Armadillidiam vulgare derived carbon quantum dots as well as preparation method and application thereof

Abstract

The invention relates to armadillidiam vulgare derived carbon quantum dots as well as a preparation method and application thereof, the armadillidiam vulgare derived carbon quantum dots are characterized in that the particle size of the armadillidiam vulgare derived carbon quantum dots is 2.2-6.0 nm, and the armadillidiam vulgare derived carbon quantum dots can inhibit pain response, reduce oxidative stress level and reduce inflammatory factor gene expression level. The armadillidiam vulgare derived analgesic carbon quantum dots inhibit irritant movement and oxidative damage in a pain model, and reduce inflammatory factors and pain pathway gene expression; the oxidative stress level is reduced, and the effects of relieving pain, resisting inflammation and resisting oxidative stress are achieved.

Description

Technical Field

[0001] The present invention relates to the field of biomaterials, and in particular to woodlice-derived carbon quantum dots for efficient pain relief, anti-oxidative stress and inflammation elimination, as well as a preparation method and application thereof. Background Art

[0002] Pain is a strong stimulus that can cause a series of severe stress reactions in the human body, causing the release of multiple stress hormones in the body, and then affecting the functions of various systems in the body. Not only will it cause the patient's condition to worsen, but it will also seriously affect the patient's quality of life. Therefore, it is urgent to find a pain relief method that has good analgesic effect and fewer adverse reactions and is more acceptable to patients. However, the currently commonly used opioid analgesics are due to patients' concerns about their addictiveness and fear of adverse reactions such as constipation, nausea and vomiting, loss of appetite, and fatigue caused by opioid analgesics. Therefore, oral painkillers are refused, compliance is poor, clinical applications are difficult, and the analgesic effect is not effective. Therefore, the new nanomaterial carbon quantum has appeared in people's field of vision due to its chemical inertness, high environmental compatibility, low toxicity, low synthesis cost and retention of raw material characteristics, and has gradually received widespread attention.

[0003] Inflammation is considered to be one of the main causes of pain in the body. Inflammation produces inflammatory factors and chemicals that stimulate nerve endings and cause pain. Inflammation stimulates oxidative stress and induces reactive oxygen species (ROS). Excessive ROS not only destroys cell proteins, lipids and DNA, causes cell damage and triggers cell apoptosis. At the same time, it leads to chronic inflammation of many neurodegenerative, cardiovascular and metabolic diseases, further promoting the body's inflammatory response. Therefore, eliminating ROS and resisting oxidative stress are the key to combating inflammation and relieving pain. In recent years, there have been a variety of carbon quantum dots with antioxidant enzyme activity, which have achieved relatively ideal results in the prevention and treatment of various diseases. Therefore, carbon quantum dots have great potential in anti-inflammatory and analgesic aspects. Summary of the invention

[0004] The present invention designs a woodlice-derived carbon quantum dot and a preparation method and application thereof, and solves the technical problem that the existing Chinese medicinal material carbon quantum dots have poor analgesic and anti-inflammatory effects.

[0005] In order to solve the above-mentioned technical problems, the present invention adopts the following solutions: A woodlice-derived carbon quantum dot, characterized in that its particle size is between 2.0 and 6.0 nm; the woodlice-derived carbon quantum dot can inhibit pain response, reduce oxidative stress level, and reduce inflammatory factor gene expression level.

[0006] Preferably, the woodlice-derived carbon quantum dots contain five elements, namely, C, N, O, H, and Na, and the element contents of C, O, N, and Na on their surfaces are 33.29%, 25.37%, 3.59%, and 17.76%, respectively.

[0007] Preferably, the pillbug-derived carbon quantum dots have a wavelength of 3291 cm -1 、2923cm -1 , 2850 cm -1 、1574 cm -1 、1406cm -1 、1048 cm -1 The characteristic infrared absorption peaks are v(-OH), v(CH), v(CH), v(C=O), v(CN) and v(C-0-C) respectively.

[0008] Preferably, a 0.1 mg / mL concentration of pillbug-derived carbon quantum dot solution can effectively reduce the stress effects caused by pain response.

[0009] A method for preparing pillbug-derived carbon quantum dots comprises the following steps: Step 1, dissolving dried pill bugs and sodium hydroxide in deionized water to obtain solution A; Step 2, solution A is placed in a reactor for hydrothermal reaction to obtain a brown solution A; Step 3: Centrifuge, dialyze, and freeze-dry the brown solution A in sequence to obtain brown analgesic and anti-inflammatory carbon quantum dots, that is, woodlice-derived carbon quantum dots.

[0010] Preferably, in step 1, the mass ratio of dried pill bugs to sodium hydroxide is 1:1 to 5:1.

[0011] Preferably, in step 2, solution A is placed in a reaction kettle and reacted in a reaction furnace for 300 minutes to obtain a brown solution A.

[0012] Preferably, in step 3, the mixture is centrifuged at 12000 rpm for 20 min and dialyzed for 48 h.

[0013] An analytical application of pillbug-derived carbon quantum dots, characterized by: Step 1, adding different concentrations of the above-mentioned pillbug-derived carbon quantum dot solutions to 0 dpf zebrafish embryos for culture, recording the effects of different time and concentrations of carbon quantum dots on the zebrafish nervous system and blood circulation system, and determining the concentration of carbon quantum dots acting on zebrafish; Step 2: According to the concentration range determined in step 1, the glacial acetic acid-induced pain model was used to detect the effects of carbon quantum dots on pain stress and movement distance, and it was analyzed that the pillbug-derived carbon quantum dots effectively reduced the stress movement phenomenon caused by exogenous irritation pain, and the pillbug-derived carbon quantum dots had a good analgesic effect; Alternatively, according to the concentration range determined in step 1, it is analyzed that the pillbug-derived carbon quantum dots inhibit the increase in ROS and MDA content induced by acetic acid and inhibit the level of oxidative stress.

[0014] Preferably, the pillbug-derived carbon quantum dots have a pain model concentration range of 50-250 µg / mL, preferably 100 µg / mL.

[0015] Preferably, the time range of the pillbug-derived carbon quantum dots pain model is 1 minute.

[0016] A highly biocompatible analgesic, characterized by comprising the above-mentioned pillbug-derived carbon quantum dots.

[0017] A highly biocompatible anti-inflammatory drug, characterized by comprising the above-mentioned woodlice-derived carbon quantum dots.

[0018] The pillbug-derived carbon quantum dots and their preparation method and application have the following beneficial effects: (1) The pillbug-derived analgesic carbon quantum dots of the present invention inhibit irritant migration and oxidative damage in the pain model, and reduce the expression of inflammatory factors and pain pathway genes; reduce the level of oxidative stress, and achieve analgesic, anti-inflammatory and anti-oxidative stress effects.

[0019] (2) The present invention uses woodlice and sodium hydroxide as reaction precursors for one-time hydrothermal reaction synthesis. Infrared spectra and XPS spectra show that the quantum dots contain rich functional groups. Woodlice-derived carbon quantum dots can inhibit pain response, reduce oxidative stress levels, reduce the gene expression level of inflammatory factors, and achieve pain relief and anti-inflammatory effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 TEM image of pillbug-derived carbon quantum dots of the present invention; Figure 2 The lattice fringes of the pillbug-derived carbon quantum dots of the present invention; Figure 3 This is an infrared spectrum of the pillbug-derived carbon quantum dots of the present invention; Figure 4 This is the XPS spectrum of the pillbug-derived carbon quantum dots of the present invention; Figure 5 The effect of pillbug-derived carbon quantum dots on the movement distance of zebrafish in the acetic acid pain model; Figure 6The effect of pillbug-derived carbon quantum dots on the action pathway map of zebrafish in the acetic acid pain model; Figure 7 The effect of the pillbug-derived carbon quantum dots of the present invention on the expression of pain factors in the zebrafish acetic acid pain model; Figure 8 The effect of pillbug-derived carbon quantum dots on the expression of inflammatory factors in zebras in the acetic acid pain model; Fig. 9 This is the effect of the pillbug-derived carbon quantum dots of the present invention on the oxidative stress level of zebras in the acetic acid pain model. DETAILED DESCRIPTION

[0021] Combine the following Figures 1 to 9 , the present invention is further described: The present invention will study the preparation and analgesic and anti-inflammatory effects of pillbug-derived carbon dots.

[0022] Example 1: The present invention provides a biosafe analgesic and anti-inflammatory carbon quantum dots and a preparation method and application thereof. The present invention uses Chinese medicinal materials as synthetic raw materials and prepares carbon quantum dots by a one-step hydrothermal method.

[0023] Weigh 1g of dried woodlice, and dissolve sodium hydroxide in 20 mL of deionized water (0.1g, 0.2g, 0.4g). Place the reaction solution in a reactor and perform hydrothermal synthesis for 5h. Cool to room temperature, filter, dialyze for 48h, and freeze-dry to obtain woodlice-derived carbon quantum dots. The morphology of woodlice-derived carbon quantum dots is as follows: Figure 1 and Figure 2 As shown, the UV absorption capacity and fluorescence spectrum are Figure 3 and Figure 4 shown.

[0024] Embodiment 2: (1) Normally developed zebrafish fry 3 days after fertilization were selected and placed in a 6-well plate. A control group, a model group, a drug treatment group, and a carbon quantum dot treatment group were set up, with 30 zebrafish fry in each group.

[0025] (2) The model group, drug group, and carbon quantum dot juvenile fish group were treated with 0.01% acetic acid solution to induce pain and inflammatory response, and the treatment time was 1 min.

[0026] (3) A 100 µg / mL solution of the carbon quantum dots obtained in Example 1 was prepared and added to the holes of the fry in the carbon quantum dots treatment group. A 200 ug / mL solution of Lesong was prepared and added to the holes of the fry in the drug group. The control group and the model group were cultured with deionized water.

[0027] (4) Collect samples of zebrafish fry from each group after incubation for 1 min.

[0028] Embodiment 3: (1) Normally developed zebrafish fry 3 days after fertilization were selected and placed in a 6-well plate. A control group, a model group, a drug treatment group, and a carbon quantum dot treatment group were set up, with 30 zebrafish fry in each group.

[0029] (2) The model group, drug group, and carbon quantum dot juvenile fish group were treated with 0.01% acetic acid solution to induce pain and inflammatory response, and the treatment time was 1 min.

[0030] (3) A 100 µg / mL solution of the carbon quantum dots obtained in Example 1 was prepared and added to the holes of the fry in the carbon quantum dots treatment group. A 200 ug / mL solution of Lesong was prepared and added to the holes of the fry in the drug group. The control group and the model group were cultured with deionized water.

[0031] (4) Tracker animal trajectory tracking software was used to record the movement trajectories of the control group, model group, drug group and carbon quantum dot fry group during the 0.01% acetic acid treatment and analyze them.

[0032] The conclusion is: Figure 5 and Figure 6 As shown, the total distance moved by the larvae in the drug group and the pillbug-derived carbon quantum dot group was significantly lower than that in the model group. This proves that pillbug quantum dots effectively reduce the stress movement phenomenon caused by exogenous irritation pain. Pillbug quantum dots have a good analgesic effect.

[0033] Embodiment 4: (1) Based on the samples from each group obtained in Experimental Example 2, qPCR was used to analyze the genes related to pain and inflammatory factors.

[0034] The conclusion is: Figure 7 and Figure 8 As shown in the figure, the expression of related genes in the group treated with pillbug-derived carbon quantum dots was significantly lower than that in the model group, indicating that pillbug-derived carbon quantum dots can inhibit pain pathways and inflammatory responses.

[0035] Embodiment 5: (1) Based on the samples obtained in Experimental Example 2, a standard curve was drawn using the Bio-Tech Enhanced BCA Protein Concentration Detection Kit, and the protein concentration of each sample was calculated using the curve.

[0036] (2) ROS and MDA detection kits (Nanjing Jiancheng) were used to detect the ROS and MDA levels in each group.

[0037] The conclusion is: Fig. 9 As shown, pillbug-derived carbon quantum dots inhibited the increase of ROS and MDA content induced by acetic acid and suppressed the level of oxidative stress.

[0038] The above animal toxicity tests and pain model tests show that pillbug-derived quantum dots can act on pain response mediators, reduce the expression of inflammatory-related factors, and achieve analgesia and anti-inflammation. Pillbug-derived carbon quantum dots can effectively activate antioxidant pathways, reduce the level of reactive oxygen in the body, and achieve antioxidant effects by reducing oxidative stress. Pillbug-derived carbon quantum dots are a highly biocompatible analgesic that can be widely used in the treatment of various types of pain and inflammatory reactions.

[0039] The present invention is described above by way of example in conjunction with the accompanying drawings. It is obvious that the implementation of the present invention is not limited to the above-mentioned method. As long as various improvements are made using the method concept and technical solution of the present invention, or the concept and technical solution of the present invention are directly applied to other occasions without improvement, they are all within the protection scope of the present invention.

Claims

1. A pillbug-derived carbon quantum dot, characterized in that The particle size is between 2.0 and 6.0 nm; the pillbug-derived carbon quantum dots can inhibit pain response, reduce oxidative stress levels, and reduce the gene expression levels of inflammatory factors.

2. The pillbug-derived carbon quantum dots according to claim 1, characterized in that The pillbug-derived carbon quantum dots contain five elements: C, N, O, H, and Na, and the element contents of C, O, N, and Na on their surfaces are 33.29%, 25.37%, 3.59%, and 17.76%, respectively; or, The pillbug-derived carbon quantum dots have a peak at 3291 cm -1 、2923cm -1 , 2850 cm -1 、1574 cm -1 、1406 cm -1 、1048cm -1 There are characteristic infrared absorption peaks of v(-OH), v(CH), v(CH), v(C=O), v(CN) and v(C-0-C) respectively; or, A 0.1 mg / mL concentration of pillbug-derived carbon quantum dots aqueous solution can effectively reduce the stress effects caused by pain response.

3. A method for preparing pillbug-derived carbon quantum dots according to claim 1 or 2, comprising the following steps: Step 1, dissolving dried pill bugs and sodium hydroxide in deionized water to obtain solution A; Step 2, solution A is placed in a reactor for hydrothermal reaction to obtain a brown solution A; Step 3: Centrifuge, dialyze, and freeze-dry the brown solution A in sequence to obtain brown analgesic and anti-inflammatory carbon quantum dots, that is, woodlice-derived carbon quantum dots.

4. The method for preparing pillbug-derived carbon quantum dots according to claim 3, characterized in that: In step 1, the mass ratio of dried pill bugs to sodium hydroxide is 1:1 to 5:1, and the added deionized water is at least enough to completely dissolve the sodium hydroxide.

5. The method for preparing pillbug-derived carbon quantum dots according to claim 3, characterized in that: In step 2, solution A is placed in a reaction kettle and reacted in a reaction furnace for 300 minutes to obtain a brown solution A.

6. The method for preparing pillbug-derived carbon quantum dots according to claim 3, characterized in that: In step 3, centrifuge at 8000-20000 rpm for 5-40 min and dialyze for 20-60 h.

7. An analytical application of pillbug-derived carbon quantum dots, characterized in that: Step 1, adding different concentrations of the solution of the pillbug-derived carbon quantum dots according to any one of claims 1 to 5 to 0 dpf zebrafish embryos for cultivation, recording the effects of the carbon quantum dots at different times and concentrations on the nervous system and blood circulation system of the zebrafish, and determining the concentration of the pillbug-derived carbon quantum dots acting on the zebrafish; Step 2. According to the concentration range determined in step 1, the glacial acetic acid-induced pain model is used to detect the effect of carbon quantum dots on pain stress and movement distance, and it is analyzed that the woodlice-derived carbon quantum dots effectively reduce the stress movement phenomenon caused by exogenous irritation pain, and the woodlice-derived carbon quantum dots have a good analgesic effect; or, according to the concentration range determined in step 1, it is analyzed that the woodlice-derived carbon quantum dots inhibit the increase of ROS and MDA content induced by acetic acid, and inhibit the level of oxidative stress.

8. The analytical application of pillbug-derived carbon quantum dots according to claim 7, characterized in that: The concentration range of the woodlice-derived carbon quantum dots pain model is 50-250µg / mL, preferably 100µg / mL; or, the time range of the woodlice-derived carbon quantum dots pain model is 1min.

9. A highly biocompatible analgesic, characterized in that: The invention comprises the woodlice-derived carbon quantum dots as described in any one of claims 1 to 6.

10. An anti-inflammatory drug with high biocompatibility, characterized in that: The invention comprises the woodlice-derived carbon quantum dots as described in any one of claims 1 to 6.

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