Gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets, their preparation methods and applications

Abstract

The application discloses a kind of gold palladium-nitrogen doped carbon ribbon decorated g-C3N4 Nanosheet and its preparation method and application, UFR-NC is doped in g-C3N4 / To form g-C3N4 / UFR-NC composite nanomaterial;With g-C3N4 / UFR-NC composite nanomaterial as carrier, gold palladium shell core particle is loaded on g-C3N4 / UFR-NC composite nanomaterial to obtain gold palladium-nitrogen doped carbon ribbon decorated g-C3N4 Nanosheet.The gold palladium-nitrogen doped carbon ribbon decorated g-C3N4 Nanosheet of the application has multiple enzyme-like activities, good photocatalytic hydrogen production and hydrogen storage and release performance;At the same time, under the enhancement of 808 nm near-infrared light, it has multiple enzyme-like activities, high hydrogen storage and release performance.Oxidative stress damage in vivo can be reduced and neuroinflammation is inhibited, and in the treatment of Parkinson's disease, Alzheimer's disease, brain trauma, cerebral apoplexy and other microglia-mediated neuroinflammatory encephalopathy, there is obvious therapeutic effect.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and particularly relates to a gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheet, its preparation method and application. Specifically, the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheet is used in drugs that synergistically enhance the treatment of microglial-mediated neuroinflammatory encephalopathy such as Parkinson's disease, Alzheimer's disease, brain trauma, and stroke. Background Technology

[0002] Current treatments for neuroinflammatory diseases such as Parkinson's disease, Alzheimer's disease, traumatic brain injury, and stroke primarily focus on symptom relief (e.g., dopamine replacement therapy, cholinesterase inhibitors). However, these treatments have limitations, including poor blood-brain barrier penetration, significant long-term side effects, and the inability to reverse neurodegenerative processes. Oxidative stress and neuroinflammation are the core pathological mechanisms of these diseases: Oxidative stress leads to excessive production of reactive oxygen species (ROS), resulting in oxidative damage to neuronal proteins / lipids, promoting Aβ plaques, α-synuclein aggregation, and mitochondrial dysfunction; neuroinflammation is driven by abnormal activation of microglia, releasing pro-inflammatory factors such as TNF-α and IL-1β, exacerbating neurotoxicity and forming a vicious cycle with oxidative stress. Therefore, developing multifunctional antioxidants that can efficiently scavenge ROS, inhibit lipid peroxidation, and penetrate the blood-brain barrier for targeted delivery, thereby reducing neuronal apoptosis and inhibiting microglial overactivation, and thus blocking the neuroinflammatory-oxidative stress positive feedback loop, can alleviate ischemia-reperfusion injury and delay neurodegeneration. This has become a promising treatment strategy for neuroinflammatory diseases such as Parkinson's disease, Alzheimer's disease, brain trauma, and stroke. Hydrogen (H2) is a selective antioxidant gas. However, research on H2 therapy in brain diseases is limited and the mechanisms are unclear. Therefore, further exploration of the medical applications of H2 therapy in brain diseases is of great significance. Although H2 therapy has many advantages, it suffers from problems such as small molecule size, easy diffusion, and low solubility in water. With the rapid development of nanotechnology in the biomedical field, nanomaterials have also received widespread attention in H2 therapy. PdNPs have good hydrogen storage and release properties, but hydrogen needs to be stored first and then released, so the amount of hydrogen released is limited, and the amount of ROS scavenged is also limited, and the therapeutic effect may not meet expectations. Therefore, when nanomaterials reach the lesion site, how to increase the amount of hydrogen and enhance the antioxidant capacity of nanomaterials to more efficiently and thoroughly remove ROS becomes an extremely challenging problem. Summary of the Invention

[0003] The technical problem this invention aims to solve is to overcome the shortcomings of existing technologies and provide a gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet, its preparation method, and its applications. The gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet prepared by this invention is a graphitic carbon nitride (g-C3N4 / UFR-NC / Au@Pd) composite nanomaterial with a gold-palladium core-shell and nitrogen-doped carbon ribbon. This gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet exhibits multiple enzyme-like activities and excellent photocatalytic hydrogen production and hydrogen storage / release performance. Furthermore, under enhanced 808 nm near-infrared irradiation, the high hydrogen storage / release performance of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial can reduce oxidative stress damage and inhibit neuroinflammation in vivo, achieving a protective effect on neurons and providing new insights for the treatment of neuroinflammatory diseases such as Parkinson's disease, Alzheimer's disease, brain trauma, and stroke.

[0004] To address the aforementioned technical problems, this invention provides a gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet (also known as gold@palladium-carbon nitride nanozyme). UFR-NC is doped into g-C3N4 to form a g-C3N4 / UFR-NC composite nanomaterial. Using the g-C3N4 / UFR-NC composite nanomaterial as a carrier, gold-palladium core-shell particles are loaded onto the g-C3N4 / UFR-NC composite nanomaterial. Further, in the aforementioned gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet, the gold-palladium core-shell particles have a gold core and a palladium shell. Further, in the aforementioned gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheet, the diameter of the gold-palladium core-shell particles is 20–30 nm. Based on a general technical concept, the present invention also provides a method for preparing the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets, the preparation method comprising the following steps: S1, preparing nitrogen-doped carbon ribbon modified graphitic carbon nitride; S2, dissolving the nitrogen-doped carbon ribbon modified graphitic carbon nitride in water, and adding sodium citrate solution, gold chloride trihydrate and Na2PdC l4The solution is stirred and mixed, ascorbic acid is added to react, and then dried to obtain gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets. Further, in the above preparation method, step S1 includes the following steps: S1-1, urea is placed in a crucible and heated, and maintained in an oil bath to obtain a transparent molten urea solution; S1-2, UFR is added to the urea solution and mixed to form a homogeneous solution, which is then condensed to obtain a solid powder; S1-3, the solid powder is calcined in a tube furnace at 600°C for 3 h at a heating rate of 5°C / min to obtain nitrogen-doped carbon ribbon modified graphitic carbon nitride. Further, in the above preparation method, step S2 includes the following steps: S2-1, PdCl2 and NaCl are dissolved, heated to 90°C, and stirred for 30 min to obtain Na2PdC. l4 Solution; S2-2, nitrogen-doped carbon ribbon-modified graphitic carbon nitride is dissolved in water, and sodium citrate solution, gold chloride trihydrate and Na2PdC are added. l4 The solution is stirred and mixed to obtain a reaction system; S2-3 and ascorbic acid are dissolved in water and added to the reaction system at a rate of 2 mL / h using a syringe pump. After centrifugation to remove the supernatant, the reaction solution is dried to obtain gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets. Based on a general technical concept, this invention also provides the application of the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets in the preparation of drugs for protecting neurons. Based on a general technical concept, this invention also provides the application of the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets in the preparation of drugs for treating neuroinflammatory diseases such as Parkinson's disease, Alzheimer's disease, brain trauma, and stroke. This invention also provides the application of the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets in the preparation of drugs for inhibiting neuroinflammatory diseases. This invention also provides the application of the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets in the preparation of drugs for reducing oxidative stress damage in vivo.

[0005] Compared with the prior art, the advantages of the present invention are as follows: (1) The present invention provides a gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheet with multiple antioxidant enzyme activities. First, it can scavenge ROS and reduce oxidative stress and inflammation levels. Second, under the enhancement effect of near-infrared light, it greatly improves the hydrogen release capacity and photocatalytic hydrogen production performance of g-C3N4 / UFR-NC / Au@Pd. Studies have shown that hydrogen has the effect of anti-oxidative stress and can efficiently scavenge ROS, thereby reducing the level of oxidative stress and inflammation in vivo. Third, g-C3N4 / UFR-NC / Au@Pd can efficiently penetrate the BBB and can be administered non-invasively through nasal administration to ultimately reach the midbrain for treatment. Based on the superposition of the above multiple effects, the oxidative stress level in vivo is finally regulated, neuronal apoptosis is inhibited, the inflammatory microenvironment is improved, and the neuronal protection effect is achieved. Furthermore, the 6-OHDA-induced Parkinson's disease (PD) mouse model is treated through nasal administration. As can be seen from the above, g-C3N4 / UFR-NC / Au@Pd, which possesses multiple antioxidant enzyme activities and hydrogen storage and release properties, combined with hydrogen-photothermal energy, can effectively scavenge ROS, reduce oxidative stress levels, decrease neuronal apoptosis, and improve the inflammatory microenvironment, thereby achieving the goal of treating Parkinson's disease. This provides a more promising new treatment strategy for the clinical treatment of brain diseases such as Parkinson's disease, Alzheimer's disease, brain trauma, and stroke, and also demonstrates the long-term development potential of nanomedicine in the field of neurological diseases.

[0006] (2) This invention provides a gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheet. g-C3N4 has a strong photocatalytic hydrogen production ability, but the band gap of g-C3N4 is about 2.7 eV, which can only absorb and utilize visible light with wavelengths less than 460 nm. In order to improve the photocatalytic effect of g-C3N4 and expand its application range, this invention uses g-C3N4 / UFR-NC composite material, which has excellent optical adsorption performance for both visible and near-infrared light, and the band gap can be easily adjusted by changing the content of UFR-NC band. g-C3N4 / UFR-NC nanosheets exhibit several advantages over pure g-C3N4 nanosheets: First, the doping with nitrogen and carbon significantly broadens the light absorption range of g-C3N4 nanosheets. Nitrogen-doped carbon-modified g-C3N4 nanosheets can absorb near-infrared light and, under near-infrared light stimulation, activate g-C3N4 photocatalytic water release of hydrogen. Second, the larger specific surface area, better crystallinity, and excellent stability provide more active sites for the loaded gold-palladium core-shell nanostructure. Attached Figure Description

[0007] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Figure 1 The results are from the HRTEM test in Example 1. Figure 2 XPS characterization images of g-C3N4 and g-C3N4 / UFR-NC. Figure 3 XPS characterization of g-C3N4 / UFR-NC / Au@Pd. Figure 4 XRD characterization images of g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd. Figure 5 The PL spectra of g-C3N4 / UFR-NC / Au@Pd and RhB@g-C3N4 / UFR-NC / Au@Pd are shown. Figure 6 The results show the SOD-like enzyme activity of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 7 The results show the detection of POD-like enzyme activity in g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 8 The results show the CAT-like enzyme activity assays for g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 9 The results show the GPx-like enzyme activity assay results for g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 10 The results show the photocatalytic hydrogen production performance of g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 11 The results show the in vitro hydrogen release capacity of g-C3N4 / UFR-NC / Au@Pd after passing hydrogen gas through it, under both near-infrared laser irradiation and without. Figure 12 The extinction coefficients of the g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials are measured. Figure 13 The results show the in vitro photothermal properties of the g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials. Figure 14 The results are from the MTT assay. Figure 15 The image shows the Calcein-AM / PI staining results of the effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on damaged neurons. Figure 16The image shows the Calcein-AM / PI staining results of the combined effects of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials and hydrogen-photothermal interaction on damaged neurons. Figure 17 The image shows the ROS staining results of the combined effects of hydrogen-photothermal interaction on damaged neurons using g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 18 The results show the expression level of iNOS / Arg1 mRNA in BV2 cells induced by 6-OHDA after treatment with g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal treatment. Figure 19 The results show the secretion of pro-inflammatory factors IL-1β, IL-6, and TNF-α by BV2 cells induced by 6-OHDA and treated with g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal treatment. Figure 20 The results of in vivo fluorescence imaging of mice after intranasal administration of RhB@g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different time points. Figure 21 The results of animal behavior experiments on PD mice using g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction. Figure 22 The figure shows the results of an immunoblotting experiment on the effects of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on PD mice. Detailed Implementation

[0008] Example 1: A gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheet of the present invention.

[0009] The preparation method includes the following steps: (1) Preparing nitrogen-doped carbon ribbon modified graphite phase carbon nitride (g-C3N4 / UFR-NC). 1.1. Place 20 g of urea in a crucible, heat at 150°C, and maintain in an oil bath for 20 min to obtain a transparent molten urea solution. 1.2. Then add 0.02 g of UFR to form a homogeneous solution and cool to room temperature. The solid powder formed after condensation is calcined in a tube furnace at 600°C for 3 h at a heating rate of 5°C / min. After cooling to room temperature, the prepared solid powder is labeled as g-C3N4 / UFR-NC.

[0010] (2) Preparation of nitrogen-doped carbon ribbons loaded with gold-palladium core and shell to modify graphitic carbon nitride (g-C3N4 / UFR-NC / Au@Pd). 2.1 Weigh 86.4 mg PdCl2 and 60 mg NaCl and dissolve them in 10 mL of ultrapure water. Heat to 90 °C and stir for 30 min to obtain Na2PdC. l4Solution. 2.2. Weigh 5 mg of the g-C3N4 / UFR-NC solid powder prepared in the previous step and dissolve it in 40 mL of ultrapure water. Then add sodium citrate solution, gold chloride trihydrate, and Na2PdC solution respectively. l4 The solution was stirred at room temperature. 2.3 Finally, 18 mg of ascorbic acid was weighed and dissolved in 10 mL of ultrapure water, and added to the reaction system using a syringe pump at a rate of 2 mL / h. After the reaction was complete, the supernatant was discarded by centrifugation, and the mixture was washed three times with ultrapure water, twice with 75% ethanol, and finally once with anhydrous ethanol. The resulting gray-black solid was dried and labeled as g-C3N4 / UFR-NC / Au@Pd. The preparation method of pure graphitic carbon nitride (g-C3N4) is as follows: 20 g of urea was weighed and placed in a tube furnace, heated to 600℃ at a heating rate of 5℃ / min, calcined for 3 h, and cooled to room temperature. The resulting solid powder was labeled as g-C3N4.

[0011] Experiment 1: Characterization of g-C3N4 / UFR-NC / Au@Pd composite material.

[0012] 1. High-resolution transmission electron microscopy (HRTEM) detection: Figure 1 The figures show the HRTEM results. A in the figure is the HRTEM image of g-C3N4 / UFR-NC, scale bar = 200 nm. B in the figure is the HRTEM image of g-C3N4 / UFR-NC, scale bar = 100 nm. C in the figure is the HRTEM image of g-C3N4 / UFR-NC / Au@Pd, scale bar = 200 nm; D in the figure is the HRTEM image of g-C3N4 / UFR-NC / Au@Pd, scale bar = 10 nm. From A and B, it can be seen that g-C3N4 / UFR-NC is a thin-layered nanosheet with a large specific surface area and irregular edges, modified with thin, elongated black ribbons. Figure C shows that a large number of gold-palladium core-shell nanoparticles are uniformly distributed on the g-C3N4 / UFR-NC composite nanomaterial. As can be seen from D in the figure, the overall size of the gold-palladium core-shell supported by the g-C3N4 / UFR-NC composite nanomaterial is about 20-30 nm, and the size of the gold nanoparticles serving as the core is about 5 nm. This preliminarily proves the successful synthesis of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial.

[0013] 2. X-ray photoelectron spectroscopy (XPS) detection: Figure 2The figures show the XPS characterization of g-C3N4 and g-C3N4 / UFR-NC. Figure A shows the full XPS spectrum of g-C3N4 and g-C3N4 / UFR-NC; Figures B, C, and D show the fine XPS spectra of C1s, N1s, and O1s for g-C3N4 and g-C3N4 / UFR-NC, respectively. The figures show that both g-C3N4 and g-C3N4 / UFR-NC contain specific peaks at C1s, N1s, and O1s. The carbon atom ratio of g-C3N4 / UFR-NC is significantly higher than that of g-C3N4. The peak area ratios of the characteristic peaks at 287.78 eV and 284.61 eV for g-C3N4 and g-C3N4 / UFR-NC are 7.54 and 1.59, respectively. These results demonstrate the successful preparation of g-C3N4 / UFR-NC.

[0014] Figure 3 The figure shows the XPS characterization of g-C3N4 / UFR-NC / Au@Pd; A in the figure is the full XPS spectrum of g-C3N4 / UFR-NC / Au@Pd; B, C, D, E, and F in the figure are the fine XPS spectra of C 1s, N 1s, O 1s, Pd 3d, and Au 4f of g-C3N4 / UFR-NC / Au@Pd, respectively. From the full XPS spectrum and the various fine spectra of g-C3N4 / UFR-NC / Au@Pd, peaks of five elements—C, N, O, Pd, and Au—can be observed. The Pd element shows absorption peaks at [insert values ​​here] at [insert values ​​here], corresponding to Pd 3d3 and Pd 3d5 absorption peaks, respectively, while the Au element shows absorption peaks at [insert values ​​here] at [insert values ​​here], corresponding to Au 4f5 and Au 4f7 absorption peaks, respectively. These results are compared with [insert values ​​here]. Figure 2 The XPS spectra of g-C3N4 / UFR-NC show a significant increase in Au and Pd elements in the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial, indicating that gold-palladium core-shell nanoparticles were successfully loaded onto g-C3N4 / UFR-NC, i.e., the successful synthesis of g-C3N4 / UFR-NC / Au@Pd.

[0015] 3. X-ray diffraction (XRD) detection: Figure 4XRD characterization images of g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd. The results show that g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd all have diffraction peaks at two positions with 2θ values ​​of approximately 13.0° and 27.6°, corresponding to the (100) and (002) crystal planes of g-C3N4, respectively. The characteristic peak intensity of g-C3N4 / UFR-NC at the (002) crystal plane is significantly higher than that of g-C3N4, indicating that the modification of g-C3N4 by UFR-NC improves the crystallinity of g-C3N4 / UFR-NC. The XRD results of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials show five characteristic peaks of metal Au and Pd, indicating that gold-palladium core-shell nanoparticles have been successfully loaded onto g-C3N4 / UFR-NC composite nanomaterials, that is, the successful synthesis of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials.

[0016] 4. Fluorescence spectroscopy (PL) detection: Figure 5 The figures show the photoluminescence (PL) spectra of g-C3N4 / UFR-NC / Au@Pd and RhB@g-C3N4 / UFR-NC / Au@Pd; A in the figure is the PL spectrum of g-C3N4 / UFR-NC / Au@Pd; and B in the figure is the PL spectrum of RhB@g-C3N4 / UFR-NC / Au@Pd. As can be seen from the figures, with 368 nm as the excitation wavelength, the emission peak of g-C3N4 / UFR-NC / Au@Pd is approximately located at 430 nm, demonstrating that g-C3N4 / UFR-NC / Au@Pd exhibits certain photoluminescence properties and emits blue fluorescence under ultraviolet light excitation. To facilitate further in vivo fluorescence imaging to determine the time of action of the material after nasal administration to the brain, we tested the fluorescence emission spectrum of the synthesized RhB@g-C3N4 / UFR-NC / Au@Pd. As can be seen from B in the figure, when RhB@g-C3N4 / UFR-NC / Au@Pd is used as the excitation wavelength of 532 nm, the emission peak in the obtained emission spectrum is approximately located at 557 nm, indicating that RhB was successfully modified on the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial.

[0017] Experiment 2: Detection of enzyme-like activity in g-C3N4 / UFR-NC / Au@Pd composite nanomaterials.

[0018] 1. Detection of SOD-like enzyme activity in g-C3N4 / UFR-NC / Au@Pd composite nanomaterials: The SOD-like enzyme activity of the two composite nanomaterials, g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd, was detected using a SOD activity assay kit (WST-8 method). Figure 6 The figures show the SOD-like enzyme activity detection results of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figures A and B represent the statistical graphs (n>3) of the inhibition rate of superoxide radicals and SOD enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different concentrations (5 μg / mL, 15 μg / mL, 45 μg / mL, and 90 μg / mL), respectively. Figures C and D represent the statistical graphs (n>3) of the inhibition rate of superoxide radicals and SOD enzyme activity of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at the same concentration (45 μg / mL), respectively. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. As shown in Figures A and B, the superoxide radical inhibition rate and SOD enzyme activity of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials increased in a concentration-dependent manner, indicating that the g-C3N4UFR-NC / Au@Pd composite nanomaterials possess strong SOD enzyme activity and can scavenge superoxide anion radicals. Furthermore, we measured the superoxide radical inhibition rate and SOD enzyme activity of the g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. The results, shown in Figures C and D, indicate that the superoxide radical inhibition rate and SOD enzyme activity of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials are approximately eight times that of the g-C3N4 / UFR-NC composite nanomaterials.

[0019] 2. Detection of POD-like enzyme activity in g-C3N4 / UFR-NC / Au@Pd composite nanomaterials: Figure 7The figures show the POD-like enzyme activity assay results for g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure A shows the UV-Vis spectra of different concentrations (5 μg / mL, 15 μg / mL, 45 μg / mL, and 90 μg / mL) of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials for POD-like enzyme activity; Figure B shows the 20... The graph shows the absorbance changes over a period of time (min) reflecting the POD-like enzyme activity of different concentrations of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure C shows the UV-Vis spectra of the POD-like enzyme activity of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure D shows the relative POD-like enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials under different pH conditions (pH=2.5, 3.5, 4.5, 5.5, 7.4). Figures A and B show that at pH 4.5, the POD-like enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials increases with increasing material concentration. Figure C shows that the POD-like enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials is significantly higher than that of g-C3N4 / UFR-NC composite nanomaterials. As shown in Figure D, the POD enzyme activity of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial is higher under acidic conditions, reaching its peak at pH 4.5. At pH 7.4, the relative enzyme activity of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial is as low as 19.77%. Therefore, the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial exhibits strong POD enzyme activity under acidic or weakly acidic conditions, while under normal physiological conditions (pH=7.4), its POD enzyme activity is very weak, generating a small amount of ·OH, thus it does not have a significant toxic effect on normal tissues and cells.

[0020] 3. Detection of CAT-like enzyme activity in g-C3N4 / UFR-NC / Au@Pd composite nanomaterials: Figure 8Figure 1 shows the CAT enzyme activity detection results of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure A shows the dissolved oxygen content curves of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different concentrations (5 μg / mL, 15 μg / mL, 45 μg / mL, and 90 μg / mL); Figure B shows digital photographs of the reaction solutions after adding the same amount of H2O2 to the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different concentrations; Figure C shows the statistical graph of CAT enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different concentrations (n>3); Figure D shows the dissolved oxygen content curves of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. ****p<0.0001. As shown in Figure A, the dissolved oxygen content of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial increases with increasing concentration of the composite nanomaterial. Figure B, a digital image, shows that under constant conditions, more and more bubbles are generated with increasing concentration of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial in the solution. These results reflect a significant concentration-dependent CAT enzyme activity of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial. Similarly, Figure C further validates this conclusion using a CAT enzyme activity assay kit. Figure D compares the dissolved oxygen content of the two composite nanomaterials, g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd, at the same concentration (45 μg / mL), indicating that the CAT enzyme activity of g-C3N4 / UFR-NC is significantly enhanced after loading with gold-palladium core-shell nanoparticles.

[0021] 4. Detection of GPx-like enzyme activity in g-C3N4 / UFR-NC / Au@Pd composite nanomaterials: The GPx-like enzyme activity of the two composite nanomaterials, g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd, was detected using a total GPx detection kit. Figure 9Figure 1 shows the GPx enzyme activity detection results of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure A shows the statistical graph of GPx enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at different concentrations (n>3); Figure B shows the statistical graph of GPx enzyme activity of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials (n>3). ***p<0.001, ****p<0.0001. Figure A shows that the GPx enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials gradually increases with increasing concentration, exhibiting a clear concentration dependence. Figure B shows that the GPx enzyme activity of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials is significantly stronger than that of g-C3N4 / UFR-NC composite nanomaterials (p<0.001).

[0022] Experiment 3: Investigating the photocatalytic hydrogen production capability of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials.

[0023] Figure 10The results show the photocatalytic hydrogen production performance of g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figures A, B, and C show the H2 unit yield (n=3), total H2 yield (n=3), and H2 production rate (n=3) of the photocatalytic hydrogen production of g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials, respectively, within 4 hours. Figure D shows the stability test results of the photocatalytic hydrogen production of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials after two cycles within 8 hours. *p<0.05, ***p<0.001, ****p<0.0001. Figures A and B compare the H2 unit yield and total H2 yield of three composite nanomaterials (g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd) over 4 hours. It can be seen that the g-C3N4 / UFR-NC composite nanomaterial has a higher H2 unit yield and total H2 yield than the g-C3N4 composite nanomaterial. Furthermore, the H2 unit yield and total H2 yield of the g-C3N4 / UFR-NC composite nanomaterial with a gold-palladium core-shell loading are both improved compared to before loading. Additionally, Figure C shows that the photocatalytic hydrogen production rates of the g-C3N4, g-C3N4 / UFR-NC, and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials are 96.3 μmol / h, 123.9 μmol / h, and 141.9 μmol / h, respectively. The above results indicate that the photocatalytic hydrogen production performance of the g-C3N4 / UFR-NC composite nanomaterial is significantly better than that of g-C3N4 alone. Furthermore, the photocatalytic hydrogen production rate of the g-C3N4 / UFR-NC composite nanomaterial with a gold-palladium core-shell loading is improved compared to that of the standard g-C3N4 / UFR-NC composite nanomaterial. As shown by D in the figure, the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial showed no significant change after two cycles within 8 hours, indicating that the photocatalytic hydrogen production performance of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial exhibits good stability.

[0024] Experiment 4: Investigating the hydrogen release performance of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. Figure 11The results show the in vitro hydrogen release capacity of g-C3N4 / UFR-NC / Au@Pd after passing hydrogen gas through it, under both near-infrared laser irradiation and without. Figure A shows the UV-Vis absorption spectra of methylene blue solutions at different concentrations; Figure B shows the linear fitting standard curve of the absorption intensity of MB at 664 nm versus MB concentration; Figure C shows the reaction process of g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials with MB without 808 nm near-infrared illumination; Figure D shows the reaction process of g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials with MB under 808 nm near-infrared illumination; Figure E shows the change in absorbance difference of MB in the reaction solution of g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials with and without 808 nm near-infrared illumination; Figure F shows the change in absorbance difference of MB in the reaction solution of g-C3N4 / UFR-NC / Au@Pd after passing hydrogen gas through it under 808 nm near-infrared illumination for 60 min (n=3). ****p<0.0001. Figure A shows that MB has a characteristic absorption peak at 664 nm, and its absorbance at 664 nm increases with increasing MB solution concentration. Figure B also shows that the absorbance of the MB solution changes with concentration, and the two have a linear correlation. According to Figures C, D, and E, in the first hour under 808 nm near-infrared light, the absorbance of MB at 664 nm in the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial aqueous solution decreased faster than that in the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial aqueous solution without 808 nm near-infrared light, and the H2 release rate was also faster in the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial aqueous solution without 808 nm near-infrared light. Subsequently, under the irradiation of 808 nm near-infrared light, the H2 release rate of the aqueous solution of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials gradually slowed down, while the hydrogen release rate of the aqueous solution of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials without 808 nm near-infrared light gradually accelerated after 1 h. This indicates that the H2 release and function of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials without 808 nm near-infrared light is significantly slower than that of the aqueous solution of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials under 808 nm near-infrared light.The difference in absorbance (MB) in the reaction solution of g-C3N4 / UFR-NC / Au@Pd after 60 min of reaction with and without 808 nm near-infrared light after hydrogen gas purging in Figure F indicates that 808 nm near-infrared light irradiation significantly enhances the hydrogen release performance of the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial (p<0.0001). These results suggest that the reducing power of g-C3N4 / UFR-NC / Au@Pd+H2 is significantly enhanced under 808 nm near-infrared light irradiation, which may facilitate faster H2 release at the lesion site in in vivo experiments, potentially leading to better hydrogen therapy effects.

[0025] Experiment 5: Investigating the in vitro photothermal properties of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials.

[0026] 1. Extinction coefficient detection of g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials: Figure 12 The extinction coefficients of the g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials are shown in the figure. Figures A, C, and E represent the UV-Vis-NIR spectra of different concentrations of the g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials, respectively. Figures B, D, and F represent the linear fitting curves of absorbance at 808 nm versus concentration for the g-C3N4 / UFR-NC composite nanomaterial, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+ H2, respectively. As shown in Figures A, C, and E, the g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd composite nanomaterials, and g-C3N4 / UFR-NC / Au@Pd+ H2 all exhibit strong absorption in the NIR-I region, showing a clear concentration dependence. Furthermore, comparisons reveal that g-C3N4 / UFR-NC / Au@Pd+ H2 demonstrates stronger absorption at 808 nm than both g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials. We then performed a linear fit between the nanomaterial concentration and the corresponding absorbance value at 808 nm (as shown in Figures B, D, and F), and calculated the extinction coefficients of different materials using the Lambert-Beer law. Finally, we obtained the extinction coefficients of g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials as 4.81, 6.96, and 7.69 L·g, respectively.-1 ·cm -1 It can be seen that the extinction coefficient of the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial is the largest. In terms of light absorption capacity, the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterial is stronger than that of the g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd composite nanomaterials.

[0027] 2. Temperature rise curve detection of g-C3N4 / UFR-NC / Au@Pd+ H2 composite nanomaterials: Figure 13 The figures show the in vitro photothermal performance of g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials. Figure A represents the performance at a power density of 0.5 W / cm². 2 The temperature rise curves of g-C3N4 / UFR-NC / Au@Pd+ H2 at different concentrations (5, 15, 45, and 90 μg / mL) were obtained after irradiation with an 808 nm near-infrared laser for 10 min. Figure B shows the temperature rise curves of g-C3N4 / UFR-NC / Au@Pd+ H2 at the same concentration under an 808 nm near-infrared laser power density of 0.5 W / cm². 2 0.75 W / cm 2 and 1 W / cm 2 The figure shows the temperature rise curves under irradiation; C in the figure represents the temperature rise curves of g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+H2 after irradiation for 10 min under 808 nm near-infrared laser with a power of 0.5 W / cm2; D in the figure represents the temperature rise curves under irradiation with 0.5 W / cm2. 2 A bar chart (n=3) showing the temperature difference between g-C3N4 / UFR-NC, g-C3N4 / UFR-NC / Au@Pd, and g-C3N4 / UFR-NC / Au@Pd+ H2 after 10 min irradiation with an 808 nm near-infrared laser. ****p<0.0001.

[0028] As shown in Figure A, under the same near-infrared illumination power (0.5 W / cm²), 2 Under irradiation, the photothermal conversion performance of aqueous solutions of g-C3N4 / UFR-NC / Au@Pd+H2 nanomaterials with different concentrations increased with increasing concentration. As shown in Figure B, different power densities (0.5, 0.75, and 1 W / cm²) were used. 2An aqueous solution of g-C3N4 / UFR-NC / Au@Pd+H2 nanomaterials was irradiated with an 808 nm laser for 10 min. Real-time near-infrared irradiation revealed that the photothermal conversion performance of the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials improved with increasing laser irradiation power density. As shown in Figure C, the temperature of the aqueous solution for each group of nanomaterials gradually increased with prolonged laser irradiation time, indicating a significant improvement in the photothermal performance of the g-C3N4 / UFR-NC / Au@Pd+H2 composite nanomaterials. Then, according to the bar chart of temperature difference of different groups of nanomaterials monitored by D infrared thermal imager within 10 minutes, it can be seen that the solution temperature of g-C3N4 / UFR-NC group increased by 7.3℃, the solution temperature of g-C3N4 / UFR-NC / Au@Pd group increased by 11.5℃, and the solution temperature of g-C3N4 / UFR-NC / Au@Pd+ H2 group increased by 13.5℃. This further illustrates that the photothermal conversion performance of g-C3N4 / UFR-NC / Au@Pd+ H2 group is significantly higher than that of g-C3N4 / UFR-NC and g-C3N4 / UFR-NC / Au@Pd groups.

[0029] Example 2: Application of Example 1 in the preparation of a drug.

[0030] Experiment 6: MTT assay to explore the synergistic protective effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on damaged neurons. (1) After culturing primary neurons for 7 days, 6-OHDA at concentrations of 0.05, 0.1, 0.2, 0.4 and 0.8 mM were added to the primary neurons and incubated in a 37℃ incubator for 24 h to explore the appropriate concentration of 6-OHDA to induce an in vitro PD model. The degree of cell damage in primary neurons was detected by the MTT assay to explore the appropriate modeling concentration of 6-OHDA for primary neurons. (2) Following the aforementioned method, aqueous solutions of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at concentrations of 2.5, 5, 15, 45, and 90 μg / mL, along with 0.1 mM 6-OHDA solution, were added to cultured primary neurons. The absorbance values ​​of each group were detected by the MTT assay to reflect the protective effect of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on damaged neurons. (3) Different materials (g-C3N4 / UFR-NC / Au@Pd, g-C3N4 / UFR-NC / Au@Pd+NIR, g-C3N4 / UFR-NC / Au@Pd+H2, g-C3N4 / UFR-NC / Au@Pd+H2+NIR) were used to treat neurons according to the aforementioned method.

[0031] Figure 14The results are shown in Figure A, which represents the effect of different concentrations of 6-OHDA on the cell viability of primary neurons as detected by the MTT assay. As shown, primary neurons modeled with 0.1 mM 6-OHDA achieved a viability of nearly 50%, suitable for use as an in vitro PD model in subsequent experiments. Figure B shows the effect of different concentrations of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on the cell viability of damaged neurons. As shown, primary neurons treated with g-C3N4 / UFR-NC / Au@Pd composite nanomaterials at concentrations of 2.5, 5, 15, 45, and 90 μg / mL all showed increased cell viability compared to the 6-OHDA group, indicating that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials have a significant protective effect on damaged neurons. Furthermore, the cell viability of the group treated with 15 μg / mL of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials was significantly higher than that of the 6-OHDA group (p<0.05). Therefore, we finally selected 15 μg / mL as the final drug treatment concentration. In the figure, C represents the effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on the cell viability of damaged neurons. As can be seen from the figure, the MTT assay results show that the cell viability of all four material treatment groups was significantly higher than that of the 6-OHDA group. Moreover, the g-C3N4 / UFR-NC / Au@Pd+H2+NIR group showed a more significant increase in cell viability than the other three material treatment groups, indicating that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction have a strong synergistic protective effect on damaged neurons.

[0032] Experiment 7: Exploring the effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on cell viability and mortality using the Calcein-AM / PI method. (1) Primary neurons were treated with 0.1 mM 6-OHDA solution and different concentrations of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials for 24 h, followed by Calcein-AM / PI staining for viability and mortality. (2) Cells were treated with 6-OHDA and different groups of materials, with 6-OHDA concentration of 0.1 mM and material concentration of 15 μg / mL.

[0033] Figure 15The image shows the Calcein-AM / PI staining results of the effects of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial on damaged neurons. Figure A shows that the Control group exhibited the strongest green fluorescence and the highest number of live cells. The 6-OHDA group showed a significantly lower number of live cells compared to the Control group, with the strongest red fluorescence and the highest number of dead cells. Furthermore, the different concentrations of the material treated significantly increased the number of live cells and significantly reduced the number of dead cells compared to the 6-OHDA group. Additionally, the statistical results in Figures B and C show that with increasing concentrations of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial, the number of green-labeled live cells gradually increased, while the number of red-labeled dead cells decreased, with statistically significant differences. This indicates that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial itself has a significant protective effect on damaged neurons.

[0034] Figure 16 The image shows the Calcein-AM / PI staining results of the effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on damaged neurons. Figure A shows that the Control group exhibited the strongest green fluorescence and the highest number of live cells, while the 6-OHDA group showed a significantly reduced number of live cells compared to the Control group, with the strongest red fluorescence and the highest number of dead cells. Figures B and C show that the different material treatment groups significantly increased the number of live cells and significantly reduced the number of dead cells compared to the 6-OHDA group. The g-C3N4 / / UFR-NC / Au@Pd+H2+NIR group had the most live cells labeled with green fluorescence and the fewest dead cells labeled with red fluorescence compared to other treatment groups. This indicates that the g-C3N4 / / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction have a significant synergistic protective effect on damaged neurons.

[0035] Experiment 8: ROS staining method to explore the effect of g-C3N4 / / UFR-NC / Au@Pd composite nanomaterials on intracellular ROS levels. (1) Primary neurons were treated with 0.1 mM 6-OHDA solution and different concentrations of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials for 24 h and then subjected to ROS fluorescent probe. (2) ROS staining was performed on each treatment group of cells, where the concentration of 6-OHDA was 0.1 mM and the concentration of material was 15 μg / mL. Figure 17Figure A shows the ROS staining results of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials on damaged neurons. As shown in Figure A, under a fluorescence microscope, enhanced green fluorescence was observed after 6-OHDA-induced damage to primary neurons, indicating increased intracellular ROS activity. Compared to the 6-OHDA group, the green fluorescence of different concentrations of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials showed varying degrees of decrease, indicating reduced intracellular ROS activity. With increasing nanomaterial concentration, ROS activity decreased accordingly, as shown in Figure B, demonstrating a statistically significant difference. This indicates that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials can reduce cellular oxidative stress and have a significant protective effect on damaged neurons. Figure C shows the ROS staining results of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on damaged neurons. As can be seen, compared with the 6-OHDA group, the intracellular ROS activity levels of all four material treatment groups were reduced, and the intracellular ROS activity level of the g-C3N4 / UFR-NC / Au@Pd+ H2+NIR group was the lowest, as shown in Figure D. This further illustrates that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction can reduce cellular oxidative stress levels and have a more significant protective effect on damaged neurons than the other three material treatment groups.

[0036] Experiment 9: Detection of the inhibitory effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal on the inflammation of activated BV2 cells by qPCR and ELISA.

[0037] 9.1 Detection of BV2 microglia polarization by qPCR: The mRNA levels of the M1 / M2 marker iNOS / Arg1 in BV2 cells (6-OHDA induced) after treatment with different materials were detected by qPCR. Power SYBR Green PCRMaster Mix was used for qPCR analysis. Figure 18The figures show the mRNA expression levels of iNOS / Arg1 in 6-OHDA-induced BV2 cells after treatment with g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal therapy. In the figures, A represents iNOS and B represents Arg1. The figures show that after treatment with each material group, the mRNA expression level of Arg1, a marker of M2 microglia, increased, while the mRNA expression level of iNOS, a marker of M1 microglia, decreased. The results were most significant with the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal therapy, indicating that this combination significantly remodeled the microglia phenotype from M1 to M2.

[0038] 9.2 Enzyme-linked immunosorbent assay (ELISA) to determine the levels of inflammatory factors in BV2 microglia: The concentrations of pro-inflammatory factors IL-1β, IL-6, and TNF-α in the culture supernatant of BV2 cells (induced by 6-OHDA) after treatment with each group of materials were determined using ELISA. BV2 cells were cultured in 6-well plates for 24 h after 6-OHDA induction, followed by co-incubation with each group of materials for another 24 h before detection. Protein concentration was determined using the BCA protein assay method. The concentrations of pro-inflammatory factors IL-1β, IL-6, and TNF-α were determined using an ELISA kit. Figure 19 The figures show the results of detecting the secretion of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α in BV2 cells induced by 6-OHDA and treated with g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal therapy. In the figures, A represents IL-1β, B represents IL-6, and C represents TNF-α. Thanks to the ideal polarization capability of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal therapy in vitro, we further detected the levels of typical pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in BV2 cells. ELISA results showed that, compared with the 6-OHDA model group, the levels of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α, were decreased in cells of all material groups. Among them, the most significant decrease was observed in the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial combined with hydrogen-photothermal treatment. This indicates that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterial combined with hydrogen-photothermal treatment can significantly reduce the secretion of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α, in BV2 cells, suggesting that it has a good anti-inflammatory effect in 6-OHDA-induced damaged cells.

[0039] Experiment 10: In vivo fluorescence imaging to explore the appropriate illumination time after intranasal administration of nanomaterials to mice. An in vivo fluorescence imager was used to explore the fluorescence intensity of the nanomaterials in the mouse brain after intranasal administration. Figure 20 This image shows the in vivo fluorescence imaging results of mice administered RhB@g-C3N4 / UFR-NC / Au@Pd composite nanomaterials via intranasal administration at different time points. Figure A shows that some nanomaterials reached the brain of mice as early as 1 hour after administration. With increasing time, the fluorescence intensity in the mouse brain increased, reflecting the increasing accumulation of nanomaterials in the brain. The fluorescence intensity was even stronger at 6 hours after administration. Subsequently, at 24 hours after administration, the fluorescence intensity in the mouse brain was significantly weaker than at 6 hours. Afterward, the fluorescence intensity gradually decreased with further time. This also indirectly indicates that the nanomaterials do not accumulate in the brain for extended periods, demonstrating high biocompatibility. Figure B shows the statistical analysis results of the fluorescence intensity in the mouse brain at different time points before and after administration.

[0040] Experiment 11: Animal behavioral experiments to explore the therapeutic effects of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on PD mice. PD mice were subjected to open field test, pole climbing test, and apomorphine (APO) rotation test. Figure 21This figure shows the results of a behavioral experiment on PD mice using g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction. Figure A shows the mice's route in an open field over 5 minutes, indicating their spontaneous activity. The figure shows that the PD group mice exhibited significantly less spontaneous activity compared to the Control group, tending to move only in the periphery of the open field or remain stationary. In contrast, the material-treated group mice showed significantly increased activity, were more active, and exhibited significantly increased activity in the central area. Figures B and D represent statistical graphs of the total distance traveled, average speed, and total resting time in the affected area (n=3), respectively. These statistical results indicate that the PD group mice, compared to the Control group, had significantly less total distance traveled, significantly lower average speed, and significantly increased total resting time in the affected area, indicating successful establishment of the PD model. Compared with the PD group mice, the total walking distance, average speed, and total resting time in the material treatment groups were significantly increased, and the overall statistical difference was significantly greater in the g-C3N4 / UFR-NC / Au@Pd+H2+NIR group than in the g-C3N4 / UFR-NC / Au@Pd and g-C3N4 / UFR-NC / Au@Pd+NIR groups. These results indicate that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction can significantly alter the spontaneous activity ability of PD mice, reduce their anxiety about new environments, and have a significant therapeutic effect on PD mice. Figure E shows the pole climbing experiment results of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on PD mice. Figure F shows the APO rotation experiment results of the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on PD mice. As shown in Figures E and F, the PD group mice exhibited significantly longer pole-climbing time and a significantly increased number of APO rotations compared to the Control group mice, indicating successful PD mouse model establishment. In contrast, the material-treated groups showed varying degrees of reduction in pole-climbing time and a significant decrease in the number of APO rotations, all with statistically significant differences. The g-C3N4 / UFR-NC / Au@Pd+ H2+NIR group showed the best results.

[0041] Experiment 12: Immunoblotting experiment to explore the protective effect of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on neurons in PD mice. The expression levels of MAP2 and Caspase 3 proteins in the brain tissues of mice in different experimental groups were detected using immunoblotting. Figure 22This figure shows the immunoblotting results of the effects of g-C3N4 / UFR-NC / Au@Pd composite nanomaterials combined with hydrogen-photothermal interaction on PD mice. Figure A shows the MAP2 and Caspase 3 protein bands in the brain tissue of mice in each treatment group; Figures B and C show the statistical levels of MAP2 / β-actin and Caspase 3 / β-actin proteins, respectively (n=3). *p<0.05, **p<0.01. As shown in Figures A and B, compared with the normal control group, the MAP2 expression level in the PD group was significantly decreased, while compared with the PD group, the MAP2 protein expression levels in the four material treatment groups showed varying degrees of increase. Among them, the PD+ g-C3N4 / UFR-NC / Au@Pd+ H2+NIR group showed the most significant decrease in MAP2 expression level compared with the PD group (P<0.01). As can be seen from Figures A and C, compared with the normal control group, the Caspase 3 expression level in the PD group was significantly increased, while compared with the PD group, the Caspase 3 protein expression levels in the four material treatment groups showed varying degrees of decrease. Among them, the PD+ g-C3N4 / UFR-NC / Au@Pd+ H2+NIR group showed the most significant decrease in Caspase 3 expression level compared to the PD group (P<0.01). This indicates that the g-C3N4 / UFR-NC / Au@Pd composite nanomaterials, in synergy with hydrogen-photothermal interaction, can inhibit neuronal apoptosis and reduce the damaging effects of 6-OHDA on neurons, demonstrating a significant therapeutic effect on PD mice.

Claims

1. The application of gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheets in the preparation of drugs for treating Parkinson's disease, characterized in that, UFR-NC is doped into g-C3N4 to form g-C3N4 / UFR-NC composite nanomaterials; using the g-C3N4 / UFR-NC composite nanomaterials as a carrier, gold-palladium core-shell particles are loaded onto the g-C3N4 / UFR-NC composite nanomaterials to obtain the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets.

2. The application according to claim 1, characterized in that, The gold-palladium core-shell particles have a gold core and a palladium shell.

3. The application according to claim 1, characterized in that, The diameter of the gold-palladium core-shell particles is 20–30 nm.

4. The application according to any one of claims 1 to 3, characterized in that, The preparation method of the gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets includes the following steps: S1. Preparation of graphitic carbon nitride modified with nitrogen-doped carbon ribbons; S2. Dissolve the nitrogen-doped carbon ribbon-modified graphitic carbon nitride in water, add sodium citrate solution, gold chloride trihydrate and Na2PdCl4 solution and stir to mix, add ascorbic acid to react, and dry to obtain gold@palladium-nitrogen-doped carbon ribbon-modified g-C3N4 nanosheets.

5. The application according to claim 4, characterized in that, S1 includes the following steps: S1-1. Place urea in a crucible and heat it, while keeping it in an oil bath to obtain a transparent, molten urea solution. S1-2, Add UFR to the urea solution and mix to form a homogeneous solution, then condense to obtain a solid powder; S1-3. The solid powder is calcined in a tube furnace at 600°C for 3 h at a heating rate of 5°C / min to obtain nitrogen-doped carbon ribbon modified graphite phase carbon nitride.

6. The application according to claim 4, characterized in that, S2 includes the following steps: After dissolving S2-1, PdCl2 and NaCl, heat to 90℃ and stir for 30 min to obtain Na2PdCl4 solution; S2-2, Nitrogen-doped carbon ribbon-modified graphitic carbon nitride was dissolved in water, and sodium citrate solution, gold chloride trihydrate and Na2PdCl4 solution were added and stirred to obtain the reaction system; S2-3, ascorbic acid was dissolved in water and added to the reaction system at a rate of 2 mL / h using a syringe pump. After centrifugation to remove the supernatant, the reaction solution was dried to obtain gold@palladium-nitrogen-doped carbon ribbon modified g-C3N4 nanosheets.

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