Carbon-based nano material for relieving ischemia reperfusion injury

Abstract

The invention discloses a carbon-based nano material for relieving ischemia reperfusion injury, and relates to the technical field of medical chemistry application. The carbon-based nano material comprises an active component and a pharmaceutically acceptable carrier, the active component is at least one of fullerene, fullerol, carbon quantum dots, carbon fibers, carbon nanotubes, carbon nanowires and graphene; the pharmaceutically acceptable carrier is at least one of water, cellulose, acrylic resin, polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, a polyoxyethylene-polyoxypropylene copolymer and a derivative thereof, hyaluronic acid and a derivative thereof, alginic acid and a derivative thereof, and chitosan and a derivative thereof. The invention also provides a preparation method and application of the carbon-based nano material. By utilizing the pH response characteristic of the alginate microspheres, the fullerol is accurately released in the small intestine, and meanwhile, by virtue of the mucosa adhesion property of the chitosan, the retention time of the fullerol in the small intestine is prolonged, and the slow release of the medicine is realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, and particularly relates to a carbon-based nanomaterial for reducing ischemia-reperfusion injury. BACKGROUND

[0002] Ischemia reperfusion injury (IRI) is a phenomenon in which reperfusion of blood to ischemic tissue or organ causes more severe damage than ischemia itself. Intestinal ischemia reperfusion injury (IIRI) is a common clinical condition in which the intestinal tissue is damaged after a certain period of ischemia followed by reperfusion of blood. It often occurs after extracorporeal circulation, small bowel transplantation, hemorrhagic shock, trauma, strangulated intestinal obstruction, and acute mesenteric ischemia. The main pathological features of IIRI are damage to the intestinal mucosa and severe dysfunction of the intestinal barrier. Specifically, this damage can cause extensive loss of intestinal villi, epithelial cell necrosis, destruction of the lamina propria structure, and reduced intestinal peristalsis. In addition, IIRI can also cause intestinal bleeding and ulceration. However, the real threat of IIRI is not limited to local intestinal damage, but also affects distant non-ischemic organs, damaging their integrity and function. In severe cases, IIRI can trigger systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS), which can ultimately lead to patient death. It is estimated that IIRI accounts for 30-40% of total hospital mortality. Although the specific mechanisms of IIRI have not been fully elucidated, current research suggests that oxidative stress, calcium overload, excessive activation of the inflammatory response, and intestinal flora translocation are the main factors leading to tissue cell damage. ROS plays a key role in IIRI, and its highly reactive nature allows it to undergo oxidation reactions with membrane phospholipids, proteins, and nucleic acids, leading to cell structure damage and dysfunction, and even triggering cell necrosis or apoptosis. It is well known that intestinal flora imbalance plays an important role in the occurrence and development of human diseases. In the case of IIRI, intestinal mucosal epithelial cells swell, and cell tight junctions are disrupted, leading to impaired intestinal barrier function and increased permeability. This allows bacteria or enterotoxins to be transferred from the intestine to lymph nodes and blood, and to continuously produce and activate inflammatory mediators. If the degree of ischemia is severe or the ischemic mesenteric tissue area is extensive, it may lead to sepsis and multiple organ failure. In the etiological analysis of IIRI, the excessive accumulation of ROS and the resulting oxidative stress damage are considered to be one of the main factors leading to pathological changes. Therefore, inhibiting the so-called 'free radical storm' is a key strategy for reducing IIRI. In previous studies, antioxidants have been shown to effectively improve the symptoms of IIRI, such as superoxide dismutase (SOD), reduced glutathione (GSH), vitamin E, ascorbic acid, melatonin, etc. However, due to their own antioxidant mechanisms and the complexity of the intestinal environment, the therapeutic effect of traditional antioxidants has been limited.First, most antioxidant molecules have limited ability to scavenge ROS, and cannot fully cope with the various ROS produced in the IIRI reaction. For example, SOD has no significant scavenging effect on other ROS except superoxide anion (O2. ·- )Second, most antioxidants are unstable and easily oxidized by the surrounding environment, resulting in a significant decrease in antioxidant effect before reaching the target site. Finally, the complexity of the intestinal environment, including the variable pH environment, the erosion of digestive enzymes, and the flushing of intestinal contents (including food, intestinal juice, intestinal flora, and their metabolites), limits the application of antioxidants. Most studies show that IIRI is administered by intravenous injection for systemic administration or intraperitoneal injection for local administration, but these methods have poor targeting and it is difficult to achieve effective drug concentration at the injury site, resulting in unsatisfactory treatment effect.

[0003] Based on the above problems, enhancing the free radical scavenging ability of the antioxidant system is one of the main defense means for preventing and reducing intestinal ischemia-reperfusion injury, and antioxidants are often supplemented to reduce oxidative stress or up-regulate antioxidant capacity. Carbon nanomaterials can become ideal ROS scavengers due to their excellent ROS scavenging ability, high antioxidant performance, biological safety, and biodegradability. Among these materials, fullerols, as fullerene derivatives, are particularly attractive, and are hailed as "free radical sponges". This is due to their rich delocalized conjugated double pi bond structure, which gives them extensive ROS scavenging ability and plays an important role in various oxidative stress injury scenarios. It not only can enhance the antioxidant capacity of cells by scavenging ROS and activating the Nrf2 / HO-1 signaling pathway, but also can alleviate colonic radiation damage by scavenging ROS and regulate intestinal flora structure to promote the recovery of colonic barrier function. In summary, nanometer fullerols have good biocompatibility, can efficiently and stably scavenge ROS, and can maintain their chemical structure and function in the gastrointestinal tract with varying pH and abundant digestive enzymes. These characteristics make fullerols have great application potential in the treatment of oxidative stress-related diseases, especially IRI. SUMMARY

[0004] To solve the above technical problems, the purpose of the present application is to provide a carbon-based nanomaterial for reducing ischemia-reperfusion injury, which utilizes the pH response characteristics of alginate microspheres to achieve precise release of fullerols in the small intestine, and at the same time, utilizes the mucosal adhesion properties of chitosan to prolong the retention time of fullerols in the small intestine, achieving sustained release of the drug.

[0005] The technical solution of the present application to solve the above technical problems is as follows: a carbon-based nanomaterial for reducing ischemia-reperfusion injury is provided, which comprises an active ingredient and a pharmaceutically acceptable carrier.

[0006] The active component is at least one of fullerene, fullerenol, carbon quantum dots, carbon fiber, carbon nanotube, carbon nanowire and graphene;

[0007] The pharmaceutically acceptable carrier is at least one of water, cellulose, acrylic resin, polyvinylpyrrolidone (such as polyvinylpyrrolidone K30, polyvinylpyrrolidone K90), polyvinyl alcohol, polyethylene glycol, polyoxyethylene polyoxypropylene copolymer and its derivatives (such as poloxamer), hyaluronic acid and its derivatives, alginic acid and its derivatives, and chitosan and its derivatives.

[0008] Further, the carbon-based nanomaterial is a nanomaterial mainly composed of carbon, which can be divided into zero-dimensional carbon nanomaterial, one-dimensional carbon nanomaterial, two-dimensional carbon nanomaterial and three-dimensional carbon nanomaterial according to dimensions; the zero-dimensional carbon nanomaterial is carbon quantum dots, fullerene and fullerenol; the one-dimensional carbon nanomaterial is carbon fiber, carbon nanotube and carbon nanowire; the two-dimensional carbon nanomaterial is graphene; and the three-dimensional carbon nanomaterial is also called bulk material, which is specifically various three-dimensional intrinsic or composite systems.

[0009] Further, the size of the carbon-based nanomaterial for alleviating ischemia-reperfusion injury is 1-500 nm.

[0010] Further, the carbon-based nanomaterial for alleviating ischemia-reperfusion injury is sodium alginate-chitosan nanogel microspheres FOH@SA-CS MS loaded with FOH, which is targeted to deliver to the target position of ischemia-reperfusion injury.

[0011] The application also provides application of the above-mentioned carbon-based nanomaterial for alleviating ischemia-reperfusion injury in preparation of a drug for ischemia-reperfusion injury.

[0012] Further, the ischemia-reperfusion injury is at least one of cerebral ischemia-reperfusion injury, myocardial ischemia-reperfusion injury, intestinal ischemia-reperfusion injury, renal ischemia-reperfusion injury and skeletal muscle ischemia-reperfusion injury.

[0013] The application also provides a preparation method of the above-mentioned carbon-based nanomaterial for alleviating ischemia-reperfusion injury, which comprises the following steps: taking fullerene C 60 as a raw material, adding sodium hydroxide and hydrogen peroxide respectively to realize hydroxylation of C 60 under the assistance of a mechanical chemical method, synthesizing nanometer fullerenol FOH with a hydroxyl number of 20-24, and then constructing SA-CS nanogel microspheres FOH@SA-CS MS based on the nanometer fullerenol FOH by using airflow control technology and ionic crosslinking method, i.e. the carbon-based nanomaterial for alleviating ischemia-reperfusion injury.

[0014] Further, the preparation method specifically comprises the following steps:

[0015] (1) adding fullerene C60 The sodium hydroxide is added to the fullerene C

[0016] (2) The sodium alginate solution and the nano-fullerol FOH are mixed, and then the calcium chloride solution is injected to form gelatinous crosslinked microspheres;

[0017] (3) The gelatinous crosslinked microspheres and the chitosan solution are incubated at room temperature, and then filtered and washed to obtain the carbon-based nanomaterial for reducing ischemia-reperfusion injury.

[0018] Further, in step (1), the sodium hydroxide is added to the fullerene C 60 The sodium hydroxide is added to the fullerene C 60 The fullerene C

[0019] Further, in step (2), the sodium alginate solution with a concentration of 2-5 wt% and the nano-fullerol FOH are mixed at a mass-volume ratio of 100-200:10 mg / mL, the mixed solution is injected into the calcium chloride solution with a concentration of 0.2-0.3 M using a coaxial needle under the accompaniment of a high-speed nitrogen gas flow of 1-8 L / min, the distance between the needle head and the liquid surface of the calcium chloride solution is 5-15 cm, and the needle head aperture is 0.16-0.6 mm, so as to form gelatinous crosslinked microspheres.

[0020] Further, in step (3), the gelatinous crosslinked microspheres and the chitosan solution with a concentration of 5-10 wt% are incubated at room temperature, and then filtered and washed with the PBS solution.

[0021] The present application has the following beneficial effects:

[0022] 1. The present application utilizes the pH response characteristics of the alginate microspheres to achieve precise release of the fullerol in the small intestine, and simultaneously utilizes the mucosal adhesion properties of chitosan to prolong the retention time of the fullerol in the small intestine, so as to achieve drug sustained release, the fullerol can more efficiently and precisely target the small intestine, effectively play the role of scavenging reactive oxygen species (ROS) and relieving ischemia-reperfusion injury (IIRI), and has high biological safety, effectiveness and stability.

[0023] 2、The carbon-based nanomaterial for alleviating ischemia-reperfusion injury has the following technical advantages: the use of fullerenol for efficient ROS removal reduces oxidative stress damage caused by ROS storm; precise control of drug release location and rate improves treatment effect and reduces side effects; the use of the mucosal adhesion of chitosan enhances the retention time of the drug at the action site, thereby improving the therapeutic effect; the carbon-based nanomaterial design takes into account the bioavailability and stability of the drug, ensuring the effectiveness and safety of the drug.

[0024] 3、The nanogel microsphere system of the application integrates intestinal targeting, adhesion and antioxidant properties, and is aimed at the "ROS storm" caused by ischemia-reperfusion and the oxidative stress damage caused thereby. FOH, a carbon nanomaterial, is used as the core due to its efficient and stable ROS removal capacity, combined with the pH responsiveness of sodium alginate and the mucosal adhesion properties of chitosan, to develop an oral nanogel microsphere suitable for treating IIRI. This innovative nanogel microsphere will achieve precise targeted drug delivery in the intestinal tract, enhance the retention effect and bioavailability of fullerenol in the intestinal tract, thereby effectively reducing intestinal mucosal damage, reducing the accompanying inflammatory response, and regulating intestinal flora imbalance, providing a new approach and method for the treatment of IIRI.

[0025] 4、The synthesis strategy, hydroxylation level and water solubility of nanometer FOH, and other key parameters will directly affect its free radical scavenging efficiency. Further, the performance of FOH@SA-CS nanogel microspheres based on nanometer FOH in the drug delivery system is also influenced by various physicochemical properties such as chemical composition, structure design, particle size and surface modification. These properties together determine the way and effect of the system interacting with the gastrointestinal tract. Therefore, studying the structure-activity relationship between the physical and chemical properties of these micro-nano materials and the treatment effect of IIRI aims to provide scientific guidance and theoretical basis for the optimization of material properties, the expansion of application range and the development of new application fields.

[0026] 5、The application utilizes the efficient and stable ROS scavenging properties of carbon nanomaterial FOH, organically combines the pH responsiveness of alginate SA and the mucosal adhesion of the positively charged CS shell, and constructs an oral nanogel microsphere suitable for IIRI, which helps to achieve efficient targeted drug delivery in the intestinal tract, thereby improving the retention and bioavailability of nanometer FOH in the intestinal tract, to relieve intestinal mucosal damage and the accompanying inflammatory response and intestinal flora disorder, and is expected to overcome many problems that restrict oral administration of IIRI, and promote the development of IIRI microsphere drugs. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1Figure for FOH nanoparticle and FOH@SA-CS nanogel microsphere IIRI treatment effect;

[0028] Figure 2 Process for establishing mouse IIRI model;

[0029] Figure 3 H&E staining results of small intestine tissue;

[0030] Figure 4 Biological safety evaluation results of FOH@SA-CS nanogel microspheres. DETAILED DESCRIPTION

[0031] The principles and features of the present application are described below, and the examples are only used to explain the present application, and are not used to limit the scope of the present application. If the specific conditions are not indicated in the examples, the conventional conditions or the conditions recommended by the manufacturer are used. If the reagents or instruments used are not indicated by the manufacturer, they are all conventional products that can be purchased on the market.

[0032] Example 1

[0033] A carbon-based nanomaterial for reducing ischemia-reperfusion injury, a preparation method thereof, comprising the following steps:

[0034] (1) Fullerene C 60 was added with sodium hydroxide at a mass ratio of 10:1, placed in a agate jar and ball milled at 150-450 r / min for 1 h, then hydrogen peroxide was added and ball milled at 300 r / min for 1 h, the molar ratio of fullerene C 60 and hydrogen peroxide was 0.139:2, 1 mg of product was dissolved in 1 mL of water, hydrolyzed with normal temperature water, and centrifuged to remove water-insoluble substances, dialyzed in deionized water for 3 days, and freeze-dried to obtain nanofullerol FOH with a hydroxyl number of 20-24;

[0035] (2) A 3wt% sodium alginate solution and nanofullerol FOH were mixed at a mass volume ratio of 150:10 mg / mL, and the mixed solution was injected into a 0.2-0.3 M calcium chloride solution using a coaxial needle under the accompaniment of a high-speed nitrogen gas flow of 1-8 L / min, the needle distance from the calcium chloride solution surface was 5-15 cm, and the needle aperture was 0.16-0.6 mm, forming gel-like cross-linked microspheres;

[0036] (3) The gel-like cross-linked microspheres and a 8wt% chitosan solution were incubated at room temperature, filtered, and washed with PBS solution to obtain a carbon-based nanomaterial for reducing ischemia-reperfusion injury.

[0037] Test Example 1

[0038] The carbon-based nanomaterial obtained in Example 1 for reducing ischemia-reperfusion injury was used for cell ROS level detection, and the process was as follows: IEC-6 cells were planted in a 24-well plate with a climbing sheet, 5-10 x 10 4 cells per well. According to different groups, the cells were treated as "cell grouping". Then the cells were treated with Hoechst33342 fluorescent solution (1:100) and 2', 7'-dichlorofluorescin diacetate (DCFH-DA) probe (1:1000), and incubated at 37°C in the dark for 20 min. PBS was washed for 3 times. Finally, the fluorescence image was observed using an upright fluorescence microscope.

[0039] The above results are shown in Figure 1 . Among them, Figure 1 , A is the ROS scavenging effect diagram of nanometer FOH in cells, H / R refers to hypoxia / reoxygenation treatment, and the scale is 100 μm; B is the fluorescence quantitative analysis of ROS scavenging by nanometer FOH.

[0040] It can be seen from Figure 1 that at the cellular level, FOH nanoparticles have the effect of reducing excess ROS and DNA damage induced by hypoxia / reoxygenation (H / R); incubation of FOH nanoparticles can significantly reduce the DCF level, indicating effective ROS scavenging.

[0041] Test Example 2

[0042] According to the process shown in Figure 3 , a mouse IIRI model was established, and the process was as follows: anesthesia fixation, separation of superior mesenteric artery, clamping of superior mesenteric artery, release of blood vessel clamp and reperfusion; after the model was successfully constructed, the mice were sacrificed, and the specimens were collected. The same intestinal segment (5 cm from the distal ileum) of the small intestine was selected, and the intestinal tissue with a length of 2 cm was cut. First, the intestinal contents were thoroughly washed, and then the intestinal tissue was fixed with 4% paraformaldehyde solution for 24 hours.

[0043] After fixation, the intestinal tissue was prepared into paraffin sections with a thickness of about 5 microns, and H&E staining was performed for subsequent histological observation. At the same time, fecal samples were taken from the cecum, stored in 1.5 milliliter dry sterile EP tubes, and placed in a -80°C refrigerator for frozen storage for subsequent experimental use. The results of FOH@SA-CS nanogel microspheres reducing IIRI (H&E staining of small intestinal tissue) are shown in Figure 3 ; wherein the scale is 50 μm. Black arrow: broken and shed intestinal villi; red arrow: bleeding; green arrow: intestinal submucosal space.

[0044] It can be seen from Figure 3It can be seen that FOH@SA-CS nanogel microspheres have the effect of reducing IIRI injury.

[0045] Test Example 3

[0046] The product obtained in Example 1 was used for biological safety detection, and the process was as follows:

[0047] Three healthy female C57BL / 6 mice aged 6 to 8 weeks were selected to evaluate the biological safety of fullerol microspheres. During the experiment, the mice were raised under controlled environmental conditions, with a temperature maintained at 25±2℃, a relative humidity controlled at 50±15%, and a normal light cycle (12 hours light / 12 hours dark). The mice were free to eat standard pellet feed and tap water, and were given fullerol@alginate chitosan microspheres by oral administration (dose of 20 mg / kg), continuously for one week. During this period, the behavior and physiological state of the mice were closely monitored and recorded. After the experiment, the mice were euthanized, and then the heart, liver, spleen, lung, kidney and intestinal organs were collected and fixed with 4% formaldehyde. These organs were then made into paraffin sections and stained with hematoxylin-eosin (H&E). Finally, the tissue sections were imaged and analyzed using a fluorescence inverted microscope to evaluate the effect of fullerol microspheres on the mouse organs. The results are shown in Figure 4 .

[0048] From Figure 4 It can be seen that, within the conventional dose range, fullerol has no toxic effect on cells, demonstrating excellent biological safety, laying a solid foundation for its application in the biomedical field. The daily behavior of the mice, including eating, drinking, weight and defecation, showed no abnormal changes. Histopathological section analysis also found no obvious signs of damage to the main organs of the mice; the results showed that the fullerol used not only has good biocompatibility, but also its nanoscale particle size helps it to be quickly cleared in the body, thereby avoiding the toxic side effects that may be caused by long-term accumulation, providing a solid safety guarantee for the further application of fullerol in the biomedical field.

[0049] In summary, carbon nanomaterial FOH can efficiently and stably scavenge ROS, and FOH-based oral nanogel microspheres suitable for IIRI can increase the concentration of nanoscale FOH in the intestinal tract, and be used to alleviate intestinal mucosal injury and the consequent inflammatory response and intestinal flora disorder.

[0050] The application of the FOH-loaded alginate chitosan nanoparticles of the application in IIRI proposes an innovative strategy for treating IIRI, and develops a nanogel microsphere system integrating intestinal targeting, adhesion and antioxidant properties. In combination with the progress of clinical research, the ROS storm and oxidative stress damage caused by ischemia-reperfusion are targeted, and FOH is used as a carbon nanomaterial due to its efficient and stable ROS scavenging properties, and a FOH-based oral nanogel microsphere suitable for IIRI is constructed. This system aims to increase the concentration of FOH in the intestine to effectively alleviate intestinal mucosal damage, accompanying inflammatory response and intestinal flora disorder.

[0051] The application proposes a new idea based on "spatial precision, time matching and activity matching" to break through the limitation of oral administration. By determining the target site of the drug and using nanogel microspheres to precisely deliver the drug to the target area, the project focuses on time matching, carefully designs the release kinetics of FOH, and ensures that it matches the in vivo process of the microsphere. At the same time, the project pays attention to the matching of activity, and uses the pH responsiveness of SA in the nanogel microsphere and the mucosal adhesion of the CS shell to enhance the retention and bioavailability of FOH in the intestine. This design idea is expected to overcome the difficulties of oral administration of IIRI and promote the development of IIRI microsphere drugs.

[0052] The above only describes the preferred embodiments of the application and is not intended to limit the application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the application shall be included in the protection scope of the application.

Claims

1. A carbon-based nanomaterial for mitigating ischemia-reperfusion injury, characterized in that, The active component and the pharmaceutically acceptable carrier; The active component is at least one of fullerene, fullerenol, carbon quantum dots, carbon fiber, carbon nanotube, carbon nanowire and graphene; The pharmaceutically acceptable carrier is at least one of water, cellulose, acrylic resin, polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, polyoxyethylene polyoxypropylene copolymer and its derivatives, hyaluronic acid and its derivatives, alginic acid and its derivatives and chitosan and its derivatives.

2. The carbon-based nanomaterial for mitigating ischemia-reperfusion injury of claim 1, wherein, The carbon-based nanomaterial for reducing ischemia-reperfusion injury has a size of 1-500 nm.

3. The carbon-based nanomaterial for reducing ischemia-reperfusion injury according to claim 1, wherein, The carbon-based nanomaterial for reducing ischemia-reperfusion injury is FOH@SA-CS MS, which is loaded with FOH and targets delivery to the ischemia-reperfusion injury target position.

4. Use of the carbon-based nanomaterial for reducing ischemia-reperfusion injury according to any one of claims 1-3 in the preparation of an ischemia-reperfusion injury drug.

5. The use according to claim 4, wherein the compound is ###0002### The ischemia-reperfusion injury is at least one of brain ischemia-reperfusion injury, myocardial ischemia-reperfusion injury, intestinal ischemia-reperfusion injury, renal ischemia-reperfusion injury and skeletal muscle ischemia-reperfusion injury.

6. The method of claim 1-3 for the preparation of carbon-based nanomaterials for attenuating ischemia-reperfusion injury, characterized in that, The method comprises the following steps: To take fullerene C 60 As raw material, by adding sodium hydroxide and hydrogen peroxide respectively, the hydroxylation of C 60 Is realized by mechanical chemical method, synthesis of nano fullerol FOH with 20-24 hydroxyl number, and then use airflow control technology and ion crosslinking method to construct SA-CS nanogel microspheres FOH@SA-CS MS based on nano fullerol FOH, namely carbon-based nanomaterial for reducing ischemia-reperfusion injury.

7. The method for preparing carbon-based nanomaterials for mitigating ischemia-reperfusion injury as described in claim 6, characterized in that, The method comprises the following steps: (1) Fullerene C 60 hydrogen peroxide ball-milling, hydrolysis, centrifugation, dialysis and freeze-drying, to obtain nano-fullerols FOH. (2) mixing the sodium alginate solution and the nanometer fullerenol FOH, then injecting into the calcium chloride solution to form gel-like cross-linked microspheres; (3) incubating the gel-like cross-linked microspheres and the chitosan solution at room temperature, filtering and washing to obtain the carbon-based nanomaterial for reducing ischemia-reperfusion injury.

8. The method for preparing carbon-based nanomaterials for mitigating ischemia-reperfusion injury as described in claim 7, characterized in that, In step (1), to fullerene C 60 was added sodium hydroxide at a mass ratio of 10:1, placed in a agate jar and ball-milled at 150-450 r / min for 1 h, then hydrogen peroxide was added and ball-milled at 150-450 r / min for 1 h, and fullerene C 60 was hydrolyzed with water at room temperature at a ratio of 1 mg product to 1 mL water, and the water-insoluble substance was removed by centrifugation, dialyzed in deionized water for 3 d, and freeze-dried to obtain nanometer fullerol FOH with a hydroxyl number of 20-24.

9. The method for preparing carbon-based nanomaterials for mitigating ischemia-reperfusion injury as described in claim 7, characterized in that, In step (2), the sodium alginate solution with a concentration of 2-5 wt% and the nanometer fullerenol FOH are mixed in a mass-volume ratio of 100-200:10 mg / mL, the mixed solution is injected into the calcium chloride solution with a concentration of 0.2-0.3 M under the accompaniment of high-speed nitrogen gas flow of 1-8 L / min, the needle distance from the calcium chloride solution is 5-15 cm, and the needle aperture is 0.16-0.6 mm, to form gel-like cross-linked microspheres.

10. The method for preparing carbon-based nanomaterials for mitigating ischemia-reperfusion injury as described in claim 7, characterized in that, In step (3), the gel-like cross-linked microspheres and the chitosan solution with a concentration of 5-10 wt% are incubated at room temperature, filtered and washed with PBS solution.

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