Application of RuO2 nano-enzyme in preparation of medicine for treating osteoarthritis

Abstract

The invention relates to the field of osteoarthritis drug treatment, in particular to application of RuO2 nano-enzyme in preparation of a drug for treating osteoarthritis, and particularly relates to knee osteoarthritis. The RuO2 nano-enzyme disclosed by the invention is prepared according to the following method: carrying out incubation deposition on Cu2O sphere nano-particles and RuCl3 to obtain Cu2O and RuO2, and etching Cu2O and RuO2 spheres to obtain the RuO2 nano-enzyme. The RuO2 nano-enzyme provided by the invention is a hollow RuO2 nano-enzyme sphere, and after the RuO2 nano-enzyme is horizontally used in an animal body for intervention, the hollow RuO2 nano-enzyme sphere can be used as a hollow RuO2 nano-enzyme sphere; the RuO2 nano-enzyme can effectively delay the development of knee osteoarthritis of a mouse knee osteoarthritis model caused by medial meniscus instability (DMM), improve the pain condition and movement function of a DMM modeling mouse, reduce the gonarthromeningitis degree and relieve knee joint cartilage degeneration and injury, and the effectiveness of the RuO2 nano-enzyme in delaying the knee osteoarthritis disease is further proved. The invention provides a novel method for treating knee osteoarthritis, and has strong practicability and application prospect.

Description

Technical Field

[0001] The present invention relates to the field of osteoarthritis drug treatment, and in particular to the application of RuO2 nanozyme in the preparation of drugs for treating osteoarthritis. Background Art

[0002] Osteoarthritis (OA) is a chronic joint disease characterized by degenerative changes in articular cartilage and secondary bone hyperplasia, affecting 7% of the world's population. More than 500 million people suffer from OA worldwide, causing a huge social and economic burden. OA lesions often involve articular cartilage, subchondral bone, synovium, joint capsule, and surrounding muscles. Cartilage degeneration, osteophyte formation, and chronic inflammatory response are the three typical pathological processes of OA. At present, the treatment of OA mainly adopts conservative treatment to relieve symptoms. Drug treatment includes non-steroidal anti-inflammatory drugs, chondroprotective agents, joint cavity injection lubricants, hormones, etc. For end-stage OA, artificial joint replacement is used to improve the quality of life. However, for early and middle-stage patients, there is still no effective way to reverse or delay the progression of OA. At present, with the rapid development of nanomaterial technology, the application of nanomaterials in the treatment of osteoarthritis provides a new way to treat OA.

[0003] Nanozymes are nanomaterials with intrinsic enzyme-like activity that can effectively catalyze enzyme substrates and exhibit catalytic efficiency and enzyme reaction kinetics similar to those of natural enzymes. Compared with ordinary biological enzymes, nanozymes have advantages such as higher catalytic stability, adjustable activity, larger surface area, and can be used as functional carriers for cell internalization and absorption. Some nanozymes have pseudo-superoxide dismutase activity and pseudo-catalase activity, which can regulate reactive oxygen in organisms. Due to their unique physical, chemical and biological properties, coupled with the advantages of large-scale production, convenient transportation and storage, nanozymes are widely used in medicine, chemical industry, food, environment and other fields.

[0004] Nanozyme RuO2 is a new type of ruthenium-based oxygen compound with good biological activity and biosafety. Studies have shown that ruthenium nanoparticles show significant therapeutic effects in animal models of myocardial ischemia-reperfusion injury, liver injury and acute kidney injury, but it has not yet been used in the preparation of drugs for the treatment of osteoarthritis. Summary of the invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to provide an application of RuO2 nanozyme in the preparation of drugs for the treatment of osteoarthritis in view of the shortcomings of the prior art. The present invention discovers for the first time that nanozyme RuO2 has significant effects on relieving synovitis, delaying cartilage degeneration and improving osteoarthritis joint pain and motor function.

[0006] To achieve the above purpose, the technical solution provided by the present invention is:

[0007] The present invention provides the use of RuO2 nanozyme in preparing a drug for treating osteoarthritis.

[0008] Wherein, the RuO2 nanozyme is a hollow RuO2 nanozyme sphere, and the diameter of the hollow RuO2 nanozyme sphere is 200nm.

[0009] The hollow RuO2 nanozyme sphere is formed by the aggregation of RuO2 nanozyme particles, and the diameter of the RuO2 nanozyme particles is 2-3nm.

[0010] Wherein, the osteoarthritis is knee osteoarthritis. Preferably, the RuO2 nanozyme is used in the preparation of a drug for inhibiting inflammatory response in knee osteoarthritis.

[0011] Wherein, the osteoarthritis is osteoarthritis induced by medial meniscus instability.

[0012] Among them, the RuO2 nanozyme alleviates cartilage damage and degeneration, reduces synovial hyperplasia, improves motor function and relieves joint pain.

[0013] Wherein, RuO2 nanozyme is used as an active ingredient in combination with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to prepare a drug for treating osteoarthritis in any dosage form. Preferably, the drug is an injection preparation composed of nanozyme RuO2 dissolved in water for injection, that is, nanozyme RuO2 is added to the drug for treating osteoarthritis or prepared into an injection preparation to treat osteoarthritis.

[0014] Wherein, in the drug, the concentration of RuO2 nanozyme is 1-25 μg / g. Preferably, when the drug is an injection preparation, the concentration of RuO2 nanozyme is 25 μg / mL.

[0015] Wherein, the RuO2 nanozyme is prepared according to the following steps:

[0016] (1) Preparation of Cu2O spherical nanoparticles;

[0017] (2) incubating and depositing the Cu2O spherical nanoparticles with RuCl3 to obtain Cu2O@RuO2 spheres;

[0018] (3) Etching the Cu2O@RuO2 spheres and centrifuging to obtain RuO2 nanozymes.

[0019] Wherein, in step (2), the mass ratio of the Cu2O spherical nanoparticles to the RuCl3 is 1:1-2; the product obtained after incubation and deposition is centrifuged, dispersed in water, and heated at 70-90°C for 20-50min; preferably, the mass ratio of the Cu2O spherical nanoparticles to the RuCl3 is 1:1.5; the product obtained after incubation and deposition is centrifuged, dispersed in water, and heated at 80°C for 30min.

[0020] In step (3), the etching solution used in the etching is an ammonia solution, and each mL of the ammonia solution is mixed with 25-35 mg of Cu2O@RuO2 spheres. Preferably, each mL of the ammonia solution is mixed with 30 mg of Cu2O@RuO2 spheres and incubated for 1 hour.

[0021] Further preferably, the Cu2O spherical nanoparticles are synthesized using polyvinyl pyrrolidone and copper nitrate. Then, Cu2O is incubated with RuCl3.xH2O at room temperature to obtain Cu2O@RuO2 spheres, and then the Cu2O@RuO2 spheres are etched with ammonium hydroxide to obtain hollow RuO2 nanozyme spheres.

[0022] Further preferably, the present invention verifies that RuO2 nanozyme has good biocompatibility and safety through in vitro and in vivo experiments. The RuO2 nanozyme preparation is then injected into the knee joint of osteoarthritis model mice by knee joint cavity injection. After 8 weeks of continuous injection treatment, the therapeutic effect is evaluated by behavioral experiments such as open field test, footstep imprint test, and Von-Frey fiber test; then the mouse knee joint is taken for Micro-CT analysis, and then the knee joint is decalcified, paraffin embedded and sectioned, and the tissue section is subjected to H&E staining, safranin fast green staining, immunohistochemistry staining and other staining experiments to evaluate the therapeutic effect of osteoarthritis.

[0023] Beneficial effects:

[0024] Compared with existing drugs for treating osteoarthritis, the beneficial effects of the present invention are:

[0025] The present invention discovers for the first time the remarkable effect of RuO2 nanozyme in treating knee osteoarthritis. The RuO2 nanozyme used in the present invention has been used in a variety of animal models such as myocardial ischemia-reperfusion injury model, acute liver and kidney injury model, and cell experiments and animal experiments have shown high biosafety; at the same time, the synthesis method of RuO2 nanozyme is mature, and intensive production can be achieved. The structure and properties of RuO2 nanozyme are stable, and the storage and transportation conditions are not demanding, which greatly reduces the production, transportation and storage costs of RuO2 nanozyme.

[0026] The present invention found that after intervention with RuO2 nanozyme at the animal level, the progression of knee osteoarthritis in a mouse knee osteoarthritis model caused by destabilization of the medial meniscus (DMM) can be effectively delayed, the pain condition and motor function of DMM modeled mice can be improved, the degree of knee synovitis can be reduced, and knee cartilage degeneration and damage can be alleviated, further confirming the effectiveness of RuO2 nanozyme in delaying knee osteoarthritis, providing a new method for the treatment of knee osteoarthritis, which has strong practicality and application prospects.

[0027] The present invention provides a dosage and frequency of RuO2 nanozyme as a drug for treating knee osteoarthritis. The method of use is to use an injection preparation for joint cavity injection, which can significantly increase the concentration and utilization rate of RuO2 nanozyme in the joint cavity and improve the therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more clear.

[0029] Figure 1 The synthesis process and characterization diagram of RuO2 nanozyme. Figure 1 A in the figure is the synthesis process diagram of RuO2 nanozyme. Figure 1 B in FIG. is a characterization diagram of RuO2 nanozyme.

[0031] Figure 2 These are the results of in vitro cell CCK-8 experiments.

[0032] Figure 3 Figure 3 shows the body weight, knee joint diameter data and Von-Frey fiber test results of each group of mice after treatment.

[0033] Figure 4 The results of the mouse footstep imprint experiment and open field experiment are shown in the figure. Figure 4 A in the figure is the result of the mouse footstep imprint experiment. Figure 4 Figure B shows the results of the open field test on mice.

[0035] Figure 5 Results of Micro-CT analysis of mouse knee joints.

[0036] Figure 6 The staining and scoring results of mouse knee joint tissue sections are shown in Figure 2. Figure 6 A in the figure is the result of H&E staining and synovitis scoring. Figure 6 B in the figure shows the results of safranin-fast green staining and osteoarthritis OARSI scoring.

[0037] Figure 7Immunohistochemical staining and statistical results of mouse knee joint tissue.

[0038] Figure 8 H&E staining of tissue sections of mouse heart, liver, spleen, lung, and kidney. DETAILED DESCRIPTION

[0039] The above contents of the present invention are further described in detail below in the form of embodiments, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following embodiments, and all technologies realized based on the above contents of the present invention belong to the scope of the present invention.

[0040] The experimental methods used in the following examples are all conventional methods unless otherwise specified, and the reagents, methods and equipment used are all conventional reagents, methods and equipment in the technical field unless otherwise specified.

[0041] Example 1 Preparation and characterization of RuO2 nanozymes

[0042] Preparation process of RuO2 nanozyme Figure 1 As shown in A in FIG. 1 , the specific steps include:

[0043] (1) Synthesis of Cu2O spheres: 0.3 g of polyvinyl pyrrolidone and 4 mmol of copper (II) nitrate trihydrate were dissolved in 50 mL of ethylene glycol; then, the solution was heated to 185°C and maintained for 30 min under vigorous magnetic stirring; the resulting product was collected by centrifugation and washed successively with ethanol and distilled water to obtain Cu2O spherical nanoparticles.

[0044] (2) Synthesis of Cu2O@RuO2 spheres: 50 mg of the product (Cu2O) synthesized in step (1) was incubated with 75 mg of RuCl3.xH2O at room temperature to deposit RuO2 nanoparticles; after stirring for 10 min, the solution was centrifuged, and the recovered brown material was dispersed in 20 mL of water and heated at 80°C for 30 min to improve the quality of RuO2 therein.

[0045] (3) Synthesis of hollow RuO2 nanozyme spheres: The etched Cu2O@RuO2 spheres were treated with NH4OH (ammonia solution). 30 mg of the copper-containing spheres prepared in step (2) were dispersed in deionized water (5 mL), and then 1 mL of NH4OH (i.e., 25% g / mL ammonia solution) was slowly added and stirred. After incubation in ammonia solution for 1 hour, the final hollow RuO2 nanozyme spheres were collected by centrifugation and washed with water and ethanol to obtain the final product.

[0046] (4) The hollow RuO2 nanozyme sphere is formed by the aggregation of single RuO2 nanozyme particles. The diameter of a single RuO2 nanozyme particle is 2-3 nanometers. Single nanozyme RuO2 particles aggregate to form a hollow RuO2 nanozyme sphere. The diameter of the hollow nanozyme RuO2 sphere is 200 nanometers. TEM, SEM, STEM, and elemental mapping of RuO2 nanozyme are shown in Figure 1 B in.

[0047] Example 2 Preparation of RuO2 Nanozyme Injection

[0048] Weigh 25 mg of the RuO2 nanozyme prepared in Example 1, dissolve it in 75% v / v alcohol, and sterilize it for 15 minutes; centrifuge at 13000g for 5 minutes, discard the supernatant, and dry it in an oven at 60°C; sterilize with ultraviolet light for 15 minutes, add 1 mL of sterile double distilled water under sterile conditions to obtain 25 mg / mL RuO2 nanozyme injection.

[0049] Example 3 Detection of cytotoxicity of RuO2 nanozyme by CCK8 method

[0050] Mouse primary chondrocytes were extracted and cultured to the second generation, then seeded into a 96-well cell culture plate. When the cell density was >90%, RuO2 nanozymes with a concentration gradient of 0 μg / mL, 1 μg / mL, 5 μg / mL, 10 μg / mL, 25 μg / mL, 50 μg / mL, and 100 μg / mL were added to the culture medium, and the cell activity was detected with a CCK8 kit after 24 hours.

[0051] In the same way, mouse-derived macrophages (RAW264.7 cells) and mouse synovial sarcoma cells (SW982 cells) were plated in a 96-well plate, and different concentration gradients of RuO2 nanozymes (0 μg / mL, 1 μg / mL, 5 μg / mL, 10 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL) were added. After culturing for 24 hours, the cell activity was detected using the CCK8 kit.

[0052] The experimental results can be found in Figure 2 , showing the changes in cell viability after adding nanozyme RuO2 of different concentration gradients to chondrocytes, RAW264.7 cells, and SW982 cells. The results showed that after using different concentrations of nanozyme RuO2 to treat mouse-derived chondrocytes, RAW264.7 cells, and SW982 cells, there was no significant decrease in cell viability at 1μg / mL, 5μg / mL, 10μg / mL, and 25μg / mL of RuO2 nanozymes, indicating that RuO2 nanozymes have no obvious toxic effects on cells and have high biosafety.

[0053] Example 4 Establishment of DMM osteoarthritis model in mice

[0054] Modeling group: 12-week-old C57BL / 6 male mice (22-24g) were selected for the experiment. After isoflurane inhalation anesthesia, the right knee joint was routinely prepared and disinfected. The medial parapatellar ligament incision was made, the fat pad was bluntly incised, and the medial meniscus tibial ligament was found under a stereoscope. The medial meniscus tibial ligament was transected, and it was found that the medial meniscus was completely unstable and could move freely. The cartilage surface was not damaged during the operation, and the wound was sutured after thorough hemostasis.

[0055] The sham group did not cut the medial meniscus tibial ligament, and the rest of the operations were the same as those in the model group.

[0056] One week after modeling, RuO2 nanozyme injection was started for knee joint cavity injection:

[0057] The RuO2 nanozyme injection prepared in Example 2 was injected into the mouse knee joint cavity. The drug concentration was 25 μg / mL, the volume of each administration was 10 μL, the frequency of administration was once a week, and the injection was continued for 8 weeks. Mouse knee joint injection method: The mouse was anesthetized with isoflurane gas, the right knee joint of the mouse was fixed with the left hand, and the needle was held in the right hand with the knee joint gap on the inner side of the patella as the insertion point, perpendicular to the joint gap, and a sense of emptiness was felt after passing through the skin. At this time, the corresponding volume of RuO2 nanozyme injection was injected, and the needle was pulled out.

[0058] Example 5 Grouping and Dosing of Mouse DMM Osteoarthritis Model

[0059] C57BL / 6 male mice were divided into 4 groups, 8 mice in each group, namely:

[0060] Group 1—sham-operated mice injected with saline;

[0061] Group 2-sham-operated mice were injected with RuO2 nanozyme injection;

[0062] Group 3—DMM osteoarthritis model mice were injected with normal saline;

[0063] Group 4-DMM osteoarthritis model mice were injected with RuO2 nanozyme injection.

[0064] The injection was performed as described in Example 4. The concentration of RuO2 nanozyme injection was 25 μg / mL; the frequency of administration was once a week; and the administration method was knee joint cavity injection. Detailed grouping is shown in Table 1.

[0065] Table 1 Experimental grouping

[0066]

[0067]

[0068] After 8 weeks of continuous administration, the therapeutic effect was evaluated by behavioral experiments such as open field test, footstep imprint test, and Von-Frey fiber test. Then the experimental animals were euthanized, and the specimens of the right knee joint, heart, liver, spleen, lung, and kidney of the mice were obtained. Micro-CT scanning of the knee joint of the mice was performed, and the sections were sliced ​​after decalcification for histological staining. Immunohistochemical staining was used to evaluate the inflammation of the knee joint and cartilage degeneration. HE staining was performed on the sections of the heart, liver, spleen, lung, and kidney of the mice to evaluate the in vivo toxicity of RuO2 nanozyme.

[0069] Example 6 RuO2 nanozymes can relieve pain and improve motor ability in DMM osteoarthritis model mice

[0070] like Figure 3 As shown, the body weight and knee diameter of mice were measured 8 weeks after the knee joint was injected with nanozyme RuO2. The results showed that the knee joints of DMM model mice were significantly swollen; while the knee diameter of DMM model mice injected with nanozyme RuO2 was significantly lower than that of DMM model mice, indicating that nanozyme RuO2 can relieve knee swelling in osteoarthritis. Mechanical pain experiments were performed using an Electronic von Frey device (IITC, Woodland Hills, USA). The mice were placed in a cage with a metal mesh at the bottom, and fibers were used to poke the bottom of the right hind paw of the mouse from the bottom of the cage. The force was gradually increased until the mouse withdrew its right hind foot, and the threshold of the peak detector was recorded at this time for scoring. The lower the threshold when the mouse's right hind paw was withdrawn, the more sensitive the response to pain was. The results showed that the withdrawal reaction threshold of DMM model mice was reduced, and the withdrawal threshold was significantly increased after treatment with nanozyme RuO2, indicating that nanozyme RuO2 can relieve the pain sensitivity of DMM model mice.

[0071] like Figure 4 As shown in A in the figure, the gait of mice was analyzed by footstep imprinting experiment. The front paws of mice were dipped in red ink and the hind paws were dipped in blue ink, and then placed on a runway covered with white paper to let them walk straight. The mouse footprints were recorded and the relative stride length, relative step length and relative distance between the front and rear paws were analyzed. The results showed that RuO2 nanozyme can increase the relative stride length and relative step length of the DMM model mice and shorten the relative distance between the front and rear paws, indicating that RuO2 nanozyme can improve the gait of DMM model mice.

[0072] like Figure 4As shown in B, the motor function of mice was analyzed by open field test. The mice were placed in an open field and moved freely for 3 minutes. The spontaneous activity trajectory was recorded with an infrared camera, and the relative activity, distance and mean speed of the mice were analyzed. The results showed that the relative activity, distance and mean speed of DMM model mice were lower than those of the sham operation group, and the situation could be improved after treatment with RuO2 nanozyme, indicating that RuO2 nanozyme can improve the motor function of DMM model mice.

[0073] Example 7 Nanozyme RuO2 can alleviate the progression of osteoarthritis in DMM osteoarthritis model mice

[0074] like Figure 5 As shown, after 8 weeks of treatment with nanozyme RuO2, the knee joints of mice were examined by Micro-CT to analyze the osteophyte situation and subchondral bone plate (SBP). The results showed that the number of osteophytes (No.ofsteophyte) in DMM mice increased, the knee bone volume (Knee Bone volume, knee BV) and bone volume / tissue volume (BV / TV) increased, and the subchondral bone thickness (SBP thickness) and trabecular thickness (Trabecular thickness, Tb.Th) increased. The treatment group used nanozyme RuO2 to significantly reduce the number of osteophytes, knee bone volume, bone volume / tissue volume, subchondral bone thickness, and trabecular thickness in the knee joints of DMM osteoarthritis model mice, indicating that RuO2 nanozyme can reduce osteophyte formation and subchondral bone sclerosis in DMM osteoarthritis model mice.

[0075] like Figure 6 As shown in A, according to the Histopathological assessment of synovitis (modifiedfrom Krenn et al. Histopathology 2006) scoring criteria, the synovitis score was performed on the H&E staining results of the knee joints of each group of mice according to the synovial lining cell layer and cell density. The results showed that nanozyme RuO2 can reduce the infiltration of inflammatory cells in the synovium of DMM osteoarthritis model mice, reduce synovial hyperplasia, and lower the synovitis score.

[0076] like Figure 6As shown in B, the mouse knee joint tissue sections were stained with safranin-fast green, and the results showed that the cartilage thickness of the DMM osteoarthritis model mice was thinner, the chondrocytes were disordered, the cartilage surface was rough, and the cartilage layer was defective. The above conditions were improved after injection of nanozyme RuO2 treatment; according to the Osteoarthritis Research Society International (OARSI) standard for animal osteoarthritis model pathological scoring, the OARSI score of the mouse articular cartilage was evaluated by evaluating the damage of the articular cartilage. The results showed that nanozyme RuO2 can reduce the OARSI score of DMM osteoarthritis model mice, indicating that nanozyme RuO2 can alleviate cartilage degeneration and damage in DMM model mice.

[0077] like Figure 7 As shown in the figure, immunohistochemical staining was performed on the mouse knee joint tissue sections to analyze the expression of cartilage anabolic index collagen II (Collagen II, Col II), aggrecan (Aggrecan) and catabolic index matrix metallopeptidase 13 (Matrix metallopeptidase 13, MMP13) in cartilage. The results showed that compared with the Sham group, the Col II of the DMM bone joint modeling mice was significantly higher than that of the Sham group. + The area of ​​the region is significantly reduced, Aggrecan + The proportion of cells decreased, while MMP13 + The cell ratio increased, and the above trend was alleviated after the DMM bone joint model mice were treated with RuO2 nanozymes, indicating that RuO2 nanozymes can promote the anabolism of chondrocytes and inhibit the catabolism of chondrocytes, thereby alleviating cartilage degeneration.

[0078] Example 8 RuO2 nanozyme has high biosafety in vivo

[0079] like Figure 8 As shown, the tissues of the heart, liver, spleen, lung, and kidney of mice were embedded and sliced ​​for H&E staining. The staining results showed that there was no significant difference in the tissue structure morphology and inflammatory cell infiltration of the heart, liver, spleen, lung, and kidney of mice in each group, proving that RuO2 nanozyme has no obvious toxicity in vivo and has high safety.

[0080] The present invention provides a method and idea for the application of RuO2 nanozyme in the preparation of a drug for treating osteoarthritis. There are many methods and approaches to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention. All components not specified in this embodiment can be implemented using existing technologies.

Claims

1. Application of RuO2 nanozymes in the preparation of drugs for the treatment of osteoarthritis.

2. The use according to claim 1, characterized in that: The RuO2 nanozyme is a hollow RuO2 nanozyme sphere, and the diameter of the hollow RuO2 nanozyme sphere is 200nm.

3. The use according to claim 2, characterized in that: The hollow RuO2 nanozyme sphere is formed by the aggregation of RuO2 nanozyme particles, and the diameter of the RuO2 nanozyme particles is 2-3nm.

4. The use according to claim 1, characterized in that: The osteoarthritis is knee osteoarthritis.

5. The use according to claim 1, characterized in that: The osteoarthritis is osteoarthritis induced by medial meniscus instability.

6. The use according to claim 1, characterized in that: The RuO2 nanozyme alleviates cartilage damage and degeneration, reduces synovial hyperplasia, improves motor function and relieves joint pain.

7. The use according to claim 1, characterized in that: RuO2 nanozyme is used as an active ingredient in combination with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to prepare a drug for treating osteoarthritis in any dosage form.

8. The use according to claim 7, characterized in that: In the medicine, the concentration of RuO2 nanozyme is 1-25 μg / g.

9. The use according to claim 1, characterized in that: The RuO2 nanozyme is prepared according to the following steps: (1) Preparation of Cu2O spherical nanoparticles; (2) incubating and depositing the Cu2O spherical nanoparticles with RuCl3 to obtain Cu2O@RuO2 spheres; (3) Etching the Cu2O@RuO2 spheres and centrifuging to obtain RuO2 nanozymes.

10. The use according to claim 9, characterized in that: In step (2), the mass ratio of the Cu2O spherical nanoparticles to the RuCl3 is 1:1-2; the product obtained after incubation and sedimentation is centrifuged, dispersed in water, and heated at 70-90°C for 20-50min to obtain the Cu2O@RuO2 spheres; In step (3), the etching solution used in the etching is an ammonia solution, and each mL of the ammonia solution is mixed with 25-35 mg of Cu2O@RuO2 spheres.

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