CN112602931A

CN112602931A - Application of seedling extract

Info

Publication number: CN112602931A
Application number: CN202011596975.7A
Authority: CN
Inventors: 郝利民; 鲁吉珂; 李雪; 崔燕; 伊娟娟; 朱家庆; 郑志强; 宋景新; 赵昌成; 张华�; 王刚; 严湘军; 余坚勇; 康彩彩
Original assignee: Zhengzhou University; Institute of Quartermaster Engineering Technology Institute of Systems Engineering Academy of Military Sciences
Current assignee: Zhengzhou University; Institute of Quartermaster Engineering Technology Institute of Systems Engineering Academy of Military Sciences
Priority date: 2020-12-28
Filing date: 2020-12-28
Publication date: 2021-04-06

Abstract

The invention relates to the field of food, in particular to application of a seedling extract in preparation of food for resisting radiation damage and preventing radiation damage. The invention discovers that the green seedling extract has obvious protective effect on the hematopoietic system and immune system of the irradiation experimental animals.

Description

Application of seedling extract

Technical Field

The invention relates to the field of food, in particular to application of a seedling extract.

Background

Ionizing radiation is a general term for all radiation capable of causing substance ionization, and is a high-energy physical injury factor seriously harming human health. With the wide application of nuclear energy and nuclear technology in the fields of military, medical treatment, agriculture and the like, ionizing radiation brings harm to the health of human beings while benefiting the human beings. Ionizing radiation induces a large number of active oxygen free radicals in the body to cause oxidative stress, which in turn causes significant damage to the hematopoietic, metabolic, immune, reproductive systems of the body. At present, the intervention on radiation injury becomes a research hotspot, the traditional radiation protective agent mainly takes artificially synthesized chemical substances as main materials, and a new health hidden danger is brought to the body due to great side effects while a certain treatment effect is achieved. The search for a novel natural radioprotectant which is efficient, free from toxic and side effects and can be taken for a long time has become the focus of current research.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide application of a seedling extract.

In order to achieve the purpose of the invention, the technical scheme is as follows:

the invention provides application of a seedling extract in preparation of foods for resisting radiation damage and preventing radiation damage.

Optionally, the radiation damage comprises radiation damage to a cancer patient and radiation damage to a normal person;

preferably, the radiation damage to the cancer patient is radiation damage in radiation therapy of the cancer patient;

more preferably, the radiation damage to a normal person is radiation damage to a person who performs a radiological task.

Optionally, the radiation damage includes damage to the hematopoietic and immune systems.

Optionally, the application includes: the application of the composition in preparing food for increasing the leukocyte level after ionizing radiation, the bone marrow DNA level after ionizing radiation, the antioxidant index level in blood serum after ionizing radiation and the antioxidant index level in spleen after ionizing radiation;

the antioxidant indicators include superoxide dismutase levels and malondialdehyde levels.

Optionally, the young plants are young plants obtained by cultivating gramineous plant seeds; the seed of the Gramineae plant is preferably at least one of seed of Coicis semen, seed of semen Avenae Nudae and seed of Avena Nudae.

Optionally, the green seedlings are obtained by harvesting when the height of the green seedlings is 20-30 cm;

preferably, the growth cycle of the highland barley is 14-16 days, the growth cycle of the coix seed is 17-19 days, and the growth cycle of the hulless oat is 18-20 days.

Optionally, the young seedling extract is obtained by adding the young seedlings into a mixed solvent of an organic solvent and water for extraction;

preferably, the organic solvent is selected from alcohols, more preferably ethanol;

the volume ratio of the organic solvent to the water in the mixed solvent of the organic solvent and the water is 5-7: 3 to 5.

Optionally, 20-40 mL of the mixed solvent is added into each gram of the young seedlings, and preferably 30mL of the mixed solvent is added into each gram of the young seedlings.

Optionally, the extraction temperature is 60-75 ℃, and preferably 70 ℃;

the extraction time is 100-150 minutes, preferably 120 minutes;

preferably, the extraction process is performed with shaking or stirring, and the rotation speed of shaking or stirring is more preferably 120-180 rpm.

Optionally, the extraction further comprises the steps of filtering, concentrating and drying; the concentration is preferably carried out at 45 ℃, and the drying is preferably freeze drying.

The invention has at least the following beneficial effects:

the research of the invention finds that the green seedling extract has obvious protective effect on the hematopoietic system and the immune system of the irradiation experimental animal.

Drawings

FIG. 1 is a graph of the effect of shoot extracts on mouse body weight;

FIG. 2 is a graph of the effect of shoot extracts on leukocytes in peripheral blood of mice after ionizing radiation;

FIG. 3 is a graph of the effect of extract of young plants on red blood cells in peripheral blood of mice after ionizing radiation;

FIG. 4 is a graph of the effect of extract of young plants on platelets in peripheral blood of mice after ionizing radiation;

FIG. 5 is a graph of the effect of extract of young plants on hemoglobin in peripheral blood of mice after ionizing radiation;

FIG. 6 is a graph of the effect of shoot extracts on bone marrow DNA content after ionizing radiation in mice;

FIG. 7 is a graph showing the effect of extract of young plants on mouse serum SOD after ionizing radiation;

FIG. 8 is a graph of the effect of shoot extracts on mouse serum MDA after ionizing radiation;

FIG. 9 is a graph of the effect of extract of young plants on mouse spleen SOD after ionizing radiation;

FIG. 10 is a graph of the effect of shoot extracts on mouse spleen MDA following ionizing radiation.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms also include the plural forms unless the context clearly dictates otherwise, and further, it is understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, devices, components, and/or combinations thereof.

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The natural gramineae plants, such as coix seed, highland barley, hulless oat, etc., contain rich nutrient substances and bioactive components, and are important cereal crops in human diet. The utilization of grain crops is mostly limited to grains, and the research utilization of seedlings is relatively less. The embodiment of the invention finds that the application of the green seedling extract in preparing the food for resisting radiation damage and preventing the radiation damage is provided through keen research. Experiments of the embodiment of the invention show that the dosage of 6Gy⁶⁰The green seedling extract has obvious protection effect on the hematopoietic system and immune system of an irradiation experimental animal and has the capability of resisting oxidative damage after the Kunming mouse is irradiated by Co-gamma rays (the dosage rate is 2 Gy/min).

Optionally, the radiation damage includes radiation damage to cancer patients and radiation damage to normal persons; preferably, the radiation damage to the cancer patient is radiation damage in radiation therapy of the cancer patient; more preferably, the radiation damage to a normal person is radiation damage to a person who is engaged in a radiological task. Radiation damage includes, among other things, damage to the hematopoietic and immune systems. The antioxidant index includes superoxide dismutase level and malondialdehyde level.

Specifically, the application of the embodiment of the present invention includes: the application of the antioxidant in the spleen and the spleen after ionizing radiation is used for preparing food for increasing the level of leucocytes after ionizing radiation, the application of the antioxidant in the spleen and the spleen after ionizing radiation, and the application of the antioxidant in the spleen and the spleen after ionizing radiation.

In the embodiment of the invention, the young seedling is a young seedling obtained by cultivating seeds of gramineous plants. Specifically, the gramineous plant seed is preferably at least one of coix seed, highland barley seed and naked oat seed, but not limited thereto.

In the embodiment of the invention, the green seedlings are obtained by harvesting when the height of the green seedlings is 20-30 cm.

Specifically, before germination, seeds of gramineous plants are disinfected: the body was sterilized with 2% NaClO solution for 10min and rinsed 3 times with distilled water. Sterilizing with 70% ethanol for 30s, and washing with distilled water for 3 times. Then adding warm water (25 ℃), and soaking for 12h at room temperature. The experimental culture conditions are constant temperature (25 ℃ +/-1) day and night, the illumination time is 12h, the illumination intensity is 3500lx, the humidity is 85%, and the seeds need to be kept wet during the seed germination. Selecting green seedlings which are not subjected to jointing, have rich meat quality and are dark green, growing to the height of 20-30 cm, cutting at the position 2.5-3 cm away from the ground, and removing yellow leaves and impurities for subsequent extraction.

Specifically, the growth cycle of the highland barley is 14-16 days, preferably 15 days; the growth cycle of the coix seeds is 17-19 days, preferably 18 days, and the growth cycle of the hulless oat is 18-20 days, preferably 19 days.

In the embodiment of the invention, the green seedling extract is obtained by adding green seedlings into a mixed solvent of an organic solvent and water for extraction. And selecting fresh young seedlings for extraction.

Specifically, the organic solvent used in the extraction is selected from alcohols, and preferably ethanol.

Specifically, the volume ratio of the organic solvent to the water in the mixed solvent of the organic solvent and the water is 5-7: 3 to 5. The extraction efficiency of effective components in the young plants can be improved by adopting the proportion.

Specifically, 20-40 mL of mixed solvent is added to each gram of young seedlings, and preferably 30 mL.

Specifically, the extraction temperature is 60-75 ℃, and preferably 70 ℃; if the extraction temperature is too low, there is a certain adverse effect on the extraction efficiency, and if it is too high, the activity of the effective ingredient may be affected.

Specifically, the extraction time is 100-150 minutes, preferably 120 minutes; preferably, the extraction process is simultaneously vibrated or stirred so as to further improve the extraction efficiency and shorten the extraction time; the rotation speed of the shaking stirring is 120-180 rpm, and preferably 150 rpm.

Specifically, the extraction also comprises the steps of filtering, concentrating and drying; wherein the filtering step is to perform rough filtration on the green seedling alcohol extract by using gauze, and then perform suction filtration to obtain the green seedling extract supernatant. Concentrating the supernatant of the green seedling extracting solution by rotary evaporation at 42-45 ℃; the drying is preferably freeze drying, and the concentrated solution is dried by a vacuum freeze dryer to avoid the damage of high temperature to the effective components.

Detailed Description

In order that those skilled in the art will better understand the technical solutions of the present invention, the present invention will be further described in detail with reference to the following detailed description.

Example 1

1. Culturing: sterilizing Coicis semen seed with 2% NaClO solution for 10min, and washing with distilled water for 3 times. Sterilizing with 70% ethanol for 30s, washing with distilled water for 3 times, adding warm water (25 deg.C), and soaking at room temperature for 12 hr. Placing in a climatic chamber for seed germination, culturing at constant temperature (25 + -1 deg.C) day and night, with illumination time of 12h, illumination intensity of 3500lx and humidity of 85%, and keeping seed moist during seed germination. Selecting green seedlings which are not subjected to jointing, have rich meat quality and are dark green, growing for 18 days until the height is 20-30 cm, cutting the seedlings at a position 2.5-3 cm away from the ground, and removing yellow leaves and impurities to perform experiments.

2. Extraction: taking pearl barley seedlings according to the weight ratio of 1: 30(g/mL) of the extract in 60% ethanol, and shaking in a water bath at 70 ℃ for 120min (150 rpm).

3. And (3) filtering: and (3) coarsely filtering the alcohol extract by using gauze, and performing suction filtration to obtain a supernatant of the green seedling extract.

4. Concentration: and (3) carrying out rotary evaporation and concentration on the supernatant of the green seedling extracting solution at the temperature of 45 ℃.

5. And (3) freeze drying: drying the concentrated solution with vacuum freeze dryer to obtain young seedling extract.

Example 2

1. Culturing: the highland barley seeds are sterilized by 2 percent NaClO solution for 10min and washed by distilled water for 3 times. Sterilizing with 70% ethanol for 30s, washing with distilled water for 3 times, adding warm water (25 deg.C), and soaking at room temperature for 12 hr. Placing in a climatic chamber for seed germination test, wherein the culture condition is constant temperature (25 + -1 deg.C) day and night, the illumination time is 12h, the illumination intensity is 3500lx, the humidity is 85%, and the seed should be kept moist during seed germination. Selecting green seedlings which are not subjected to jointing, have rich meat quality and are dark green, growing for 15 days until the height is 20-30 cm, cutting the seedlings at a position 2.5-3 cm away from the ground, and removing yellow leaves and impurities to perform experiments.

2. Extraction: taking highland barley seedlings according to the proportion of 1: 30(g/mL) of the extract in 60% ethanol, and shaking in a water bath at 70 ℃ for 120min (150 rpm).

Example 3

1. Culturing: hulless oat seed is sterilized with 2% NaClO solution for 10min, and washed with distilled water for 3 times. Sterilizing with 70% ethanol for 30s, washing with distilled water for 3 times, adding warm water (25 deg.C), and soaking at room temperature for 12 hr. Placing in a climatic chamber for seed germination test, wherein the culture condition is constant temperature (25 ℃ + -1) day and night, the illumination time is 12h, the illumination intensity is 3500lx, the humidity is 85%, and the seed is required to be kept moist during seed germination. Selecting green seedlings which are not subjected to jointing, have rich meat quality and are dark green, growing for 19 days until the height is 20-30 cm, cutting the seedlings at a position 2.5-3 cm away from the ground, and removing yellow leaves and impurities to perform experiments.

2. Extraction: taking naked oats seedlings according to the ratio of 1: 30(g/mL) of the extract in 60% ethanol, and shaking in a water bath at 70 ℃ for 120min (150 rpm).

Experimental example 1 Effect of the extract of the young plants on the body weight of mice

1. Drugs and agents

Test drugs: the green seedling extract of example 1;

positive drugs: likejun tablet.

2. Experimental animals: male Kunming mice bred by an animal-grade SPF-grade barrier system are 6-8 weeks old and 18-20 g in weight. Placing in dry and clean plastic cage, feeding with standard experimental animal feed, and feeding with free water.

3. The experimental method comprises the following steps: animals were randomly grouped into groups of 12 animals each. The experiment is provided with a blank control group, an irradiation control group, a positive control group and three high, medium and low dose groups of the green seedling extract.

Blank control (NC): 10mL/kg bw normal saline;

radiation control (IR): 10mL/kg bw normal saline;

positive control group (PC): 12mg/kg bw Likejun tablet;

shoot extract low dose group (CSS-L): 100mg/kg bw;

young shoot extract medium dose group (CSS-M): 200mg/kg bw;

shoot extract high dose group (CSS-H): 400mg/kg bw.

The gavage is continued for 30 days. After 30 days of gavage, the weight change of the mice was recorded. The results of the detection are shown in FIG. 1.

As can be seen from FIG. 1, the weight growth of mice receiving the extract of young plants was normal. I.e., no obesity and no toxic side effects.

Experimental example 2 Effect of extract of young plants on organ index after ionizing radiation in mice

Animal experiments were carried out in the manner of reference example 1 using the extract of young plants of example 1. After 30 days of gastric lavage, radiation treatment is carried out under the irradiation condition⁶⁰Co-gamma rays are irradiated on the whole body at one time, the irradiation dose rate is 2Gy/min, and the total radiation dose of the mouse is 6 Gy. The mice were dislocated and sacrificed 2 days after the irradiation treatment, the heart, liver, spleen and kidney were immediately removed, fat was removed, blood was washed away with sterile ice-cold physiological saline, surface water was blotted with filter paper, and the organ index of the mice was calculated.

Organ index is organ weight/mouse weight x 100%.

The results of the experiment are shown in table 1:

TABLE 1

Group of	Index of heart	Liver index	Spleen index	Renal index
					NC	0.0060±0.0013	0.0460±0.0044	0.0026±0.0006	0.0134±0.0020
IR	0.0056±0.0012	0.0437±0.0034	0.0011±0.0001****	0.0126±0.0012
					PC	0.0061±0.0010	0.0454±0.0038	0.0012±0.0003	0.0136±0.0012
CSS-L	0.0065±0.0013	0.0465±0.0041	0.0013±0.0001^##	0.0134±0.0013
					CSS-M	0.0061±0.0012	0.0438±0.0027	0.0013±0.0002^#	0.0131±0.0010
CSS-H	0.0054±0.0010	0.0436±0.0045	0.0011±0.0001	0.0133±0.0013

Note: p<0.0001 is compared with an NC group,^#P<0.05 was compared to the IR group,^##P<0.01 in comparison with the IR group (n 10).

According to the experimental data in table 1, the effect of the young sprout extract on the organ index of the spleen is remarkable, which indicates that the young sprout extract has a protective effect on the immune system damage caused by ionizing radiation.

Experimental example 3 Effect of extract of young plants on peripheral blood after ionizing radiation in mice

Using the extract of young plants of example 2, an animal experiment was conducted in the same manner as in example 1, and after 30 days of gastric lavage, irradiation was conducted under the irradiation conditions⁶⁰Co-gamma rays are irradiated on the whole body at one time, the irradiation dose rate is 2Gy/min, and the total radiation dose of the mouse is 6 Gy. And (3) taking blood from eyeballs of the mice on the 2 nd day after the radiation treatment, and detecting the quantity of white blood cells, red blood cells and platelets and the content of hemoglobin in peripheral blood. The obtained experimental results are shown in fig. 2 to 5. FIGS. 2 to 5 show the effect of the extract of the young plants on white blood cells, red blood cells, platelets and hemoglobin in peripheral blood of mice after ionizing radiation.

As shown in fig. 2 to 5, the leukocyte content in the peripheral blood of the mice is significantly reduced after the ionizing radiation, and the content of the leukocyte in the peripheral blood of the mice after the ionizing radiation can be significantly increased by the young sprout extract, and the effect is particularly optimal in the medium-dose group.

Experimental example 4 Effect of extract of young plants on the DNA content of bone marrow after ionizing radiation in mice

Using the extract of young plants of example 3, an animal experiment was conducted in the same manner as in example 1, and after 30 days of gastric lavage, irradiation was conducted under the irradiation conditions⁶⁰Co-gamma rays are irradiated on the whole body at one time, the irradiation dose rate is 2Gy/min, and the total radiation dose of the mouse is 6 Gy. Mice were sacrificed by dislocation on day 2 after irradiation, and the left femur was treated with 5mL of 5mmol/L CaCl₂The solution washes out all bone marrow cells in femur, stands at 4 deg.C for 30min, centrifugates at 2500rpm for 10min, removes supernatant, and adds 5mL of 0.2mol/L HClO₄Heating at 90 deg.C for 15min, cooling, centrifuging at 2500rpm for 10min, and collecting supernatant; diluting 3 times, measuring absorbance at 260nm, and calculating bone marrow cell DThe NA content and the light absorption value are the DNA content of the bone marrow cells. The results obtained are shown in FIG. 6, in which<0.0001 is compared with an NC group,^#P<0.05 was compared to the IR group,^##P<0.01 is compared to the IR group,^###P<0.001 vs IR group (n ═ 6).

As shown in fig. 6, the DNA content of mouse bone marrow cells after ionizing radiation was significantly reduced, and the young sprout extract significantly increased the DNA content of mouse bone marrow cells after ionizing radiation, and the effect was best particularly in the medium dose group.

Experimental example 5 Effect of extract of young plants on antioxidant index in serum after ionizing radiation in mice

Using the extract of young plants of example 1, an animal experiment was conducted in the same manner as in example 1, and after 30 days of gastric lavage, irradiation was conducted under the irradiation conditions of⁶⁰Co-gamma rays are irradiated on the whole body at one time, the irradiation dose rate is 2Gy/min, and the total radiation dose of the mouse is 6 Gy. The experimental results of the blood sampling of the eyeballs of the mice on the 2 nd day after the radiation treatment, the blood sampling in a refrigerator at 4 ℃ for 4 hours until the blood coagulation, the low-temperature centrifugation (4000rpm) for 10min, the supernatant liquid being taken and stored at-80 ℃ for later use, and the contents of SOD and MDA in the blood serum of the mice being detected are shown in fig. 7 and 8. FIG. 7 shows the effect of extract of young plants on mouse serum SOD after ionizing radiation, and FIG. 8 shows the effect of extract of young plants on mouse serum MDA after ionizing radiation, wherein<0.001 vs. NC groups<0.0001 compared with the NC group,^##P<0.01 compared to the IR group,^###P<0.001 was compared with the IR group,^####P<0.0001 compared to the IR group (n ═ 7).

As shown in figure 7, the mouse serum SOD content is significantly reduced after ionizing radiation, and the Qingmiao extract can significantly improve the mouse serum SOD content after ionizing radiation, and particularly has the best effect in a medium-dose group. As shown in fig. 8, the mouse serum MDA content is significantly increased after the ionizing radiation, and the young sprout extract can significantly reduce the mouse serum MDA content after the ionizing radiation, and particularly, the effect is best in the medium-dose group.

Experimental example 6 Effect of the extract of the young plants on antioxidant index in spleen after ionizing radiation in mice

Reference experiment using the extract of the young plants of example 2Example 1 an animal experiment was carried out in the manner that after gavage for 30 days, irradiation was carried out under irradiation conditions of⁶⁰Co-gamma rays are irradiated on the whole body at one time, the irradiation dose rate is 2Gy/min, and the total radiation dose of the mouse is 6 Gy. Mice were sacrificed by dislocation on day 2 after irradiation treatment, spleens were immediately removed, fat was removed, blood was washed off with sterile ice-cold physiological saline, surface water was blotted with filter paper, and tissue homogenization was performed. The contents of SOD and MDA in mouse tissues were determined, and the experimental results are shown in FIGS. 9 and 10. FIG. 9 is the effect of extract of young plants on mouse spleen SOD after ionizing radiation, and FIG. 10 is the effect of extract of young plants on mouse spleen MDA after ionizing radiation, wherein<0.001 vs. NC groups<0.001 vs. NC groups<0.0001 compared with the NC group,^#P<0.05 compared to the IR group,^##P<0.01 compared to the IR group,^###P<0.001 was compared with the IR group,^####P<0.0001 compared to the IR group (n ═ 7).

As shown in fig. 9, the spleen SOD content of the mice after the ionizing radiation was significantly reduced, and the blue-green plantlet extract significantly increased the spleen SOD content of the mice after the ionizing radiation, and the effect was most excellent particularly in the medium dose group. As shown in fig. 10, the spleen MDA content of the mice is significantly increased after the ionizing radiation, and the young sprout extract can significantly reduce the spleen MDA content of the mice after the ionizing radiation, and particularly has the best effect in the medium-dose group.

Although the present application has been described with reference to preferred embodiments, it is not intended to limit the scope of the claims, and many possible variations and modifications may be made by one skilled in the art without departing from the spirit of the application.

Claims

1. The application of the extract of the young sprout is characterized in that the application is the application in preparing food for resisting radiation damage and preventing the radiation damage.

2. The use of claim 1, wherein the radiation damage comprises radiation damage to a cancer patient and radiation damage to a normal human;

more preferably, the radiation damage to a normal person is radiation damage to a radiation worker.

3. The use of claim 1 or 2, wherein the radiation damage comprises damage to the hematopoietic system and the immune system.

4. The application according to claim 1 or 2, characterized in that it comprises: the application of the composition in preparing food for increasing the leukocyte level after ionizing radiation, the bone marrow DNA level after ionizing radiation, the antioxidant index level in blood serum after ionizing radiation and the antioxidant index level in spleen after ionizing radiation;

the antioxidant indicators include superoxide dismutase (SOD) level and Malondialdehyde (MDA) level.

5. The use according to claim 1, wherein the young plants are young plants grown from seeds of gramineous plants; the seed of the Gramineae plant is preferably at least one of seed of Coicis semen, seed of semen Avenae Nudae and seed of Avena Nudae.

6. The application of claim 5, wherein the young plants are harvested after being cultivated to a height of 20-30 cm;

7. The use of claim 5, wherein the young sprout extract is obtained by adding the young sprout into a mixed solvent of an organic solvent and water, and extracting;

8. The use according to claim 7, wherein 20-40 mL of the mixed solvent is added per gram of the young plants, preferably 30 mL.

9. Use according to claim 7, characterized in that the temperature of the extraction is 60-75 ℃, preferably 70 ℃;

the extraction time is 100-150 minutes, preferably 120 minutes;

10. The use according to claim 7, wherein the extraction further comprises the steps of filtration, concentration and drying; the concentration is preferably carried out at 45 ℃, and the drying is preferably freeze drying.