Selenium-carrying system for cabbage mustard seedlings as well as preparation method and application of selenium-carrying system

Abstract

The invention provides a safe and effective selenium supplement, namely a cabbage mustard seedling selenium-carrying system and a preparation method thereof. The invention also provides application of the selenium-loaded system of the cabbage mustard in an anti-tumor immunomodulator, the selenium-loaded system of the cabbage mustard has excellent characteristics of the cabbage mustard and selenium, and the excellent pharmacological activity of the selenium-loaded system is enhanced under the action of the selenium. The compound provided by the invention has good NK92 cell enhancement effect and tumor sensitization effect, has good sensitization effect on anti-tumor immunoregulation of NK92 cells, has small adverse reaction, and provides a new scientific basis for developing a safe and efficient immunomodulator.

Description

Technical Field

[0001] The invention belongs to the technical field of innovative drug research and development, and particularly relates to kale seedling-loaded selenium, i.e. selenium enriched and biotransformed in kale seedlings, a preparation method of the kale seedling-loaded selenium system, and application of the system in anti-tumor immunomodulators. Background Art

[0002] Selenium, as an indispensable trace element for the human body, has shown a wide range of biological effects. Numerous epidemiological, preclinical and clinical intervention studies have confirmed the key role of selenium compounds in cancer prevention and treatment. Selenium in nature exists in two forms: organic selenium and inorganic selenium. Among them, inorganic selenium (such as selenate and selenite) has high toxicity and a narrow safety range, so it is risky to use it directly as a source of selenium supplementation. In contrast, organic selenium (such as selenomethionine, selenocysteine, and selenium yeast) is less toxic and has multiple biological activities. Among organic selenium, selenocysteine ​​(SeCys) is the most important. Organic selenium usually acts as selenoproteins, and the core of the biological function of most selenoproteins is selenocysteine. However, selenocysteine ​​has poor water solubility, which greatly limits the body's intake of organic selenium. How to improve its water solubility and biocompatibility is still a technical problem that needs to be solved urgently.

[0003] With the advancement of nanotechnology, nano-selenium (SeNPs) has shown significant advantages over traditional inorganic and organic selenium due to its low toxicity, high bioavailability, high yield and easy surface modification, and has received widespread attention. China is a country of herbal medicine. Traditional Chinese medicine uses various Chinese herbal medicines to treat diseases and strengthen the body. Inspired by this, we consider using natural plants to enrich selenium and convert it into an organic selenium source. In fact, converting selenium into highly active organic selenium through biotransformation technology is one of the safest and most effective ways to develop selenium products. In this context, it is particularly important to select highly safe selenium sources for biotransformation enrichment.

[0004] Although sodium selenite has high biological activity, its high toxicity also poses a serious safety hazard. Currently, sodium selenite is the commonly used selenium source in selenium-enriched technology. Given its narrow safety window, exploring selenium sources with higher biocompatibility and safety is crucial to the development of selenium-enriched technology.

[0005] The biological function of selenium is mainly achieved by embedding selenocysteine ​​(SeCys) into proteins to form selenoproteins. However, the poor water solubility of selenocysteine ​​has greatly limited its application in the biological field. How to improve its water solubility and biocompatibility is still a technical problem that needs to be solved urgently. Summary of the invention

[0006] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to solve the problems of low safety and poor biocompatibility of existing selenium supplements and biotransformation selenium sources.

[0007] The present application provides a method for preparing a kale seedling selenium-loaded system, comprising the following steps:

[0008] (1) Germinating kale seeds under appropriate temperature, humidity and light to allow them to germinate and grow; after roots grow, adding lentinan nano-selenium to the water used to cultivate kale so that the roots are immersed in the water;

[0009] (2) Harvest on the 5th to 9th day, wash the plants, remove excess Lentinan nano-selenium in the water, and dry the whole kale seedlings in the sun to remove moisture, and grind them into powder using a ball mill; dissolve the powder in water and boil it to dissolve the active ingredients in the water; filter out insoluble matter to obtain a clear aqueous solution; dry the aqueous solution in a high-temperature dry atmosphere to obtain a kale seedling selenium-loaded system.

[0010] Specifically, the water in step (1) is deionized water or ultrapure water.

[0011] Specifically, the kale seedlings are harvested on the 7th day.

[0012] A kale seedling selenium-loaded system prepared by any of the above preparation methods.

[0013] Specifically, the selenium content in the kale seedling selenium-loaded system is 319±34 μg / g, for example, 280 μg / g, 290 μg / g, 300 μg / g, 310 μg / g, 320 μg / g, 330 μg / g, 340 μg / g, 350 μg / g, etc.; wherein the content of selenocystine is 26.5%-38.5%, for example, 27%, 28%, 29%, 30%, 31%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%, 35% , 36%, 37%, 38% and the like; the content of selenomethionine is 9.5%-21.5%, for example, 10%, 11%, 12%, 13%, 14%, 15%, 15.5%, 16%, 17%, 18%, 19%, 20%, 21% and the like; the content of methylselenocysteine ​​is 24.5%-36.5%, for example, 25%, 26%, 27%, 28%, 29%, 29.5%, 30%, 31.5%, 32%, 33%, 34%, 35%, 36% and the like.

[0014] Specifically, the selenium content of the Chinese kale seedling selenium-loaded system is 319 μg / g, wherein the content of selenocysteine ​​is 32.68%, the content of selenomethionine is 15.68%, and the content of methylselenocysteine ​​is 30.63%.

[0015] Specifically, the concentration of the kale seedling selenium-loaded system is 200-400 μg / mL.

[0016] Application of any of the above-mentioned Chinese kale seedling selenium-loaded systems in anti-tumor immunomodulators.

[0017] The improvements of this application bring the following advantages:

[0018] The present invention uses nano-selenium with excellent biocompatibility as a selenium source, combined with the excellent selenium absorption capacity of kale, and innovatively develops a highly safe, highly active oral system obtained through the biotransformation of kale seedlings. This system is rich in selenocysteine ​​(SeCys), which can effectively enhance the recognition ability of NK cells to tumor cells. At the same time, it can also synergize with indole substances in kale to further enhance the immune activity of NK cells and their immune killing effect, and has no toxic side effects on NK cells and normal cells.

[0019] Selenium-enriched Cordyceps developed based on Cordyceps can also enrich selenium, but the selenium content is 120.9±17.92, which is lower than the selenium content of the Chinese kale seedling selenium-carrying system provided by the present invention, which is 319.18±34.22 μg / g. In addition, selenium-enriched Cordyceps is relatively weak in enhancing the ability of immune cells to kill tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is the HPLC-ICP-MS (high performance liquid chromatography-inductively coupled plasma emission spectrometry-mass spectrometry) spectrum of kale water extract, kale seedlings loaded with selenium, and standard samples of selenium in different forms.

[0021] From top to bottom, the first spectrum line represents selenium in Chinese kale seedlings; the second spectrum line represents water extract of Chinese kale seedlings; the third spectrum line represents standard samples of different forms of selenium, and the specific selenium forms represented by each peak are marked in the figure, among which SeCys2 represents selenocysteine, MetSeCys represents methylselenocysteine, Se 4+ represents selenium with a valence of +4, SeMet represents selenomethionine, Se 6+ Represents the selenium element with a valence of +6.

[0022] Figure 2 This is the direct toxicity of selenium-loaded kale seedlings to NK92 cells and A549 cells, as well as the enhancement of NK cell killing of A549 cells.

[0023] A represents the toxicity of different concentrations of Chinese kale water extract and Chinese kale seedling selenium-loaded system to NK92 cells; B represents the toxicity of different concentrations of Chinese kale water extract and Chinese kale seedling selenium-loaded system to A549 cells; C represents the killing of A549 cells by NK92 cells after being treated with different concentrations of Chinese kale water extract and Chinese kale seedling selenium-loaded system.

[0024] Figure 3 It is based on the attached Figure 1 The contents of selenocystine and selenomethionine were compared with those of the selenium-loaded system in Chinese kale seedlings.

[0025] The groups include: selenocystine alone group, selenomethionine alone group, selenocysteine ​​and selenomethionine mixed group and kale seedling selenium-loaded system group, in order to determine the contribution of selenocysteine ​​and selenomethionine in the kale seedling selenium-loaded system.

[0026] Figure 4 It is the change of tumor volume after tumor-bearing mice are divided into different groups and treated separately.

[0027] A, B, C, and D represent the control group (no treatment), NK92 group (1 million NK92 cells were injected into the tail vein of mice every 3 days), kale seedling selenium system group (the mice were gavaged with nano-selenium kale seedlings biotransformed selenium every 2 days, with a concentration of 2 mg / mL), and combined treatment group (1 million NK92 cells were injected into the tail vein of mice every 3 days, and the kale seedling selenium system was gavaged into the mice every 2 days, with a concentration of 2 mg / mL). The horizontal axis represents time in days; the vertical axis represents the volume of the tumor in cubic millimeters (mm 3 ). Each small graph has 7 lines, each representing a mouse.

[0028] Figure 5 yes Figure 4 The weight changes of the mice during the progress of the experiment.

[0029] It includes data from 4 groups, presented with different data points.

[0030] Figure 6 This is the proportion of NK cells in the spleen of mice in different groups after the treatment.

[0031] The four flow cytometry images on the left are from left to right the control group, NK92 group, Chinese kale seedlings selenium system group and combined treatment group. The rightmost bar graph is the statistics of the former, which can more intuitively show the changes in the proportion of NK cells infiltrating the spleen of mice in different treatment groups.

[0032] Figure 7 This is the situation of NK cells killing target cells in the control group mice and the kale seedling bio-fermentation nano-selenium group mice after a period of treatment.

[0033] The two classic flow cytometry images on the left represent the proportion of target cells in mouse peritoneal cells, and the bar graph on the right is a statistical analysis of the former, which allows you to intuitively see the changes in the proportion of target cells in peritoneal cells.

[0034] Figure 8 It is a comparison between the selenium-loaded system of kale seedlings and another selenium-rich substance, selenium-rich Cordyceps.

[0035] A represents the change in selenium content, and B represents the ability of the two to enhance NK cells' killing of A549 cells. DETAILED DESCRIPTION

[0036] The following describes the embodiments of the present invention through specific examples. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0037] Different specific implementation methods may be implemented or applied, and the details in this specification may also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the invention.

[0038] In the acute toxicity assessment, the LD50 of inorganic selenium, organic selenium and nano-selenium are 15 mg / kg, 35 mg / kg and 113 mg / kg respectively. In biological enrichment, nano-selenium has the lowest toxicity and the best safety. It has a larger safety window and can enrich selenium in larger quantities. Even if there are trace amounts of residues, it is safer than inorganic and organic selenium. Therefore, using nano-selenium as a source of selenium enrichment is the best choice.

[0039] Oral administration remains the preferred method for drug delivery. To overcome biological barrier challenges, the development of biomimetic and bio-inspired strategies is gaining increasing attention. Plant-based drug delivery platforms have shown great potential in improving drug bioavailability, optimizing drug distribution, and enhancing therapeutic effects. These systems are not only renewable and sustainable, but also highly biocompatible, non-toxic, and can minimize the risk of adverse reactions.

[0040] Kale belongs to the class Magnoliaceae, the subclass Rosaceae, the superorder Rosaceae, the order Cruciferae, the family Cruciferae, the genus Brassica, and the species Brassica. As a cruciferous plant, its tocopherols, flavonoids, sulforaphane, and indole substances are the main components of its function. Its water extract has a variety of physiological activities, including antiviral, anti-inflammatory, and anti-tumor. The "Dietary Guidelines for Chinese Residents" also emphasizes that cruciferous plants can inhibit cancers induced by various carcinogens; epidemiological surveys show that the rates of gastric cancer and esophageal cancer are reduced in areas where cauliflower, broccoli, and cabbage are often eaten. A 2009 study showed that the accumulated selenium concentration of cruciferous plants is much higher than the selenium concentration in the soil. Selenium will be converted into organic selenium in the plant body to exert its effects, enhancing its health properties.

[0041] Considering the excellent selenium-enriching capacity and excellent physiological activity of kale, kale is the best selenium-enriching carrier. In addition, due to its extremely high biosafety, using kale to convert nano-selenium into plant-derived organic selenium is the best selenium supplementation strategy.

[0042] The present invention proposes an innovative application, namely, the use of a selenium-loaded system of kale seedlings to demonstrate unique value in the field of anti-tumor immune regulation. This application focuses on how water extracts of natural plants containing selenium can effectively enhance the ability of immune cells, especially NK cells, to kill tumor cells. Specifically, the transformation product can significantly enhance the killing effect of NK cells (such as NK92 cells) on lung cancer cells A549, while ensuring that there is no significant damage to the immune cells themselves, thereby achieving precise and safe immune regulation function.

[0043] Example 1

[0044] A method for preparing a kale seedling selenium-loaded system comprises the following steps:

[0045] (1) Germinating kale seeds under appropriate temperature, humidity and light to allow them to germinate and grow; after roots grow, adding lentinan nano-selenium to the water used to cultivate kale, so that the roots are immersed in deionized water.

[0046] (2) Harvest on the 7th day, wash the plants, remove excess Lentinan nano-selenium in the water, and dry the whole kale seedlings in the sun to remove moisture, and grind them into powder using a ball mill. Dissolve the powder in water and boil it to dissolve the active ingredients in the water. Filter to remove insoluble matter to obtain a clear aqueous solution. Dry the aqueous solution in a high-temperature dry atmosphere to obtain a kale seedling selenium-loaded system.

[0047] (3) When used, the powder was dissolved in ultrapure water at a concentration of 20 mg / mL, ultrasonically assisted, and filtered through a 0.22 μm filter membrane in an ultra-clean bench to obtain a sterile extract solution. The extract solution was stored at 4°C and used in various subsequent experiments.

[0048] At present, there is no technology to convert nano-selenium into biogenic selenium using kale seedlings. The present invention innovatively enriches the proven safe lentinan nano-selenium in natural plants, providing a new and safe option for selenium supplementation. By selecting nano-selenium with high biocompatibility and safety as a selenium source additive, kale with high selenium enrichment can be obtained by utilizing the selenium enrichment conversion ability of kale, and at the same time, the ability of selenium to be converted into selenocysteine ​​in the organism can be improved.

[0049] Example 2

[0050] A method for preparing a kale seedling selenium-loaded system comprises the following steps:

[0051] (1) Germinating kale seeds under appropriate temperature, humidity and light to allow them to germinate and grow; after roots grow, adding lentinan nano-selenium to the deionized water used to cultivate kale, so that the roots are immersed in the deionized water.

[0052] (2) Harvest on the 6th day, wash the plants, remove excess Lentinan nano-selenium in the water, and dry the whole kale seedlings in the sun to remove moisture, and grind them into powder using a ball mill. Dissolve the powder in water and boil it to dissolve the active ingredients in the water. Filter to remove insoluble matter to obtain a clear aqueous solution. Dry the aqueous solution in a high-temperature dry atmosphere to obtain a kale seedling selenium-loaded system.

[0053] (3) When used, the powder was dissolved in ultrapure water at a concentration of 19 mg / mL, ultrasonically assisted, and filtered through a 0.20 μm filter membrane in an ultra-clean bench to obtain a sterile extract solution. The extract solution was stored at 4°C and used in various subsequent experiments.

[0054] Example 3

[0055] A method for preparing a kale seedling selenium-loaded system comprises the following steps:

[0056] (1) Germinating kale seeds under appropriate temperature, humidity and light to allow them to germinate and grow; after roots grow, adding lentinan nano-selenium to the water used to cultivate kale, so that the roots are immersed in deionized water.

[0057] (2) Harvest on the 9th day, wash the plants, remove excess Lentinan nano-selenium in the water, and dry the whole kale seedlings in the sun to remove moisture, and grind them into powder using a ball mill. Dissolve the powder in water and boil it to dissolve the active ingredients in the water. Filter to remove insoluble matter to obtain a clear aqueous solution. Dry the aqueous solution in a high-temperature dry atmosphere to obtain a kale seedling selenium-loaded system.

[0058] (3) When used, the powder was dissolved in ultrapure water at a concentration of 21 mg / mL, ultrasonically assisted, and filtered through a 0.24 μm filter membrane in an ultra-clean bench to obtain a sterile extract solution. The extract solution was stored at 4°C and used in various subsequent experiments.

[0059] Example 4

[0060] Selenium content and selenium forms in the selenium-loaded system of Chinese kale seedlings

[0061] Preparation of kale seedling water extract: weigh a certain amount of the kale seedling or kale seedling selenium-loaded system, crush it, add water and then perform ultrasonic extraction to obtain the kale seedling or kale seedling selenium-loaded water extract, and freeze-dry or spray-dry the water extract before storage.

[0062] The selenium content in the kale seedling water extract and the nano kale seedling selenium-loaded system was measured respectively, and it was found that the selenium content in the kale seedling water extract was only 3.710±1.136μg / g, while the selenium content in the kale seedling selenium-loaded system was 319.176±34.216μg / g, which was a huge increase. Figure 1 The results showed that the content of selenocysteine ​​was 32.68%, the content of selenomethionine was 15.68%, and the content of methylselenocysteine ​​was 30.63%.

[0063] Example 5

[0064] The kale water extract and kale seedling selenium-loaded system had no obvious toxicity to NK92 cells and A549 cells, but could enhance the tumor-killing ability of NK92.

[0065] NK92 cells were cultured in α-MEM containing 12.5% ​​fetal bovine serum, 12.5% ​​inactivated horse serum, 200U / mL hIL-2, 0.2mM inositol, 0.1mM β-mercaptoethanol, 0.02mM folic acid, 100U / mL penicillin and 50U / mL streptomycin in a 37°C constant temperature incubator (5% CO2). A549 cells were cultured in DMEM containing 10% fetal bovine serum, 100U / mL penicillin and 50U / mL streptomycin in a 37°C constant temperature incubator. 20mg of kale seedling water extract and kale seedling selenium system were weighed and dissolved in 1mL ultrapure water, and sterile solutions were obtained by filtering through a 0.22μm filter membrane in a clean bench.

[0066] The concentration of NK92 cells was 8×10 4 The cells were inoculated at a density of 100 μg / mL in a 96-well plate, and different concentrations and types of drugs were added at the same time. After the drugs were co-incubated with NK92 cells for 24 hours, 20 μL of CCK8 solution was added to each well, and the cells were incubated in a cell culture incubator for another 4 hours. The absorbance at 450 nm was detected with an ELISA reader, and the cell survival rate was calculated according to the following formula:

[0067] Cell survival rate (% control) = (Ai-Ab) / (A0-Ab)*100%

[0068] Among them, Ai represents the absorbance value of the drug-added group, Ab represents the absorbance value of the blank control group, and A0 represents the absorbance value of the cell group without drug treatment. Figure 2 A shows that different concentrations of Chinese kale water extract and Chinese kale seedling selenium-loaded system have no toxicity to NK92 cells.

[0069] A549 cells in the logarithmic growth phase were taken and 3×10 4The cells were inoculated at a density of 100 μL / mL in a 96-well plate. After culturing in a cell culture incubator for 24 hours to adhere to the wall, different concentrations of Chinese kale seedling water extract and Chinese kale seedling selenium-loaded system were added, with 3 replicate wells in each group and a blank control group. After the cells were incubated with the drug for 24 hours, 20 μL of MTT stock solution (5 mg / mL) was added to each well. After continuing to incubate in the cell culture incubator for 4 hours, the supernatant was discarded, and 150 μL of DMSO was added to each well. After fully dissolving the generated formazan, the absorbance value at 570 nm was detected with an enzyme marker. According to Figure 2 The results of B showed that different concentrations of Chinese kale water extract and Chinese kale seedling selenium-loaded system had no direct killing effect on A549 cells.

[0070] Example 6

[0071] A549 cells in the logarithmic growth phase were taken and 3×10 4 The cells were inoculated at a density of 100 / mL in a 96-well plate. After culturing in a cell culture incubator for 24 hours, different concentrations of Chinese kale seedling water extract and Chinese kale seedling selenium-loaded system were added, with 3 replicates in each group and a blank control group. After the cells were co-incubated with the drug for 24 hours, each well was incubated with 7.5×10 4 NK92 cells were added at a density of 100 μL / mL. After A549 cells and NK92 cells were co-incubated for 24 hours, the supernatant was discarded and gently washed with PBS to remove the NK92 cells. 100 μL of DMEM medium and 20 μL of MTT stock solution (5 mg / mL) were added to each well. After continuing to incubate in the cell culture incubator for 4 hours, the supernatant was discarded and 150 μL of DMSO was added to each well. After fully dissolving the generated formazan, the absorbance value at 570 nm was detected with an enzyme marker, and the cell survival rate was calculated according to the formula. Figure 2 C shows that the kale seedling selenium-loaded system at concentrations of 200 and 400 μg / mL can effectively improve the killing ability of NK92 cells against A549 cells. The addition of NK92 cells alone reduced the survival rate of A549 to 75.76%, but after treatment with the kale seedling selenium-loaded system at concentrations of 200 and 400 μg / mL, the survival rate dropped to 39.05% and 38.98%, respectively. There was no significant change in the survival rate of A549 cells pretreated with kale water extract.

[0072] Example 7

[0073] According to the attached Figure 1 The ratio of selenomethionine to selenocysteine ​​is shown, with Figure 3The experiment compared the control group, single selenium, mixed selenium and nano-selenium kale seedling biotransformed selenium, and found that a single organic selenium could not effectively enhance the NK cell killing of A549, while the mixture of the two seleniums had a certain effect, but it was weaker than the kale seedling selenium system. Therefore, it can be seen that the kale seedling selenium system is not only effective in the organic selenium in it, but kale itself also has a certain immune regulation effect.

[0074] Example 8

[0075] Evaluation of antitumor effects in tumor-bearing mice.

[0076] A549 cells were implanted subcutaneously in 4-6 week old Balb / c-Nude mice and the tumors were grown to a volume of 100 cm. 3 The mice were randomly divided into 4 groups: control group, NK92 group, kale seedling selenium-loaded system group, and combined treatment group. The NK92 group was injected with NK92 cells via tail vein every 3 days at a concentration of 1×10 6 The kale seedlings-selenium-loaded system group was given 2 mg / kg of kale seedlings-selenium-loaded system by gavage every two days; the combination group was given both administration methods at the same time. The experiment lasted for 30 days, during which the weight, tumor volume and other indicators of the mice were measured. After the experiment, the changes in the proportion of NK cells in the spleen of the mice were measured.

[0077] Attached Figure 4 The results showed that the tumor volume of mice was inhibited after the administration of the kale seedling selenium system and NK92 cells, and the combined treatment had the best therapeutic effect. Compared with the untreated group, the kale seedling selenium system treatment could inhibit tumor growth to a certain extent. When evaluated on the 30th day, the volume of the control group increased from 1117.894±239.058mm 3 Reduced to 615.993±182.107mm in the combined treatment group 3 (. In addition, when the kale seedlings selenium system was combined with NK92 cells for treatment, it showed a stronger tumor inhibition effect. Figure 5 The results showed that there was no significant change in body weight between the treatment groups and the control group, which proved the safety of the selenium-loaded kale seedling system.

[0078] Example 9

[0079] The mice were evaluated on day 30, and the spleens were collected to prepare single cell suspensions. The proportion of NK cells in the suspensions was detected by flow cytometry. Figure 6 It shows that the proportion of NK cells in the spleen of mice increased after gavage with nano-selenium kale seedlings biotransformed selenium, indicating that the kale seedlings selenium-loaded system can increase the number of NK cells in the mouse body to fight tumors and has the function of immune regulation.

[0080] Example 10

[0081] The NK cell killing ability of mice treated with the Chinese kale seedling selenium system was enhanced.

[0082] Mouse lymphoma Yac-1 cells were stained with CFSE and then injected into the mouse peritoneal cavity. After 24 hours, the peritoneal cells were removed and detected using flow cytometry. Figure 7 It showed that the NK1.1 cells in mice treated with the selenium-loaded kale seedling system had stronger killing ability against cells and Yac-1 cells, which was reflected in the fact that the proportion of Yac-1 cells remaining in the mouse peritoneal cavity decreased from 5.213±0.726% to only 1.595±0.837%, which further illustrates the immunoregulatory and NK cell activation function of the selenium-loaded kale seedling system.

[0083] Embodiment 11

[0084] The kale seedling selenium-loaded system has a strong ability to enrich selenium and a unique immune regulatory function.

[0085] According to the attached Figure 8 A, selenium-enriched Cordyceps developed based on Cordyceps can also enrich selenium, but the selenium content therein is 120.9±17.92, which is lower than the 319.18±34.22 μg / g of the kale seedling selenium-carrying system provided by the present invention. Figure 8 B shows that selenium-rich Cordyceps does not have the function of immune regulation and cannot enhance the ability of immune cells to kill tumors.

[0086] Nano-selenium kale seedlings biotransform selenium. Nano-selenium kale seedlings biotransform selenium.

[0087] Currently, selenium-based immunomodulators are usually artificially synthesized nanometers or complex materials, and are used for direct anti-tumor or anti-inflammatory purposes. The present invention not only innovatively enriches selenium through natural plants, converts it into organic selenium such as selenocysteine, and uses it for anti-tumor immune regulation, enhancing NK cell killing of tumors, but also has high biosafety.

[0088] The present invention provides a safe and effective selenium supplement: a method for preparing bioconversion nano-selenium. The present invention also provides an application of a kale seedling selenium-loaded system in anti-tumor immunomodulation, which combines the excellent properties of kale and selenium, and enhances its excellent pharmacological activity under the action of selenium. It has good NK92 cell enhancement effect and tumor sensitization effect, has a good sensitization effect on the anti-tumor immunomodulation of NK92 cells, has little adverse reaction, and provides a new scientific basis for the development of safe and efficient immunomodulators.

[0089] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for preparing a selenium-loaded system of kale seedlings, characterized in that: The following steps are involved: (1) Germinating kale seeds under appropriate temperature, humidity and light to allow them to germinate and grow; after roots grow, adding lentinan nano-selenium to the water used to cultivate kale so that the roots are immersed in the water; (2) Harvest on the 6th to 9th day, wash the plants, remove excess Lentinan nano-selenium in the water, and dry the whole kale seedlings in the sun to remove moisture, and grind them into powder using a ball mill; dissolve the powder in water and boil it to dissolve the active ingredients in the water; filter out insoluble matter to obtain a clear aqueous solution; dry the aqueous solution in a high-temperature dry atmosphere to obtain a kale seedling selenium-loaded system.

2. The preparation method according to claim 1, characterized in that: The water described in step (1) is deionized water or ultrapure water.

3. The preparation method according to claim 1, characterized in that: The harvest time of kale seedlings is the 7th day.

4. A kale seedling selenium-loaded system prepared by the preparation method according to any one of claims 1 to 3.

5. The Chinese kale seedling selenium loading system according to claim 4, characterized in that: The selenium content of the kale seedlings is 319±34 μg / g, wherein the content of selenocysteine ​​is 26.5%-38.5%, the content of selenomethionine is 9.5-21.5%, and the content of methylselenocysteine ​​is 24.5%-36.5%.

6. The selenium-loading system for Chinese kale seedlings according to claim 5, characterized in that: The selenium content of the Chinese kale seedling selenium-carrying system is 319 μg / g, wherein the content of selenocysteine ​​is 32.68%, the content of selenomethionine is 15.68%, and the content of methylselenocysteine ​​is 30.63%.

7. The selenium-loading system for Chinese kale seedlings according to claim 4, characterized in that: The concentration of the Chinese kale seedling selenium-carrying system is 200-400 μg / mL.

8. Use of the kale seedling selenium-loaded system according to any one of claims 4 to 7 in anti-tumor immune regulation.

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