Application of Lactobacillus paracasei IOB413 and Its Postbiotics in Improving Radiation-Induced Hematopoietic Injury

Abstract

The present invention provides an application of Lactobacillus paracasei IOB413 and its postbiotics in the preparation of a medicament for improving radiation-induced hematopoietic injury, which relates to the field of microorganisms. Lactobacillus paracasei ( Lacticaseibacillus paracasei ) IOB413 was deposited at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms on June 29, 2018, with the deposit number of CGMCC No. 16022. Lactobacillus paracasei IOB413 and its postbiotics can improve radiation-induced hematopoietic injury by improving the damage of immune organs, increasing the number of lymphocytes, raising the number of red blood cells and platelets, and restoring the immune function and hematopoietic function of the body.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, and specifically relates to the application of Lactobacillus paracasei IOB413 and its postbiotics in improving radiation-induced hematopoietic injury. Background Art

[0002] Ionizing radiation is a form of energy released by atoms and propagated in the form of electron waves such as γ and X-rays or particles such as neutrons, c particles, and particles. Ionizing radiation has important applications in fields such as medicine and industry and is closely related to people's lives. In the medical field, ionizing radiation has become an indispensable means for the 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 treatment methods such as interventional procedures and radiotherapy are the main sources. Ionizing radiation damage caused by medical radiation and potential radioactive contamination has attracted extensive attention from the international community and has become an urgent problem to be solved. Exposure of the human body to a large dose of ionizing radiation in a short period of time can lead to acute radiation sickness, causing damage to multiple organs. According to the different damaged organs, acute radiation sickness can be divided into radiation-induced hematopoietic injury, radiation-induced intestinal injury, etc. After irradiation with a relatively large dose, both hematopoietic parenchymal cells and the hematopoietic microenvironment in the hematopoietic organs will be damaged, and hematopoietic function will be inhibited. Hematopoietic stem cell transplantation is currently the main method for treating radiation-induced hematopoietic injury in clinical practice, but its practical application 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 protectants with good efficacy and low toxicity and side effects.

[0003] The intestinal microecology is closely related to host health. Host genetic characteristics determine its intestinal microecological composition, but the intestinal microecology is simultaneously affected by various environmental factors such as climate, diet, exercise, and work and rest and living habits. Based on the biological relationship between the intestinal microecology and host health, more and more studies have confirmed that regulating the intestinal microecology can treat or contribute to the treatment of 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 the application of Lactobacillus paracasei IOB413 and its postbiotics in improving radiation-induced hematopoietic injury.

[0005] One of the purposes of the present invention is to provide the application of Lactobacillus paracasei IOB413 in the preparation of drugs for improving radiation-induced hematopoietic injury. The taxonomic name of Lactobacillus paracasei IOB413 is Lactobacillus paracasei ( Lacticaseibacillus paracasei ), and the deposit number is CGMCC No. 16022.

[0006] Preferably, the method for preparing the bacterial powder of Lactobacillus paracasei IOB413 includes:

[0007] Inoculate the strain into a liquid medium at an inoculation amount of 1%, and culture it statically and airtightly at 35-37°C for 30-36 hours to obtain an activated seed solution. Then inoculate the activated seed solution into an activation medium at an inoculation amount of 3%, and culture it statically and airtightly at 35-37°C for 30-36 hours to obtain a fermentation seed solution;

[0008] Inoculate the fermentation seed solution into a fermentation medium at an inoculation amount of 2.5-3.5%, and culture it at a temperature of 34-38°C statically and airtightly for 10-14 hours to obtain a fermentation broth;

[0009] Vacuum freeze-dry the fermentation broth to obtain the bacterial powder of Lactobacillus paracasei IOB413.

[0010] Preferably, the liquid medium and the activation medium used are modified MRS medium, and its formula is: peptone 10.0±2.0 g, beef extract 10.0±2.0 g, yeast extract 4.5±0.5 g, glucose 20.0±2.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H 2 O 2.0±0.5 g, sodium acetate·3H 2 O 5.0±0.5 g, ammonium citrate 1.8±0.5 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2±0.2, and the water addition is 1 L

[0011] Preferably, the fermentation medium used is modified MRS medium, and its formula is: peptone 10.0±2.0 g, beef extract 10.0±2.0 g, yeast extract 6±0.5 g, glucose 23.0±2.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H 2 O 2.0±0.5 g, sodium acetate·3H 2 O 5.0±0.5 g, ammonium citrate 1.8±0.5 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2±0.2, and the water addition is 1 L.

[0012] The second object of the present invention is to provide an application of postbiotics of Lactobacillus paracasei IOB413 in the preparation of drugs for improving radiation-induced hematopoietic injury. The classification and naming of Lactobacillus paracasei IOB413 is Lactobacillus paracasei( Lacticaseibacillus paracasei ), and the preservation number is CGMCC No. 16022.

[0013] Preferably, the preparation method of the postbiotic of Lactobacillus paracasei IOB413 includes:

[0014] Inoculate the strain into a liquid medium at an inoculation amount of 1%, and culture it statically and airtightly at 35-37 °C for 30-36 h to obtain an activated seed liquid. Then inoculate the activated seed liquid into an activation medium at an inoculation amount of 3%, and culture it under the conditions of 35-37 °C, statically and airtightly for 30-36 h to obtain a fermentation seed liquid;

[0015] Inoculate the fermentation seed liquid into a fermentation medium at an inoculation amount of 2.5-3.5%, and culture it at a temperature of 34-38 °C, statically and airtightly for 10-14 h to obtain a fermentation broth;

[0016] Inactivate and dry the fermentation broth to obtain the postbiotic fermented from Lactobacillus paracasei IOB413.

[0017] Preferably, the liquid medium and the activation medium used are modified MRS medium, and its formula is: peptone 10.0±2.0 g, beef extract 10.0±2.0 g, yeast extract 4.5±0.5 g, glucose 20.0±2.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H 2 O 2.0±0.5 g, sodium acetate·3H 2 O 5.0±0.5 g, ammonium citrate 1.8±0.5 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2±0.2, and the water addition amount is 1 L

[0018] Preferably, the fermentation medium used is modified MRS medium, and its formula is: peptone 10.0±2.0 g, beef extract 10.0±2.0 g, yeast extract 6±0.5 g, glucose 23.0±2.0 g, Tween 80 1.0 mL, dipotassium hydrogen phosphate·7H 2 O 2.0±0.5 g, sodium acetate·3H 2 O 5.0±0.5 g, ammonium citrate 1.8±0.5 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2±0.2, and the water addition amount is 1 L.

[0019] Advantages of the present invention:

[0020] The present invention provides an application of Lactobacillus paracasei IOB413 and its postbiotics in the preparation of a medicament for improving radiation-induced hematopoietic injury. Lactobacillus paracasei IOB413 and its postbiotics can improve radiation-induced hematopoietic injury by improving the damage of immune organs, increasing the number of lymphocytes, raising the number of red blood cells and platelets, and restoring the immune function and hematopoietic function of the body. Description of the Drawings

[0021] Figure 1 shows the change in body weight of mice;

[0022] Figure 2 shows the change in thymus index of mice;

[0023] Figure 3 shows the change in lymphocyte percentage of mice;

[0024] Figure 4 shows the change in red blood cell count of mice;

[0025] Figure 5 shows the change in platelet count of mice.

[0026] The Lactobacillus paracasei Lacticaseibacillus paracasei ) IOB413 used in the present invention was deposited at the China General Microbiological Culture Collection Center on June 29, 2018. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 16022. Detailed Embodiments

[0027] The following further describes the present application in detail with reference to the drawings and embodiments. It can be understood that the specific embodiments described herein are only used to explain the relevant invention, rather than limiting the invention. Additionally, it should be noted that for the sake of convenience of description, only the parts related to the invention are shown in the drawings.

[0028] It should be noted that, without conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The following will describe the present application in detail with reference to the drawings and embodiments.

[0029] The Lactobacillus paracasei Lacticaseibacillus paracasei ) IOB413 of the present invention was deposited at the China General Microbiological Culture Collection Center on June 29, 2018. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 16022.

[0030] The Lactobacillus paracasei Lacticaseibacillus paracasei ) IOB413 has the following biological characteristics:

[0031] (1)Lactobacillus paracasei IOB413 was isolated from a naturally fermented sourdough at a resident's home in Tianjin.

[0032] (2)Bacterial identification and detection: The detection methods were FMIC-QO01-001-2015 microbiological detection and polyphasic bacterial identification detection. The detection result was Lactobacillus ( Lacticaseibacillus sp. ).

[0033] (3)Cell characteristics: The cells are rod-shaped, arranged singly or in pairs, and Gram-positive.

[0034] (4)Colony characteristics: The colonies are white, round, with a moist surface, opaque, and regular edges.

[0035] (5)Growth characteristics: The minimum growth temperature of this strain is 15°C, the maximum growth temperature is 45°C, and it grows best at 35 - 37°C. The optimal growth pH is 6.0 - 6.4, and it enters the stationary phase after 18 h of cultivation. The strain was cultured in MRS medium at 36°C for 48 h.

[0036] (6)Acute oral toxicity test: The test items and basis were GB15193.3-2014 National Food Safety Standard Acute Oral Toxicity Test. The test result was actually non-toxic level.

[0037] The Lactobacillus paracasei IOB413 and its postbiotics have the following biological characteristics:

[0038] (1)The results of the thymus index determination experiment showed that it could significantly improve the impaired immune function.

[0039] (2)The results of the lymphocyte count determination experiment showed that it could significantly increase the lymphocyte count and restore the body's immune function.

[0040] (3)The results of the lymphocyte count determination experiment showed that it could significantly increase the red blood cell and platelet counts and restore the body's hematopoietic function.

[0041] Example 1

[0042] Preparation method of Lactobacillus paracasei IOB413 bacterial powder and its postbiotics:

[0043] The strain was inoculated into a liquid medium at an inoculation amount of 1%, and cultured at 35 - 37°C under static and airtight conditions for 30 - 36 h to obtain an activated seed solution. The activated seed solution was inoculated into an activated medium at an inoculation amount of 3%, and the culture conditions were 35 - 37°C, static and airtight culture for 30 - 36 h to obtain a fermentation seed solution.

[0044] Inoculate the fermented seed liquid into the fermentation medium at an inoculation amount of 2.5 - 3.5%, culture at a temperature of 34 - 38°C, and statically culture in a sealed environment for 10 - 14 h to obtain the fermentation broth;

[0045] Vacuum freeze-dry the fermentation broth to obtain the powder of Lactobacillus paracasei IOB413;

[0046] Inactivate and dry the fermentation broth to obtain the postbiotics of Lactobacillus paracasei IOB413 after fermentation.

[0047] The liquid medium and the activation medium used are the modified MRS medium, and its formula is: 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·7H 2 O 2.0 g, sodium acetate·3H 2 O 5.0 g, ammonium citrate tribasic 1.8 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2 ± 0.2, and the water addition is 1 L.

[0048] The fermentation medium used is the modified MRS medium, and its formula is: 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·7H 2 O 2.0 g, sodium acetate·3H 2 O 5.0 g, ammonium citrate tribasic 1.8 g, magnesium sulfate·7H 2 O 0.18 g, manganese sulfate·4H 2 O 0.03 g, pH = 6.2 ± 0.2, and the water addition is 1 L.

[0049] Example 2

[0050] The powder of Lactobacillus paracasei IOB413 and the postbiotics can improve radiation-induced hematopoietic injury

[0051] Animal experiment

[0052] Use 28 male C57BL / 6 mice at the SPF level, 6 - 8 weeks old, with a body weight of 20 ± 5 g. The relative humidity of the mouse breeding environment is 40 - 70%, and the temperature is 22 ± 2°C.

[0053] Before constructing the model, all mice were weighed and randomly divided into 4 groups: the control group, the model group, the IOB413 live bacteria group, and the IOB413 postbiotics group. There are 6 mice in each of the control group and the model group, and 8 mice in each of the IOB413 live bacteria group and the IOB413 postbiotics group.

[0054] Specific modeling process: Three days after the mice adapted to the new environment, the mice in the model group, the IOB413 live bacteria group, and the IOB413 postbiotics group were subjected to whole-body irradiation with 5 Gy of γ-rays to construct an irradiation model. Irradiator model: A Gammacell® 40 Exactor (Atomic Energy of Canada Lim, Chalk River, ON, Canada), dose rate: 0.77 Gy / min.

[0055] Specific administration process: The control group and the model group were gavaged with 0.2 mL of normal saline, and the mice in the IOB413 live bacteria group and the IOB413 postbiotics group were gavaged with 0.2 mL of Lactobacillus paracasei powder / postbiotics (8×10 7 CFU / 20 g (mouse weight)). It was gavaged once a day from Monday to Friday for three consecutive weeks. The total experimental duration was 24 days, and the administration was carried out 15 times in total.

[0056] Specific sampling process: Sampling was carried out on the 24th day of the experiment. Whole blood of the mice was taken by eye bleeding into an anticoagulation tube, and 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 calculated by thymus weight / body weight.

[0057] 1. Effects of γ-rays on the general condition of mice

[0058] No mice died after whole-body irradiation with 5 Gy of γ-rays; as Figure 1 shown, the body weight of the mice gradually decreased after whole-body irradiation with 5 Gy of γ-rays and then gradually recovered on the 7th day; after whole-body irradiation with 5 Gy of γ-rays, on the 4th day, the mice also showed listlessness, reduced activity, disordered hair, and reduced food intake and defecation.

[0059] 2. Effects of IOB413 powder / postbiotics intervention on the immune function of mice

[0060] 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 thymic radiation damage can lead to impaired immune function of the body. 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 cells produced by lymphoid organs and are important cellular components of the body's immune response function. The lymphocyte percentage refers to the percentage of lymphocyte number in the total number of white blood cells.

[0061] After whole-body irradiation with 5 Gy of γ-rays, the thymus index and lymphocyte count of the mice decreased significantly; after whole-body irradiation of mice with 5 Gy of γ-rays and intervention with IOB413 powder / postbiotics, as Figure 2 and Figure 3As shown, the thymus index and the number of lymphocytes in mice were increased to varying degrees. This indicates that the impaired immune function of mice irradiated with 5 Gy γ-rays was improved after intervention with IOB413 bacterial powder / postbiotics.

[0062] 3. Effects of IOB413 bacterial powder / postbiotics intervention on the hematopoietic function of mice

[0063] The hematopoietic system refers to the entire system in the body that manufactures blood, consisting of hematopoietic organs, hematopoietic cells, and the hematopoietic microenvironment. Among them, 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 relatively large dose of radiation will damage the body's bone marrow hematopoietic system, resulting in a decrease in blood cells, etc. Detecting the peripheral blood cells of mice is one of the ways to verify the radioactive damage of the bone marrow hematopoietic system.

[0064] The peripheral blood cells of the control group mice were normal, and the number of blood cells was within the normal range; as Figure 4 and Figure 5 shown, after whole-body irradiation with 5 Gy γ-rays, the numbers of red blood cells and platelets in the peripheral blood of mice decreased significantly; in mice irradiated with 5 Gy γ-rays and intervened with IOB413 bacterial powder / postbiotics, the numbers of red blood cells and platelets in mice were increased to varying degrees.

[0065] The above description is only the preferred embodiment of the present application and the explanation of the applied technical principles. Those skilled in the art should understand that the scope of the invention involved in the present application is not limited to the technical solutions formed by the specific combination of the above technical features, and 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 technical solutions formed by mutually replacing the above features with the (but not limited to) technical features with similar functions disclosed in the present application.

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 preparing 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; 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.

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 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.

7. 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 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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