steps are involved: The sodium selenite solution is mixed with the LET solution, water and VC solution respectively for reaction, and then the reaction solution is separated and purified, and filtered to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
5. The preparation method according to claim 4, characterized in that: The concentrations of the sodium selenite solution, the LET solution and the VC solution are 7.5-10 mg / ml, 10.5-14.5 mg / ml and 33-37 mg / ml respectively.
6. The preparation method according to claim 4, characterized in that: The volume portions of the sodium selenite solution, the LET solution, water and the VC solution added during the mixed reaction are 0.9-1.1 parts, 1.8-2.2 parts, 5-7 parts and 0.9-1.1 parts respectively.
7. The preparation method according to claim 4, characterized in that: The following steps are involved: Slowly add LET solution to the sodium selenite solution, stir after adding until completely mixed; slowly add water and stir until completely mixed; slowly add VC solution at a uniform speed, stir after adding until completely mixed; stir and react for a certain period of time after standing; put into a dialysis bag for dialyzing, recover into a centrifuge tube, centrifuge, and then filter with a filter to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
8. The preparation method according to claim 4, characterized in that: The following steps are involved: Slowly add LET solution to the sodium selenite solution, stir for 4-6 minutes after adding until completely mixed; slowly add water and stir for 4-6 minutes until completely mixed; slowly add VC solution at a uniform speed, one drop every 15-25 seconds, stir for 4-6 minutes after adding until completely mixed; let stand at 3℃-5℃ for 25-40 minutes, then stir and react for 8-12 hours; put into a dialysis bag with a molecular weight of 9000-11000 for dialyzing, recover into a centrifuge tube, centrifuge at 2500-3500rpm for 8-12 minutes, and then filter with a 0.15-0.25μm filter head to obtain a nano-selenium preparation for targeted regulation of GPx3 expression.
9. The preparation method according to claim 8, characterized in that: The following steps are involved: Slowly add 2-4 ml of 12.5-25 mg / ml LET solution to 1-2 ml of 8.64-17.28 mg / ml sodium selenite solution, stir for 3-5 minutes after adding, until completely mixed; slowly add 4-8 ml of water and stir for 5-10 minutes until completely mixed; slowly add 1-2 ml of 70.4 mg / ml VC solution at a rate of one drop per 15-25 seconds, stir for 5-10 minutes after adding, until completely mixed; 3 After standing at -5°C for 20-60 minutes, stir and react for 8-12 hours; put into dialysis bag with molecular weight of 9000-11000 for dialyzing, replace ultrapure water every 1.5-3 hours for the first 10-14 hours; replace ultrapure water every 10-14 hours thereafter, and the total dialysis is 3-4 days; recover into centrifuge tube, centrifuge at 2500-3500rpm for 8-12 minutes, and filter with 0.15-0.20μm filter head to obtain nano-selenium preparation for targeted regulation of GPx3 expression.
10. A nano-selenium preparation for targeted regulation of GPx3 expression, characterized in that: Prepared according to the preparation method according to any one of claims 4 to 9.