Application of Lactobacillus paracasei IOB413 and metagen thereof in improvement of radioactive hematopoietic injury

Abstract

The invention provides application of lactobacillus paracasei IOB413 and a metagen thereof in preparation of a medicine for improving radioactive hematopoietic injury, and relates to the field of microorganisms. The lactobacillus paracasei IOB413 is preserved in the China General Microbiological Culture Collection Center on June 29, 2018, and the preservation number of the lactobacillus paracasei IOB413 is CGMCC (China General Microbiological Culture Collection Center) No.16022. The lactobacillus paracasei IOB413 has the advantages that the lactobacillus paracasei IOB413 is preserved in the China General Microbiological Culture Collection Center; the lactobacillus paracasei IOB413 and the metaplasm thereof can improve the damage condition of immune organs, increase the number of lymphocytes, increase the number of red blood cells and blood platelets and recover the immune function and hematopoietic function of the organism, so that the radioactive hematopoietic damage is improved.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to application of Lactobacillus paracasei IOB413 and postbiotics thereof in improving radiation hematopoietic damage. Background Art

[0002] Ionizing radiation is a type of energy released by atoms and propagated in the form of electron waves such as gamma and X-rays or particles such as neutrons, c particles, and particles. Ionizing radiation has important applications in the fields of medicine and industry, and is closely related to people's lives. In the field of medicine, ionizing radiation has become an indispensable means of diagnosis and treatment of a large number of clinical diseases. Previous studies have shown that the main source of human radiation exposure is medical radiation sources, among which imaging examinations such as X-rays and CT scans and interventional and radiotherapy treatments are the main sources. Ionizing radiation damage caused by medical radiation and potential radioactive contamination has attracted widespread attention from the international community and has become an urgent problem to be solved. Exposure of the human body to large doses of ionizing radiation in a short period of time can cause acute radiation sickness, causing damage to multiple organs. Depending on the damaged organs, acute radiation sickness can be divided into radiation hematopoietic injury, radiation intestinal injury, etc. After a large dose of irradiation, the hematopoietic parenchymal cells and hematopoietic microenvironment in the hematopoietic organs will be damaged, and the hematopoietic function will be suppressed. Hematopoietic stem cell transplantation is currently the main method for the clinical treatment of radiation hematopoietic injury, but in actual application it is still restricted by factors such as the small source of hematopoietic stem cells. Therefore, it is of great practical significance to develop and reserve hematopoietic system radiation protectors with good efficacy and few toxic side effects.

[0003] The intestinal microecology is closely related to the health of the host. The host's genetic characteristics determine the composition of its intestinal microecology, but the intestinal microecology is also affected by a variety of environmental factors and living habits such as climate, diet, exercise, and work and rest. Based on the biological relationship between the intestinal microecology and the health of the host, more and more studies have confirmed that regulating the intestinal microecology can treat or help treat metabolic diseases. Therefore, maintaining a healthy and stable intestinal microecology is of great significance for resisting the occurrence of diseases and promoting the treatment of diseases. Summary of the invention

[0004] In view of the technical problems existing in the prior art, the present invention aims to provide an application of Lactobacillus paracasei IOB413 and its postbiotics in improving radiation hematopoietic damage.

[0005] One of the objects of the present invention is to provide an application of Lactobacillus paracasei IOB413 in the preparation of a medicine for improving radiation hematopoietic damage. The classification name of Lactobacillus paracasei IOB413 is Lactobacillus paracasei ( Lacticaseibacillus paracasei ), the deposit number is CGMCC No.16022.

[0006] Preferably, the method for preparing the bacterial powder of Lactobacillus paracasei IOB413 comprises: The strain is inoculated into a liquid culture medium at a rate of 1%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain an activated seed solution. The activated seed solution is inoculated into an activated culture medium at a rate of 3%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain a fermentation seed solution. The fermentation seed liquid is inoculated into the fermentation medium at an inoculum rate of 2.5-3.5%, the culture temperature is 34-38°C, and the culture is kept in a closed state for 10-14 hours to obtain a fermentation liquid; The fermentation liquid is vacuum freeze-dried to obtain Lactobacillus paracasei IOB413 bacterial powder.

[0007] Preferably, the liquid culture medium and the activated culture medium use a modified MRS culture medium, the formula of which is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 4.5±0.5g, glucose 20.0±2.0g, Tween 801.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L Preferably, the fermentation medium uses a modified MRS medium, whose formula is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 6±0.5g, glucose 23.0±2.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

[0008] The second object of the present invention is to provide an application of a postbiotic of Lactobacillus paracasei IOB413 in the preparation of a drug for improving radiation hematopoietic damage. The classification name of the Lactobacillus paracasei IOB413 is Lactobacillus paracasei ( Lacticaseibacillus paracasei ), the deposit number is CGMCC No.16022.

[0009] Preferably, the preparation method of the Lactobacillus paracasei IOB413 postbiotics comprises: The strain is inoculated into a liquid culture medium at a rate of 1%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain an activated seed solution. The activated seed solution is inoculated into an activated culture medium at a rate of 3%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain a fermentation seed solution. The fermentation seed liquid is inoculated into the fermentation medium at an inoculum rate of 2.5-3.5%, the culture temperature is 34-38°C, and the culture is kept in a closed state for 10-14 hours to obtain a fermentation liquid; The fermentation liquid is inactivated and dried to obtain the post-fermentation biomass of Lactobacillus paracasei IOB413.

[0010] Preferably, the liquid culture medium and the activated culture medium use a modified MRS culture medium, the formula of which is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 4.5±0.5g, glucose 20.0±2.0g, Tween 801.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L Preferably, the fermentation medium uses a modified MRS medium, whose formula is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 6±0.5g, glucose 23.0±2.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

[0011] Beneficial effects of the present invention: The invention provides an application of Lactobacillus paracasei IOB413 and postbiotics thereof in preparing a medicine for improving radiation hematopoietic damage. Lactobacillus paracasei IOB413 and postbiotics thereof can improve the damage of immune organs, increase the number of lymphocytes, increase the number of red blood cells and platelets, and restore the body's immune function and hematopoietic function, thereby improving radiation hematopoietic damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 is the weight change of mice; Figure 2 is the change of mouse thymus index; Figure 3 is the change in percentage of lymphocytes in mice; Figure 4It is the change in the number of red blood cells in mice; Figure 5 It is the change of platelet count in mice.

[0013] The Lactobacillus paracasei used in the present invention is Lacticaseibacillus paracasei )IOB413, deposited on June 29, 2018 in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.16022. DETAILED DESCRIPTION

[0014] The present application is further described in detail below in conjunction with the accompanying drawings and embodiments. It is to be understood that the specific embodiments described herein are only used to explain the relevant invention, rather than to limit the invention. It is also necessary to explain that, for ease of description, only the parts related to the invention are shown in the accompanying drawings.

[0015] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present application will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0016] Lactobacillus paracasei of the present invention ( Lacticaseibacillus paracasei )IOB413, deposited on June 29, 2018 in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.16022. The Lactobacillus paracasei ( Lacticaseibacillus paracasei )IOB413 has the following biological properties: (1) Lactobacillus paracasei IOB413 was isolated from naturally fermented sourdough in a resident’s home in Tianjin.

[0017] (2) Bacterial identification test: The detection method is FMIC-QO01-001-2015 microbiological test, bacterial multiphase identification test, and the test results are Lactobacillus ( Lacticaseibacillus sp. ).

[0018] (3) Bacterial characteristics: The bacteria are rod-shaped, arranged singly or in pairs, and are Gram-positive.

[0019] (4) Colony characteristics: The colonies are white, round, moist, opaque, and have neat edges.

[0020] (5) Growth characteristics: The minimum growth temperature of the strain is 15°C, the maximum growth temperature is 45°C, the best growth temperature is 35-37°C, the optimal growth pH is 6.0-6.4, and it enters the stable phase after 18 hours of cultivation. The strain was cultured in MRS medium at 36°C for 48 hours.

[0021] (6) Acute oral toxicity test: The test items and basis are the acute oral toxicity test of the national food safety standard GB15193.3-2014, and the test results are actually non-toxic.

[0022] The Lactobacillus paracasei IOB413 and its postbiotics have the following biological characteristics: (1) The results of the thymus index measurement experiment showed that it can significantly improve the impaired immune function.

[0023] (2) The results of the lymphocyte count measurement experiment showed that it can significantly increase the lymphocyte count and restore the body's immune function.

[0024] (3) The results of the lymphocyte count measurement experiment showed that it can significantly increase the number of red blood cells and platelets and restore the body's hematopoietic function.

[0025] Example 1 Preparation method of Lactobacillus paracasei IOB413 bacterial powder and its postbiotics: Inoculate the strain into liquid culture medium at a rate of 1%, culture it at 35-37°C in a closed environment for 30-36 hours to obtain activated seed solution. Inoculate the activated seed solution into activation culture medium at a rate of 3%, culture it at 35-37°C in a closed environment for 30-36 hours to obtain fermentation seed solution.

[0026] The fermentation seed liquid is inoculated into the fermentation medium at an inoculum rate of 2.5-3.5%, the culture temperature is 34-38°C, and the culture is kept in a closed state for 10-14 hours to obtain a fermentation liquid; The fermentation liquid is vacuum-freeze-dried to obtain Lactobacillus paracasei IOB413 bacterial powder; The fermentation liquid is inactivated and dried to obtain the post-fermentation biomass of Lactobacillus paracasei IOB413.

[0027] The liquid culture medium and activated culture medium used were modified MRS culture medium, whose formula was: peptone 10.0 g, beef extract 10.0 g, yeast extract 4.5 g, glucose 20.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H2O 2.0 g, sodium acetate·3H2O 5.0 g, triammonium citrate 1.8 g, magnesium sulfate·7H2O 0.18 g, manganese sulfate·4H2O 0.03 g, pH=6.2±0.2, and the amount of water added was 1 L.

[0028] The fermentation medium used was modified MRS medium, whose formula was: peptone 10.0 g, beef extract 10.0 g, yeast extract 6 g, glucose 23.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H2O 2.0 g, sodium acetate·3H2O 5.0 g, triammonium citrate 1.8 g, magnesium sulfate·7H2O 0.18 g, manganese sulfate·4H2O 0.03 g, pH = 6.2±0.2, and the amount of water added was 1 L.

[0029] Example 2 Lactobacillus paracasei IOB413 powder and postbiotics can improve radiation-induced hematopoietic damage Animal experiments 28 SPF grade, 6-8 week old male C57BL / 6 mice, weighing 20±5g, were used. The relative humidity of the mice was 40-70% and the temperature was 22±2°C.

[0030] Before model construction, all mice were weighed and randomly divided into 4 groups: control group, model group, IOB413 live bacteria group and IOB413 postbiotic group. There were 6 mice in each of the control group and model group, and 8 mice in each of the IOB413 live bacteria group and IOB413 postbiotic group.

[0031] Specific modeling process: After the mice adapted to the new environment for 3 days, the mice in the model group, IOB413 live bacteria group, and IOB413 metabiotic group were irradiated with 5Gy gamma rays to construct the irradiation model. Irradiator model: A Gammacell® 40 Exactor (Atomic Energy of Canada Lim, Chalk River, ON, Canada), dose rate: 0.77 Gy / min.

[0032] Specific administration process: The control group and the model group were gavaged with 0.2 mL of normal saline, and the IOB413 live bacteria group and IOB413 postbiotic group were gavaged with 0.2 mL of Lactobacillus paracasei powder / postbiotic (8×10 7 CFU / 20g (mouse weight). The mice were gavaged once a day from Monday to Friday for three weeks. The experiment lasted for 24 days in total, with a total of 15 administrations.

[0033] Specific sampling process: Samples were collected on the 24th day of the experiment. Blood was collected from the eyeballs of mice and placed in anticoagulant tubes. White blood cells, lymphocytes, etc. in the blood were detected by a blood analyzer. The thymus of the mice was taken, and the thymus index of the mice was obtained by calculating the thymus mass / body weight.

[0034] 1. Effects of γ-rays on the general condition of mice No mice died after whole-body irradiation with 5 Gy γ-rays; Figure 1As shown in the figure, after whole-body irradiation with 5Gy γ rays, the weight of mice gradually decreased, and then gradually recovered on the 7th day; after whole-body irradiation with 5Gy γ rays, the mice also showed mental depression, decreased activity, disordered hair, and reduced eating and defecation on the 4th day.

[0035] 2. Effects of IOB413 powder / postbiotics intervention on the immune function of mice The thymus is a key site for the development of T lymphocytes and is also one of the central immune organs. The thymus is highly sensitive to ionizing radiation, and radiation damage to the thymus can lead to impaired immune function. Therefore, it is particularly important to enhance the protection of immune organs, especially the thymus, in ionizing radiation environments such as radiotherapy. Lymphocytes are a type of white blood cell produced by lymphoid organs and are an important cellular component of the body's immune response function. The lymphocyte percentage refers to the percentage of lymphocytes to the total number of white blood cells.

[0036] After 5Gy whole-body irradiation, the thymus index and lymphocyte count of mice decreased significantly. After 5Gy whole-body irradiation, mice were treated with IOB413 powder / postbiotics. Figure 2 and Figure 3 As shown in the figure, the thymus index and lymphocyte count of mice were increased to varying degrees, indicating that the immune function impairment of mice irradiated with 5Gy γ-rays was improved after intervention with IOB413 bacterial powder / postbiotics.

[0037] 3. Effects of IOB413 powder / postbiotic intervention on hematopoietic function in mice The hematopoietic system refers to the entire system that produces blood in the body, which consists of hematopoietic organs, hematopoietic cells and hematopoietic microenvironment, among which the bone marrow is the main site for hematopoiesis and blood cell formation. The bone marrow hematopoietic system is the most sensitive system to radiation. A large dose of radiation will damage the body's bone marrow hematopoietic system, leading to a decrease in blood cells, etc. Detecting mouse peripheral blood cells is one of the ways to verify radiation damage to the bone marrow hematopoietic system.

[0038] The peripheral blood cells of mice in the control group were normal, and the number of blood cells was within the normal range; Figure 4 and Figure 5 As shown in the figure, after whole-body irradiation with 5Gy γ-rays, the number of peripheral red blood cells and platelets in mice decreased significantly; after whole-body irradiation with 5Gy γ-rays and intervention with IOB413 bacterial powder / postbiotics, the number of red blood cells and platelets in mice increased to varying degrees.

[0039] The above description is only a preferred embodiment of the present application and an explanation of the technical principles used. Those skilled in the art should understand that the scope of the invention involved in the present application is not limited to the technical solution formed by a specific combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the above features are replaced with the technical features with similar functions disclosed in this application (but not limited to) by each other.

Claims

1. An application of Lactobacillus paracasei IOB413 in preparing a medicine for improving radiation hematopoietic damage, characterized in that: The classification of the Lactobacillus paracasei IOB413 is named as Lactobacillus paracasei ( Lacticaseibacillus paracasei ), the deposit number is CGMCC No.16022.

2. the application of Lactobacillus paracasei IOB413 as claimed in claim 1 in preparing a medicine for improving radiation hematopoietic damage, is characterized in that, The preparation method of the bacterial powder of the Lactobacillus paracasei IOB413 comprises: The strain is inoculated into a liquid culture medium at a rate of 1%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain an activated seed solution. The activated seed solution is inoculated into an activated culture medium at a rate of 3%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain a fermentation seed solution. The fermentation seed liquid is inoculated into the fermentation medium at an inoculum rate of 2.5-3.5%, the culture temperature is 34-38°C, and the culture is kept in a closed state for 10-14 hours to obtain a fermentation liquid; The fermentation liquid is vacuum freeze-dried to obtain Lactobacillus paracasei IOB413 bacterial powder.

3. the application of Lactobacillus paracasei IOB413 as claimed in claim 2 in the preparation of a medicine for improving radioactive hematopoietic damage, is characterized in that, The liquid culture medium and the activated culture medium use a modified MRS culture medium, and the formula thereof is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 4.5±0.5g, glucose 20.0±2.0g, Tween 801.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

4. the application of Lactobacillus paracasei IOB413 as claimed in claim 2 in preparing a medicine for improving radioactive hematopoietic damage, is characterized in that, The fermentation medium used is a modified MRS medium, and its formula is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 6±0.5g, glucose 23.0±2.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

5. An application of a Lactobacillus paracasei IOB413 postbiotic in the preparation of a drug for improving radiation hematopoietic damage, characterized in that: The classification of the Lactobacillus paracasei IOB413 is named as Lactobacillus paracasei ( Lacticaseibacillus paracasei ), the deposit number is CGMCC No.16022.

6. The application of the postbiotic of Lactobacillus paracasei IOB413 as claimed in claim 5 in preparing a medicine for improving radiation hematopoietic damage, characterized in that, The preparation method of the Lactobacillus paracasei IOB413 postbiotics comprises: The strain is inoculated into a liquid culture medium at a rate of 1%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain an activated seed solution. The activated seed solution is inoculated into an activated culture medium at a rate of 3%, and cultured in a closed state at 35-37°C for 30-36 hours to obtain a fermentation seed solution. The fermentation seed liquid is inoculated into the fermentation medium at an inoculum rate of 2.5-3.5%, the culture temperature is 34-38°C, and the culture is kept in a closed state for 10-14 hours to obtain a fermentation liquid; The fermentation liquid is inactivated and dried to obtain the post-fermentation biomass of Lactobacillus paracasei IOB413.

7. The application of the postbiotic of Lactobacillus paracasei IOB413 as claimed in claim 6 in preparing a medicine for improving radiation hematopoietic damage, characterized in that, The liquid culture medium and the activated culture medium use a modified MRS culture medium, and the formula thereof is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 4.5±0.5g, glucose 20.0±2.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O 0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

8. The application of the postbiotic of Lactobacillus paracasei IOB413 as claimed in claim 6 in preparing a medicine for improving radiation hematopoietic damage, characterized in that, The fermentation medium used is a modified MRS medium, and its formula is: peptone 10.0±2.0g, beef extract 10.0±2.0g, yeast extract 6±0.5g, glucose 23.0±2.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate·7H2O 2.0±0.5g, sodium acetate·3H2O 5.0±0.5g, triammonium citrate 1.8±0.5g, magnesium sulfate·7H2O0.18g, manganese sulfate·4H2O 0.03g, pH=6.2±0.2, and the amount of water added is 1L.

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