A degradable block copolymer and its application in the prevention and control of atmospheric particulate pollution

Abstract

The invention discloses an application of a degradable block copolymer in the prevention and control of atmospheric particulate pollution. The block copolymer can effectively adsorb harmful substances in atmospheric particulate matter, especially can effectively adsorb negatively charged harmful substances in atmospheric particulate matter, thereby playing the role of adsorption and detoxification, and reducing the damage of atmospheric particulate pollutants to human body; it can also significantly reduce the toxic effect of atmospheric particulate pollutants on human myocardial cells or human bronchial epithelial cells, alleviate the damage caused by atmospheric particulate pollutants to human myocardial cells or human bronchial epithelial cells, and alleviate the excessive generation of ROS in human myocardial cells or human bronchial epithelial cells caused by PM. Specifically, the cell activity of the PM+PHB-P group is at least 10% higher than that of the PM group, the cell apoptosis rate of the PM+PHB-P group is significantly lower than that of the PM group by at least 43%, and the cell ROS of the PM+PHB-P group is significantly lower than that of the PM group by at least 13%.

Description

Technical Field

[0001] The present invention relates to the technical field of pollution prevention and control, and in particular to an application of a degradable block copolymer in the prevention and control of atmospheric particulate pollution. Background Art

[0002] Atmospheric particulate matter (PM) is the main factor causing air pollution. Long-term exposure to high concentrations of PM will bring great risks to human health. In addition, PM is very easy to absorb harmful substances in the environment, including inorganic substances such as heavy metals, organic substances such as polycyclic aromatic hydrocarbons, and some pathogens such as viruses and bacteria. These PMs that absorb harmful substances can induce diseases such as respiratory diseases, cardiovascular diseases, nervous system diseases, and cancer.

[0003] Patent document CN111760384A discloses an anti-haze and antibacterial filter material, which includes a flexible base fabric and a composite adhesive coated on at least one side of the flexible base fabric. A trace amount of adhesive with adsorption and filtration functions is coated on the flexible base fabric to form a barrier to block pollutant particles, bacteria, and viruses, prevent the inhalation of pollutants such as haze particles, and the spread of bacterial and viral droplets. The raw materials of the composite adhesive include functional particles with adsorption and / or retention capabilities, and the functional particles with adsorption and / or retention capabilities include at least one of activated carbon, sepiolite, natural zeolite, molecular sieve, porous silica gel, nano silicon dioxide, nano titanium dioxide, nano zirconium dioxide, nano aluminum oxide, nano zinc oxide, nano phosphorus pentoxide, nano calcium carbonate, graphene oxide, diatomaceous earth, montmorillonite, kaolin, synthetic hectorite, bentonite, Laponite clay, nano chitin, starch nanoparticles and cellulose nanocrystals. However, the aforementioned anti-haze and antibacterial filter materials cannot specifically and efficiently adsorb negatively charged pollutant particles in PM, and there is no evidence that they can effectively reduce the toxicity of PM to human cardiomyocytes or human bronchial epithelial cells.

[0004] Patent document CN102089039B discloses a method for producing a permeable mask worn on the nose and mouth of a user, the mask being impermeable to harmful pollutants, the method comprising:

[0005] a) coating the mask with a formulation comprising:

[0006] - at least one quaternary ammonium conditioning agent;

[0007] - wetting agents;

[0008] -preservative;

[0009] - biocides; and

[0010] - a neutralizing agent, which is added to adjust and achieve a pH in the range of 5.0 to 6.8;

[0011] b) establishing an electrostatic field using the at least one quaternary ammonium conditioning agent in the vicinity of the mask; and

[0012] c) using the at least one quaternary ammonium conditioning agent to repel similarly charged contaminants away from the mask and to attract and capture oppositely charged contaminants;

[0013] Thereby less of said pollutants reach the nose and mouth of the user.

[0014] However, the quaternary ammonium regulator in the aforementioned mask is selected from the group consisting of: polyquaternium-6, polyquaternium-7, polyquaternium-10, polyquaternium-22, polyquaternium-70, Merquat280, Merquat10, Merquat5, coconut dimethyl ammonium hydroxypropyl hydrolyzed keratin, quaternized oats, hydrolyzed oat protein and benzalkonium chloride, which is difficult to be degraded in actual application and will cause great harm to the ecological environment.

[0015] In summary, there is no report in the prior art on using the degradable block copolymer hydroxybutyric acid-polymethacrylic acid-dimethylamino ester to adsorb pollutant particles in PM. In view of this, the present invention is proposed. Summary of the invention

[0016] The present invention overcomes the deficiencies in the prior art. In a first aspect, the present invention provides an application of a degradable block copolymer in the prevention and control of atmospheric particulate pollution.

[0017] Furthermore, the application includes:

[0018] 1) Application in adsorption of atmospheric particulate matter;

[0019] 2) Application in reducing the toxicity of atmospheric particulate matter.

[0020] Furthermore, the specific application method is: spraying the degradable block copolymer directly into the air; or placing the degradable block copolymer at the pollutant exhaust port; or placing the degradable block copolymer on a pollutant protection device that filters the air.

[0021] In a second aspect, the present invention provides an application of a degradable block copolymer in preparing a product for adsorbing atmospheric particulate matter.

[0022] Furthermore, the product is an adsorbent, a mask, a filter or a protective device.

[0023] In one embodiment of the present invention, the product is an adsorbent.

[0024] In one embodiment of the present invention, the product is a mask.

[0025] Furthermore, when the product is an adsorbent, the amount of the degradable block copolymer in the adsorbent is 2.5 mg / m 3 .

[0026] Furthermore, when the product is a mask, the amount of the degradable block copolymer distributed in the adsorption interlayer of the mask is 2.5 mg / m 3 .

[0027] In a third aspect, the present invention provides an application of a degradable block copolymer in the preparation of a product for reducing the toxicity of atmospheric particulate matter.

[0028] Furthermore, the product is an adsorbent, a mask, a filter or a protective device.

[0029] In one embodiment of the present invention, the product is an adsorbent.

[0030] In one embodiment of the present invention, the product is a mask.

[0031] Furthermore, when the product is an adsorbent, the amount of the degradable block copolymer in the adsorbent is 2.5 mg / m 3 .

[0032] Furthermore, when the product is a mask, the amount of the degradable block copolymer distributed in the adsorption interlayer of the mask is 2.5 mg / m 3 .

[0033] In a fourth aspect, the present invention provides an application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on cell activity, wherein the cells are human cardiomyocytes and / or human bronchial epithelial cells.

[0034] In a fifth aspect, the present invention provides an application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on cell apoptosis, wherein the cells are human cardiomyocytes and / or human bronchial epithelial cells.

[0035] In a sixth aspect, the present invention provides an application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on active oxygen in cells, wherein the cells are human cardiomyocytes and / or human bronchial epithelial cells.

[0036] In the present invention, the degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester (PHB-PDMAEMA), and its chemical structure is as follows:

[0037] wherein m is an integer selected from 100-120 (e.g., 100, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120); and n is an integer selected from 10-20 (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20).

[0038] Furthermore, the m is an integer selected from 105-115, and preferably, the m is 111.

[0039] Furthermore, the n is an integer selected from 10-15, and preferably, the n is 13.

[0040] In the present invention, the atmospheric particulate matter includes PM particulate matter. Preferably, the atmospheric particulate matter includes PM polluted particulate matter. Further preferably, the atmospheric particulate matter includes negatively charged polluted particulate matter among PM polluted particulate matter.

[0041] In one embodiment of the present invention, the PM particles include PM 2.5 、PM 10 , TSP, one or more, preferably PM 2.5 .

[0042] In one embodiment of the present invention, the PM polluted particles include PM 2.5 、PM 10 , TSP, one or more, preferably PM 2.5 .

[0043] The beneficial effects of the present invention are:

[0044] 1. The degradable block copolymer PHB-PDMAEMA of the present invention can effectively adsorb harmful substances in atmospheric particulate matter, especially negatively charged harmful substances in atmospheric particulate matter, thereby playing the role of adsorption and detoxification, and reducing the damage of atmospheric particulate pollutants to the human body;

[0045] 2. The degradable block copolymer PHB-PDMAEMA of the present invention can significantly reduce the toxic effects of atmospheric particulate pollutants on human myocardial cells, alleviate the damage caused by atmospheric particulate pollutants to human myocardial cells, and alleviate the excessive generation of ROS in human myocardial cells caused by PM. Specifically, the cell activity of the PM+PHB-P group is at least 10% higher than that of the PM group, the cell apoptosis rate of the PM+PHB-P group is significantly lower than that of the PM group by at least 56%, and the cell ROS of the PM+PHB-P group is significantly lower than that of the PM group by at least 13%;

[0046] 3. The degradable block copolymer PHB-PDMAEMA of the present invention can also significantly reduce the toxic effects of atmospheric particulate pollutants on human bronchial epithelial cells, alleviate the damage caused by atmospheric particulate pollutants to human bronchial epithelial cells, and alleviate the excessive generation of ROS in human bronchial epithelial cells caused by PM. Specifically, the cell activity of the PM+PHB-P group was at least 23% higher than that of the PM group, the cell apoptosis rate of the PM+PHB-P group was significantly lower than that of the PM group by at least 43%, and the cell ROS of the PM+PHB-P group was significantly lower than that of the PM group by at least 76%.

[0047] Description of the accompanying drawings

[0048] Figure 1 This is the NMR spectrum of PHB-P.

[0049] Figure 2 The effect of PM on the activity of human cardiomyocytes (AC16).

[0050] Figure 3 The effect of PHB-P on the activity of human cardiomyocytes (AC16).

[0051] Figure 4 The effects of PHB-P and PDMAEMA on the activity of human cardiomyocytes (AC16) exposed to PM.

[0052] Figure 5 The effect of PHB-P on the apoptosis level of human cardiomyocytes (AC16) exposed to PM (Figure A is the flow cytometric analysis image of AC16, and Figure B is the quantitative analysis of the apoptosis level of AC16).

[0053] Figure 6 The effect of PHB-P on the reactive oxygen levels of human cardiomyocytes (AC16) exposed to PM (Figure A is the AC16 flow cytometric analysis image; Figure B is the quantitative analysis of AC16 ROS fluorescence intensity).

[0054] Figure 7 The effect of PM on the activity of human bronchial epithelial cells (BEAS-2B).

[0055] Figure 8 The effect of PHB-P on the activity of human bronchial epithelial cells (BEAS-2B).

[0056] Fig. 9 The effects of PHB-P and PDMAEMA on the activity of human bronchial epithelial cells (BEAS-2B) exposed to PM.

[0057] Fig.10The effect of PHB-P on the apoptosis level of human bronchial epithelial cells (BEAS-2B) exposed to PM (Figure A is the flow cytometric analysis image of BEAS-2B; Figure B is the quantitative analysis of the apoptosis level of BEAS-2B).

[0058] Fig.11 The effect of PHB-P on the level of reactive oxygen species in PM-exposed human bronchial epithelial cells (BEAS-2B) (Figure A is a flow cytometric analysis image of BEAS-2B; Figure B is a quantitative analysis of BEAS-2B ROS fluorescence intensity). DETAILED DESCRIPTION

[0059] In the present invention, the term "PHB" refers to polyhydroxybutyrate, the full name of which is poly[(R)-3-hydroxybutyrate]

[0060] In the present invention, the term "PDMAEMA" is poly(2-(dimethylamino)ethyl methacrylate)

[0061] In the present invention, the term "PHB-PDMAEMA" is polyhydroxybutyrate-polymethacrylate N,N-dimethylamino ester, and its full English name is Poly[(R)-3-hydroxybutyrate]-Poly(2-(dimethylamino)ethyl methacrylate)

[0062] In the present invention, the term "HMTETA" is 1,1,4,7,10,10-hexamethyltriethylenetetramine.

[0063] In the present invention, the term "Br-PHB-Br" refers to bromine-substituted polyhydroxybutyric acid.

[0064] In the present invention, the term "CuBr" means copper bromide.

[0065] In order to more clearly understand the technical content of the present invention, the following embodiments are given in detail, the purpose of which is only to better understand the content of the present invention but not to limit the protection scope of the present invention.

[0066] Experimental materials:

[0067] Reagents: PHB was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); dichloromethane (purity 99.9%) was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); CuBr was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); DMEMA was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); ether was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); ethylene glycol was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); 2-bromoisobutyryl bromide was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); 1,4-dioxane was purchased from Aldrich-Sigma ma company (St. Louis, Missouri, USA); triethylamine was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); methanol was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); hexane was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); tetrahydrofuran was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); acetone was purchased from Aldrich-Sigma (St. Louis, Missouri, USA); culture medium used for AC16 cells: DMEM / F-12 culture medium, vivacell, C3130-0500; culture medium used for BEAS-2B cells: DMEM (high glucose), CORNING, 10-013-CVR; PM: NIST 1648a, Urban Particulate Matter; PDMAEMA: TCL P2558, Poly[2-(Dimethylamino)ethyl Methacrylate]; Trypsin: Trypsin-EDTA digestion solution (PBS), Keygen, KGY0012; Human bronchial epithelial cell line (BEAS-2B), Cell Resource Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences; CCK-8 solution: Cell Counting Kit-8, Japan Tongren Chemical, CK04; Human cardiomyocytes (AC16) were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences; Binding Buffer and Annexin V-FITC / PI staining solution: AnnexinV-FITC / PI double staining cell apoptosis kit, Keygen, KGA1102-50;.

[0068] Example 1 Preparation and Characterization of PHB-P

[0069] 1. Preparation of PHB-P:

[0070] First, add 8 g of PHB into a dichloromethane solution containing 20 mmol of triethylamine, stir, and then add

[0071] 10 mmol of 2-bromoisobutyryl bromide was stirred at room temperature for 24 hours;

[0072] Then, the above solution was added dropwise to an ether-methanol (4:1) solution to precipitate a crude Br-PHB-Br product, which was dissolved in tetrahydrofuran and precipitated in ether-methanol (4:1) for 3 times. The product was finally purified by vacuum drying.

[0073] Then Br-PHB-Br, DMAEMA, CuBr and HMTETA were dissolved in 15 mL of dioxane and reacted under nitrogen for 24 hours, and the reaction temperature was controlled at 45°C. The reaction solution was then diluted with tetrahydrofuran and exposed to air for 4 hours to stop the reaction. The product was passed through an alumina adsorption column and dissolved in tetrahydrofuran, tetrahydrofuran was added to hexane, the product was redissolved in tetrahydrofuran, and then added to ether to obtain purified PHB-P.

[0074] 2. Characterization of PHB-P:

[0075] Using deuterated dichloromethane as solvent, take 5.0 mg of the freeze-dried polymer sample and dissolve it in a 5 mm NMR test tube. 1 H-NMR spectrum. Figure 1 This is the NMR spectrum of the synthesized PHB-PDMAEMA. The corresponding characteristic peaks of various hydrogen atoms in the compound are marked in each figure, which confirms the successful synthesis of PHB-PDMAEMA.

[0076] Example 2 Effects of PHB-P on Cell Viability, Cell Apoptosis and Cellular Reactive Oxygen Species

[0077] 1. Experimental methods:

[0078] 1. Cell activity analysis

[0079] (1) Human cardiomyocyte (AC16) activity analysis

[0080] Experimental steps:

[0081] 1) Human cardiomyocytes (AC16) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension. The cell density was adjusted to 5×10 4 / mL. Inoculate in a 96-well culture plate, 100 μL per well, and place at 37°C.

[0082] The cells were cultured in an incubator with 5% CO2.

[0083] 2) Remove the culture medium, add the poison solution, and continue culturing in an incubator at 37°C and 5% CO2.

[0084] PM staining solution: Use serum-free culture medium to prepare PM staining solution with concentrations of 0, 25, 50, 100, and 200 μg / mL.

[0085] PHB-P staining solution: PHB-P staining solutions with concentrations of 0, 0.625, 1.25, 2.5, and 5 ng / mL were prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0086] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing 50 μg / mL PM and 0.625 ng / mL PHB-P was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0087] PDMAEMA staining solution: Prepare PDMAEMA staining solution at a concentration of 0.625 ng / mL using serum-free culture medium.

[0088] PM+PDMAEMA staining solution: Use serum-free culture medium to prepare PM+PDMAEMA staining solution containing 50 μg / mL PM and 0.625 ng / mL PDMAEMA.

[0089] 3) 24 hours after exposure, add 10 μL of CCK-8 solution to each well. Return the culture plate to the incubator for 0.5-1 hour, measure the absorbance at 450 nm with an ELISA reader, and calculate the cell survival rate.

[0090] (2) Activity analysis of human bronchial epithelial cells (BEAS-2B)

[0091] Experimental steps:

[0092] 1) Human bronchial epithelial cells (BEAS-2B) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension. The cell density was adjusted to 5×10 4 / mL. Inoculate into 96-well culture plates, 100 μL per well, and place in an incubator at 37°C and 5% CO2 for adherent culture.

[0093] 2) Remove the culture medium, add the poison solution, and continue culturing in an incubator at 37°C and 5% CO2.

[0094] PM staining solution: Use serum-free culture medium to prepare PM staining solution with concentrations of 0, 100, 200, 300, and 400 μg / mL.

[0095] PHB-P staining solution: PHB-P staining solutions with concentrations of 0, 1.25, 2.5, 5, and 10 ng / mL were prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0096] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing a PM concentration of 300 μg / mL and a PHB-P concentration of 2.5 ng / mL was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0097] PDMAEMA staining solution: Prepare PDMAEMA staining solution at a concentration of 2.5 ng / mL using serum-free culture medium.

[0098] PM+PDMAEMA staining solution: Use serum-free culture medium to prepare PM+PDMAEMA staining solution containing 300 μg / mL PM and 2.5 ng / mL PDMAEMA.

[0099] 3) 24 hours after exposure, add 10 μL of CCK-8 solution to each well. Return the culture plate to the incubator for 0.5-1 hour, measure the absorbance at 450 nm with an ELISA reader, and calculate the cell survival rate.

[0100] 2. Cell apoptosis analysis

[0101] (1) Apoptosis analysis of human cardiomyocytes (AC16)

[0102] Experimental steps:

[0103] 1) Human cardiomyocytes (AC16) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension. The cell density was adjusted to 1×10 5 / mL. Inoculate into 6-well culture plates, 2 mL per well, and place in an incubator at 37°C and 5% CO2 for adherent culture.

[0104] 2) Remove the culture medium, wash the cells once with PBS, add the poison solution, and culture in a 37°C, 5% CO2 incubator for 24 hours.

[0105] PM staining solution: Use serum-free culture medium to prepare PM staining solution at a concentration of 50 μg / mL.

[0106] PHB-P staining solution: A PHB-P staining solution with a concentration of 0.625 ng / mL was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0107] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing 50 μg / mL PM and 0.625 ng / mL PHB-P was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0108] 3) Remove the staining solution, wash the cells twice with PBS, and add 200 μL of EDTA-free trypsin to each well to digest and collect the cells. Wash the cells twice with PBS (800 g centrifugal force, 5 min) and collect 1-5×10 5 Cells. Add 300 μL of Binding Buffer and gently blow to form a single cell suspension, add 3 μL of Annexin V-FITC and PI staining solution, and mix well. Stain for 5-10 minutes at room temperature in the dark, and detect with flow cytometry within 1 hour.

[0109] (2) Apoptosis analysis of human bronchial epithelial cells (BEAS-2B)

[0110] Experimental steps:

[0111] 1) Human bronchial epithelial cells (BEAS-2B) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension and the cell density was adjusted to 1×10 5 / mL. Inoculate into 6-well culture plates, 2 mL per well, and place in an incubator at 37°C and 5% CO2 for adherent culture.

[0112] 2) Remove the culture medium, wash the cells once with PBS, add the poison solution, and culture in a 37°C, 5% CO2 incubator for 24 hours.

[0113] PM staining solution: Use serum-free culture medium to prepare PM staining solution at a concentration of 300 μg / mL.

[0114] PHB-P staining solution: A PHB-P staining solution with a concentration of 2.5 ng / mL was prepared using a serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0115] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing a PM concentration of 300 μg / mL and a PHB-P concentration of 2.5 ng / mL was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0116] 3) Remove the staining solution, wash the cells twice with PBS, and add 200 μL of EDTA-free trypsin to each well to digest and collect the cells. Wash the cells twice with PBS (800 g centrifugal force, 5 min) and collect 1-5×10 5Cells. Add 300 μL of Binding Buffer and gently blow to form a single cell suspension, add 3 μL of Annexin V-FITC and PI staining solution, and mix well. Stain for 5-10 minutes at room temperature in the dark, and detect with flow cytometry within 1 hour.

[0117] 3. Cellular Reactive Oxygen Species Analysis

[0118] (1) Analysis of reactive oxygen species in human cardiomyocytes (AC16)

[0119] Experimental steps:

[0120] 1) Human cardiomyocytes (AC16) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension. The cell density was adjusted to 1×10 5 / mL. Inoculate into 6-well culture plates, 2 mL per well, and place in an incubator at 37°C and 5% CO2 for adherent culture.

[0121] 2) Remove the culture medium, wash the cells once with PBS, add the poison solution, and culture in a 37°C, 5% CO2 incubator for 24 hours.

[0122] PM staining solution: Use serum-free culture medium to prepare PM staining solution at a concentration of 50 μg / mL.

[0123] PHB-P staining solution: A PHB-P staining solution with a concentration of 0.625 ng / mL was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0124] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing 50 μg / mL PM and 0.625 ng / mL PHB-P was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0125] 3) Remove the poison solution by aspirating, wash the cells twice with PBS, add 1 mL of DCFH-DA working solution to each well, shake gently to mix, and place in the incubator for 30 minutes. After the incubation, wash the cells once with PBS, collect the cell suspension with trypsin-EDTA in a 1.5 mL centrifuge tube, put it into a centrifuge at 1200 rpm, centrifuge for 3 minutes, discard the supernatant, wash twice with PBS, add 500 μL of PBS to each sample, gently resuspend the cells, select the corresponding channel on the flow cytometer to detect the fluorescence intensity, and perform quantitative analysis.

[0126] (2) Analysis of reactive oxygen species in human bronchial epithelial cells (BEAS-2B)

[0127] Experimental steps:

[0128] 1) Human bronchial epithelial cells (BEAS-2B) in the logarithmic growth phase were obtained, digested with trypsin, and then the cells were blown into a single cell suspension and the cell density was adjusted to 1×10 5 / mL. Inoculate into 6-well culture plates, 2 mL per well, and place in an incubator at 37°C and 5% CO2 for adherent culture.

[0129] 2) Remove the culture medium, wash the cells once with PBS, add the poison solution, and culture in a 37°C, 5% CO2 incubator for 24 hours.

[0130] PM staining solution: Use serum-free culture medium to prepare PM staining solution at a concentration of 300 μg / mL.

[0131] PHB-P staining solution: A PHB-P staining solution with a concentration of 2.5 ng / mL was prepared using a serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0132] PM+PHB-P poisoning solution: A PM+PHB-P poisoning solution containing a PM concentration of 300 μg / mL and a PHB-P concentration of 2.5 ng / mL was prepared using serum-free culture medium (wherein PHB-P was synthesized in Example 1).

[0133] 3) Remove the poison solution by aspirating, wash the cells twice with PBS, add 1 mL of DCFH-DA working solution to each well, shake gently to mix, and place in the incubator for 30 minutes. After the incubation, wash the cells once with PBS, collect the cell suspension with trypsin-EDTA in a 1.5 mL centrifuge tube, put it into a centrifuge at 1200 rpm, centrifuge for 3 minutes, discard the supernatant, wash twice with PBS, add 500 μL of PBS to each sample, gently resuspend the cells, select the corresponding channel on the flow cytometer to detect the fluorescence intensity, and perform quantitative analysis.

[0134] 2. Experimental Results

[0135] 1. Effects of PM, PHB-P alone and in combination, and PDMAEMA alone, a commercially available material, on the cell viability, apoptosis level, and cellular reactive oxygen species level of human cardiomyocytes (AC16)

[0136] The product PHB-P proposed by the present invention was used for comparative research, and human cardiomyocytes (AC16) were used for comparison of cell activity, cell apoptosis and cell reactive oxygen species. The experimental results are as follows:

[0137] (1) Effects of PM exposure on the activity of human cardiomyocytes (AC16)

[0138] The present invention uses CCK-8 to detect the cell activity of human cardiomyocytes (AC16) after exposure to different concentrations of PM (0, 25, 50, 100, 200 μg / mL) for 24 hours. Figure 2 The results showed that when the PM concentration was between 50 and 200 μg / mL, the cell activity decreased in a dose-dependent manner and was significantly lower than that of the control group. In the present invention, a PM concentration of 50 μg / mL was selected for subsequent experiments. * Statistically significant, p<0.05.

[0139] (2) Effects of PHB-P exposure on the activity of human cardiomyocytes (AC16)

[0140] The present invention uses CCK-8 to detect the cell activity of human cardiomyocytes (AC16) after exposure to different concentrations of PHB-P (0, 0.625, 1.25, 2.5, 5 ng / mL) for 24 hours. Figure 3 The results showed that when the PHB-P concentration was between 1.25 and 5 ng / mL, the cell activity decreased in a dose-dependent manner and was significantly lower than that of the control group. In the present invention, the PHB-P concentration of 0.625 ng / mL was selected for subsequent experiments. * Statistically significant, p<0.05.

[0141] (3) Effects of PHB-P on the activity of human cardiomyocytes (AC16) exposed to PM

[0142] CCK-8 was used to analyze the activity of AC16 cells. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Figure 4 The results showed that the cell activity in the PM group was significantly lower than that in the control group, and the cell activity in the PM+PHB-P group was higher than that in the PM group, and the differences were statistically significant, indicating that PHB-P can reduce the toxic effects of PM on human cardiomyocytes. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0143] (4) Effect of PHB-P on apoptosis of human cardiomyocytes (AC16) exposed to PM

[0144] Flow cytometry was used to analyze the apoptosis level of AC16. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Figure 5 The results showed that compared with the control group, the total apoptosis rate of cells in the PM group increased, while the apoptosis rate of cells in the PM+PHB-P group was significantly lower than that in the PM group, indicating that PHB-P can alleviate the damage caused by PM to human cardiomyocytes. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0145] (5) Effects of PHB-P on reactive oxygen species levels in human cardiomyocytes (AC16) exposed to PM

[0146] Flow cytometry was used to analyze the level of ROS produced by AC16. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Figure 6 The results showed that compared with the control group, the cellular ROS content in the PM group increased, while the cellular ROS in the PM+PHB-P group decreased and was significantly lower than that in the PM group, indicating that PHB-P can alleviate the excessive generation of ROS in human cardiomyocytes caused by PM. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0147] (6) Effect of PDMAEMA on AC16 activity after PM exposure

[0148] CCK-8 was used to analyze the activity of AC16 cells. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Figure 4 The results showed that the cell activity in the PM group was significantly lower than that in the control group, and there was no statistical difference in cell activity between the PM+PDMAEMA group and the PM group, indicating that at the same concentration as PHB-P, PDMAEMA could not reduce the toxic effect of PM on human cardiomyocytes, and PHB-P had a better attenuation effect. * There was a statistical difference compared with the Control group, # There was a statistical difference compared with the PM group, p<0.05.

[0149] 2. Effects of PM, PHB-P alone and in combination, and PDMAEMA alone, a commercially available material, on the cell viability, apoptosis, and reactive oxygen species levels of human bronchial epithelial cells (BEAS-2B)

[0150] The product PHB-P proposed by the present invention was used for comparative study, and human bronchial epithelial cells (BEAS-2B) were used for comparison of cell activity, cell apoptosis and cell reactive oxygen species. The experimental results are as follows:

[0151] (1) Effects of PM exposure on the activity of human bronchial epithelial cells (BEAS-2B)

[0152] The present invention uses CCK-8 to detect the cell activity of human bronchial epithelial cells (BEAS-2B) after exposure to different concentrations of PM (0, 100, 200, 300, 400 μg / mL) for 24 hours. Figure 7 The results showed that when the PM concentration was between 200 and 400 μg / mL, the cell activity decreased in a dose-dependent manner and was significantly lower than that of the control group. In the present invention, a PM concentration of 300 μg / mL was selected for subsequent experiments. * Statistically significant, p<0.05.

[0153] (2) Effects of PHB-P exposure on the activity of human bronchial epithelial cells (BEAS-2B)

[0154] The present invention uses CCK-8 to detect the cell activity of human bronchial epithelial cells (BEAS-2B) after exposure to different concentrations of PHB-P (0, 1.25, 2.5, 5, 10 ng / mL) for 24 hours. Figure 8 The results showed that when the PHB-P concentration was between 5 and 10 ng / mL, the cell activity decreased in a dose-dependent manner and was significantly lower than that of the control group. In the present invention, a PHB-P concentration of 2.5 ng / mL was selected for subsequent experiments. * Statistically significant, p<0.05.

[0155] (3) Effects of PHB-P on the activity of PM-exposed human bronchial epithelial cells (BEAS-2B)

[0156] CCK-8 was used to analyze the activity of BEAS-2B cells. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Fig. 9 The results showed that the cell activity in the PM group was significantly lower than that in the control group, and the cell activity in the PM+PHB-P group was higher than that in the PM group, and the differences were statistically significant, indicating that PHB-P can reduce the toxic effects of PM on human bronchial epithelial cells. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0157] (4) Effect of PHB-P on apoptosis of PM-exposed human bronchial epithelial cells (BEAS-2B)

[0158] Flow cytometry was used to analyze the apoptosis level of BEAS-2B. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Fig.10 The results showed that compared with the control group, the total apoptosis rate of cells in the PM group increased, while the apoptosis rate of cells in the PM+PHB-P group was significantly lower than that in the PM group, indicating that PHB-P can alleviate the damage caused by PM to human bronchial epithelial cells. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0159] (5) Effects of PHB-P on reactive oxygen species levels in PM-exposed human bronchial epithelial cells (BEAS-2B)

[0160] Flow cytometry was used to analyze the level of ROS produced by BEAS-2B. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Fig.11The results showed that compared with the control group, the cellular ROS content in the PM group increased, while the cellular ROS in the PM+PHB-P group decreased and was significantly lower than that in the PM group, indicating that PHB-P can alleviate the excessive generation of ROS in human bronchial epithelial cells caused by PM. * There is a statistical difference compared with the Control group, # There is a statistical difference compared with the PM group, p<0.05.

[0161] (6) Effects of PDMAEMA on the activity of PM-exposed human bronchial epithelial cells (BEAS-2B)

[0162] CCK-8 was used to analyze the activity of BEAS-2B cells. For specific experimental steps, please refer to the aforementioned “Experimental Methods” section. Fig. 9 The results showed that the cell activity in the PM group was significantly lower than that in the control group, and there was no statistical difference in the cell activity between the PM+PDMAEMA group and the PM group, indicating that at the same concentration as PHB-P, PDMAEMA could not reduce the toxic effect of PM on human bronchial epithelial cells, and PHB-P had a better attenuation effect. * There was a statistical difference compared with the Control group, # There was a statistical difference compared with the PM group, p<0.05.

Claims

1. Application of a degradable block copolymer in the prevention and control of atmospheric particulate pollution, characterized in that: The applications described include: 1) Application in adsorption of atmospheric particulate matter; 2) Application in reducing the toxicity of atmospheric particulate matter; The degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester, and its chemical structure is as follows: , wherein m is an integer selected from 100-120; n is an integer selected from 10-20; The atmospheric particulate matter includes PM particulate matter.

2. The use according to claim 1, characterized in that The degradable block copolymer is directly sprayed into the air; or, the degradable block copolymer is placed at the pollutant exhaust port; or, the degradable block copolymer is placed on protective equipment for filtering air pollutants.

3. Use of a degradable block copolymer in preparing a product for absorbing atmospheric particulate matter or preparing a product for reducing the toxicity of atmospheric particulate matter, characterized in that: The product is an adsorbent, a mask or a filter; the degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester, and its chemical structure is as follows: , wherein m is an integer selected from 100-120; n is an integer selected from 10-20.

4. The use according to claim 3, characterized in that When the product is an adsorbent, the amount of the degradable block copolymer in the adsorbent is 2.5 mg / m 3 When the product is a mask, the amount of the degradable block copolymer distributed in the mask adsorption interlayer is 2.5 mg / m 3 .

5. Application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on cell activity, wherein the degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester, and its chemical structure is as follows: ,in, m is an integer selected from 100-120; n is an integer selected from 10-20; The cells are human myocardial cells and / or human bronchial epithelial cells.

6. Application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on cell apoptosis, wherein the degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester, and its chemical structure is as follows: ,in, m is an integer selected from 100-120; n is an integer selected from 10-20; The cells are human myocardial cells and / or human bronchial epithelial cells.

7. Application of a degradable block copolymer in reducing the effect of atmospheric particulate matter on cell active oxygen, wherein the degradable block copolymer is hydroxybutyric acid-polymethacrylic acid-dimethylamino ester, and its chemical structure is as follows: ,in, m is an integer selected from 100-120; n is an integer selected from 10-20; The cells are human myocardial cells and / or human bronchial epithelial cells.

8. The use according to any one of claims 1 to 7, characterized in that: The atmospheric particulate matter includes negatively charged polluted particulate matter among PM polluted particulate matter.

9. The use according to claim 8, characterized in that The PM pollution particles include PM 2.5 、PM 10 , one or more of TSP.

10. The use according to claim 9, characterized in that The PM pollution particles are PM 2.5 .

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