CN117045607A - Preparation method and application of bionic tea polyphenol nano-particles - Google Patents
Preparation method and application of bionic tea polyphenol nano-particles Download PDFInfo
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- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
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Abstract
The invention discloses a preparation method and application of bionic tea polyphenol nano-particles, and belongs to the technical field of biological medicines. The invention discloses a preparation method of bionic tea polyphenol nano-particles, which comprises the steps of firstly preparing the tea polyphenol nano-particles by adopting a high-pressure homogenizer emulsification-volatilization method, then adding vesicles containing PRP, and modifying platelet vesicles on the surfaces of the tea polyphenol nano-particles by ultrasound to obtain the bionic tea polyphenol nano-particles; the bionic tea polyphenol nano-particles remarkably improve the cellular absorption, bioavailability and targeting of inflammatory lesions of tea polyphenol.
Description
Technical Field
The invention relates to the technical field of biological medicine, in particular to a preparation method and application of bionic tea polyphenol nano-particles.
Background
Respiratory diseases are common diseases in China and even worldwide, and have the characteristics of high morbidity, high mortality and high economic burden. In China, along with the development of industrialization, town and population aging and the change of ecological environment and life behavior, respiratory diseases become main death reasons and disease burden of residents.
Among respiratory diseases in our country, sepsis-induced pneumonia infections are most common, often leading to acute lung injury (acute lung injury, ALI) and more severe acute respiratory distress syndrome (aute respiratory distress syndrome, ARDS). In sepsis patients, bacterial infection causes the release of a large number of cytokines, leading to an excessive inflammatory response, ultimately leading to the occurrence of inflammatory factor storms (cytokine release syndrome, CRS). CRS not only results in an imbalance in immune response homeostasis, CRS occurring in the lungs can also lead to increased alveolar capillary permeability, pulmonary edema, and inflammation, leading to lung injury and respiratory failure. How to deal with inflammatory factor storms remains a great clinical challenge.
Bronchial asthma (asthma for short) is the most common chronic airway inflammatory disease. Asthma is one of the diseases with the fastest rising global morbidity. At present, the glucocorticoid comprehensive treatment is mainly used clinically, and symptoms can be effectively controlled, but serious toxic and side effects such as femoral head necrosis, liver and kidney injury and the like can be caused after long-term use. Therefore, finding effective therapeutic drugs and strategies for preventing and treating asthma is a problem which is urgent to be solved clinically.
Tea polyphenols are natural compounds, and widely exist in foods such as tea leaves and wine. It has various biological activities including antioxidant, antiinflammatory, anticancer, blood pressure lowering, blood sugar lowering, and blood lipid improving effects. However, due to the complex mechanism of action and low bioavailability, the characteristics of off-target property and difficult cell entry exist, and the clinical curative effect and application of the drug are greatly limited. Based on the above, if the modern nano technology is adopted, the tea polyphenol is subjected to nano modification, a new formulation is developed to improve the bioavailability, enhance the slow release property, and the targeted accurate administration of the disease part is adopted, so that the preparation has important significance for improving the curative effect of the medicine and expanding the clinical application.
Therefore, providing a preparation method and application of the bionic tea polyphenol nano-particles is a problem to be solved by the person skilled in the art.
Disclosure of Invention
In view of the above, the invention provides a preparation method and application of bionic tea polyphenol nano-particles, which can improve a series of problems of off-target, low selectivity, short half-life, low bioavailability and the like of small molecular medicine tea polyphenol, improve the curative effect and expand the clinical application.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a preparation method of bionic tea polyphenol nano-particles comprises the following specific steps:
1) Preparing a nano Chinese medicinal preparation:
(1) According to 0.3:1, weighing tea polyphenol and PLGA-PEG according to the weight ratio, and dispersing in dichloromethane; homogenizing for 3-5 minutes under high pressure under ice bath condition to obtain primary nanocrystalline suspension; the weight-volume ratio of PLGA-PEG to dichloromethane is 1:20 (1 g:20 mL);
(2) Taking 0.1-3% PVA water solution as external water phase; the volume ratio of the aqueous solution of 0.1-3% PVA to the dichloromethane is 3:1, a step of;
(3) Rapidly adding the primary nanocrystalline suspension prepared in the step (1) into the external water phase in the step (2) for shaking, and homogenizing for 3-5 minutes under high pressure under ice bath conditions to obtain nanocrystalline suspension;
(4) Adding ultra-pure water and 0.1-3% PVA water solution into the nanocrystalline suspension obtained in the step (3), and magnetically stirring for 4-6 hours in a fume hood (to volatilize dichloromethane in an organic phase and enable nanoparticles to form and harden) to obtain nanoemulsion; the volume ratio of the ultrapure water to the aqueous solution of 0.1-3% PVA is 1:1, a step of; the volume ratio of the ultrapure water to the dichloromethane is 3:2;
(5) Centrifuging the nanoemulsion obtained in the step (4) at a high speed, discarding the supernatant after centrifuging, and washing the precipitate with ultrapure water for three times to obtain tea polyphenol nanoparticles;
2) PRP (PlateletRichPlasma) was extracted to prepare PRP-containing vesicles;
3) Preparing bionic tea polyphenol nano particles:
dissolving tea polyphenol nano-particles into PBS buffer solution, adding PRP-containing vesicles, performing ultrasonic treatment in ice bath, and performing freeze drying to obtain bionic tea polyphenol nano-particles; the weight-to-volume ratio of the tea polyphenol nano-particles to the PBS buffer solution is 0.01:1 (0.01 g:1 mL); the volume ratio of the PRP-containing vesicles to the PBS buffer was 0.05:1.
further, the PVA in the step (2) is PVA with a deacetylation degree of 85-95%.
Further, the high-speed centrifugation in the step (5) is performed at 4 ℃, and 12000g is performed for 20min.
Further, the specific steps of extracting PRP and preparing PRP-containing vesicles in the step 2) are as follows:
centrifuge the centrifuge tube with blood (anticoagulant added) (blood from hospital boots or healthy volunteers donation) into a centrifuge at 2500-3500rpm for 4-6 minutes; extracting supernatant containing PRP, centrifuging at 12000-18000rpm for 8-12 min, removing supernatant with total volume of 3/4-4/5 in the centrifuge tube, and collecting the rest PRP; and (3) placing the obtained PRP on liquid nitrogen or dry ice, repeatedly freezing and thawing for three times, homogenizing for 1 minute on ice by using a handheld homogenizer, and fully breaking the platelet cells in the liquid nitrogen or dry ice into small vesicles to obtain the vesicles containing the PRP.
Further, the ultrasound time of step 3) is 1 minute.
Furthermore, the bionic nano-Chinese medicinal preparation is in a form of freeze-dried powder (for treatment by nasal drip or atomization administration).
Further, the bionic tea polyphenol nano-particles prepared by the preparation method of the bionic tea polyphenol nano-particles.
Furthermore, the bionic tea polyphenol nano-particles are applied to preparing medicines for targeted treatment of lung inflammation.
Compared with the prior art, the invention discloses a preparation method and application of the bionic tea polyphenol nano-particles, wherein the tea polyphenol is subjected to nanoemulsion, and platelet vesicles are modified on the surfaces of the tea polyphenol nano-particles by ultrasound to obtain the bionic tea polyphenol nano-particles; the bionic tea polyphenol nano-particles remarkably improve cellular absorption, bioavailability and targeting to inflammatory lesions.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a drawing showing a flow chart of the preparation of the bionic tea polyphenol nano-particles;
FIG. 2 is a scanning electron microscope image of the tea polyphenol nano-particles and the bionic tea polyphenol nano-particles of the invention;
FIG. 3 is a graph showing particle size distribution of the nano-sized Chinese medicinal preparation and the bionic nano-sized Chinese medicinal preparation according to the present invention;
FIG. 4 is a graph showing the surface potential of the nano-and biomimetic nano-chinese herbal preparation of the present invention;
in fig. 2 to 4, a: tea polyphenol nanoparticles; b: bionic tea polyphenol nano particles;
FIG. 5 is a graph showing the stability of the biomimetic tea polyphenol nanoparticles of the present invention after being dispersed in water for 1 month;
wherein A: tea polyphenols; b: tea polyphenol nanoparticles; c: bionic tea polyphenol nano particles;
FIG. 6 is a graph showing the particle size change of the bionic tea polyphenol nanoparticles of the present invention after being dispersed in water for 4 weeks;
FIG. 7 is a graph showing the change of each histopathological section after the bionic tea polyphenol nano-particles are administered;
FIG. 8 is a drawing showing the modeling time axis of in vivo imaging and tissue imaging of a mouse in accordance with the present invention;
FIG. 9 is a drawing showing an in vivo image of a mouse of the present invention;
FIG. 10 is a drawing showing an image of the tissue of a mouse in accordance with the present invention;
FIG. 11 is a graph showing the time axis of modeling and treatment of a mouse asthma model in accordance with the present invention;
FIG. 12 is a graph showing the change in inflammatory factors (IgE, IL-4, IL-13, IL-5) in alveolar lavage fluid of the present invention;
wherein A: igE; b: IL-4; c: IL-13; d: IL-5;
FIG. 13 is a drawing of a tissue imaging of a mouse in accordance with the present invention;
FIG. 14 is a graph showing fluorescence expression levels of lung tissue in accordance with the present invention;
FIG. 15 is a graph showing the time axis of a model of lung injury induced by pneumonia in mice in accordance with the present invention;
FIG. 16 is a graph showing IL-6 expression levels in plasma according to the invention;
FIG. 17 is a graph showing IL-1. Beta. Expression levels in plasma according to the invention;
FIG. 18 is a view showing pathological sections of lung tissue according to the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
House Dust Mite (HDM) extract was purchased from geer laboratories-rie, usa; aluminum hydroxide (AlOH) 3 ) Purchased from Sigma, usa; tea polyphenols were purchased from Shaanxi Xishan Jinhe chemical Co.
Example 1
A preparation method of a lung inflammation targeted bionic tea polyphenol nanoparticle comprises the steps of preparing the tea polyphenol nanoparticle by adopting a high-pressure homogenizer emulsification-volatilization method, then adding a PRP-containing vesicle, and modifying a platelet vesicle on the surface of the tea polyphenol nanoparticle by ultrasonic (a flow chart is shown in figure 1), wherein the preparation method comprises the following specific steps:
1) Preparing a nano Chinese medicinal preparation:
(1) 300mg of tea polyphenol and 1g of PLGA-PEG are weighed and dispersed in 20ml of dichloromethane, and added into a high-pressure homogenizer for emulsification, so as to obtain primary nanocrystalline suspension;
(2) 60ml of PVA (deacetylation degree is 85% -95%) with 0.1-3% is dissolved and used as an external water phase;
(3) Rapidly adding the primary nanocrystalline suspension prepared in the step (1) into the external water phase in the step (2) for shaking, and homogenizing for 3-5 minutes under high pressure under ice bath conditions to obtain nanocrystalline suspension;
(4) Adding 30ml of ultrapure water and 30ml of 0.1-3% PVA water solution into the nanocrystalline suspension obtained in the step (3), and magnetically stirring for 4-6 hours in a fume hood (to volatilize dichloromethane in an organic phase and enable nanoparticles to form and harden) to obtain a nanoemulsion;
(5) Centrifuging the nanoemulsion obtained in the step (4) at a high speed, wherein the temperature is 4 ℃, and the centrifugation is 12000g for 20min; centrifuging, discarding the supernatant, and washing the precipitate with ultrapure water for three times to obtain tea polyphenol nano particles;
2) PRP was extracted to prepare PRP-containing vesicles:
centrifuge the centrifuge tube with blood (anticoagulant added) (blood from hospital boots or healthy volunteers donation) in a centrifuge at 3000rpm for 5 minutes at 4 ℃; extracting supernatant containing PRP, centrifuging at 12000rpm and 4deg.C for 12 min, removing supernatant with total volume of 3/4-4/5 in the centrifuge tube, and collecting the rest PRP; and (3) placing the obtained PRP on liquid nitrogen or dry ice, repeatedly freezing and thawing for three times, homogenizing for 1 minute on ice by using a handheld homogenizer, and fully breaking the platelet cells in the liquid nitrogen or dry ice into small vesicles to obtain the vesicles containing the PRP.
3) Bionic tea polyphenol nano-particles:
weighing 10mg of nano traditional Chinese medicine preparation, dissolving into 1ml of PBS buffer solution, adding 50 microliter of PRP-containing vesicles, performing ultrasonic treatment in ice bath for 1 minute, and performing freeze drying to obtain the bionic tea polyphenol nano particles.
Characterization of the prepared tea polyphenol nano-particles and bionic tea polyphenol nano-particles comprises electron microscope scanning, particle size distribution and surface potential, and is shown in fig. 2-4: the tea polyphenol nano-particles are uniform in shape, the particle size is about 280nm, the particle size is increased to about 400nm after being modified by platelet vesicles, the surface potential of the bionic tea polyphenol nano-particles shows negative electricity due to the encapsulation of the platelet vesicles, and the absolute value of the surface potential is obviously increased.
Example 2 stability test
(1) The tea polyphenol, the tea polyphenol nano-particles and the bionic tea polyphenol nano-particles are respectively dissolved by water, and are observed after being placed for 1 month at room temperature in a dark place, and the result is shown in figure 5; it can be seen that the raw tea polyphenol is oxidized and has dark brown precipitate, and the tea polyphenol nano-particles and the bionic tea polyphenol nano-particles are relatively stable. Nanoemulsion can significantly improve the stability of tea polyphenols.
(2) The prepared tea polyphenol nano-particles and bionic tea polyphenol nano-particles are respectively dispersed in water, and are placed at room temperature for four weeks, and the particle size change of the tea polyphenol nano-particles is detected by sampling every week, and the result is shown in figure 6. The results of fig. 6 show that both the tea polyphenol nanoparticles and the biomimetic tea polyphenol nanoparticles are relatively stable, and the particle size is not significantly changed; the tea polyphenol nano-particles and the bionic tea polyphenol nano-particles are good in stability.
Example 3 toxicity test
Toxicity detection: normal healthy female C57/6J mice of 8 weeks old were purchased from beijing Bei Fu biotechnology limited, and after one week of adaptive feeding, the mice were divided into physiological saline groups, tea polyphenol nanoparticle groups, and bionic tea polyphenol nanoparticle groups, 5 mice per group, each group was given once daily by nasal drip (dose 10 mg/kg), for 7 consecutive days, euthanized mice 24 hours after the last administration, major organs (heart, liver, spleen, kidney) were collected, tissue fixative was fixed, paraffin sections were made, and sappan-eosin (HE) staining was performed, and changes in each histopathological section were observed, and the results are shown in fig. 7.
Because the tea polyphenol raw medicine has no toxicity, the detection is not used as a tea polyphenol raw medicine group.
The results of fig. 7 show that neither the tea polyphenol nanoparticles nor the bionic tea polyphenol nanoparticles cause damage to tissues, indicating that the biological safety in vivo is good.
Example 4 targeting of biomimetic tea polyphenol nanoparticles to pulmonary inflammation
(1) Mice were divided into three groups (tea polyphenol/ICG, tea polyphenol/ICG nanoparticles, biomimetic tea polyphenol/ICG nanoparticles), 5 each. On day 0 and day 2, mice asthma model (100 μg/mouse) was induced first with dust mite antigen HDM, inducing respiratory inflammation in mice; nasal administration (tea polyphenol/ICG, tea polyphenol/ICG nanoparticles, biomimetic tea polyphenol/ICG nanoparticles; 10mg/kg body weight) was performed simultaneously the following day; after nasal administration for 2 hours, the mice were anesthetized (modeling time axis is shown in fig. 8), and living imaging was performed on the mice, and the results are shown in fig. 9; mice were euthanized and major organs (kidney, lung, spleen, liver, heart) were collected for tissue imaging and the results are shown in fig. 10.
Since tea polyphenol itself is weak in fluorescence, the fluorescent dye, indocyanine green (ICG), is simultaneously loaded into nanoparticles as fluorescent markers for the nanoparticles. Fluorescence is a marker for nanoparticles, the stronger the fluorescence, indicating more nanoparticles. The results of fig. 9 and 10 show that the tea polyphenol/ICG nanoparticles and the bionic tea polyphenol/ICG nanoparticles can be aggregated to the lung of the mouse, and the bionic tea polyphenol/ICG nanoparticles target the lung inflammation part to the greatest extent.
(2) The mouse asthma model is induced by dust mites (HDM), and the specific modeling method is as follows:
on day 0 and day 2, eight week old female C57BL/6 mice were placed in a small animal gas anesthesia device, and after 10min the mice entered a stable anesthesia state, and 50 μl of sensitization HDM suspension A (containing 100 μg HDM,1mg of aluminum hydroxide in saline) or equivalent saline blank was given to each mouse by nasal drip; on days 14-18, each mouse was given 30. Mu. LHDM suspension B (containing 10. Mu. G HDM in saline) or an equivalent saline blank, and the mice were placed in a small animal gas anesthesia apparatus, and after 10min, were put into a stable anesthesia state, and then were nasal-dropped.
Tea polyphenol drug (tea polyphenol, tea polyphenol nanoparticles, biomimetic tea polyphenol nanoparticles) treatment time: nasal drops (10 mg/kg body weight) were administered once daily on days 0,2, 14-20. On day 21, mice were euthanized and the material was tested. The time axis for modeling and administration is shown in FIG. 11.
The preparation method of the alveolar lavage fluid comprises the following steps: 1ml of physiological saline is pumped into the lung from the tracheal cannula by the injector, lavage liquid is pumped after 3 times of repeated lavage, alveolar lavage liquid is centrifuged for 15min at 3000r/min at 4 ℃, and the supernatant is stored at-80 ℃ for subsequent detection.
After treatment, changes in inflammatory factors (IgE, IL-4, IL-13, IL-5) in alveolar lavage fluid were examined and the results are shown in FIG. 12. The results of fig. 12 show: the tea polyphenol can obviously reduce the level of inflammatory factors in alveoli, and the tea polyphenol nano-particles, particularly the bionic tea polyphenol nano-particles, are most obvious in inhibiting the inflammatory factors and have statistical differences.
Example 5 application of bionic tea polyphenol nanoparticles in pneumonia
(1) Evaluation of cell targeting by biomimetic nanoparticles: since tea polyphenol itself is very weak in fluorescence, the fluorescent dye, indocyanine green (ICG), is co-loaded into the nanoparticle as a fluorescent marker of the nanoparticle. Lipopolysaccharide LPS (7.5 mg/kg body weight) was injected intraperitoneally for 20 hours to induce pneumonia and acute lung injury in mice, then nasal administration (10 mg/kg body weight) was performed, after 4 hours, mice were euthanized, and major organs (brain, kidney, lung, spleen, liver, heart) were collected for tissue imaging, and the results are shown in FIG. 13. The fluorescence expression level of lung tissue is shown in FIG. 14.
After nasal administration, the tea polyphenol nano-particles, particularly the bionic tea polyphenol nano-particles, can be more enriched and detained in lung inflammation parts.
(2) Healthy female C57/6J mice were divided into a healthy group, a physiological saline group, a tea polyphenol nanoparticle group and a bionic tea polyphenol nanoparticle group, each group being 5. The healthy group was not treated, and except for the healthy group, mice of each group were given 10mg/kg body weight by nasal drip, after 3 hours, mice were euthanized by injecting LPS (7.5 mg/kg body weight) to induce pneumonic lung injury model, after 21 hours, mice were euthanized (time axis see FIG. 15), whole blood was collected to anticoagulate, centrifuged at 3000rpm at 4℃for 10 minutes, upper plasma was separated, IL-6 and IL-1β levels were detected, and the detection results were shown in FIG. 16 and FIG. 17 (P <0.05, P <0.001, compared with saline group). Lung tissue was collected, fixed with tissue fixative for 48 hours, paraffin block specimens were prepared, sectioned, and stained with Hematoxylin and Eosin (HE), and the results are shown in fig. 18.
The results in fig. 16 and 17 show that the tea polyphenol nano-particles can significantly inhibit the production of inflammatory factors in the lung compared with the tea polyphenol raw medicine, and particularly the effect of the bionic tea polyphenol nano-particles is most significant, and the three are statistically different.
Pathological sections of lung tissues (figure 18) show that LPS can successfully induce the lung inflammation of mice, and after different drug treatment, the tea polyphenol nano-particles, in particular the bionic tea polyphenol nano-particles, can remarkably inhibit the infiltration of inflammatory cells in the lung and relieve the secretion of inflammatory factors in the lung.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (8)
1. The preparation method of the bionic tea polyphenol nano-particles is characterized by comprising the following specific steps:
1) Preparing a nano Chinese medicinal preparation:
(1) According to 0.3:1, weighing tea polyphenol and PLGA-PEG according to the weight ratio, and dispersing in dichloromethane; homogenizing for 3-5 minutes under high pressure under ice bath condition to obtain primary nanocrystalline suspension; the weight-volume ratio of PLGA-PEG to dichloromethane is 1:20, a step of;
(2) Taking 0.1-3% PVA water solution as external water phase; the volume ratio of the aqueous solution of 0.1-3% PVA to the dichloromethane is 3:1, a step of;
(3) Rapidly adding the primary nanocrystalline suspension prepared in the step (1) into the external water phase in the step (2) for shaking, and homogenizing for 3-5 minutes under high pressure under ice bath conditions to obtain nanocrystalline suspension;
(4) Adding ultra-pure water and 0.1-3% PVA water solution into the nanocrystalline suspension obtained in the step (3), and magnetically stirring for 4-6 hours in a fume hood to obtain nanoemulsion; the volume ratio of the ultrapure water to the aqueous solution of 0.1-3% PVA is 1:1, a step of; the volume ratio of the ultrapure water to the dichloromethane is 3:2;
(5) Centrifuging the nanoemulsion obtained in the step (4) at a high speed, discarding the supernatant after centrifuging, and washing the precipitate with ultrapure water for three times to obtain tea polyphenol nanoparticles;
2) Extracting PRP, and preparing vesicles containing PRP;
3) Preparing bionic tea polyphenol nano particles:
dissolving tea polyphenol nano-particles into PBS buffer solution, adding PRP-containing vesicles, performing ultrasonic treatment in ice bath, and performing freeze drying to obtain bionic tea polyphenol nano-particles; the weight-to-volume ratio of the tea polyphenol nano-particles to the PBS buffer solution is 0.01:1, a step of; the volume ratio of the PRP-containing vesicles to the PBS buffer was 0.05:1.
2. the method for preparing the bionic tea polyphenol nano-particles according to claim 1, wherein the PVA in the step (2) is PVA with a deacetylation degree of 85% -95%.
3. The method for preparing the bionic tea polyphenol nano-particles according to claim 1, wherein the high-speed centrifugation in the step (5) is performed at 4 ℃, and the centrifugation is performed at 12000g for 20min.
4. The method for preparing the bionic tea polyphenol nano-particles according to claim 1, wherein the specific steps of extracting PRP and preparing vesicles containing PRP in the step 2) are as follows:
placing the centrifuge tube filled with blood into a centrifuge, and centrifuging for 4-6 minutes at the rotating speed of 2500-3500 rpm; extracting supernatant containing PRP, centrifuging at 12000-18000rpm for 8-12 min, removing supernatant with total volume of 3/4-4/5 in the centrifuge tube, and collecting the rest PRP; and (3) placing the obtained PRP on liquid nitrogen or dry ice, repeatedly freezing and thawing for three times, homogenizing for 1 minute on ice by using a handheld homogenizer, and fully breaking the platelet cells in the liquid nitrogen or dry ice into small vesicles to obtain the vesicles containing the PRP.
5. The method for preparing the bionic tea polyphenol nano-particles according to claim 1, wherein the ultrasonic time in the step 3) is 1 minute.
6. The method for preparing the bionic tea polyphenol nano-particles according to claim 1, wherein the preparation of the bionic nano-Chinese medicinal preparation is freeze-dried powder.
7. The bionic tea polyphenol nanoparticle prepared by the preparation method of the bionic tea polyphenol nanoparticle of any one of claims 1 to 6.
8. Use of the biomimetic tea polyphenol nano-particles of claim 7 in the preparation of a drug for targeted treatment of pulmonary inflammation.
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