Light-operated nano-reactor as well as preparation method and application thereof

Abstract

The invention provides a light-operated nano-reactor and a preparation method and application thereof, and belongs to the technical field of bionic nano-reactors, the light-operated nano-reactor comprises manganese-based nano-particles and a hybrid membrane coating the surfaces of the manganese-based nano-particles, the hybrid membrane is formed by fusing plant capsule-like bodies, egg yolk lecithin and cholesterol, and the plant capsule-like bodies, the egg yolk lecithin and the cholesterol are mixed to form the light-operated nano-reactor. The photosynthetic phosphorylation performance is realized. The nanoreactor provided by the invention can deliver an immunologic adjuvant and activate the anti-tumor response of immune cells, and can generate ATP through light control to supply energy to the immune response activity of the immune cells, so that the response efficiency and response time of the immune cells are improved. The invention has important significance for improving the response efficiency of various immune cells.

Description

Technical Field

[0001] The present invention relates to the technical field of bionic nanoreactors, and specifically to a light-controlled nanoreactor and a preparation method and application thereof, wherein the light-controlled nanoreactor uses a hybrid membrane obtained by the fusion of thylakoids and egg yolk phosphatidylcholine as a carrier to load an immune adjuvant, thereby effectively activating immune cells and providing energy to the immune cells under light conditions, thereby improving the response efficiency of the immune cells. Background Art

[0002] Malignant tumors have long been a serious threat to human life and health. The tumor microenvironment is composed of cells, extracellular matrix, soluble factors and signaling molecules, and the cells include lymphatic endothelial cells, immune cells, fibroblasts, angiogenic cells, etc. The tumor microenvironment is characterized by nutrient deficiency, tissue hypoxia, tumor angiogenesis and decreased pH.

[0003] As one of the most important physiological and biochemical processes in nature, green plant photosynthesis transforms inorganic matter into organic matter. It transforms light energy into electrical energy, bioenergy, and chemical energy, providing people with ideas for energy conversion and is a research hotspot in the field of molecular bionics.

[0004] In an oxygen-deficient environment, tumor cells achieve unlimited proliferation through anaerobic glycolysis. The competitive consumption of glucose by tumor cells and the hypoxic microenvironment of the tumor not only leads to changes in the polarity and phenotype of immune cells, but also causes immune cells to be in a metabolic deficiency and high oxidative stress environment for a long time, destroying the activation of immune cells and their tumor immune surveillance function, thereby achieving tumor immune escape. Therefore, it is particularly important to achieve the activation of immune cells in the tumor microenvironment and improve the efficiency of immune response. Summary of the invention

[0005] In view of the technical problems existing in the background technology, the present invention provides a light-controlled nanoreactor and a preparation method and application thereof, aiming to provide a new solution for improving the efficiency of immune cell response in the tumor microenvironment.

[0006] In a first aspect, the present invention provides a light-controlled nanoreactor, comprising manganese-based nanoparticles and a hybrid membrane coated on the surface of the manganese-based nanoparticles, wherein the hybrid membrane is formed by the fusion of plant thylakoids, egg yolk lecithin and cholesterol, and the mass ratio of plant thylakoids, egg yolk lecithin and cholesterol in the hybrid membrane is 1:(40~60):(10~20).

[0007] In some embodiments of the present invention, the manganese-based nanoparticles are hollow mesoporous manganese dioxide in the shape of sea urchins. In the light-controlled nanoreactor of the present invention, the manganese-based nanoparticles have at least the following functions: as nanoscale immune adjuvants, they support the hybrid membrane and can improve the electron conduction efficiency and ATP yield. Preferably, the particle size of the manganese-based nanoparticles is 220-260 nm.

[0008] In the above-mentioned light-controlled nanoreactor, plant thylakoids can be extracted from plants such as spinach, Arabidopsis, pea, Physcomitrella patens, etc. For example, in some embodiments of the present invention, the thylakoids used are extracted from spinach.

[0009] Preferably, in the above-mentioned light-controlled nanoreactor, the hybrid membrane is loaded with a small molecule immune adjuvant, including but not limited to a STING agonist, etc. For example, in some embodiments of the present invention, the hybrid membrane is loaded with DMXAA. It is understood that those skilled in the art can change the type of small molecule immune adjuvant according to actual needs.

[0010] Preferably, in the above-mentioned light-controlled nanoreactor, mannose is also modified; more preferably, the mass ratio of mannose to chlorophyll protein on plant thylakoids is (5-10): 1. Experimental data show that mannose modification has better cell uptake ability, while too low a modification amount will affect targeting, and too high a modification amount will affect the photosynthetic performance of chlorophyll protein on thylakoids.

[0011] In a second aspect, the present invention provides a method for preparing a light-controlled nanoreactor, comprising the following steps: S1, preparing liposome solution by egg yolk lecithin and cholesterol; S2, ultrasonically fusing the liposome solution in step S1 with the plant thylakoids in a dark environment; S3, combining the product obtained in step S2 with manganese-based nanoparticles through electrostatic adsorption to obtain a light-controlled nanoreactor.

[0012] In some embodiments of the present invention, plant thylakoids are extracted from spinach by osmosis, and the extraction operation is as follows: (a) Cut the spinach into 1-2 cm pieces 2 The cells were sized and placed in a low-temperature pretreated chloroplast extract for cell wall disruption, wherein the chloroplast extract is an aqueous solution containing 400 mM sucrose, 20 mM tris(hydroxymethyl)methylglycine, 2.5 mM anhydrous magnesium chloride, 15 mM sodium chloride, 5 mM sodium ascorbate and 0.2 wt% BSA; (d) filtering and collecting the filtrate, centrifuging the filtrate to obtain a chloroplast precipitate, and adding a thylakoid extract to the precipitate for extraction reaction, wherein the thylakoid extract is an aqueous solution containing 20 mM tris(hydroxymethyl)methylglycine, 5 mM anhydrous magnesium chloride, 10 mM sodium chloride and 0.2 wt% BSA; (c) Centrifuge the reaction solution, wash the precipitate, and obtain the thylakoids, which are then preserved.

[0013] Preferably, in the above method, the liposome solution prepared in step S1 contains a small molecule immune adjuvant. Specifically, egg yolk lecithin, cholesterol and a small molecule immune adjuvant are dissolved in an organic solvent and then rotary evaporated to obtain a liposome membrane, which is then hydrated and ultrasonicated to obtain a liposome solution.

[0014] Preferably, in the above method, the ultrasonic conditions of step S2 are: the temperature is controlled at 37±3°C, the ultrasonic operating frequency is selected to be 40 KHz, the ultrasonic power is set to 200~300 W, and the reaction time is 25~45 min.

[0015] Preferably, in the above method, step S3 is specifically: adding positively charged manganese-based nanoparticles to the product obtained in step S2, and ultrasonicating. In some embodiments of the present invention, the manganese-based nanoparticles are hollow mesoporous manganese dioxide in the shape of sea urchins, and polyacrylamine hydrochloride (PAH) is used to reverse its potential to positive charge, so as to better react electrostatically with the negatively charged membrane fusion thylakoids prepared in step S2.

[0016] More preferably, in the above method, the ultrasonic conditions of step S3 are: the temperature is controlled at 25±5°C, the ultrasonic operating frequency is selected to be 40 KHz, the ultrasonic power is set to 200 W to 300 W, and the reaction is carried out for 25 to 45 min.

[0017] Preferably, in step S3 of the above method, after the electrostatic adsorption is completed, mannose is added to modify the surface of the nanoreactor.

[0018] In a third aspect, the present invention provides a preservation solution for a light-controlled nanoreactor, which contains 380-420 mM sucrose, 13.5-16.5 mM sodium chloride and 18-22 mM 2-morpholineethanesulfonic acid. The preservation solution can effectively delay the morphological changes and sedimentation of the thylakoid and the hybrid membrane in the light-controlled nanoreactor during the preservation process, and can well maintain the stability of the light-controlled nanoreactor, laying a foundation for its sales, preservation, transportation, etc.

[0019] In a fourth aspect, the present invention provides the use of a light-controlled nanoreactor in the preparation of the following products: Anti-tumor products; Products that provide energy to immune cells; Products that improve the efficiency and duration of immune cell responses; Among them, the products can be chemical reagents, drugs, medical devices, etc.

[0020] In the above applications, immune cells include but are not limited to macrophages, DC cells, T cells, neutrophils, and NK cells, etc. For example, in some embodiments of the present invention, RAW264.7 macrophages are used as cell models, and macrophages internalize a large number of light-controlled nanoreactors loaded with STING agonists, produce ATP under light conditions, and within a certain period of time, the effect is enhanced over time, and the macrophage M1 polarization index is significantly improved, while the ATP content of the non-light-treated group is increased relative to the group, and the macrophage M1 polarization index is slightly improved, but the secretion intensity and the duration of the time window are much weaker than those of the light-treated group.

[0021] In a fifth aspect, the present invention provides a method for improving the response efficiency of immune cells, specifically: after the immune cells take up the light-controlled nanoreactor provided by the present invention, they are irradiated with red light; the immune adjuvant loaded in the light-controlled nanoreactor first acts on the relevant immune targets to activate the anti-tumor response of the immune cells, and at the same time, the ATP generated by the light-controlled nanoreactor can continuously supply energy to the immune cells, thereby improving the response efficiency and response time of the immune cells.

[0022] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention fuses thylakoids with liposomes, which greatly improves the biocompatibility and in vitro stability of thylakoids, so that the bionic nanoreactor obtained by the present invention has strong stability under 4°C, providing a new idea for bionic nanoreactors.

[0023] (2) The biomimetic nanoreactor obtained by the transformation strategy of the present invention activates immune cells by delivering small molecule immune adjuvants (such as STING agonist, DMXAA) and manganese-based nanoparticles. At the same time, under light conditions, thylakoids simulate photosynthesis to produce ATP, which provides energy for immune cells and improves the efficiency of immune response. Therefore, it has better tumor treatment efficacy.

[0024] (3) The bionic nanoreactor obtained by the transformation strategy of the present invention not only has the function of producing ATP and stimulating the activation of immune cells, but also has the performance of promoting the polarization of macrophages M1. Through the stimulation of heterologous substances in the thylakoid protein itself and the combined action of ATP, it improves the adverse factors such as insufficient metabolism and high oxidative stress environment in the tumor microenvironment. M2 macrophages are reversed into M1 macrophages in a pro-inflammatory state, producing a large number of inflammatory factors and activating adaptive immunity, further improving the efficiency of the immune response and achieving tumor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solution of the present invention, the following is a brief introduction to the drawings used in the present invention. Obviously, the drawings described below are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained based on these drawings without creative work.

[0026] Figure 1 TEM images of the hollow mesoporous manganese dioxide and the light-controlled nanoreactor prepared in the embodiments of the present invention; Figure 2 The endocytosis efficiency of RAW264.7 cells under different treatment conditions of the light-controlled nanoreactor in the embodiment of the present invention; Figure 3 This is a graph showing the experimental results of ATP expression in a light-controlled nanoreactor under different time conditions in an embodiment of the present invention; Figure 4 This is a graph showing the experimental results of the expression of TNF-α protein under different time conditions in the light-controlled nanoreactor according to an embodiment of the present invention; Figure 5 Graph showing the particle size changes at different times in the light-controlled nanoreactor according to an embodiment of the present invention. DETAILED DESCRIPTION

[0027] The following embodiments of the technical solution of the present invention are described in detail in conjunction with the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore only used as examples, and cannot be used to limit the protection scope of the present invention.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by technicians in the technical field to which this application belongs; the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention; the terms "including" and "having" and any variations thereof in this document are intended to cover non-exclusive inclusions.

[0029] In order to solve the technical problem of immunosuppression in the tumor microenvironment, the present invention provides a light-controlled nanoreactor and a preparation method and application thereof. The bionic nanoreactor includes manganese-based nanoparticles and a hybrid membrane coated on the surface of the manganese-based nanoparticles, wherein the hybrid membrane is formed by the fusion of plant thylakoids, egg yolk lecithin and cholesterol, has photosynthetic phosphorylation properties, and the hybrid membrane can be loaded with anti-tumor active small molecules. When the nanoreactor is internalized by immune cells, ATP is generated under light conditions to provide energy for the immune cells to perform immune response activities, thereby enhancing the intensity and duration of the immune cell response.

[0030] Some specific examples are listed below. It should be noted that the examples described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be obtained commercially.

[0031] Example 1 This example provides a light-controlled nanoreactor, including a hollow mesoporous manganese dioxide in the shape of a sea urchin and a hybrid membrane coated on the surface of the hollow mesoporous manganese dioxide, wherein the hybrid membrane is constructed by fusion of thylakoids, egg yolk phosphatidylcholine and cholesterol, and a small molecule adjuvant DMXAA is loaded on the hybrid membrane. Specifically, the preparation process of the light-controlled nanoreactor is as follows: (1) Extraction and processing of thylakoids.

[0032] Cut the spinach into 1-2 cm 2 The size of the cells was determined and the chloroplast extract solution pretreated at 4°C was added to break the cell wall. The filtrate was filtered through medical gauze folded into eight layers. The cells were centrifuged at 4°C and 6000 rpm for 15 min, and the supernatant was discarded to obtain the precipitate as chloroplasts. The thylakoid extract solution was added and ultrasonically dispersed, and the cells were placed on ice for reaction for 20 min. The cells were centrifuged at 4°C and 10000 rpm for 15 min, and the supernatant was discarded to obtain the precipitate as thylakoids. The thylakoid preservation solution was added and ultrasonically dispersed to obtain the concentrated solution.

[0033] Among them, the chloroplast extract is an aqueous solution containing 400 mM sucrose, 20 mM tri(hydroxymethyl)methylglycine, 2.5 mM anhydrous magnesium chloride, 15 mM sodium chloride, 5 mM sodium ascorbate and 0.2 wt% BSA; the thylakoid extract is an aqueous solution containing 20 mM tri(hydroxymethyl)methylglycine, 5 mM anhydrous magnesium chloride, 10 mM sodium chloride and 0.2 wt% BSA; the thylakoid preservation solution is an aqueous solution containing 400 mM sucrose, 15 mM sodium chloride and 20 mM 2-morpholineethanesulfonic acid.

[0034] (2) Manganese-based nanoparticles MnO 2 synthesis.

[0035] Weigh Triton X-100, cyclohexane and n-hexanol into a flask, stir at room temperature to dissolve, and immediately add ammonia and ddH 2 O mixed solution was stirred for 1 hour. Tetraethoxysilane and 3-aminopropyltriethoxysilane were added to the flask and the reaction was continued at room temperature for 24 hours. The reaction solution was collected by centrifugation, washed with water, dispersed in distilled water, and stored at 4°C. Weigh KMnO 4 Add ddH 2O, stirred at room temperature to dissolve. The above reaction solution was added at a uniform rate and the reaction was continued for 6 hours. The nanoparticles were collected by centrifugation to obtain sSiO 2 @MnO 2 , collected and washed with water to make up to volume, and Na 2 CO 3 solution, adding sSiO at a reaction temperature of 60 °C 2 @MnO 2 , etching for 12 hours to obtain nano H-MnO 2 The above mixture was centrifuged and washed with water to obtain hollow mesoporous manganese dioxide (MnO 2 ), and an equal volume of polyacrylamine hydrochloride (PAH) solution were ultrasonically reacted for 30 min.

[0036] (3) Preparation of light-controlled nanoreactor.

[0037] Weigh 75 mg of egg yolk lecithin, 20 mg of cholesterol, and 300 μg of DMXAA, dissolve in 1 mL of chloroform, add into a 25 mL round-bottom flask, and perform rotary evaporation in a 40°C water bath to obtain a single-layer liposome membrane. Add 5 mL of double distilled water and hydrate in a 50°C water bath. Set the ultrasonic emulsification instrument to on for 2 s and off for 3s, and perform ultrasonic emulsification for 2 min to make the solution clear and transparent to obtain a liposome solution. The diluted thylakoids were further nanosized by a pneumatic extruder. The concentration of the nanosized thylakoids was determined by a UV spectrophotometer, and 300 μg of thylakoids were added to 1 mL of pre-synthesized liposome solution loaded with DMXAA (each 1 ml of liposomes contained 15 mg of egg yolk phosphatidylcholine, 4 mg of cholesterol, and 60 μg of DMXAA) and subjected to ultrasonic reaction at 37°C for 30 min in the dark to obtain membrane fusion thylakoids (TLD).

[0038] Add PAH-modified MnO to TLD 2 The ultrasonic reaction was carried out for 30 min, and the photo-controlled nanoreactor TLDM was obtained by centrifugation and washing.

[0039] Transmission electron microscopy was used to characterize the morphology of hollow mesoporous manganese dioxide and light-controlled nanoreactor TLDM. Figure 1 As shown in the results, the hollow mesoporous manganese dioxide prepared in this example presents a sea urchin shape, while the surface of the manganese-based bionic nanoreactor after membrane fusion modification shows an obvious membrane structure.

[0040] Example 2 Different from Example 1, the light-controlled nanoreactor provided in this example is also modified with mannose, and its preparation is specifically as follows: Steps (1) and (2) are the same as those in Example 1; The preparation of TLDM in step (3) is the same as that in Example 1; then, the synthesized TLDM is added to the pre-prepared mannose aqueous solution, ultrasonicated at 25°C for 30 min, and centrifuged and washed with water to obtain the mannose-modified light-controlled nanoreactor TLDM@Man, wherein the mass ratio of mannose to chlorophyll protein is 5:1.

[0041] RAW264.7 cells with good growth status were inoculated into confocal dishes and cultured overnight. After the cells attached to the wall, one dish was pretreated with mannose and TLDM@Man modified with mannose was added. The other two dishes were not pretreated with mannose, and TLDM without mannose modification was added to one dish, and TLDM@Man modified with mannose was added to the other dish. After incubation for four hours, the cells were stained with Hoechst and tested on the microscope. The results are shown in Figure 2. Figure 2 As shown: blue is the cell nucleus, and red is the material luminescence. The results show that TLDM@Man exhibits higher drug enrichment than TLDM, indicating that it has good cellular uptake ability after modification with mannose.

[0042] Example 3 This example demonstrates the ability of the nanoreactor TLDM to produce ATP under different conditions. The specific experiments and results are as follows: The ATP detection kit was used for detection. The detection principle is: when firefly luciferase catalyzes luciferin to produce fluorescence, ATP is required to provide energy. When luciferase and luciferin are in excess, within a certain concentration range, the generation of fluorescence is proportional to the concentration of ATP, and the ATP concentration in the solution can be detected with high sensitivity. The detection method is: RAW264.7 cells are planted in a six-well plate, and then TLDM (chlorophyll protein quantitative 80 μg mL -1 ) were co-cultured with cells for 4 hours, and the culture medium was replaced. After 8 hours, the illumination group was irradiated with red light, and the control group was not illuminated, and then co-cultured for 4 hours, 8 hours, and 12 hours respectively; the illumination group was treated with light for the second time, and then cultured for 1 hour, digested and resuspended in DMEM culture medium, and then centrifuged, and the supernatant was discarded; ATP lysis buffer was added, resuspended and centrifuged to obtain the supernatant, and the ATP concentration was detected according to the steps of the ATP detection kit.

[0043] Test results such as Figure 3 As shown: compared with the non-illumination treatment group, the ATP content of the bionic nanoreactor loaded with membrane fused thylakoids under illumination increased within the 8-16 hour window, and decreased after 20 hours; after illumination again, the ATP content increased again. This result shows that the bionic nanoreactor provided by the present invention simulates photosynthesis and produces ATP under illumination conditions.

[0044] Example 4 This example tests the ability of the biomimetic nanoreactor TLDM to promote macrophage M1 polarization under different conditions. The specific experiments and results are as follows: M1 macrophages secrete a large number of pro-inflammatory factors such as TNF-α, so measuring the TNF-α content can indirectly reflect the degree of M1 macrophage polarization.

[0045] Mouse TNF-α enzyme-linked immunosorbent assay kit was used for detection. The detection principle is as follows: using the double antibody sandwich ELISA method, anti-mouse TNF-α antibody is coated on the enzyme-labeled plate. During the experiment, mouse TNF-α in the sample (or standard) will bind to the coated antibody; then biotinylated anti-mouse TNF-α antibody and horseradish peroxidase-labeled avidin are added in sequence, and the anti-mouse TNF-α antibody binds to the mouse TNF-α bound to the coated antibody. Biotin and avidin specifically bind to form an immune complex, and the free components are washed away; add a color substrate (TMB), TMB appears blue under the catalysis of horseradish peroxidase, and turns yellow after adding the stop solution; use an enzyme reader to measure the OD value at a wavelength of 450 nm. The TNF-α concentration is proportional to the OD450 value. The concentration of TNF-α in the sample is calculated by drawing a standard curve. The detection method is as follows: take RAW264.7 cells and plant them in a six-well plate, then use TLDM (chlorophyll protein quantification 80 μg mL -1 ) and cells for 4 hours; the culture medium was replaced, and after 8 hours, the illumination group was irradiated with red light, and the control group was not irradiated with light and co-cultured for 4 hours, 8 hours, and 12 hours respectively; the illumination group was treated with light for the second time, and then cultured for 1 hour, digested and resuspended in DMEM medium, centrifuged, and the supernatant was discarded; the cell lysis buffer was added, resuspended and centrifuged to obtain the supernatant, and the detection was performed according to the steps of the mouse TNF-α enzyme-linked immunosorbent assay kit. Test results such as Figure 4 As shown: Compared with the non-illumination treatment group, the TNF-α content of the bionic nanoreactor TLDM under illumination conditions increased significantly within the 8-16 hour window, and began to decline after 16 hours; after 20 hours of illumination again, the TNF-α content increased again. The above results show that the bionic nanoreactor provided by the present invention simulates photosynthesis under illumination conditions to produce ATP, supply energy to immune cells, and promote the polarized expression of macrophage M1.

[0046] Comparative Example 1 Different from Example 1, this example directly combines thylakoids with hollow mesoporous manganese dioxide to prepare a light-controlled nanoreactor, and the preparation method is as follows: Steps (1) and (2) are the same as those in Example 1.

[0047] (3) Preparation of light-controlled nanoreactor.

[0048] The diluted thylakoids were further nanosized by a pneumatic extruder, and the concentration of the nanosized thylakoids was measured by a UV spectrophotometer. MnO was added dropwise to the thylakoid solution. 2 After ultrasonic reaction for 30 min, the cells were centrifuged and washed with water to obtain the light-controlled nanoreactor TM without liposome membrane fusion.

[0049] An aqueous solution containing 400 mM sucrose, 15 mM sodium chloride and 20 mM 2-morpholineethanesulfonic acid was used as the storage solution, and the hollow mesoporous manganese dioxide, TM without liposome membrane fusion and TLDM with liposome membrane fusion at different storage times were analyzed using a Malvern potential meter to determine the stability of the nanoreactor.

[0050] Test results such as Figure 5 As shown in the figure, the particle size of TM began to change gradually and flocculated after 2 days of storage, while the particle size of TLDM remained relatively stable, and the particle size of TLDM remained stable until the 12th day. The above results show that compared with directly combining thylakoids with manganese dioxide nanoparticles, the bionic nanoreactor obtained by coating thylakoids on manganese dioxide through membrane fusion technology in the present invention has strong stability.

[0051] In summary, the present invention constructs a bionic nanoreactor by fusing thylakoids with liposomes, simulates photosynthesis to produce ATP, supplies energy to immune cells, improves the efficiency of immune response, explains and verifies its mechanism of action, and lays a theoretical and experimental foundation for the application of thylakoids in bionic nanoreactors; at the same time, the bionic nanoreactor obtained by the transformation strategy of the present invention not only has the function of producing ATP and stimulating the activation of immune cells, but also has the performance of promoting the polarization of macrophages M1. Through the stimulation of heterologous substances in the thylakoid protein itself and the joint action of ATP, the adverse factors such as insufficient metabolism and high oxidative stress environment in the tumor microenvironment are improved, and M2 macrophages are reversed to M1 macrophages in a pro-inflammatory state, producing a large number of inflammatory factors and activating adaptive immunity; it can be seen that the bionic nanoreactor provided by the present invention can improve the efficiency of immune response and achieve tumor treatment.

[0052] It should be noted that the present invention is not limited to the above-mentioned embodiments. The above-mentioned embodiments are only examples, and the embodiments having the same structure as the technical idea and exerting the same effect within the scope of the technical solution of the present application are all included in the technical scope of the present invention. In addition, without departing from the scope of the main purpose of the present invention, various modifications that can be thought of by those skilled in the art to the embodiments and other methods of combining some of the constituent elements in the embodiments are also included in the scope of the present invention.

Claims

1. A light-controlled nanoreactor, characterized in that: The invention comprises manganese-based nanoparticles and a hybrid membrane coated on the surface of the manganese-based nanoparticles, wherein the hybrid membrane is formed by fusing plant thylakoids, egg yolk lecithin and cholesterol, and the mass ratio of the plant thylakoids, egg yolk lecithin and cholesterol is 1:(40-60):(10-20).

2. The light-controlled nanoreactor according to claim 1, characterized in that: The manganese-based nanoparticles are hollow mesoporous manganese dioxide in the shape of sea urchins, and / or; The plant is one of spinach, Arabidopsis thaliana, pea and Physcomitrella patens.

3. The light-controlled nanoreactor according to claim 1, characterized in that: The hybrid membrane is loaded with small molecule immune adjuvants, and / or; The light-controlled nanoreactor is modified with mannose, and the mass ratio of the mannose to the chlorophyll protein on the plant thylakoid is (5-10):

1.

4. A method for preparing the light-controlled nanoreactor as claimed in claim 1, characterized in that: The following steps are involved: Firstly, egg yolk lecithin and cholesterol are prepared into liposome solution, then the liposome solution and plant thylakoid are ultrasonically fused in the dark, and finally the obtained product is combined with manganese-based nanoparticles through electrostatic adsorption to obtain a nanoreactor.

5. The method according to claim 4, characterized in that The plant thylakoids were extracted from spinach by osmosis.

6. The method according to claim 4, characterized in that The liposome solution contains small molecule immune adjuvant.

7. The method according to claim 4, characterized in that After the electrostatic adsorption is completed, mannose is added to modify the surface of the nanoreactor.

8. A storage solution for storing the light-controlled nanoreactor according to claim 1, characterized in that: Contains 380-420 mM sucrose, 13.5-16.5 mM sodium chloride, and 18-22 mM 2-morpholineethanesulfonic acid.

9. Use of the light-controlled nanoreactor according to claim 1 in product preparation, wherein the product comprises any of the following: Products that provide energy to immune cells; Products that improve the efficiency and duration of immune cell responses; Anti-tumor products; The product is any one of a chemical reagent, a drug, and a medical device, and the immune cell is at least one of a macrophage, a DC cell, a T cell, a neutrophil, and a NK cell.

10. A method for improving immune cell response efficiency without the purpose of disease diagnosis and treatment, characterized in that: After the immune cells take up the light-controlled nanoreactor as claimed in claim 1, they are irradiated with red light.

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