Liposome coated nano-selenium as well as preparation method and application thereof

Abstract

The invention relates to the technical field of nano-drug preparation. The invention provides lipidosome coated nano-selenium and a preparation method thereof, and the preparation method comprises the following steps: (1) dissolving chitosan in an acetic acid aqueous solution, sequentially adding sodium selenite and ascorbic acid, and mixing and stirring to obtain chitosan nano-selenium; (2) adding TW-80 into chitosan nano-selenium, and mixing and stirring to obtain a solution A; and (3) mixing lecithin dissolved in the ethanol solution with the solution A, and stirring to obtain the liposome-coated nano-selenium. The liposome coated nano-selenium prepared by the technical scheme of the invention has good biocompatibility, and can be better absorbed by organisms; meanwhile, the stability and the slow release property in intestines and stomach are better. In addition, the liposome coated nano-selenium prepared by the invention can also improve the problems of low testosome rate, low sperm activity and in-vivo hormone disorder caused by microplastics in the environment.

Description

Technical Field

[0001] The present invention relates to the technical field of nano-drug preparation, and particularly relates to a liposome-coated nano-selenium, a preparation method thereof, and an application thereof. Background Art

[0002] Selenium has a very wide range of effects on organisms, including antioxidant, regulating hormone levels, promoting growth and development, etc. Selenium is an important part of the antioxidant system. It helps to scavenge peroxides and free radicals in the body by forming antioxidant enzymes such as glutathione peroxidase (GSH-Px), protecting cell membranes from oxidative damage. Selenium is involved in the metabolism of thyroid hormones, affects the reproductive functions of humans and animals, and at the same time has a positive impact on reproductive health, can improve sperm quality and enhance fertility. Selenium helps to protect the heart, lower cholesterol levels, prevent cardiovascular diseases, and also has a hepatoprotective effect. Selenium has a strong affinity for metals, can combine with heavy metals such as mercury, cadmium, and lead in the body and be excreted from the body, having a natural detoxifying effect against heavy metals. Selenium is an essential trace element for growth and reproduction and is involved in the body's metabolism.

[0003] The synthesis methods of nano-selenium mainly include three categories: physical method, chemical method, and biological method, and generally the chemical method is used for synthesis. When preparing nano-selenium by the chemical method, generally a reducing agent (such as ascorbic acid, glutathione, sodium sulfite, etc.) is used to reduce selenium salts (sodium selenite or selenious acid) to synthesize nano-selenium, but the synthesized nano-selenium has general stability and biocompatibility, and its application performance needs to be improved. Summary of the Invention

[0004] The purpose of the present invention is to provide a liposome-coated nano-selenium and a preparation method thereof. The liposome-coated nano-selenium has good biocompatibility, stability, and sustained-release properties, and can significantly improve the problems of low testicular rate, low sperm activity, and in vivo hormone disorder caused by microplastics in the environment.

[0005] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides a preparation method of liposome-coated nano-selenium, including the following steps:

[0007] (1) Dissolve chitosan in an acetic acid aqueous solution, then sequentially add sodium selenite and ascorbic acid, and mix and stir to obtain chitosan nano-selenium;

[0008] (2) Add TW-80 to the chitosan nano-selenium, mix and stir to obtain solution A;

[0009] (3) Mix the lecithin dissolved in an ethanol solution with solution A and stir to obtain the liposome-coated nano-selenium.

[0010] Preferably, the mass-volume ratio of chitosan, sodium selenite, ascorbic acid, and TW-80 is (0.05-0.15) g:(8-12) mL:(8-12) mL:(1500-1900) μL.

[0011] Preferably, in step (1), the volume concentration of the acetic acid aqueous solution is 0.5-1.5%, and the mass-volume ratio of chitosan to the acetic acid aqueous solution is (0.05-0.15) g:(40-60) mL.

[0012] Preferably, in step (1), the concentration of sodium selenite is 10-30 mmol / L, the concentration of ascorbic acid is 70-85 mmol / L, and the mass concentration of TW-80 is 4-6%.

[0013] Preferably, after dissolving chitosan in the acetic acid aqueous solution in step (1), stirring treatment is further included. The rotation speed of the stirring is 500-700 r / min, and the stirring time is 5-7 h.

[0014] Preferably, in step (3), the mass-volume ratio of lecithin to the ethanol solution is 1 g:(30-50) mL, and the volume concentration of the ethanol solution is 70-80%.

[0015] Preferably, the volume ratio of chitosan nano-selenium to the ethanol solution is 1:(0.2-1).

[0016] The present invention also provides liposome-coated nano-selenium prepared by the above preparation method.

[0017] The present invention also provides the application of the liposome-coated nano-selenium in the preparation of drugs for treating male reproductive system damage caused by microplastics.

[0018] Preferably, the action concentration of the liposome-coated nano-selenium, calculated by selenium content, is 0.2-1 mg / kg.

[0019] By adopting the above technical solutions, the present invention has the following beneficial effects:

[0020] The liposome-coated nano-selenium prepared by the technical solution of the present invention has good biocompatibility and can be better absorbed by organisms; at the same time, it has better stability and sustained release in the stomach and intestines. Moreover, the liposome-coated nano-selenium prepared by the present invention can also improve the problems of low testicular rate, low sperm activity, and in vivo hormone disorder caused by microplastics in the environment. Experiments also show that: the use of the liposome-coated nano-selenium significantly increases the testicular rate and sperm activity of mice, significantly reduces the level of estradiol hormone (E 2 ) in the serum of mice, and significantly increases the level of testosterone hormone (T) in the serum of mice. Description of the Drawings

[0021] Figure 1 For the static state of the nano-liquid in different treatment groups (numbered test tubes 1-4 from left to right in the figure);

[0022] Figure 2 It is the morphological map of nano-selenium in the C-SeNPS + alcohol ester (1:0.5) group under the condition of 4°C;

[0023] Figure 3 It is the transmission electron microscope image of lecithin chitosan nano-selenium in the C-SeNPS + alcohol ester (1:0.25) + 5% TW group;

[0024] Figure 4 It is the elemental mapping result image of lecithin chitosan nano-selenium in the C-SeNPS + alcohol ester (1:0.25) + 5% TW group ( Figure 4 A and B in it are transmission electron microscope images, C is the carbon elemental mapping result image, D is the nitrogen elemental mapping result image, E is the oxygen elemental mapping result image, F is the phosphorus elemental mapping result image, and G is the selenium elemental mapping result image);

[0025] Figure 5 It is the wavelength situation of the phospholipid chitosan nano-selenium prepared by the present invention under different pH and different solvent conditions ( Figure 5 A in it is Nacl with different concentrations, B represents different pH values, C represents different concentrations of ethanol, D represents different concentrations of Cacl 2 , E represents different types of organic solvents, and F represents different concentrations of H 2 O 2 );

[0026] Figure 6 It is the particle size change situation of the phospholipid chitosan nano-selenium prepared by the present invention under in vitro gastrointestinal simulation conditions ( Figure 6 A in it represents the particle size change situation only under simulated gastric digestion for different times; B represents the particle size change situation at different intestinal digestion times when gastric digestion is 0 h; C represents the particle size change situation at different intestinal digestion times when gastric digestion is 1 h; D represents the particle size change situation at different intestinal digestion times when gastric digestion is 2 h; E represents the particle size change situation at different intestinal digestion times when gastric digestion is 3 h);

[0027] Figure 7 It is the testicular body rate situation of mice in different treatment groups;

[0028] Figure 8 It is the sperm activity situation of mice in different treatment groups;

[0029] Figure 9 It is the content situation of estradiol hormone and testosterone hormone in the serum of mice in different treatment groups ( Figure 9 A in it is the content of estradiol hormone, and B is the content of testosterone hormone). Detailed implementation mode

[0030] The present invention provides a preparation method of liposome-coated nano-selenium, which comprises the following steps:

[0031] (1) Dissolve chitosan in an acetic acid aqueous solution, then sequentially add sodium selenite and ascorbic acid, and mix and stir to obtain chitosan nano-selenium;

[0032] (2) Add TW-80 to the chitosan nano-selenium, and mix and stir to obtain solution A;

[0033] (3) Mix the lecithin dissolved in an ethanol solution with solution A and stir to obtain liposome-coated nano-selenium.

[0034] In the present invention, chitosan is dissolved in an acetic acid aqueous solution. The mass-volume ratio of chitosan to the acetic acid aqueous solution is preferably (0.05-0.15) g:(40-60) mL, more preferably (0.08-0.13) g:(45-55) mL, and even more preferably 0.1 g:50 mL; the volume concentration of the acetic acid aqueous solution is preferably 0.5-1.5%, more preferably 0.8-1.2%, and even more preferably 1%. Then carry out stirring treatment. The stirring preferably uses a magnetic stirrer, and the stirring speed is preferably 500-700 r / min, more preferably 550-650 r / min, and even more preferably 600 r / min.

[0035] In the present invention, after the stirring is completed, sodium selenite and ascorbic acid are sequentially added, and mixed and stirred to obtain chitosan nano-selenium (C-SeNPS). Preferably, stirring is carried out after each addition. After adding sodium selenite, stir for 0.5-1.5 h, more preferably 0.8-1.2 h, and even more preferably 1 h; after adding ascorbic acid, stir for 10-15 h, more preferably 11-14 h, and even more preferably 12 h.

[0036] In the present invention, TW-80 is added to the chitosan nano-selenium, and mixed and stirred to obtain solution A. In the present invention, after adding TW-80, stir until it is completely dissolved.

[0037] In the present invention, the mass-volume ratio of chitosan, sodium selenite, ascorbic acid, and TW-80 is preferably (0.05 - 0.15) g : (8 - 12) mL : (8 - 12) mL : (1500 - 1900) μL, more preferably (0.07 - 0.12) g : (9 - 11) mL : (9 - 11) mL : (1600 - 1850) μL, and even more preferably 0.1 g : 10 mL : 10 mL : 1800 μL. The concentration of sodium selenite in the present invention is preferably 10 - 30 mmol / L, more preferably 15 - 25 mmol / L, and even more preferably 20 mmol / L; the concentration of ascorbic acid is preferably 70 - 85 mmol / L, more preferably 75 - 82 mmol / L, and even more preferably 80 mmol / L; the mass concentration of TW-80 is 4 - 6%, more preferably 4.5 - 5.5%, and even more preferably 5%.

[0038] In the present invention, lecithin is dissolved in an ethanol solution. The mass-volume ratio of lecithin to the ethanol solution is preferably 1 g : (30 - 50) mL, more preferably 1 g : (35 - 45) mL, and even more preferably 1 g : 40 mL. The volume concentration of the ethanol solution in the present invention is preferably 70 - 80%, more preferably 72 - 78%, and even more preferably 75%. Then, it is mixed with solution A and stirred to obtain liposome-coated nano-selenium. The stirring time is preferably 1 - 3 h, more preferably 1.5 - 2.5 h, and even more preferably 2 h.

[0039] In the present invention, the volume ratio of chitosan nano-selenium to the ethanol solution is preferably 1 : (0.2 - 1), more preferably 1 : (0.23 - 0.5), and even more preferably 1 : 0.25.

[0040] The present invention also provides liposome-coated nano-selenium prepared by the above preparation method.

[0041] The present invention also provides the application of the liposome-coated nano-selenium in the preparation of a drug for treating microplastic-induced damage to the male reproductive system.

[0042] In the present invention, the action concentration of the liposome-coated nano-selenium in terms of selenium content is preferably 0.2 - 1 mg / kg.

[0043] The technical solutions provided by the present invention will be described in detail below with reference to the examples, but they should not be construed as limiting the protection scope of the present invention.

[0044] Example 1

[0045] A preparation method of liposome-coated nano-selenium is as follows:

[0046] (1) Dissolve 0.1 g of chitosan in 50 mL of 1% aqueous acetic acid solution, stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium;

[0047] (2) Add 1800 μL of 5% TW-80 to the chitosan nano-selenium and stir until completely dissolved to obtain Solution A;

[0048] (3) Dissolve 1 g of lecithin in 40 mL of 75% ethanol, then add it to Solution A and stir for 2 h to obtain the product.

[0049] Example 2

[0050] A method for preparing liposome-coated nano-selenium is as follows:

[0051] (1) Dissolve 0.1 g of chitosan in 40 mL of 1% aqueous acetic acid solution, stir with a magnetic stirrer at 500 r / min for 7 h; then add 8 mL of 20 mmol / L sodium selenite and stir for 1 h; add 12 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium;

[0052] (2) Add 1500 μL of 5% TW-80 to the chitosan nano-selenium and stir until completely dissolved to obtain Solution A;

[0053] (3) Dissolve 1 g of lecithin in 30 mL of 75% ethanol, then add it to Solution A and stir for 2 h to obtain the product.

[0054] Example 3

[0055] A method for preparing liposome-coated nano-selenium is as follows:

[0056] (1) Dissolve 0.15 g of chitosan in 60 mL of 1% aqueous acetic acid solution, stir with a magnetic stirrer at 700 r / min for 5 h; then add 12 mL of 20 mmol / L sodium selenite and stir for 1 h; add 8 mL of 70 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium;

[0057] (2) Add 1900 μL of 5% TW-80 to the chitosan nano-selenium and stir until completely dissolved to obtain Solution A;

[0058] (3) Dissolve 1 g of lecithin in 50 mL of 75% ethanol, then add it to Solution A and stir for 2 h to obtain the product.

[0059] Test Example 1

[0060] Different treatment groups were set up based on the preparation method of liposome-coated nano-selenium to explore the stability of nano-selenium prepared in different treatment groups.

[0061] C-SeNPS group: Dissolve 0.1 g of chitosan in 50 mL of 1% acetic acid aqueous solution and stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain.

[0062] C-SeNPS + alcohol lipid (1:1) group: Dissolve 0.1 g of chitosan in 50 mL of 1% acetic acid aqueous solution and stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium; dissolve 1 g of lecithin in 40 mL of 75% ethanol and then add it to chitosan nano-selenium, where the volume ratio of chitosan nano-selenium to ethanol is 1:1, and stir for 2 h to obtain.

[0063] C-SeNPS + alcohol lipid (1:0.5) group: Dissolve 0.1 g of chitosan in 50 mL of 1% acetic acid aqueous solution and stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium; dissolve 1 g of lecithin in 40 mL of 75% ethanol and then add it to chitosan nano-selenium, where the volume ratio of chitosan nano-selenium to ethanol is 1:0.5, and stir for 2 h to obtain.

[0064] C-SeNPS + alcohol (1:0.5) group: Dissolve 0.1 g of chitosan in 50 mL of 1% acetic acid aqueous solution and stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium; add chitosan nano-selenium to 75% ethanol according to a volume ratio of 1:0.5 and stir for 2 h to obtain.

[0065] C-SeNPS + Alcohol Lipid (1:0.25) + 5% TW Group: Dissolve 0.1 g of chitosan in 50 mL of 1% acetic acid aqueous solution and stir with a magnetic stirrer at 600 r / min for 6 h; then add 10 mL of 20 mmol / L sodium selenite and stir for 1 h; add 10 mL of 80 mmol / L ascorbic acid and stir for 12 h to obtain chitosan nano-selenium; add 1800 μL of 5% TW-80 to the chitosan nano-selenium and stir until completely dissolved to obtain Solution A; dissolve 1 g of lecithin in 40 mL of 75% ethanol and then add it to Solution A, where the volume ratio of chitosan nano-selenium to ethanol is 1:0.25, and stir for 2 h to obtain the product. (I)

[0067] Pour the nano-liquids prepared in each group above into centrifuge tubes, seal them, place one group in a low-temperature environment at 4°C for static settlement, and place the other group in a room-temperature environment for static settlement. Observe the state of the nano-liquids in each group every day to see if they are clear or turbid.

[0068] Table 1 Static Settlement States of Nano-Liquids in Different Treatment Groups

[0069]

[0070] The results are as Figure 1 shown. Test tubes 1 - 3 are all opaque, with stratification and precipitation or turbidity; test tube 4 is transparent, that is, the sample in test tube 4 is a sample with good stability (from left to right are test tubes 1 - 4, and test tubes 1 - 4 correspond to the other four groups except the C-SeNPS group in turn). In addition, the alcohol content will affect the activity and morphology of nano-selenium. When the alcohol content is low, lecithin will not dissolve and cannot be coated. The nano-selenium morphology in the C-SeNPS + Alcohol Lipid (1:0.5) group is rod-shaped when statically settled at 4°C, as Figure 2 . (II)

[0072] Use a nano-particle size analyzer to measure the particle size and other properties of the lecithin-chitosan nano-selenium sample prepared in the C-SeNPS + Alcohol Lipid (1:0.25) + 5% TW group. The transmission electron microscope image is as Figure 3 shown. The results show that the particle size of this lecithin-chitosan nano-selenium sample is 273.6 nm, and the PDI value is 0.251. Compared with conventional chitosan nano-selenium (particle size is 107.9 nm, PDI value is 0.572), the particle size increases, which may be due to the coating of liposomes, and the PDI value decreases, which also indicates that the lecithin-chitosan nano-selenium prepared by the present invention is more stable. (III)

[0074] Weigh 1 ml of the lecithin-chitosan nano-selenium sample prepared with the C-SeNPS + alcohol-fat (1:0.25) + 5% TW group (accurate to 0.0001 g, sample mass 0.9035 g). Add 5 ml of nitric acid and react on a hot plate at 220 °C until no obvious phenomenon occurs. Continue heating until the volume is less than 0.5 ml. Let it stand at room temperature and make up the volume to 10 ml with high-purity water. Then use an X-ray energy spectrometer for testing.

[0075] The results are as Figure 4 shown. The selenium elements are aggregated into nano-spherical shapes, indicating the successful synthesis of nano-selenium particles. Through calculation, it can be determined that the selenium element content is 73.733 mg / kg.

[0076] Test Example 2 (1)

[0078] Respectively take the lecithin-chitosan nano-selenium (L / C-SeNPS) prepared by the technical solution of the present invention under different conditions (solvent, pH), and observe the stability of the lecithin-chitosan nano-selenium.

[0079] Take a 96-well plate. Mix 100 μL of the lecithin-chitosan nano-selenium prepared by the technical solution of the present invention with 100 μL of Nacl, ethanol, Cacl 2 and H 2 O 2 with different concentrations respectively. Mix 100 μL of the lecithin-chitosan nano-selenium prepared by the technical solution of the present invention with 100 μL of different organic solvents respectively. After incubating at room temperature for 1 h, use an enzyme-linked immunosorbent assay (ELISA) reader to perform a full-spectrum scan at wavelengths of 300 - 900 nm every 4 nm.

[0080] Similarly, take a 96-well plate, set five groups, and respectively take 100 μL of the lecithin-chitosan nano-selenium prepared by the technical solution of the present invention and place it in the wells. Use sodium hydroxide solution or hydrochloric acid solution to adjust the pH of the lecithin-chitosan nano-selenium in the wells. The pH values of the five groups are adjusted to 1, 4, 7, 10, and 13 respectively.

[0081] The results are as Figure 5 shown. Under different pH and different solvent conditions, the lecithin-chitosan nano-selenium maintains a relatively stable state. (2)

[0083] Take a 96-well plate. Place 100 μL of the lecithin-chitosan nano-selenium (L / C-SeNPS) prepared by the technical solution of the present invention in 100 μL of simulated gastric juice and 100 μL of simulated intestinal juice respectively to carry out the simulated digestion process. Then use an ELISA reader to measure at wavelengths of 410 and 490 nm, and observe the stability of the lecithin-chitosan nano-selenium.

[0084] Simulated gastric juice: Add 3.1 g of sodium chloride, 1.1 g of potassium chloride, 0.6 g of sodium bicarbonate, and 0.15 g of calcium chloride to 1 L of pure water, and adjust the pH to 3 with hydrochloric acid solution; take 150 mL of the above solution, add 35.4 mg of pepsin, and adjust the pH to 3 with hydrochloric acid solution to obtain simulated gastric juice.

[0085] Simulated intestinal juice: Add 5.4 g of sodium chloride, 0.65 g of potassium chloride, and 0.33 g of calcium chloride to 1 L of pure water; adjust the pH to 7 with sodium bicarbonate solution; take 200 mL of the above solution, add 26 mg of trypsin and 5 mg of bile salts, and adjust the pH to 7 with sodium bicarbonate solution to obtain simulated intestinal juice.

[0086] The results are as Figure 6 shown. Compared with the initial state, after 3 h of simulated gastric digestion, the change in the particle size of lecithin-chitosan nanoselenium was not significant. When simulated intestinal juice digestion was carried out for 3 h after 2 - 3 h of simulated gastric juice digestion, the particle size of the nanoparticles increased significantly.

[0087] Test Example 3

[0088] Select 5-week-old (SPF) ICR male mice (purchased from Beijing Speefoo Biotechnology Co., Ltd.), with 9 mice in each group as replicates, and divide them into the following 5 groups: CON group, BPA group, BPA + chitosan nanoselenium group prepared in Example 1, BPA + lecithin-chitosan nanoselenium (low dose) group, BPA + lecithin-chitosan nanoselenium (high dose) group. Place 3 mice in each cage, raise them for 1 week in advance to adapt to the environment, and observe the state of the mice.

[0089] Grind BPA into fine particles, then pre-dissolve it with 0.5% DMSO, and then add physiological saline to prepare a BPA solution with a concentration of 1 mg / mL.

[0090] CON group: Continuously intragastrically administer physiological saline for 3 weeks, once a day, 0.2 mL each time.

[0091] BPA group: Continuously administer BPA solution for 3 weeks, with a gavage dose of 5 mg / kg for each mouse, once a day, 0.2 mL each time.

[0092] BPA + chitosan nanoselenium group: Continuously administer BPA solution for 3 weeks, and at the same time intragastrically administer chitosan nanoselenium. Calculated by selenium content, the selenium content is 0.2 mg / kg, once a day, 0.2 mL each time.

[0093] BPA + lecithin-chitosan nanoselenium (low dose) group: Continuously administer BPA solution for 3 weeks, and at the same time intragastrically administer lecithin-chitosan nanoselenium prepared in Example 1. Calculated by selenium content, the selenium content is 0.2 mg / kg, once a day, 0.2 mL each time.

[0094] BPA + Lecithin Chitosan Nanoscale Selenium (High Dose) Group: Continuously administer BPA solution for 3 weeks, and simultaneously administer the lecithin chitosan nanoscale selenium prepared in Example 1 by gavage. Calculated by selenium content, the selenium content is 1 mg / kg, once a day, 0.2 mL each time.

[0095] After the above groups stopped administering drugs, they were stabilized for 3 days. During this period, mouse samples of each group were collected for analysis of the testis-body ratio, sperm motility, and in vivo sex hormone levels.

[0096] Weigh the testis weight of the dissected mice and the body weight of the mice on the day of dissection. The testis weight / body weight is the testis-body ratio. This data is one of the bases for judging the degree of damage to the reproductive organs of male mice. The results are as Figure 7 shown, indicating that microplastics (BPA) in the environment can reduce the testis-body ratio of mice, while the testis-body ratio increases after treatment with lecithin chitosan nanoscale selenium, and the effect of lecithin chitosan nanoscale selenium in increasing the testis-body ratio of mice is greater than that of ordinary chitosan nanoscale selenium.

[0097] After dissection, remove the epididymis of the mice, cut it into pieces in preheated physiological saline at 37°C, suck this liquid onto a glass slide, and analyze the sperm motility with a sperm analysis system. The results are as Figure 8 shown, microplastics (BPA) in the environment can reduce the sperm motility and motility of mice, while the sperm motility increases after treatment with lecithin chitosan nanoscale selenium.

[0098] Collect the blood of mice by enucleation of the eyeballs, let it stand and centrifuge to separate the serum, and use ELISA method to determine the contents of estradiol hormone (E 2 ) and testosterone hormone (T) in the serum of mice. This detection index can judge the impact of microplastics on hormone disorders in mice and the improvement after treatment with nanoscale selenium. The results are as Figure 9 shown, microplastics (BPA) in the environment can increase the estradiol hormone (E 2 ) in the serum of mice, while the hormone level decreases after treatment with lecithin chitosan nanoscale selenium; microplastics in the environment can reduce the testosterone hormone (T) in the serum of mice, while the hormone level increases after treatment with lecithin chitosan nanoscale selenium.

[0099] In summary, it can be seen that the liposome-coated nanoscale selenium prepared by the technical solution of the present invention has good biocompatibility and can be better absorbed by organisms; at the same time, it has better stability and sustained release properties in the gastrointestinal tract. Moreover, the liposome-coated nanoscale selenium prepared by the present invention can also improve the problems of low testis-body ratio, low sperm motility, and in vivo hormone disorders caused by microplastics in the environment.

[0100] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A method for preparing liposome-encapsulated nano-selenium, characterized in that: The steps include: (1) dissolving chitosan in an acetic acid aqueous solution, and then sequentially adding sodium selenite and ascorbic acid, mixing and stirring to obtain chitosan nano-selenium; (2) adding TW-80 to chitosan nano-selenium, mixing and stirring to obtain solution A; (3) Mix the lecithin dissolved in the ethanol solution with solution A and stir to obtain liposome-coated nano-selenium.

2. The preparation method according to claim 1, characterized in that: The mass volume ratio of the chitosan, sodium selenite, ascorbic acid and TW-80 is (0.05-0.15) g: (8-12) mL: (8-12) mL: (1500-1900) μL.

3. The preparation method according to claim 1, characterized in that: The volume concentration of the acetic acid aqueous solution in step (1) is 0.5-1.5%, and the mass volume ratio of the chitosan to the acetic acid aqueous solution is (0.05-0.15) g: (40-60) mL.

4. The preparation method according to claim 1, characterized in that: The concentration of the sodium selenite is 10-30 mmol / L, the concentration of the ascorbic acid is 70-85 mmol / L, and the mass concentration of the TW-80 is 4-6%.

5. The preparation method according to claim 1, characterized in that: After dissolving chitosan in the acetic acid aqueous solution in step (1), the step further includes stirring, wherein the stirring speed is 500-700 r / min, and the stirring time is 5-7 hours.

6. The preparation method according to claim 1, characterized in that: The mass volume ratio of the lecithin to the ethanol solution in step (3) is 1 g: (30-50) mL, and the volume concentration of the ethanol solution is 70-80%.

7. The preparation method according to claim 1, characterized in that: The volume ratio of the chitosan nano-selenium to the ethanol solution is 1:(0.2-1).

8. Liposome-coated nano-selenium prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the liposome-encapsulated nano-selenium prepared by the preparation method according to any one of claims 1 to 7, or the liposome-encapsulated nano-selenium according to claim 8 in the preparation of drugs for treating damage to the male reproductive system caused by microplastics.

10. The use according to claim 9, characterized in that: The effective concentration of the liposome-encapsulated nano-selenium is 0.2-1 mg / kg in terms of selenium content.

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