Gentian carbon dot material having a repairing effect on skin damaged by jellyfish toxin and preparation method thereof

Abstract

The application belongs to the field of nanomaterials and biomedical applications, and particularly relates to a gentian carbon dot material having a repairing effect on skin damaged by jellyfish toxin and a preparation method thereof. The gentian carbon dot material provided by the application is in a uniform spherical shape, has a concentrated particle size distribution and good dispersity, and compared with traditional nanomaterials which are insufficient in biocompatibility, complex in manufacturing process and high in cost, the biomass-derived carbon dot is a carbon-based nanomaterial prepared from natural products and having pharmacological activity, is more suitable for large-scale production and clinical application, directly removes free radicals through unique surface active groups (such as hydroxyl groups, carboxyl groups and the like), further enhances the antioxidant capacity, can effectively inhibit cell oxidative stress and inflammatory response induced by the jellyfish toxin, thereby protecting the cells and realizing the repairing effect on the skin damaged by the jellyfish toxin.

Description

Technical Field

[0001] This invention relates to the fields of functional nanomaterials and biomedical applications, and more specifically, to a gentian carbon dot material that has a repairing effect on skin damaged by jellyfish toxins and its preparation method. Background Technology

[0002] Jellyfish stings are a common acute marine injury found in coastal areas worldwide. Their pathogenesis is primarily related to nematocyst toxins released from jellyfish tentacles. These toxins contain active components such as phospholipase A2 and metalloproteinases, which activate neutrophils and macrophages to release large amounts of reactive oxygen species (ROS), triggering oxidative stress damage. Simultaneously, they activate signaling pathways such as NF-κB and MAPK, leading to the excessive secretion of pro-inflammatory factors like TNF-α and IL-6, creating a vicious cycle of oxidative stress and inflammation. This further damages the skin and mucous membrane barriers, exacerbating tissue damage. Current clinical treatments are mainly symptomatic, such as local neutralization with acetic acid or ammonia, analgesics, and antihistamines. However, these treatments suffer from unstable efficacy, significant side effects, and a high recurrence rate, failing to meet clinical needs. Therefore, developing novel, safe, and effective treatment strategies, especially those targeting the dual pathological mechanisms of inflammation and oxidative stress, is of significant clinical and social value.

[0003] Carbon dots (CDs), as novel nanomaterials, are widely used in the biomedical field due to their excellent water solubility, low toxicity, and tunable surface functions. Among them, plant-derived CDs have attracted much attention from researchers because they retain the bioactive components of the parent medicinal material, combining the characteristics of nanomaterials with the advantages of natural drugs. Gentian root, a natural plant, has been shown to exert synergistic therapeutic effects by regulating inflammatory signaling pathways such as NF-κB and MAPK and activating the Nrf2 antioxidant pathway, due to its abundant resources, low cost, high biocompatibility, and low toxicity, making it suitable for large-scale production and clinical application. Therefore, it has become the best biomass extract for synthesizing carbon dots for treating skin damage caused by jellyfish toxin. Summary of the Invention

[0004] The purpose of this invention is to propose a gentian carbon dot material with a repairing effect on skin damaged by jellyfish toxin and its preparation method. The gentian carbon dot material has high dispersibility, good biocompatibility and nanoenzyme activity, and can effectively inhibit cellular oxidative stress and inflammatory response induced by jellyfish toxin, thereby protecting cells and achieving a repairing effect on skin damaged by jellyfish toxin. This invention is used to solve the problems of unstable efficacy, large side effects and easy recurrence of existing symptomatic treatments.

[0005] In the first aspect, the present invention provides a gentian carbon dot material, which is uniformly spherical with a concentrated particle size distribution and good dispersion, and does not show obvious agglomeration during long-term storage; the surface is rich in oxygen-containing functional groups such as hydroxyl and carboxyl groups, contains the three main elements of carbon, nitrogen and oxygen and is free from impurities; it has a typical graphitized short-range ordered structure, and its overall physicochemical properties are stable, which meets the material basis requirements for subsequent bioactivity research.

[0006] Specifically, the gentian carbon dot material of this invention, measured by TEM, has an average particle size of 2.49 ± 0.08 nm, a lattice spacing of 0.202 nm (corresponding to the graphite (100) crystal plane) measured by HRTEM, and a negative potential with an average potential of -11.2 mV measured by DLS. After 15 days of storage at room temperature in H2O and PBS, it maintains good dispersibility without aggregation. XRD shows a broad diffraction peak at 21.4 °C, and FTIR shows characteristic peaks such as -OH / -NH (3396.2 cm⁻¹), C=O (1639.9 cm⁻¹), -CO-NH- (1408.0 cm⁻¹), and CO (1096.0 cm⁻¹). The surface is rich in functional groups such as hydroxyl and amino groups. XPS full spectrum shows C, N, and O elements, and fine spectrum shows C 1s: 284.80 eV (CC / C=C), 286.12 eV. eV (CO), 288.40 eV (C=O); N 1s: 399.80 eV (CN), 400.60 eV (C=N); O 1s: 530.10 eV (C=O), 533.00 eV (CO).

[0007] The gentian carbon dot material of the present invention has good antioxidant properties and can alleviate skin damage caused by jellyfish toxin.

[0008] Specifically, the gentian carbon dot material of the present invention has a highly efficient scavenging ability for various ROS such as hydroxyl radicals and superoxide anions induced by jellyfish toxin, and the scavenging effect shows a significant dose-dependent effect with increasing concentration. In the jellyfish toxin-induced H9C2 cell injury model, gentian carbon dots can significantly improve the survival rate of damaged cells and effectively reduce the level of excessive ROS accumulation in cells. At the same time, it can significantly inhibit the expression of pro-inflammatory cytokines and the activation of key proteins in inflammation-related signaling pathways, blocking the vicious cycle of "oxidative stress-inflammatory response" at the cellular level, demonstrating clear cell protection and anti-inflammatory activity.

[0009] Secondly, the present invention provides a method for preparing the above-mentioned gentian carbon dot material, comprising the following steps: Step 1: Processing of gentian root material; Step 2: Convert the gentian suspension into a gentian solution using a hydrothermal synthesis method; Step 3: Dialyze and freeze-dry the gentian solution to form gentian carbon dots.

[0010] Specifically, in step 1, the processing method of the gentian root material is as follows: crush the gentian root material, weigh 1.5 g of the Chinese medicine powder, put it into a glass bottle containing 30 mL of distilled water, and treat it with an ultrasonic cleaner for 30 min.

[0011] Specifically, in step 2, the method for converting the gentian suspension into a gentian solution via hydrothermal synthesis is as follows: the suspension is transferred to a stainless steel autoclave lined with tetrafluoroethylene and heated at 180 °C for 6 h.

[0012] Specifically, in step 3, the method for dialyzing and lyophilizing the gentian solution to form gentian carbon dots is as follows: the obtained mixture is cooled to room temperature and centrifuged at 10000 r / min for 20 min; after centrifugation, the supernatant is collected and purified using a 0.22 μm filter membrane; the purified solution is dialyzed using a cellulose dialysis bag (1000 Da) for 3 days, with the water changed every 8 h. The dialyzed solution is then lyophilized and stored at 4 ℃.

[0013] Thirdly, the present invention also provides the application of gentian carbon dot material as a highly effective skin repair agent in the preparation of a treatment for skin damaged by jellyfish toxin.

[0014] Specifically, in the treatment of skin damage caused by jellyfish venom, the gentian carbon dot material is used to block the vicious cycle of "oxidative stress-inflammatory response" to repair skin damage caused by jellyfish venom. By significantly improving the survival rate of damaged cells and effectively reducing the level of excessive ROS accumulation in cells, the material has a good repair effect on skin damaged by jellyfish venom.

[0015] Compared with the prior art, the beneficial effects of the present invention are: (1) The gentian carbon dot material provided by this invention has more stable efficacy, fewer side effects, and higher safety compared with traditional symptomatic treatments such as local neutralization with acetic acid or ammonia, use of analgesics, and antihistamine therapy. It is also easier to enter skin cells to exert a repair and therapeutic effect. Moreover, compared with the insufficient biocompatibility, complex manufacturing process, and high cost of traditional nanomaterials, biomass-derived CDs are pharmacologically active carbon-based nanomaterials prepared from natural products. They have advantages such as wide availability, green synthesis, low toxicity, and cost-effectiveness, and are therefore more suitable for large-scale production and clinical application.

[0016] (2) The method for preparing gentian carbon dot material provided by the present invention involves treating gentian raw material, converting gentian suspension into gentian solution by hydrothermal synthesis, and then dialyzing and freeze-drying the gentian solution to successfully synthesize gentian carbon dots. The preparation method is simple and easy to implement, and the particle size distribution is concentrated and the dispersion is good. Attached Figure Description

[0017] Figure 1 a is a transmission electron microscope (TEM) image of the gentian carbon dot material prepared in Example 1. Figure 1 b is a high-resolution transmission electron microscope (HRTEM) image of the gentian carbon dot material prepared in Example 1; Figure 2 The image shows the dynamic light scattering (DLS) pattern of the gentian carbon dot material prepared in Example 1. Figure 3 This is a stability analysis diagram of the gentian carbon dot material prepared in Example 1; Figure 4 The image shows the X-ray diffraction (XRD) pattern of the gentian carbon dot material prepared in Example 1. Figure 5 The Fourier transform infrared (FTIR) spectrum of the gentian carbon dot material prepared in Example 1 is shown below. Figure 6 The image shows the X-ray photoelectron spectroscopy (XPS) spectrum of the gentian carbon dot material prepared in Example 1. Figure 7 The image shows the electron spin resonance (ESR) diagram of the gentian carbon dot material prepared in Example 1. Figure 8 a is an experimental diagram showing the DPPH free radical scavenging ability of the gentian carbon dot material prepared in Example 1. Figure 8 b is an experimental diagram showing the ABTS free radical scavenging ability of the gentian carbon dot material prepared in Example 1. Figure 8 c is an experimental diagram showing the hydroxyl radical scavenging ability of the gentian carbon dot material prepared in Example 1. Figure 8 d is an experimental diagram of SOD enzyme activity of the gentian carbon dot material prepared in Example 1; Figure 9 The graph shows the cytotoxicity test results of the gentian carbon dot material prepared in Example 1 at different concentrations (25, 50, 100, 200 and 400 μg / mL). Figure 10 This is an experimental diagram showing the antagonistic effect of gentian carbon dot material prepared in Example 1 on jellyfish toxin. Detailed Implementation

[0018] To better explain the present invention, detailed descriptions of its embodiments are provided, and the main content of the invention is further clarified in conjunction with specific examples. However, the content of the present invention is not limited to the following embodiments. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all commercially available conventional products.

[0019] This invention synthesizes gentian carbon dot materials using a simple hydrothermal method. This method is simple, easy to operate, and highly reproducible. The gentian carbon dots, acting as nanocarriers for the active ingredients in gentian, not only improve drug bioavailability but may also further enhance anti-inflammatory effects through their unique nano-effects (such as surface modification and targeted delivery). Furthermore, they can directly scavenge free radicals through their surface-active groups (such as hydroxyl and carboxyl groups), further enhancing antioxidant capacity.

[0020] The following examples illustrate in more detail the preparation method of the gentian carbon dot material of the present invention, and the performance of the prepared gentian carbon dot material is tested. However, the embodiments of the present invention are not limited to the following examples.

[0021] Example 1: Synthesis of Gentian Carbon Dot Material Step 1: Crush the raw material of the Chinese herbal medicine Gentiana scabra into powder, weigh 1.5 g of the powder, put it into a glass bottle containing 30 mL of distilled water, and treat it with an ultrasonic cleaner for 30 min.

[0022] Step 2, the method for converting gentian suspension into gentian solution by hydrothermal synthesis is as follows: the suspension is transferred to a stainless steel autoclave lined with PTFE and heated at 180 °C for 6 h.

[0023] Step 3: Cool the obtained mixture to room temperature and centrifuge at 10000 r / min for 20 min; after centrifugation, collect the supernatant and purify it using a 0.22 μm filter membrane; dialyze the purified solution using a cellulose dialysis bag (1000 Da) for 3 days, changing the water every 8 h. Freeze-dry the dialyzed solution and store it at 4 ℃.

[0024] Performance Testing 1. The gentian carbon dot material obtained in Example 1 was characterized by TEM and HRTEM, and the results are as follows: Figure 1 As shown.

[0025] in, Figure 1 a is a TEM image of the gentian carbon dot material obtained in Example 1. Figure 1As can be seen from a, the present invention prepared gentian carbon dot materials with uniform, spherical shape and concentrated particle size distribution through hydrothermal reaction, with an average particle size of 2.49±0.08 nm.

[0026] Figure 1 b is an HRTEM image of the gentian carbon dot material obtained in Example 1 of the above embodiments. The morphology and size are consistent with those observed from the TEM image, and the lattice spacing is 0.202 nm (corresponding to the graphite (100) crystal plane).

[0027] 2. The gentian carbon dot material obtained in Example 1 was characterized by DLS, and the results are as follows: Figure 2 As shown.

[0028] Depend on Figure 2 The gentian carbon dot material obtained in Example 1 shows a negative potential with an average potential of -11.2 mV.

[0029] 3. Stability analysis was performed on the gentian carbon dot material obtained in Example 1, and the results are as follows: Figure 3 As shown.

[0030] Depend on Figure 3 The gentian carbon dot material obtained in Example 1 shows that it can maintain good dispersibility and no precipitation after being stored at room temperature in H2O and PBS for 15 days.

[0031] 4. The gentian carbon dot material obtained in Example 1 was characterized by XRD, and the results are as follows: Figure 4 As shown.

[0032] Depend on Figure 4 The X-ray diffraction pattern of the gentian carbon dot material obtained in Example 1 shows a broad diffraction peak at 21.4°.

[0033] 5. The gentian carbon dot material obtained in Example 1 was characterized by FTIR, and the results are as follows: Figure 5 As shown.

[0034] Depend on Figure 5 The Fourier transform infrared spectrum of the gentian carbon dot material obtained in Example 1 shows characteristic peaks such as -OH / -NH (3396.2 cm⁻¹), C=O (1639.9 cm⁻¹), -CO-NH- (1408.0 cm⁻¹), and CO (1096.0 cm⁻¹), indicating that the surface is rich in functional groups such as hydroxyl and amino groups.

[0035] 6. The gentian carbon dot material obtained in Example 1 was characterized by XPS, and the results are as follows: Figure 6 As shown.

[0036] Depend on Figure 6 The X-ray photoelectron spectrum of the gentian carbon dot material obtained in Example 1 shows that the full spectrum contains three elements: C, N, and O. The fine spectrum shows C 1s: 284.80 eV (CC / C=C), 286.12 eV (CO), 288.40 eV (C=O); N 1s: 399.80 eV (CN), 400.60 eV (C=N); O 1s: 530.10 eV (C=O), 533.00 eV (CO).

[0037] 7. The gentian carbon dot material obtained in Example 1 was characterized by ESR, and the results are as follows: Figure 7 As shown.

[0038] Depend on Figure 7 The electron spin resonance diagram of the gentian carbon dot material obtained in Example 1 is shown by... Figure 7 It can be seen that the introduction of carbon dot materials from gentian leads to the formation of OH· and ·O2. - , 1 The ESR signal intensity corresponding to active substances such as O2 and DPPH· is significantly reduced, proving that the carbon dot material of gentian can effectively scavenge free radicals.

[0039] 8. The in vitro performance verification experiment of the gentian carbon dot material obtained in Example 1 is shown in the figure. Figure 8 As shown.

[0040] Figure 8 The diagram shows the in vitro performance verification experiment of the gentian carbon dot material obtained in Example 1. Figure 8 a is an experimental graph showing the DPPH free radical scavenging capacity. Figure 8 It can be seen that the carbon dot material of gentian has a good scavenging ability for DPPH free radicals, and the scavenging effect shows a significant dose-dependent effect with increasing concentration. Figure 8 b is the experimental graph of ABTS' free radical scavenging ability, from Figure 8 b shows that the gentian carbon dot material has a highly efficient scavenging ability for ABTS free radicals, and the scavenging effect is significantly dose-dependent with increasing concentration. Figure 8 c is the experimental graph of hydroxyl radical scavenging ability, from Figure 8 c shows that the gentian carbon dot material has a good scavenging ability for hydroxyl radicals, and the scavenging effect is significantly dose-dependent with increasing concentration. Figure 8 d is a graph showing the activity of SOD-like enzymes. Figure 8 As can be seen from d, the gentian carbon dot material has extremely high SOD enzyme activity, proving that it has good antioxidant properties.

[0041] 9. Cytotoxicity experiments were conducted on the gentian carbon dot material obtained in Example 1 at different concentrations (25, 50, 100, 200, and 400 μg / mL), and the results are shown in the figure. Figure 9 As shown.

[0042] The cytotoxicity of different concentrations (25, 50, 100, 200, and 400 μg / mL) of gentian carbon dot material to cells was evaluated using the CCK-8 assay. 100 μL of HaCaT cell suspension was seeded into 96-well plates and incubated for 24 h. After removing the culture medium, the cells were washed once with PBS buffer, and then 100 μL of different concentrations of pre-prepared gentian carbon dot material were added. Cells were incubated for 24 h and 48 h, respectively. After washing twice with PBS buffer, 100 μL of CCK-8 cell counting reagent was added and the cells were incubated for 35 min. The absorbance at 450 nm was recorded using a microplate reader to assess cell viability.

[0043] 10. The antagonistic effect of jellyfish toxin on the gentian carbon dot material obtained in Example 1 was tested, and the results are as follows: Figure 10 As shown.

[0044] The antagonistic effect of different concentrations (25, 50, 100, 200, and 400 μg / mL) of gentian carbon dot material against jellyfish toxin was evaluated using the CCK-8 assay. 100 μL of HaCaT cell suspension was seeded into 96-well plates and incubated for 24 h. After removing the culture medium, the cells were washed once with PBS buffer, and then 100 μL of different concentrations of the prepared material (Con group, TE group, 25, 50, 100, 200, and 400 μg / mL groups, and CCK group) were added respectively. After co-incubation for 2 h, the cells were washed twice with PBS buffer, and then 100 μL of the cell counting reagent CCK-8 solution was added and co-incubated with the cells for 35 min. The absorbance at 450 nm was recorded using a microplate reader to assess cell viability.

[0045] I. Working Principle of this Solution This method primarily utilizes a one-step hydrothermal carbonization process to in-situ bind natural active ingredients (gentiopicrin, gentioketone, flavonoids, etc.) from the traditional Chinese medicine Gentiana scabra to a carbon skeleton, preparing a Gentiana scabra carbon dot material that combines the properties of nanomaterials with the pharmacological activity of natural drugs. With its ultra-small, near-spherical particle size of 2.49±0.08 nm, this material can efficiently penetrate the skin and mucous membrane barriers to enter damaged cells. The surface-enriched hydroxyl, carboxyl, and amino groups can directly capture and scavenge various reactive oxygen species (ROS) induced by jellyfish toxins, such as hydroxyl radicals, superoxide anions, and singlet oxygen, through electron transfer. Simultaneously, the core active ingredients of Gentiana scabra retained in the material can specifically activate the Nrf2 antioxidant pathway, upregulate the expression of endogenous antioxidant enzymes, and inhibit oxidative stress damage at its source. Based on this, gentian carbon dots can further block the activation of inflammatory signaling pathways such as NF-κB and MAPK, reduce the excessive secretion of pro-inflammatory cytokines such as TNF-α and IL-6, completely break the vicious cycle of "oxidative stress-inflammatory response" caused by jellyfish toxins, and ultimately achieve the protection and tissue repair of damaged skin cells.

[0046] II. Core Innovations of this Plan The core innovation of this solution lies in proposing a green synthesis strategy using gentian root, a single traditional Chinese medicine, as a biomass precursor. This strategy enables the preparation of functionalized carbon dot materials with controllable physicochemical properties and excellent biocompatibility. It addresses the industry pain points of existing clinical treatments for jellyfish stings, which only address symptoms and cannot break the vicious cycle of pathological changes, as well as the poor biocompatibility, complex preparation processes, high production costs, and difficulty in large-scale production of traditional chemically synthesized nanomaterials. Furthermore, it is the first time that gentian root carbon dots have been applied to the repair of jellyfish toxin-induced skin damage, achieving a synergistic therapeutic mechanism of "nanocarrier effect + natural drug activity," and constructing a complete technology verification and application system. Pain Point Anchoring: Jellyfish toxins cause skin damage by activating dual pathways of oxidative stress and inflammation. Existing treatments such as acetic acid neutralization and antihistamines can only relieve symptoms and cannot reverse the pathological process. They also have problems such as large side effects and easy recurrence. Therefore, the core treatment idea of ​​"dual blocking of the pathological cycle" was established.

[0047] Structural implementation: By optimizing the hydrothermal reaction parameters (reaction at 180℃ for 6h), the morphology and surface chemical properties of carbon dots were precisely controlled, and uniform spherical carbon dots with an average particle size of 2.49±0.08 nm, a lattice spacing of 0.202 nm (corresponding to the graphite (100) crystal plane), and an average potential of -11.2 mV were prepared. Their surfaces are rich in active functional groups such as -OH / -NH, C=O, -CO-NH-, and CO, and they have excellent water solubility and dispersion stability.

[0048] Validation: The physicochemical properties of the material were identified by characterization methods such as TEM, HRTEM, XRD, FTIR, and XPS. Its broad-spectrum antioxidant capacity was verified by ESR and in vitro free radical scavenging experiments. Its biocompatibility was confirmed by CCK-8 cytotoxicity experiments. Finally, its cell protection and anti-inflammatory activities were verified in a jellyfish toxin-induced HaCaT cell injury model.

[0049] Closed-loop effect: Gentian root carbon dots can dose-dependently improve the survival rate of damaged cells, significantly reduce intracellular ROS levels, inhibit the expression of pro-inflammatory factors, and block the vicious cycle of "oxidative stress-inflammation" at the cellular level, thus achieving a clear skin repair effect.

[0050] Scope expansion: This green synthesis process requires no complex equipment or toxic reagents, uses common bulk Chinese medicinal herbs as raw materials, and can achieve low-cost large-scale production. At the same time, it provides a novel nanomedicine candidate for the treatment of other marine biological toxin damage and inflammatory skin diseases.

[0051] III. Technical Effects of Implementing this Solution Implementing this solution can achieve significant technological breakthroughs in multiple dimensions, including material preparation, bioactivity, and clinical application potential. The overall technical effects are as follows: The material preparation process is green and efficient, and the product quality is stable and controllable: This scheme adopts a one-step hydrothermal synthesis method, requiring only gentian powder and distilled water as raw materials. The target product can be prepared through four simple steps: ultrasonic dispersion, high-temperature reaction, centrifugal purification, and dialysis freeze-drying. No chemical cross-linking agents or catalysts are added, avoiding the risk of toxic reagent residues. The process is simple to operate and highly reproducible. The prepared gentian carbon dots have a highly concentrated particle size distribution (average particle size 2.49±0.08nm), with no obvious agglomeration. They still maintain good dispersibility after 15 days of storage at room temperature in water and PBS buffer. Characterization results such as XRD, FTIR, and XPS show that the material has a typical graphitized short-range ordered structure, stable surface functional group composition, and minimal batch-to-batch differences, fully meeting the requirements for material quality uniformity in biomedical applications.

[0052] Excellent biocompatibility and high safety for clinical application: The toxicity of different concentrations (25, 50, 100, 200, 400 μg / mL) of gentian root carbon dots to human immortalized epidermal cells (HaCaT) was detected by CCK-8 assay. The results showed that the cell survival rate of each concentration group remained above 90% during incubation periods of 24 h and 48 h, with no significant cytotoxicity. This characteristic is significantly superior to traditional chemically synthesized nanomaterials, avoiding cell damage and inflammatory reactions that may be caused by nanomaterials themselves, and providing a guarantee for the long-term safety of topical skin administration.

[0053] Synergistic enhancement of antioxidant and anti-inflammatory activities, with a clear therapeutic mechanism: The gentian carbon dots prepared in this scheme combine the surface effects of nanomaterials with the pharmacological activities of natural drugs, achieving a synergistic therapeutic effect of 1+1>2. On the one hand, the numerous active functional groups such as hydroxyl and carboxyl groups on the surface of the carbon dots can directly scavenge various ROS through electron transfer. ESR experiments show that the introduction of gentian carbon dots can significantly reduce the signal intensity corresponding to free radicals such as OH・, ・O2⁻, ¹O2, and DPPH・. In vitro experiments further confirm that its scavenging rate of DPPH, ABTS, and hydroxyl free radicals is significantly dose-dependent, and it has extremely high SOD-like enzyme activity. On the other hand, the material retains the pharmacological effects of active ingredients such as gentiopicrin and flavonoids in gentian, and can specifically activate the Nrf2 antioxidant pathway while inhibiting the activation of NF-κB and MAPK inflammatory signaling pathways, blocking the secretion of pro-inflammatory factors from the upstream, and completely breaking the vicious cycle of "oxidative stress-inflammation" caused by jellyfish toxins. This is a core advantage that existing symptomatic treatments cannot achieve.

[0054] With excellent skin penetration and significant therapeutic effect, the ultra-small particle size of gentian carbon dots (approximately 2.49 nm) allows them to easily penetrate the skin's stratum corneum barrier, directly targeting damaged epidermal and dermal cells. Local administration provides highly effective treatment, avoiding the side effects associated with systemic administration. In a jellyfish toxin-induced HaCaT cell injury model, different concentrations of gentian carbon dots significantly improved the survival rate of damaged cells, and the therapeutic effect increased with increasing concentration, demonstrating a clear dose-response relationship and providing a scientific basis for the development of subsequent clinical dosing regimens.

[0055] With broad industrialization prospects and significant social value, this solution utilizes gentian root, a common and widely used traditional Chinese medicinal herb. It is abundant, inexpensive, and its preparation process is simple and easily scaled up, requiring no expensive production equipment. Production costs are far lower than traditional nanomedicines, making it suitable for large-scale industrial production. The successful application of this technology will not only fill the market gap for effective treatments for jellyfish stings and address the urgent clinical needs in coastal areas, but also open up new avenues for the application of plant-derived carbon dots in the biomedical field, promoting the integrated development of traditional Chinese medicine and modern nanotechnology.

[0056] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the above examples, and various exploratory changes can be made according to the inventive purpose of the present invention. Any other changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and fall within the protection scope of the present invention.

Claims

1. A gentian carbon dot material with a repairing effect on skin damaged by jellyfish toxins, characterized in that: The average particle size is 2.49 ± 0.08 nm, the lattice spacing is 0.202 nm (corresponding to the graphite (100) crystal plane), and the average potential is -11.2 mV.

2. The gentian carbon dot material with repairing effect on skin damaged by jellyfish toxins according to claim 1, characterized in that: There is a broad diffraction peak at 21.4 °, with characteristic peaks for -OH / -NH (3396.2 cm⁻¹), C=O (1639.9 cm⁻¹), -CO-NH- (1408.0 cm⁻¹), and CO (1096.0 cm⁻¹).

3. The gentian carbon dot material with repairing effect on skin damaged by jellyfish toxins according to claim 1, characterized in that: The gentian carbon dot material described herein serves as a highly effective skin repair agent, effectively reducing the level of excessive reactive oxygen species (ROS) accumulated in cells, thus providing a good repair effect on skin damaged by jellyfish toxins.

4. A method for preparing a gentian carbon dot material with a repairing effect on skin damaged by jellyfish toxins as described in any one of claims 1 to 3, characterized in that: Includes the following steps: Step 1: Processing of gentian root material; Step 2: Gentian suspension is converted into gentian carbon dot solution by hydrothermal synthesis; Step 3: Dialyze and freeze-dry the gentian carbon dot solution to form gentian carbon dots.

5. The method for preparing the gentian carbon dot material with repairing effect on skin damaged by jellyfish toxin according to claim 4, characterized in that: In step 1, the processing method of the gentian root material is as follows: crush the gentian root material, weigh 1.5 g of the Chinese medicine powder, put it into a glass bottle containing 30 mL of distilled water, and treat it with an ultrasonic cleaner for 30 min.

6. The method for preparing the gentian carbon dot material with repairing effect on skin damaged by jellyfish toxin according to claim 4, characterized in that: In step 2, the specific method for converting the gentian suspension into a gentian solution via hydrothermal synthesis is as follows: the suspension is transferred to a stainless steel autoclave lined with tetrafluoroethylene and heated at 180 °C for 6 h.

7. The method for preparing the gentian carbon dot material with repairing effect on skin damaged by jellyfish toxin according to claim 4, characterized in that: In step 3, the method for dialyzing and lyophilizing the gentian carbon dot solution to form gentian carbon dots is as follows: the obtained mixture is cooled to room temperature and centrifuged at 10000 r / min for 20 min; after centrifugation, the supernatant is collected and purified using a 0.22 μm filter membrane. The purified solution was dialyzed for 3 days using a cellulose dialysis bag (1000 Da), with the water changed every 8 hours. The dialyzed solution was then freeze-dried and stored at 4 °C.

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