Application of bacillus coagulans JA845 in preparation of health care product or food for assisting in lowering blood pressure

Abstract

The invention provides application of bacillus coagulans JA845 in preparation of health-care products or food for assisting in lowering blood pressure, and belongs to the technical field of health-care products. The bacillus coagulans JA845 has unique physiological activity, the bacillus coagulans JA845 is found and proved to have the effect of assisting in lowering blood pressure for the first time, the application blank of the bacillus coagulans in the field of hypertension adjuvant therapy is filled, the action mode of improving the vascular function by taking antioxidation as a core mechanism has uniqueness, and the bacillus coagulans JA845 can be applied to the field of hypertension adjuvant therapy. And a new microbial preparation selection is provided for adjuvant therapy of hypertension.

Description

Technical Field

[0001] This invention relates to the field of health product technology, and in particular to the application of Bacillus coagulans JA845 in the preparation of health products or foods that help lower blood pressure. Background Technology

[0002] Hypertension is a chronic disease characterized by persistently elevated arterial blood pressure. It is the most significant risk factor for cardiovascular and cerebrovascular diseases, leading to serious complications such as stroke, myocardial infarction, heart failure, and chronic kidney disease, posing a substantial threat to human health. With changing lifestyles and an aging population, the incidence of hypertension is increasing year by year. Therefore, developing safe, effective, and clearly defined mechanisms for the prevention and treatment of hypertension has significant clinical and social value.

[0003] Probiotics, as a class of live microorganisms beneficial to host health, have had their effects on regulating gut microbiota balance and improving metabolic diseases widely confirmed. In recent years, the potential of probiotics in the prevention and treatment of cardiovascular diseases has been gradually explored. Some lactobacilli and bifidobacteria have been reported to have certain antihypertensive effects, the mechanisms of which are mostly related to regulating gut microbiota and inhibiting angiotensin-converting enzyme activity. However, the physiological functions of different probiotic strains are highly specific, and the mechanisms of action and efficacy among strains vary significantly; not all probiotics possess antihypertensive activity. Bacillus coagulans is a facultative anaerobic Gram-positive bacterium with resistance to gastric acid and bile. It has a strong colonization ability in the intestine and can produce beneficial metabolites such as lactic acid and antimicrobial peptides, making it widely used in improving intestinal function and enhancing immunity.

[0004] However, there are currently no reports on the use of Bacillus coagulans strains with unique antioxidant activity as adjunctive therapy for hypertension. Summary of the Invention

[0005] The purpose of this invention is to provide the application of Bacillus coagulans JA845 in the preparation of health products or foods that help lower blood pressure. This strain, with its core mechanism of antioxidation, fills the gap in the application of Bacillus coagulans in the field of adjunctive treatment of hypertension.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides the application of Bacillus coagulans JA845 in the preparation of health products that help lower blood pressure, wherein the effective viable count of Bacillus coagulans JA845 is 1×10⁻⁶. 8 -1×10 10 CFU / mL.

[0007] Preferably, the effective viable count of the Bacillus coagulans JA845 is 1×10⁻⁶. 9 CFU / mL.

[0008] Preferably, the dosage form of the health product includes one of tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

[0009] This invention also provides the application of Bacillus coagulans JA845 in the preparation of food products that help lower blood pressure.

[0010] Preferably, the effective viable count of the Bacillus coagulans JA845 is 1×10⁻⁶. 8 -1×10 10 CFU / mL.

[0011] More preferably, the effective viable count of the Bacillus coagulans JA845 is 1×10⁻⁶. 9 CFU / mL.

[0012] Preferably, the dosage form of the food includes one of tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

[0013] The beneficial effects of this invention compared to the prior art are as follows: (1) Filling the technological gap: This invention is the first to discover and confirm that Bacillus coagulans JA845 has the effect of assisting in lowering blood pressure, filling the technological gap in the existing technology that Bacillus coagulans has not been applied to the field of adjunctive treatment of hypertension, providing a new idea and method for adjunctive treatment of hypertension, and providing a new direction for the research on probiotics to assist in lowering blood pressure.

[0014] (2) High safety: Bacillus coagulans JA845 is a probiotic that is tolerable to the human body. It has the advantages of being resistant to gastric acid and bile and having strong colonization ability. There are no obvious adverse reactions after taking it. It is suitable for long-term use, especially for the adjunctive treatment of patients with hypertension and mild symptoms.

[0015] (3) Broad application prospects: It can be used to prepare health products and foods that help lower blood pressure, which can meet the needs of different groups of people and has significant application value and market potential. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1To investigate the effect of Bacillus coagulans JA845 on systolic blood pressure (SBP) in hypertensive rats, the experimental results are as follows: Control represents the control group; Model represents the hypertension model group; JA845 represents the Bacillus coagulans JA845 intervention group. Figure 2 The analysis results of the thoracic aortic pathological sections of hypertensive rats in each experimental group are as follows: Figure 2 In the table, A represents the H&E staining results of aortic sections from the control group observed at the original magnification (×3); B represents the H&E staining results of aortic sections from the control group observed at the original magnification (×10); C represents the H&E staining results of aortic sections from the hypertension model group observed at the original magnification (×3); D represents the H&E staining results of aortic sections from the hypertension model group observed at the original magnification (×10); E represents the H&E staining results of aortic sections from the Bacillus coagulans JA845 intervention group observed at the original magnification (×3); F represents the H&E staining results of aortic sections from the Bacillus coagulans JA845 intervention group observed at the original magnification (×10); G represents the statistical results of aortic media thickness; H represents the statistical results of lumen diameter; and I represents the statistical results of the media thickness / lumen diameter ratio. Figure 3 To investigate the effects of Bacillus coagulans JA845 on adhesion factors in the serum of hypertensive rats, Figure 3 In the diagram, A represents the ICAM-1 level, and B represents the VCAM-1 level. Figure 4 To investigate the effects of Bacillus coagulans JA845 on serum TNF-α inflammatory factors in hypertensive rats; Figure 5 To investigate the effects of Bacillus coagulans JA845 on angiotensin in the serum of hypertensive rats; Figure 6 To investigate the effects of Bacillus coagulans JA845 on oxidative stress in hypertensive rats; Figure 6 In the table, A represents ROS level, B represents MDA level, C represents SOD level, and D represents CAT level. Detailed Implementation

[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0019] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0020] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0021] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0022] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0023] This invention provides the application of Bacillus coagulans JA845 in the preparation of health products that help lower blood pressure, wherein the effective viable count of Bacillus coagulans JA845 is 1×10⁻⁶. 8 -1×10 10 CFU / mL.

[0024] In this invention, the effective viable count of the Bacillus coagulans JA845 is further preferably 1×10⁻⁶. 9 CFU / mL; the preferred dosage form of the health product includes one of the following: tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

[0025] This invention also provides the application of Bacillus coagulans JA845 in the preparation of food products that help lower blood pressure.

[0026] In this invention, the effective viable count of the Bacillus coagulans JA845 is preferably 1×10⁻⁶. 8 -1×10 10 CFU / mL, further preferably 1×10⁻⁶ 9CFU / mL; the preferred dosage form of the food includes tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

[0027] In this invention, the Bacillus coagulans JA845, with its core mechanism of antioxidant activity, can achieve adjunctive treatment of hypertension. Its specific functions and effects are as follows: (a) Reduction of systolic blood pressure (SBP): After oral administration of this strain to a hypertensive model rat, the systolic blood pressure level of the rats was significantly reduced, and the core symptoms of hypertension were improved. (b) Improves arterial vascular pathological characteristics: It can effectively reduce the thickness of arterial vessels in hypertensive model rats, increase the diameter of the vessel lumen, reverse vascular remodeling, and restore normal vascular physiological function; (c) Reduced secretion of adhesion factors: Significantly reduced the levels of intercellular adhesion molecules (ICAM) and vascular cell adhesion molecules (VCAM) in serum, inhibiting vascular endothelial cell damage and inflammatory cell infiltration; (d) Inhibit the inflammatory response: Reduce the level of the inflammatory cytokine tumor necrosis factor-α (TNF-α) in serum, alleviate vascular and systemic inflammation, and reduce the damage of inflammation to blood vessels; (e) Inhibition of angiotensin secretion: By regulating the renin-angiotensin system, the secretion of angiotensin is reduced, thereby decreasing vasoconstriction and thus achieving the effect of lowering blood pressure; (f) Antioxidant damage: As the core mechanism of action, this strain can significantly improve the antioxidant capacity of vascular tissue, scavenge excess reactive oxygen species (ROS), and reduce the damage of oxidative stress to vascular endothelial cells.

[0028] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were all purchased from conventional biochemical reagent stores. All quantitative experiments in the following examples were performed in triplicate, and the results were averaged.

[0029] Solid LB medium: solvent is water, with 10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride, and 15 g / L agar; unless otherwise specified, pH=7.0.

[0030] Liquid LB medium: The only difference between liquid LB medium and solid LB medium is that liquid LB medium does not contain agar.

[0031] Bacillus coagulans ( Bacillus coagulans)JA845 was deposited on April 14, 2020 at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, abbreviated as CGMCC, with the address at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing (Institute of Microbiology, Chinese Academy of Sciences), and the deposit number is: CGMCC No. 19576.

[0032] Example 1 Preparation of Bacillus coagulans JA845 bacterial suspension Bacillus coagulans JA845 was inoculated into liquid LB medium and cultured in a shaker at 50 °C at 180 rpm for 10 h. Then it was centrifuged at 3000 r / min at 4 °C for 10 min, and the supernatant was discarded to obtain a cell precipitate. The cell pellet was prepared into a bacterial suspension with sterile physiological saline. According to its OD600 absorbance and plate counting results, the number of bacteria was adjusted to 1×10 9 CFU / mL to obtain the Bacillus coagulans JA845 bacterial suspension.

[0033] Example 2 Effects of Bacillus coagulans JA845 on blood pressure, arterial pathological characteristics, anti-inflammation, antioxidant, and reduction of adhesion factors in hypertensive rats 2.1 Experimental grouping and treatment Thirty SPF-grade healthy male SD rats (purchased from Changchun Yisi Experimental Animal Technology Co., Ltd., production license number: SCXK (Ji)-2011-0004) were fed adaptively for one week. The temperature of the animal room was 21±2 °C, the relative humidity was 40±10%, and food and water were provided freely. After the adaptive feeding, they were randomly divided into 3 groups, with 10 rats in each group. The specific grouping and treatment are as follows: Control group (Control): Given basal diet (purchased from Changchun Yisi Experimental Animal Technology Co., Ltd.) from week 1 to week 8, with free food and water intake; from week 5 to week 8, gavaged with physiological saline once a day (the single-dose for each rat was 2 mL), the blood pressure was measured weekly, fasted but not water-deprived after the last experiment, and the experimental animals were sacrificed the next day to collect serum and arterial tissues.

[0034] Model group (Model): Induced hypertension by giving NG-nitro-L-arginine methyl ester (L-NAME, purity 98%, purchased from Yuanye, Shanghai, China) at a dose of 40 mg / kg / day in drinking water from week 1 to week 8; from week 5 to week 8, gavaged with physiological saline once a day (the single-dose for each rat was 2 mL), the blood pressure was measured weekly, fasted but not water-deprived after the last experiment, and the experimental animals were sacrificed the next day to collect serum and arterial tissues.

[0035] Bacillus coagulans JA845 intervention group: From week 1 to week 8, NG-nitro-L-arginine methyl ester (L-NAME, purity 98%, purchased from Shanghai Yuanye, China) was administered to drinking water at a dose of 40 mg / kg / day to induce hypertension; from week 5 to week 8, Bacillus coagulans JA845 bacterial suspension prepared in Example 1 was administered by gavage once a day (the single dose per animal was 2 mL), and blood pressure was measured weekly. After the last experiment, the animals were fasted but allowed to drink water. The experimental animals were sacrificed the next day, and blood was collected. The blood samples were placed at 4°C and centrifuged at 3000 r / min for 10 min to separate serum and arterial tissue.

[0036] 2.2 Detection of relevant physiological and biochemical indicators in hypertensive rats 2.2.1 Rat blood pressure detection Systolic blood pressure (SBP) in rats was measured using a non-invasive tail-cuff blood pressure measurement system (SNSEK type, purchased from Shenzhen Medicon Packaging Technology Co., Ltd.). Before the experiment, rats were placed in the measurement chamber for 15 minutes to acclimatize and reduce blood pressure fluctuations caused by stress. To ensure data stability, three replicate measurements were performed on each rat, with at least five valid measurements completed in each group. During measurement, a 37°C heating pad was used to warm the rat's tail to maintain stable systolic and diastolic blood pressure. After the tail cuff was inflated and deflated, the device automatically recorded the blood pressure values. Systolic blood pressure (SBP) in rats in the control group, hypertension model group, and Bacillus coagulans JA845 intervention group was monitored for eight consecutive weeks. Results are as follows: Figure 1 As shown.

[0037] Depend on Figure 1 It was found that the SBP of the control group rats remained stable at a normal level of 115-120 mmHg; the SBP of the model group rats continued to increase over time, reaching about 205 mmHg by week 8, which is consistent with the pathological characteristics of a hypertension model; and after 1×10 9 In the intervention group treated with CFU / mL Bacillus coagulans JA845 via gavage, the upward trend of systolic blood pressure (SBP) was significantly suppressed after week 1, and began to decline continuously after week 4, reaching approximately 130 mmHg by week 8. This difference was statistically significant compared to the model group (P<0.05). These results clearly demonstrate that Bacillus coagulans JA845 can effectively reduce systolic blood pressure in hypertensive model rats, exhibiting a significant antihypertensive effect.

[0038] 2.2.2 Case Analysis of the Aorta Histological observation and photography were performed using an optical microscope (Olympus, Japan). Olympus ImageViewer software was used to measure the lumen and wall thickness of the arterial tissue. The results are as follows: Figure 2 As shown.

[0039] Depend on Figure 2 As shown in A and B, the arterial vascular structure of the rats in the control group was normal, with a medial thickness of approximately 0.10 mm, a lumen diameter of approximately 1.5 mm, and a media / lumen ratio of approximately 7%. Figure 2 As shown in C and D, the arterial vascular structure of the rats in the model group exhibited typical characteristics of hypertensive vascular remodeling: the vascular media thickness significantly increased to 0.16 mm, the lumen diameter decreased to 1.1 mm, and the media / lumen ratio increased to 15%; Figure 2 As shown in Figures E and F, the vascular pathological damage in rats treated with Bacillus coagulans JA845 was significantly improved: the tunica media thickness decreased to approximately 0.10 mm, the lumen diameter recovered to 1.4 mm, and the tunica media / lumen ratio decreased to 8%, with statistically significant differences compared to the model group (P < 0.01). These results confirm that Bacillus coagulans JA845 can effectively improve the arterial vascular pathological characteristics of hypertensive model rats and reverse vascular remodeling.

[0040] 2.2.3 Effect of Bacillus coagulans JA845 on serum adhesion factors in hypertensive rats The levels of intercellular adhesion molecule-1 (ICAM-1, catalog number JL21331) and vascular cell adhesion molecule-1 (VCAM-1, catalog number JL11762) in the serum of rats in each group were detected using an enzyme-linked immunosorbent assay (ELISA) kit (purchased from Shanghai Jianglai Biotechnology Co., Ltd.). The results are as follows: Figure 3 As shown.

[0041] Depend on Figure 3 The results showed that the serum ICAM-1 level in the control group was approximately 600 pg / mL, and the VCAM-1 level was approximately 500 ng / mL. In the model group, serum ICAM-1 levels significantly increased to over 800 pg / mL, and VCAM-1 levels increased to approximately 780 ng / mL, indicating a marked high expression of adhesion factors. After intervention with Bacillus coagulans JA845, serum ICAM-1 levels decreased to approximately 650 pg / mL, and VCAM-1 levels decreased to approximately 580 ng / mL, with statistically significant differences compared to the model group (P < 0.01). These results indicate that Bacillus coagulans JA845 can effectively reduce the secretion of adhesion factors in the serum of hypertensive model rats and alleviate the risk of inflammatory damage to the vascular endothelium.

[0042] 2.2.4 Experiment on the effects of Bacillus coagulans JA845 on hypertension and inflammation The levels of tumor necrosis factor-α (TNF-α) in the serum of rats in each group were detected using an enzyme-linked immunosorbent assay (ELISA) kit (purchased from Shanghai Jianglai Biotechnology Co., Ltd.). The results are as follows: Figure 4 As shown.

[0043] Depend on Figure 4 The results showed that the serum TNF-α level in the control group rats remained stable at around 330 pg / mL; the serum TNF-α level in the model group rats significantly increased to over 530 pg / mL, exhibiting a marked state of inflammatory activation; after intervention with Bacillus coagulans JA845, TNF-α decreased to around 400 pg / mL, with a statistically significant difference compared to the model group (P<0.01). These results indicate that Bacillus coagulans JA845 can effectively reduce the levels of inflammatory factors in hypertensive model rats and alleviate vascular and systemic inflammatory responses.

[0044] 2.2.5 Experiment on the effect of Bacillus coagulans JA845 on angiotensin in hypertension The levels of angiotensin II in the serum of rats in each group were detected using an enzyme-linked immunosorbent assay (ELISA) kit (purchased from Shanghai Jianglai Biotechnology Co., Ltd.) for the detection of angiotensin II (catalog number JL11637). The results are as follows: Figure 5 As shown.

[0045] Depend on Figure 5 The results showed that the serum angiotensin II level in the control group rats remained at around 15 pg / mL; while the serum angiotensin II level in the model group rats significantly increased to over 45 pg / mL, indicating excessive activation of the renin-angiotensin system. After intervention with Bacillus coagulans JA845, angiotensin II levels decreased to around 25 pg / mL, a statistically significant difference compared to the model group (P<0.01). These results indicate that Bacillus coagulans JA845 can effectively inhibit angiotensin secretion and reduce vasoconstriction in hypertensive model rats, thereby assisting in achieving an antihypertensive effect.

[0046] 2.2.6 Effects of Bacillus coagulans JA845 on Oxidative Stress in Hypertensive Rats Oxidative stress markers in the vascular tissues of rats in each group were detected using an enzyme-linked immunosorbent assay (ELISA) kit (purchased from Shanghai Jianglai Biotechnology Co., Ltd.) to detect superoxide dismutase (SOD, catalog number JL22893), malondialdehyde (MDA, catalog number JL53632), reactive oxygen species (ROS, catalog number JLT3037), and catalase (CAT, catalog number JL21028). Results are as follows: Figure 6 As shown.

[0047] Depend on Figure 6It was found that the ROS level in the vascular tissue of the control group rats was about 650 pg / mL and the MDA level was about 0.8 nmol / mL; the ROS level in the vascular tissue of the model group rats increased to over 1000 pg / mL and the MDA level increased to 2.0 nmol / mL, showing significant oxidative damage; after JA845 intervention, the ROS level in the vascular tissue of rats decreased to about 700 pg / mL and the MDA level decreased to 1.2 nmol / mL, and the generation of oxidative products was significantly inhibited (P<0.01).

[0048] In the control group, the SOD activity in the vascular tissue of rats was approximately 65 U / mL, and the CAT activity was approximately 10 μmol / min / mL. In the model group, the SOD activity in the vascular tissue of rats decreased to 40 U / mL, and the CAT activity decreased to 5 μmol / min / mL, indicating a decline in antioxidant capacity. After intervention with JA845, the SOD activity in the vascular tissue of rats recovered to approximately 55 U / mL, and the CAT activity recovered to 9 μmol / min / mL, with a significant increase in antioxidant enzyme activity (P<0.01). These results confirm that Bacillus coagulans JA845 can exert its core hypotensive and vascular protective effects by enhancing antioxidant capacity and reducing oxidative stress damage.

[0049] In summary, Bacillus coagulans JA845 can achieve adjunctive prevention and treatment of hypertension by regulating blood pressure, improving vascular pathology, and inhibiting inflammation and adhesion factor secretion through multiple targets with antioxidant as the core mechanism.

[0050] As can be seen from the above embodiments, the present invention provides the application of Bacillus coagulans JA845 in the preparation of health products or foods that help lower blood pressure. Bacillus coagulans JA845 has unique physiological activity, and the above experiments show that it has a significant effect on helping lower blood pressure at a specific concentration.

[0051] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The application of Bacillus coagulans JA845 in the preparation of health products for assisting in lowering blood pressure, characterized in that, The effective viable count of the Bacillus coagulans JA845 was 1×10⁻⁶. 8 -1×10 10 CFU / mL.

2. The application according to claim 1, characterized in that, The effective viable count of the Bacillus coagulans JA845 was 1×10⁻⁶. 9 CFU / mL.

3. The application according to claim 1, characterized in that, The dosage forms of the health products include one of the following: tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

4. Application of Bacillus coagulans JA845 in the preparation of foods that help lower blood pressure.

5. The application according to claim 4, characterized in that, The effective viable count of the Bacillus coagulans JA845 was 1×10⁻⁶. 8 -1×10 10 CFU / mL.

6. The application according to claim 5, characterized in that, The effective viable count of the Bacillus coagulans JA845 was 1×10⁻⁶. 9 CFU / mL.

7. The application according to claim 4, characterized in that, The dosage form of the food includes one of the following: tablets, capsules, ointments, granules, powders, oral solutions, soft candies, and beverages.

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