CN107412280B - Nano-selenium hydrosol with anti-tumor activity, preparation and preservation method and application - Google Patents
Nano-selenium hydrosol with anti-tumor activity, preparation and preservation method and application Download PDFInfo
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Abstract
The invention relates to the technical field of nano-selenium hydrosol, in particular to nano-selenium hydrosol with anti-tumor activity, and a preparation method, a preservation method and application thereof. The hydrosol at least comprises the following components in concentration: nano selenium of 0.5mmol/L to 5.0 mmol/L; the coriolus versicolor polysaccharide protein is 100.0 mg/L-600.0 mg/L. The preparation method of the hydrosol at least comprises the step S01, adding a solution containing selenium ions and/or selenious ions into the coriolus versicolor polysaccharide protein aqueous solution; s02, dropwise adding a reducing agent solution into the mixed solution obtained in the step S01, and oscillating; and S03, carrying out constant volume and dialysis treatment on the product obtained by the reduction reaction in the step S02. The hydrosol exists stably at the temperature of 2-10 ℃. Because the coriolus versicolor polysaccharide protein has hydrophilic hydroxyl and amino groups, the affinity of the nano selenium and cancer cells can be enhanced by the two groups, the intake of the nano selenium by the tumor cells is improved, the overall treatment effect of reducing the dosage, increasing the curative effect and reducing toxic and side effects is further achieved, and a more effective scheme is provided for clinical combined chemotherapy of cancer.
Description
Technical Field
The invention relates to the technical field of nano-selenium hydrosol, in particular to nano-selenium hydrosol with anti-tumor activity, and a preparation method, a preservation method and application thereof.
Background
Selenium is one of fifteen essential trace elements, and has important biological activities of resisting tumor, resisting oxidation, resisting aging, improving immunity, protecting and repairing cells, antagonizing and reducing harmful heavy metals and the like in a human body. Selenium is closely related to the occurrence and development of up to 40 diseases, including: AIDS, liver cancer, keshan disease, Kaschin-Beck disease, cardiovascular and cerebrovascular diseases, etc.
The people with low selenium or selenium deficiency can achieve the effects of improving the immunity of the organism and maintaining the normal functions of important organs such as heart, liver, lung, stomach and the like by supplementing selenium in a proper amount, and can also play a role in preventing tumors, liver diseases, cardiovascular and cerebrovascular diseases.
At present, more than 40 countries or regions around the world have the problem of selenium deficiency, and China also has a plurality of provinces belonging to the selenium deficiency or low-selenium regions. Areas with selenium deficiency or low selenium are areas with high incidence rate of tumor, liver disease, cardiovascular disease and the like, and selenium supplement is needed. However, as a nutritional supplement or a cancer preventive agent, the beneficial dose range and toxic dose range of selenium are extremely narrow, and selenium poisoning is easily caused if the selenium is not noticed, so that the potential of selenium in disease control, especially cancer control, is severely limited. However, the toxicity of selenium depends on its chemical form. Research results show that the toxicity of the inorganic selenium compound is stronger than that of the organic selenium compound, and the toxicity of the selenocysteine is similar to that of sodium selenite and is stronger than that of nano-selenium; compared with the selenium compound, the nano-selenium has the advantages of high bioavailability, strong bioactivity and low toxicity.
At present, the preparation method of nano elemental selenium is generally a reduction method, specifically, elemental selenium is obtained by using oxysalt or oxide of selenium through various reducing agents, and meanwhile, a modifier or a regulator is used for modifying and regulating the particle size and morphology, so that ideal nano elemental selenium is finally obtained. However, in the existing preparation methods of various nano elemental selenium, a regulating agent or a modifying agent is adopted to regulate or modify the appearance, and the application requirements are not considered. In addition, the regulating agent or the modifying agent is a material without biological activity, which cannot effectively improve the biological activity of the nano elemental selenium, especially cannot improve the anti-tumor activity of the nano elemental selenium, namely, the product obtained by the conventional method is not the functionalized nano elemental selenium.
Disclosure of Invention
The invention aims to provide a nano selenium hydrosol with anti-tumor activity and a preparation method thereof, aiming at the problem that the biological activity of nano elemental selenium cannot be effectively improved by a modifier or a regulator in the existing preparation method of nano elemental selenium.
The invention also aims to provide a method for preserving the nano-selenium hydrosol with the anti-tumor activity.
Another purpose of the present invention is to limit the application field of the nano selenium hydrosol with anti-tumor activity.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a nano-selenium hydrosol with anti-tumor activity at least comprises the following components in concentration:
nano selenium of 0.5mmol/L to 5.0 mmol/L;
the coriolus versicolor polysaccharide protein is 100.0 mg/L-600.0 mg/L.
The preparation method of the nano-selenium hydrosol with anti-tumor activity at least comprises the following steps:
step S01, adding a solution containing selenium ions and/or selenious ions into a coriolus versicolor polysaccharide protein aqueous solution;
s02, dropwise adding a reducing agent solution into the mixed solution obtained in the step S01, and oscillating;
and S03, carrying out constant volume and dialysis treatment on the product obtained by the reduction reaction in the step S02.
And correspondingly, the nano-selenium hydrosol with the anti-tumor activity is stored at the temperature of 2-10 ℃.
And further correspondingly, the application of the nano-selenium hydrosol with anti-tumor activity in the anti-tumor field.
The nano-selenium hydrosol with anti-tumor activity provided by the embodiment of the invention takes the medicinal coriolus versicolor polysaccharide protein as a nano-selenium functionalized molecule, and is mutually promoted in a synergistic way with the physiological effect of nano-selenium so as to jointly exert the anti-tumor activity; specifically, by means of the polyhydroxy structure of polysaccharide part in polysaccharose protein of coriolus versicolor, the strong physical adsorption effect on nano selenium is exerted, further aggregation and precipitation of nano selenium are avoided, and meanwhile, the surface of nano selenium is effectively modified, so that the particle size of nano selenium is regulated and controlled, and the nano selenium hydrosol is kept stable.
According to the preparation method of the nano-selenium hydrosol with the anti-tumor activity, provided by the embodiment of the invention, the nano-selenium hydrosol with the biological activity, especially the anti-tumor activity can be successfully prepared only by combining the coriolus versicolor polysaccharide protein and the solution containing the selenium ions/the selenious ions with the reducing agent at normal temperature and under normal pressure, and particularly, the polyhydroxy structure of the polysaccharide part in the coriolus versicolor polysaccharide protein plays a strong physical adsorption role on the nano-selenium, so that the particle size of the nano-selenium can be well regulated and controlled, the nano-selenium is stabilized, and the nano-selenium is prevented from further aggregation and precipitation; meanwhile, the method does not need to add any other template agent, thereby avoiding the possible adverse effect in practical application. Therefore, the method has the characteristics of simple preparation steps, simple preparation conditions, simple and feasible process and capability of large-scale production.
The preservation method of the nano-selenium hydrosol with anti-tumor activity does not need harsh conditions, and can obtain the nano-selenium hydrosol with good stability, almost no change of the grain diameter of the nano-selenium and no attenuation of the biological activity only by the preservation temperature of 2-10 ℃.
The physiological effects of Coriolus versicolor polysaccharide protein and nano-selenium in the nano-selenium hydrosol with anti-tumor activity are mutually promoted to exert the synergistic anti-tumor activity, because Coriolus versicolor polysaccharide protein has hydrophilic hydroxyl (-OH) and amino (-NH)2) The two groups can enhance the affinity of the nano-selenium in the nano-selenium hydrosol with cancer cells, improve the intake of the tumor cells to the nano-selenium, further achieve the overall treatment effects of reducing the dosage of the medicine, improving the curative effect and reducing the toxic and side effects, and provide a more effective scheme for the combined chemotherapy of the cancer in clinic.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a particle size distribution diagram of a nano-selenium hydrosol with anti-tumor activity of example 1 of the present invention;
FIG. 2 is a statistical graph showing the particle stability of nano-selenium hydrosol having anti-tumor activity according to the time variation in example 1 of the present invention;
FIG. 3 is a TEM image of the nano-selenium hydrosol with anti-tumor activity of example 1 of the present invention;
FIG. 4 is a TEM image of the nano-selenium hydrosol with anti-tumor activity of example 1 of the present invention;
FIG. 5 is a TEM image of the nano-selenium hydrosol with anti-tumor activity of example 1 of the present invention;
FIG. 6 is Zeta potential diagram of nano-selenium hydrosol with anti-tumor activity of example 1 of the present invention;
FIG. 7 is an EDX elemental analysis chart of nano-selenium hydrosol having anti-tumor activity of example 1 of the present invention;
FIG. 8 is a comparison of the main functional groups of the nano-selenium hydrosol with anti-tumor activity and Coriolus versicolor polysaccharide protein in example 1 of the present invention;
FIG. 9 is a statistical chart of the growth of AGS cells of EBV-infected gastric epithelium inhibited by nano-selenium hydrosol with anti-tumor activity in example 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The embodiment of the invention provides a nano-selenium hydrosol with anti-tumor activity, which at least comprises the following components in concentration:
nano selenium of 0.5mmol/L to 5.0 mmol/L;
the coriolus versicolor polysaccharide protein is 100.0 mg/L-600.0 mg/L.
In any embodiment, the coriolus versicolor proteoglycan protein is a coriolus versicolor water-soluble proteoglycan protein, which is ultimately present in the form of a hydrosol.
In a preferred embodiment, the total sugar content of the coriolus versicolor water-soluble polysaccharide protein is 70-80%, the total protein content is 15-25%, wherein the polysaccharide part mainly comprises glucose, and simultaneously comprises mannose and arabinose; the main amino acids of the protein part comprise aspartic acid, glutamic acid, threonine, serine, glycine, alanine and the like. The polyhydroxy structure contained in the polysaccharide part of the coriolus versicolor polysaccharide protein has strong physical adsorption effect on the nano selenium, and can effectively avoid the further aggregation and precipitation of the nano selenium so as to realize the modification of the surface of the nano selenium, regulate and control the particle size of the nano selenium and ensure the stability of hydrosol; in addition, the coriolus versicolor water-soluble polysaccharide protein has hydrophilic hydroxyl (-OH) and amino (-NH) groups, and the two groups can enhance the affinity of selenium and cancer cells, improve the uptake of tumor cells to nano-selenium, and achieve the overall treatment effects of reducing the dosage, increasing the curative effect and reducing the toxic and side effects.
The nano-selenium hydrosol with anti-tumor activity provided by the embodiment of the invention takes the medicinal coriolus versicolor polysaccharide protein as a nano-selenium functionalized molecule, and the anti-tumor activity of the coriolus versicolor polysaccharide protein and the physiological effect of nano-selenium are mutually coordinated and promoted to jointly play a synergistic anti-tumor effect, thereby providing a good scheme for clinical combined treatment of cancers.
Correspondingly, the invention further provides a preparation method of the nano-selenium hydrosol with anti-tumor activity on the basis of providing the nano-selenium hydrosol with anti-tumor activity. In a specific embodiment, the preparation method of the nano-selenium hydrosol with anti-tumor activity at least comprises the following steps:
step S01, adding a solution containing selenium ions and/or selenious ions into a coriolus versicolor polysaccharide protein aqueous solution;
s02, dropwise adding a reducing agent solution into the mixed solution obtained in the step S01, and oscillating;
and S03, carrying out constant volume and dialysis treatment on the product obtained by the reduction reaction in the step S02.
In any embodiment, the coriolus versicolor proteoglycan protein in the coriolus versicolor proteoglycan protein aqueous solution is coriolus versicolor water-soluble proteoglycan protein.
Preferably, in the embodiment of the invention, the concentration of coriolus versicolor proteoglycan in the nano-selenium hydrosol obtained after dialysis treatment is ensured to be 100.0 mg/L-600.0 mg/L, and the average particle size of the nano-selenium hydrosol can be kept below 100nm under the concentration of coriolus versicolor glucoprotein in the nano-selenium hydrosol, so that the particle size is not changed greatly within a certain time, and the stability is good.
In a preferred embodiment, the total sugar content of the coriolus versicolor water-soluble polysaccharide protein is 70-80%, the total protein content is 15-25%, wherein the polysaccharide part mainly comprises glucose, and simultaneously comprises mannose and arabinose; the main amino acids of the protein part comprise aspartic acid, glutamic acid, threonine, serine, glycine, alanine and the like. The polyhydroxy structure contained in the polysaccharide part of the coriolus versicolor polysaccharide protein has strong physical adsorption effect on the nano selenium, and can effectively avoid the further aggregation and precipitation of the nano selenium so as to realize the modification of the surface of the nano selenium, regulate and control the particle size of the nano selenium and ensure the stability of hydrosol; in addition, the coriolus versicolor water-soluble polysaccharide protein has hydrophilic hydroxyl (-OH) and amino (-NH), the two groups can enhance the affinity of selenium and cancer cells, improve the intake of tumor cells to nano-selenium, and achieve the overall treatment effects of reducing the dosage, increasing the curative effect and reducing the toxic and side effects.
In any embodiment, when the solution containing selenium ions or selenite ions is added, the molar ratio of the selenium ions or selenite ions to the reducing agent is (0.7-1.3): 3.0-9.0, and a slight excess of the reducing agent is ensured; when the mixed solution containing selenium ions and selenite ions is added, the molar ratio of the sum of the selenium ions and the selenite ions to the reducing agent is also (0.7-1.3): (3.0-9.0), and a slight excess of the reducing agent is ensured. At this ratio, it is ensured that the selenium ions and/or the selenite ions are all reduced by the reducing agent.
In a preferred embodiment, the concentration of the selenium ion-containing solution, the selenite ion-containing solution or the mixed solution of the selenium ion and the selenite ion is 0.5 mmol/L-2.0 mmol/L. Under the concentration, the average particle size of the prepared nano-selenium hydrosol can be kept below 100nm, the particle size is not changed greatly within a certain time, and the stability is good.
As a preferred embodiment, the selenium ion-containing solution is formulated with selenate.
As another preferred embodiment, the solution containing selenite ions is formulated with selenium dioxide and/or selenite. Furthermore, sodium selenite is adopted as a solute for preparation because sodium selenite is a component of health food and has higher safety.
In any embodiment, the concentration of the reducing agent solution is 2.0mmol/L to 8.0 mmol/L. Preferably, vitamin C is a reducing agent commonly used in the food industry, and is highly safe as well as having high reducing activity, and thus is preferred as a reducing agent.
In step S03, after the volume is fixed, dialysis is performed for 24 hours or more after the product is no longer dark in red.
In a preferred embodiment, the preparation is carried out in an environment of 15 ℃ to 35 ℃ and 1 standard atmospheric pressure, so that the external environment of the reaction is stable. In addition, the preparation conditions are energy-saving and environment-friendly, and the safety is high, thereby being beneficial to large-scale production.
The preparation method of the nano-selenium hydrosol with the anti-tumor activity provided by the embodiment of the invention can successfully prepare the nano-selenium hydrosol with the biological activity, particularly the anti-tumor activity by combining the coriolus versicolor polysaccharide protein and the solution containing the selenium ions/sub-ions with the reducing agent at normal temperature and normal pressure.
The nano-selenium hydrosol with anti-tumor activity prepared by the method has strong physical adsorption effect on nano-selenium by virtue of the polyhydroxy structure of the polysaccharide part in the coriolus versicolor polysaccharide protein, well regulates and controls the particle size of the nano-selenium, stabilizes the nano-selenium and prevents the nano-selenium from further aggregation and precipitation; meanwhile, the method does not need to add any other template agent, thereby avoiding the possible adverse effect in practical application. Therefore, the method has the characteristics of simple preparation steps, simple preparation conditions, simple and feasible process and capability of large-scale production.
Correspondingly, the embodiment of the invention further provides a preservation method of the nano-selenium hydrosol with the anti-tumor activity on the basis of providing the preparation method of the nano-selenium hydrosol with the anti-tumor activity.
In one embodiment, the nano-selenium hydrosol with anti-tumor activity provided by the embodiment of the invention should be stored at 2-10 ℃.
The preservation method provided by the embodiment of the invention can obtain the nano-selenium hydrosol which has good stability, almost no change in the particle size of the nano-selenium, no attenuation of biological activity and can exist in a sol form for a long time only by the preservation temperature of 2-10 ℃ without harsh conditions.
Correspondingly, the embodiment of the invention further provides the application of the nano-selenium hydrosol with the anti-tumor activity in the anti-tumor field on the basis of providing the nano-selenium hydrosol with the anti-tumor activity, the preparation method and the preservation method.
In one embodiment, the nano-selenium hydrosol with anti-tumor activity provided by the embodiment of the invention can be used for preparing anti-tumor drugs.
When the hydrosol is used in the anti-tumor field, the physiological effects of the coriolus versicolor polysaccharide protein and the nano-selenium are mutually promoted to play a role in synergistic anti-tumor activity, because the coriolus versicolor polysaccharide protein has hydrophilic hydroxyl (-OH) and amino (-NH)2) The two groups can enhance the affinity of the nano-selenium in the nano-selenium hydrosol with cancer cells, improve the intake of the tumor cells to the nano-selenium, further achieve the overall treatment effects of reducing the dosage of the medicine, improving the curative effect and reducing the toxic and side effects, and provide a more effective scheme for the combined chemotherapy of the cancer in clinic.
In order to better illustrate the nano-selenium hydrosol with anti-tumor activity provided by the embodiment of the invention, the following examples further illustrate the nano-selenium hydrosol with anti-tumor activity.
Example 1
(1) Adding 0mL, 0.5mL, 1.0mL, 3.0mL and 6.0mL of 2.5g/L coriolus versicolor water-soluble polysaccharide protein into 5 25mL volumetric flasks filled with 10.0mL of double distilled water respectively at 25 ℃ and 101.325 kPa;
(2) adding 1.0mL of sodium selenite solution with the concentration of 0.025mol/L into the 25mL volumetric flasks respectively, and shaking up gently to mix the sodium selenite solution and the sodium selenite solution fully to obtain mixed solutions;
(3) and (3) respectively dropwise adding 1.0mL of vitamin C solution with the concentration of 0.1mol/L into the mixed solution in the step (2), shaking up gently while dropwise adding, adding water to fix the volume to 25.0mL after dropwise adding is finished, standing until the red color is not deepened any more, and then dialyzing (with the molecular weight cut-off of 8000) for 24 hours to obtain a product. The selenium content is measured by a nitration ICP method, and the obtained product has the nano-selenium concentration of about 1.0-1.5 mmol/L and the coriolus versicolor water-soluble polysaccharide protein concentrations of 0.0mg/L, 50.0mg/L, 100.0mg/L, 300.0mg/L and 600.0mg/L respectively.
The obtained product is placed in an environment with the temperature of 2-10 ℃, and the product exists in the form of hydrosol.
The product obtained in example 1 was characterized by a detection instrument, and the tests mainly included the following:
the average particle size of the nanoparticles in the product, the Standard Deviation (SD) and the stability of the product with time were measured by a Nanosight NS3000 particle tracking Analyzer (Malvern), the test results are detailed in Table 1, the particle size distribution is detailed in the description attached figure 1, and the stability is detailed in the description attached figure 2.
The morphology of the product of example 1 is characterized by a JEM-2010 high-resolution projection electron microscope (JEOL), which is described in the attached figures 3, 4 and 5 of the specification.
The potential diagram of the product of example 1 was characterized by Nano-ZS (Malvern) and the results are detailed in FIG. 6 of the specification.
EDX elemental analysis of the product of example 1 was characterized by a JEM-2010 high resolution projection Electron microscope (JEOL) and a light energy Source diffusion Analyzer (Horiba) model EX-250, the results of which are detailed in FIG. 7 of the specification.
The main functional groups of the product of example 1 and the coriolus versicolor proteoglycan were characterized and compared by Equinox55 Fourier transform infrared spectrometer (Bruker), the results are detailed in FIG. 8 of the specification.
Table 1 the average particle size and SD of the nano-selenium particles in the product obtained in the example
| CV(mg/L) | 0 | 50 | 100 | 300 | 600 |
| Size(nm) | Precipitation of | 120.67 | 97.67 | 96.00 | 98.00 |
| SD | -- | 9.80 | 3.06 | 1.00 | 5.29 |
(Note: CV in any of the examples of the present invention, is an abbreviation for coriolus versicolor water-soluble polysaccharide protein)
As can be seen from Table 1 and FIG. 1, when Coriolus versicolor water-soluble polysaccharide protein (CV) is added as a stabilizer, the generated nano-selenium is extremely unstable and finally precipitates; the product obtained by adding the coriolus versicolor water-soluble polysaccharide protein has the particle size of 96-120 nm, which shows that the coriolus versicolor polysaccharide protein has a good regulation and control effect on the particle size of the nano-selenium. In addition, as the concentration of the polysaccharide is increased to 300.0mg/L, the particle size is reduced in a concentration effect, and SD values are small, which shows that the particle size distribution of the nano-selenium is narrow; as can be seen from FIG. 2, the particle size of the product of example 1 was maintained at about 110nm for 13 weeks, which indicates that the product has good stability, and the particle size does not change much within a certain period of time, and no particle aggregation or precipitation occurs; as can be seen from FIGS. 3, 4 and 5, the product of example 1 has good dispersibility, and is spherical nano-selenium; as can be seen from FIG. 6, the absolute value of the potential of the nano-selenium hydrosol of the product of example 1 is-16.1 mV, which indicates that the nano-selenium hydrosol system is relatively stable; as can be seen from fig. 7, the obtained product contains selenium as a main element; as can be seen from fig. 8, the polyhydroxy structure contained in the polysaccharide part of the coriolus versicolor polysaccharide protein can effectively modify the surface of the nano-selenium and regulate the particle size of the nano-selenium.
In summary, the nano selenium hydrosol obtained by reacting the coriolus versicolor polysaccharide protein, the sodium selenite and the vitamin C is particularly characterized in that a polyhydroxy structure of a polysaccharide part in the coriolus versicolor polysaccharide protein is utilized, a strong physical adsorption effect is exerted on the nano selenium, the nano selenium is prevented from being further aggregated and precipitated, meanwhile, the surface of the nano selenium is effectively modified, the particle size of the nano selenium is regulated, and the nano selenium hydrosol is kept stable.
Example 2
(1) Adding 3.0mL of Coriolus versicolor water-soluble polysaccharide protein aqueous solution with mass concentration of 2.5g/L into 4 25mL volumetric flasks filled with 10mL of double distilled water at 20 ℃ under 101.325 kPa;
(2) adding 0.1mL, 0.5mL, 1.0mL and 2.0mL of sodium selenite solution with the concentration of 0.025mol/L into the 25mL volumetric flask respectively, and shaking up lightly to mix thoroughly to obtain a mixed solution;
(3) and (3) respectively dropwise adding 0.1mL, 0.5mL, 1.0mL and 2.0mL of vitamin C (Vc) solution with the concentration of 0.1mol/L into the mixed solution in the step (2), gently shaking while dropwise adding, adding water to a constant volume of 25mL after dropwise adding is finished, standing until the red color is not deepened any more, and then dialyzing (with the molecular weight cutoff of 8000) for 24 hours to obtain a product. The content of selenium is measured by a nitration ICP method, and the concentration of nano selenium in the obtained product is basically 1.0-1.5 mmol/L.
The obtained product is placed in an environment with the temperature of 2-10 ℃, and the product exists in the form of hydrosol.
The product obtained in example 2 was characterized by a detection instrument, and the tests mainly included the following:
the mean particle size and Standard Deviation (SD) of the nanoparticles in the product were determined using a Nanosight NS3000 particle tracking Analyzer (Malvern) and the results are detailed in Table 2.
TABLE 2 average particle size and SD of nano-selenium particles in the product obtained in the example
| Se(IV)(mmol/L) | 0.1 | 0.5 | 1.0 | 2.0 |
| Vc(mmol/L) | 0.4 | 2.0 | 4.0 | 8.0 |
| Size(nm) | 151.33 | 98.00 | 96.00 | 98.00 |
| SD | 21.22 | 4.36 | 1.00 | 6.08 |
As can be seen from Table 2, the nano-selenium hydrosol product obtained by using sodium selenite solution and vitamin C as selenite solution and reducing agent has an average particle size of 96-151 nm. The average grain diameter can be kept below 100nm when the concentration of sodium selenite solution is 0.5 mmol/L-2.0 mmol/L and the concentration of vitamin C solution is 2.0 mmol/L-8.0 mmol/L. In addition, with the increase of the concentration of the sodium selenite solution to 1.0mmol/L and the increase of the concentration of the vitamin C solution to 4.0mmol/L, the particle size is reduced in a concentration effect, and the SD value is smaller, which indicates that the particle size distribution of the nano-selenium is narrower.
Example 3
(1) Adding 3.0mL of coriolus versicolor water-soluble polysaccharide protein aqueous solution with the mass concentration of 2.5g/L into a 25mL volumetric flask filled with 10mL of double distilled water at the temperature of 20 ℃ and under 101.325 kPa;
(2) adding 1.0mL of sodium selenite solution with the concentration of 0.025mol/L into the 25mL volumetric flask, and shaking up gently to mix the solution fully to obtain a mixed solution;
(3) and (3) dropwise adding 1.0mL of vitamin C (Vc for short in English) solution with the concentration of 0.1mol/L into the mixed solution in the step (2), gently shaking while dropwise adding, adding water to a constant volume of 25mL after dropwise adding is finished, standing until the red color is not deepened any more, dialyzing (molecular weight cut-off of 8000) for 24 hours to obtain a product, and measuring the selenium content by a nitration ICP method, wherein the concentration of nano-selenium in the obtained product is about 1.0-1.5 mmol/L.
Performing anti-tumor cell activity test on the product, and detecting the growth of AGS cells in EBV-infected gastric epithelium by using MTS cell proliferation detection method (BMG Labtech, Clariostar) modified by coriolus versicolor polysaccharide protein.
As can be seen from FIG. 9, the Coriolus versicolor polysaccharein-modified nano-selenium can well inhibit EBV-infected gastric epithelial AGS cell proliferation activity, the cell proliferation inhibition rate and the concentration of the Coriolus versicolor polysaccharein-modified nano-selenium show a dose effect, IC50About 5.0. mu.M.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A nano-selenium hydrosol with gastric cancer cell proliferation inhibition activity is characterized in that: the composition consists of the following components in concentration:
0.5 mmol/L-5.0 mmol/L of nano selenium;
100.0 mg/L-600.0 mg/L Coriolus versicolor polysaccharide protein;
the coriolus versicolor polysaccharide protein is coriolus versicolor water-soluble polysaccharide protein, wherein the total sugar content is 70% -80%, and the total protein content is 15% -25%.
2. The nano-selenium hydrosol of claim 1, wherein: the polysaccharide comprises at least one of glucose, mannose and arabinose; the amino acid of the protein comprises at least one of aspartic acid, glutamic acid, threonine, serine, glycine and glutamic-alanine.
3. The method for preparing nano selenium hydrosol with gastric cancer cell proliferation inhibiting activity as claimed in claim 1, comprising at least the following steps:
step S01, adding a solution containing selenium ions and/or selenious ions into a coriolus versicolor polysaccharide protein aqueous solution;
s02, dropwise adding a reducing agent solution into the mixed solution obtained in the step S01, and oscillating;
and S03, carrying out constant volume and dialysis treatment on the product obtained by the reduction reaction in the step S02.
4. The method of claim 3, wherein: in the reduction reaction, the selenium ions and/or the selenite ions are fed according to the molar ratio of (0.7-1.3) to (3.0-9.0) of the reducing agent.
5. The method of claim 3, wherein: the solution containing selenium ions is selenate solution, and the solution containing selenite ions is selenium dioxide and/or selenite solution; and/or the reducing agent is vitamin C.
6. The method of claim 3, wherein: the concentration of the solution containing selenium ions and/or selenite ions is 0.5 mmol/L-2.0 mmol/L; and/or the concentration of the reducing agent solution is 2.0 mmol/L-8.0 mmol/L.
7. The method of claim 3, wherein: in the dialysis product, the concentration of the coriolus versicolor polysaccharide protein is 100.0 mg/L-600.0 mg/L.
8. The method for preserving the nano-selenium hydrosol having gastric cancer cell proliferation inhibiting activity according to claim 1 or 2 or the method for preparing the nano-selenium hydrosol having gastric cancer cell proliferation inhibiting activity according to any one of claims 3 to 7, wherein the method comprises the following steps: the nano-selenium hydrosol is stored at the temperature of 2-10 ℃.
9. The use of the nano-selenium hydrosol having gastric cancer cell proliferation inhibiting activity according to claim 1 or 2 or the nano-selenium hydrosol having gastric cancer cell proliferation inhibiting activity prepared by the method for preparing the nano-selenium hydrosol having gastric cancer cell proliferation inhibiting activity according to any one of claims 3 to 7 in the preparation of a drug for inhibiting gastric cancer cell proliferation.
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| CN110876754B (en) * | 2019-12-02 | 2022-04-15 | 香港理工大学深圳研究院 | Nano-selenium hydrosol and preparation method and application thereof |
| CN110801438A (en) * | 2019-12-07 | 2020-02-18 | 安徽克菱保健科技有限公司 | Nano-selenium effervescent tablet and preparation method thereof |
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