Heart rate reducing composition

Abstract

[Problem] To provide a composition for lowering the heart rate, the composition including lactic acid bacteria belonging to the genus Lactobacillus. [Solution] The heart rate in a subject is lowered by causing the subject to ingest a composition that includes lactic acid bacteria belonging to the genus Lactobacillus.

Description

REFERENCE TO RELATED APPLICATIONS

[0001] This patent application claims priority to Japanese Patent Application No. 2019-225685, filed on December 13, 2019, the entire disclosure of which is incorporated herein by reference. [Technical Field]

[0002] The present invention relates to compositions for reducing heart rate. [Background technology]

[0003] Resting heart rate (pulse rate) is known to sensitively reflect physical and mental stress. It has also been reported that higher resting heart rates tend to be associated with higher mortality rates from cardiovascular and coronary diseases, as well as higher overall mortality rates (e.g., Non-Patent Document 1). It is also known that while the resting heart rate of a healthy individual is approximately 80 beats per minute (bpm), a heart rate exceeding 80 bpm increases cardiovascular complications, morbidity, and mortality, and that a heart rate of 80-85 bpm or higher should be considered the criterion for tachyarrhythmia (tachycardia). Therefore, it is appropriate to set the criterion for tachycardia at 80 bpm or higher. In such subjects, lowering the resting heart rate may potentially reduce the risk of cardiovascular and coronary diseases and may also reduce mortality.

[0004] It is also known that heart rate is closely related to blood pressure, and that an increase in heart rate increases the risk of developing diseases such as arteriosclerosis and hypertension complications (e.g., myocardial infarction, cerebral infarction, etc.).

[0005] Incidentally, it has been known that certain lactic acid bacteria affect various physiological functions, for example, they are known to improve sugar and lipid metabolism in mice (e.g., Patent Document 1, Non-Patent Documents 2 and 3).

[0006] However, nothing has been known about the ability of lactic acid bacteria intake to lower the heart rate of subjects. A high resting heart rate increases the risk of cardiovascular disease and coronary disease, and can result in a high overall mortality rate, so there is a need for a method to effectively lower the heart rate. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2012 / 014971 [Non-patent literature]

[0008] [Non-Patent Document 1] HEART's Selection (Thinking about Heart Rate) "Learning from Epidemiology: Heart Rate and Cardiovascular Disease", Vol. 43, No. 11, 2011, pp. 1397-1401 [Non-patent document 2] T. Toshimitsu et al. “Identification of Lactobacillus plantarum strain that meliorates chronic inflammation and metabolic disorders in obese and type 2 diabetic mice” J Dairy Sci, 99:933-946, 2016 [Non-patent document 3] Tohru Sakai et al. “Lactobacillus plantarum OLL2712 regulates glucose metabolism in C57BL / 6 mice fed a high-fat diet” J Nutr Sci Vitaminol, 59, 144-147, 2013

[0009] An object of the present invention is to provide a composition for effectively reducing heart rate in a subject.

[0010] As a result of extensive research, the present inventors have found that when lactic acid bacteria belonging to the genus Lactobacillus are ingested by a subject, the heart rate of the subject can be reduced. The present invention is based on these findings.

[0011] According to the present invention, the following inventions are provided. [1] A composition for lowering heart rate, comprising lactic acid bacteria belonging to the genus Lactobacillus. [2] The composition described in [1], which is for human use. [3] The composition described in [1] or [2], wherein the lactic acid bacteria are lactic acid bacteria having the activity of reducing the amount of interleukin-6 in the blood. [4] The composition according to any one of [1] to [3], wherein the lactic acid bacterium is capable of inducing the production of interleukin-10 in mammalian cells. [5] The composition described in [4], wherein the mammalian cells are selected from bone marrow-derived dendritic cells and peritoneal macrophages. [6] The composition according to any one of [1] to [5], wherein the lactic acid bacterium has a 16S rRNA gene that has 90% or more homology with the base sequence represented by SEQ ID NO: 1. [7] The composition according to any one of [1] to [6], wherein the lactic acid bacterium is Lactobacillus plantarum. [8] The composition described in any one of [1] to [7], wherein the lactic acid bacterium is Lactobacillus plantarum OLL2712 strain deposited under accession number FERM BP-11262. [9] The composition according to any one of [1] to [8], wherein the lactic acid bacteria comprise heat-killed lactic acid bacteria.

[10] The composition according to any one of [1] to [9], wherein the lactic acid bacteria comprise heat-killed lactic acid bacteria.

[11] The composition according to any one of [1] to

[10] , wherein the composition is a food composition.

[12] The composition according to any one of [1] to

[10] , wherein the composition is a pharmaceutical composition.

[13] A method for lowering the heart rate of a subject, comprising having the subject ingest a composition containing lactic acid bacteria belonging to the genus Lactobacillus.

[14] Use of lactic acid bacteria belonging to the genus Lactobacillus for lowering heart rate or for producing a composition for lowering heart rate.

[15] Lactobacillus genus lactic acid bacteria for lowering heart rate.

[0012] According to the present invention, the heart rate can be effectively reduced in subjects, including humans. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 is a graph showing the correlation between blood IL-6 levels and heart rate in all subjects in the test and control groups before the start of intake of each composition (week 0). Specific Description of the Invention

[0014] Heart rate reducing composition According to one embodiment of the present invention, there is provided a composition for reducing the heart rate of a subject, comprising lactic acid bacteria belonging to the genus Lactobacillus (hereinafter also simply referred to as "lactic acid bacteria").

[0015] According to one embodiment, the subject to which the composition of the present invention is applied is not particularly limited as long as the effects of the present invention are achieved, and may be, for example, mammals such as humans, monkeys, chimpanzees, cattle, horses, sheep, and goats, preferably humans. The subject may also be a subject with a normal heart rate, or a subject with a heart rate higher than the normal heart rate (tachyarrhythmia) (i.e., a subject with tachycardia).

[0016] As used herein, "normal heart rate" refers to the resting heart rate of a subject who is not suffering from any disease (e.g., respiratory disease, cardiac disease, mental disease, fever, etc.) and has a low risk of death, and may vary depending on the subject's species, age, sex, etc. On the other hand, if the subject's heart rate is higher than the normal heart rate range, it can be said to be tachycardia. For example, the normal heart rate in humans is 50 to 80 beats per minute, and if it exceeds this range, it can be said to be tachycardia.

[0017] As used herein, "reducing" a subject's heart rate means that when lactic acid bacteria or a composition containing lactic acid bacteria is ingested or administered to a subject, the heart rate of the subject receiving the composition is reduced statistically significantly (i.e., beyond the margin of error) compared to a control subject not receiving the composition, or the increase in heart rate in a subject with a tendency toward elevated heart rate is suppressed and maintained at a statistically significantly lower level. Statistical analysis of heart rate can be performed using statistical analysis methods known to those skilled in the art. Comparisons between groups can be performed using the Mann-Whitney U test, and comparisons within the same group (e.g., comparisons between before and after intake of the composition within the same group) can be performed using the Wilcoxon signed-rank test. The presence or absence of a heart rate-reducing effect can be specifically determined using the methods described in the Examples.

[0018] The compositions of the present invention exhibit a significant heart rate-lowering effect in subjects with tachycardia (e.g., hypertension or tachyarrhythmia) and subjects with an elevated heart rate, lowering the heart rate by, for example, 0.5 to 35%, preferably 0.5 to 20%, more preferably 0.5 to 15%, even more preferably 1 to 10%, and even more preferably 1 to 5% compared to the heart rate observed without the composition of the present invention. The compositions of the present invention also exhibit a heart rate-lowering effect in subjects with normal heart rates, but this is a gentle effect, rather than a sudden decrease or an abnormal decrease such as bradycardia, lowering the heart rate by, for example, 0.5 to 12%, preferably 1 to 10%, and more preferably 1 to 4%. Because the compositions of the present invention can lower or maintain a subject's heart rate within a normal range, they can be used to improve symptoms caused by cardiovascular diseases (e.g., heart failure, myocardial infarction, angina pectoris, arteriosclerosis, etc.), coronary artery disease, and hypertensive complications (hypertensive cardiac hypertrophy, congestive heart failure, cerebral hemorrhage, cerebral infarction, hypertensive nephropathy), etc. According to a preferred embodiment of the present invention, the improvement of symptoms caused by cardiovascular disease comprises suppressing an increase in heart rate or decreasing the heart rate. In this specification, "improvement" encompasses treatment or prevention. Furthermore, "treatment" includes not only stopping, alleviating, or delaying the progression or worsening of an abnormality or disease through medical intervention, but also stopping, alleviating, or delaying the progression or worsening of an abnormality or disease through non-medical intervention. Furthermore, "prevention" includes preparing in advance for anticipated worsening of an abnormality or disease, and preventing the occurrence or recurrence of an abnormality or disease through non-medical intervention or medical intervention.

[0019] According to one embodiment of the present invention, the lactic acid bacteria belonging to the genus Lactobacillus contained in the composition of the present invention are preferably lactic acid bacteria having the activity of reducing the blood level of interleukin-6 (IL-6).It is a surprising fact for those skilled in the art that lactic acid bacteria having the activity of reducing the blood level of IL-6 can be advantageously used to lower the heart rate.

[0020] As used herein, "reducing" the amount of IL-6 means that, when lactic acid bacteria or a composition containing lactic acid bacteria are ingested or administered to a subject, the amount of IL-6 in the subject receiving the composition is statistically significantly (i.e., beyond the margin of error) reduced compared to a subject not receiving the composition, or the increase in IL-6 in a subject whose IL-6 level tends to increase is suppressed and maintained at a statistically significantly lower level, or a statistically significant increase in IL-6 is suppressed. Therefore, in one preferred embodiment, "the activity of reducing blood levels of IL-6" refers to "the activity of suppressing an increase in blood levels of IL-6." Statistical analysis of blood levels of IL-6 can be performed using statistical analysis methods known to those skilled in the art. Comparisons between groups can be performed using the Mann-Whitney U test, and comparisons within the same group (e.g., comparisons between before and after intake of the composition within the same group) can be performed using the Wilcoxon signed-rank test. The presence or absence of the activity of reducing blood levels of IL-6 can be determined specifically by the methods described in the Examples.

[0021] According to one embodiment of the present invention, the lactic acid bacteria belonging to the genus Lactobacillus are preferably lactic acid bacteria that can induce the production of interleukin-10 (IL-10) in mammalian cells. The use of lactic acid bacteria that can induce the production of IL-10 is advantageous in reducing the heart rate and suppressing inflammation in mammals that ingest them. The mammalian cells are preferably selected from bone marrow-derived dendritic cells and peritoneal macrophages. The IL-10 production-inducing activity of lactic acid bacteria belonging to the genus Lactobacillus can be confirmed by the method described in the Examples.

[0022] The lactic acid bacteria contained in the composition of the present invention are not particularly limited as long as they belong to the genus Lactobacillus. Examples of lactic acid bacteria belonging to the genus Lactobacillus include Lactobacillus delbrueckii subsp. burgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus amylovorus, Lactobacillus gallinarum, Lactobacillus gasseri, and Lactobacillus oris. oris, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus fermentum, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus paracollinoides, Lactobacillus hammesii, etc. As the lactic acid bacterium, Lactobacillus plantarum is preferably used, and more preferably the Lactobacillus plantarum OLL2712 strain is used.

[0023] The Lactobacillus plantarum OLL2712 strain was deposited at the International Patent Organism Depositary of the National Institute of Advanced Industrial Science and Technology (6-1 Central, Higashi 1-chome, Tsukuba City, Ibaraki Prefecture, Japan) on July 2, 2010, and was subsequently transferred to international deposition and assigned the accession number FERM BP-11262. As stated in Budapest Notification No. 282 (http: / / www.wipo.int / treaties / en / notifications / budapest / treaty_budapest_282.html), the National Institute of Technology and Evaluation (IPOD, NITE) has taken over the patent microorganism deposit business from the National Institute of Advanced Industrial Science and Technology (AIST), and Lactobacillus plantarum OLL2712 strain is currently deposited at the National Institute of Technology and Evaluation (IPOD, NITE) (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) under the accession number FERM BP-11262.

[0024] The lactic acid bacteria contained in the composition of the present invention can also be strains substantially equivalent to the deposited strain. Substantially equivalent strains refer, for example, to strains of lactic acid bacteria belonging to the genus Lactobacillus, whose 16S rRNA gene nucleotide sequence has 90% or more, preferably 95% or more, more preferably 98% or more, and even more preferably 99% or more homology with the 16S rRNA gene nucleotide sequence (SEQ ID NO: 1) of the deposited strain, and preferably have the same bacteriological properties as the deposited strain. A strain having the same bacteriological properties is preferably a strain that has the same level of activity as the deposited strain in terms of reducing the blood level of IL-6 in a subject (e.g., human). Furthermore, a strain having the same bacteriological properties is preferably a strain that has the same level of activity as the deposited strain in terms of inducing IL-10 production in mammalian cells, more preferably in mammalian bone marrow-derived dendritic cells or peritoneal macrophages. Furthermore, the lactic acid bacteria contained in the composition of the present invention may be a strain bred from a deposited strain or a strain substantially equivalent thereto by mutation treatment, genetic recombination, selection of natural mutant strains, etc., as long as the effects of the present invention are achieved.

[0025] The lactic acid bacteria contained in the composition of the present invention include, for example, cultures containing lactic acid bacteria cells, as well as the lactic acid bacteria cells themselves. While either live or killed lactic acid bacteria cells can be used as the lactic acid bacteria cells, killed cells are preferred, and killed cells obtained by heat-treating live cells (heat-killed cells) are more preferred. That is, the lactic acid bacteria contained in the composition of the present invention preferably include killed cells, and more preferably heat-killed cells. The number of passages of the lactic acid bacteria is not particularly limited as long as the effects of the present invention are achieved, but is, for example, 1 to 30, preferably 5 to 15, and more preferably 11 to 13.

[0026] The conditions for culturing lactic acid bacteria are not particularly limited as long as the effects of the present invention are achieved, and can be the conditions typically used for culturing lactic acid bacteria. For example, a medium can be prepared by dissolving whey powder or whey protein concentrate in sterilized water, digesting the mixture with protease A, adding yeast extract, fish extract, and MnSO4, and then adding various nutrients (vitamins, minerals, fatty acid esters), adjusting the pH to 6.7 with NaOH, and then sterilizing the mixture in an autoclave. The pH during culture can be adjusted to 4.8 to 6.8. K2CO3 can be used to adjust the pH. The temperature during culture can be 29 to 40°C.

[0027] The heat treatment for obtaining heat-killed bacteria is not particularly limited as long as the effects of the present invention are achieved, and the heat treatment is carried out under conditions that are normally used to sterilize lactic acid bacteria.

[0028] In addition to the above-mentioned heat treatment, lactic acid bacteria that have been subjected to treatments such as concentration, dilution, freezing, drying, and powdering can also be used.

[0029] The lactic acid bacteria contained in the composition of the present invention may be those prepared by the above-mentioned culture or various treatments, or commercially available compositions containing lactic acid bacteria may be used.

[0030] In the composition of the present invention, the number of lactic acid bacteria per mass of the composition is not particularly limited as long as the effects of the present invention are exhibited. 6 ~10 14 pieces / g, more preferably 10 7 ~10 13 pieces / g, more preferably 10 8 ~10 12 pieces / g, particularly preferably 10 8 ~10 10 In the composition of the present invention, the dry mass of bacterial cells per mass of solid content in the composition is preferably 0.01 to 100% by mass, more preferably 1 to 80% by mass, and even more preferably 10 to 40% by mass.

[0031] The composition of the present invention may contain components other than lactic acid bacteria, as long as they do not impair the effects of the present invention. Examples of components other than lactic acid bacteria include medium components, additives suitable for oral tube administration, solvents such as water, carbohydrates, proteins, lipids, vitamins, minerals, biologically essential trace metals (manganese sulfate, zinc sulfate, magnesium chloride, potassium carbonate, etc.), flavorings, hygienically or pharmaceutically acceptable carriers, food additives, etc.

[0032] Examples of carbohydrates include sugars, modified starch (dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like.

[0033] Examples of proteins include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, α-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, egg protein, meat protein and other animal and plant proteins, their hydrolysates, butter, milk minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, lactose and other milk-derived components.

[0034] Examples of lipids include animal fats and oils such as lard, fish oil, and fractionated, hydrogenated, and interesterified oils thereof; and vegetable fats and oils such as palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated, hydrogenated, and interesterified oils thereof.

[0035] Examples of vitamins include vitamin A, carotenes, B vitamins, vitamin C, D vitamins, vitamin E, K vitamins, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid.

[0036] Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium.

[0037] The composition of the present invention can be produced by blending, in addition to lactic acid bacteria belonging to the genus Lactobacillus, pharmaceutically acceptable carriers and / or additives, food hygienically acceptable carriers and / or additives, etc. Thus, according to another aspect of the present invention, there is provided a method for producing a composition for lowering heart rate, which comprises blending lactic acid bacteria belonging to the genus Lactobacillus.

[0038] According to one embodiment of the present invention, the heart rate-lowering composition of the present invention can be provided as a food composition. The food composition of the present invention may be used for promoting a decrease in heart rate, suppressing an increase in heart rate, maintaining a normal heart rate, or for preventing and / or treating diseases such as cardiovascular diseases (e.g., heart failure, myocardial infarction, angina pectoris, arteriosclerosis, etc.), coronary diseases, and hypertensive complications (hypertensive cardiac hypertrophy, congestive heart failure, cerebral hemorrhage, cerebral infarction, hypertensive nephropathy).

[0039] The food composition of the present invention may be in any form as long as it contains lactic acid bacteria, such as a solution, suspension, emulsion, powder, paste, semi-solid product, or solid product that can be taken orally or through a tube. Specific examples of foods include milk, milk drinks, soft drinks, fermented milk, lactic acid bacteria drinks, dairy drinks, yogurt, cheese, ice cream, frozen desserts, chocolate, tablets, gummies, candy, bread, biscuits, crackers, pizza crust, infant formula, liquid milk, liquid diet, food for the sick, nutritional foods, frozen foods, processed foods, seasonings, and other commercially available foods.

[0040] The food compositions of the present invention can be made into foods and beverages labeled with uses such as promoting a decrease in heart rate, suppressing an increase in heart rate, maintaining a normal heart rate, promoting a reduction in the risk of cardiovascular disease, promoting a reduction in the risk of coronary disease, promoting a reduction in the risk of hypertensive complications, promoting IL-10 production, suppressing IL-6 production, and activating the parasympathetic nervous system. Specifically, the foods and beverages can be labeled with such uses as "promoting a decrease in heart rate," "suppressing an increase in heart rate," "maintaining a normal heart rate," "preventing cardiovascular disease," "preventing coronary disease," "preventing hypertensive complications," "promoting IL-10 production," "suppressing IL-6 production," "activating the parasympathetic nervous system," and "helping to relax." Labels other than these can also be used, as long as they describe the effects achieved by promoting IL-10 production or suppressing IL-6 production.

[0041] In this specification, "indication" means any act intended to inform consumers of the above-mentioned uses, and any indication that can recall or infer the above-mentioned uses falls under the "indication" of the present invention, regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. However, it is preferable to display the use in an expression that allows consumers to directly recognize the above-mentioned uses.

[0042] The labeling is preferably one approved by the government or the like (e.g., labeling approved under various government systems and made in a manner based on such approval). Examples include labeling as a health food, functional food, functional food with function claims, enteral nutritional food, special dietary use food, food for the sick, functional nutrient food, and quasi-drug. Other examples include labeling approved by the Ministry of Health, Labor and Welfare, such as a food for specified health uses (FOSHU) or labeling approved under similar systems. Examples of the latter include labeling as a food for specified health uses, a conditional food for specified health uses (FOSHU), a label indicating an effect on the structure or function of the body, and a label indicating disease risk reduction. More specifically, labeling as a food for specified health uses (particularly a label indicating health uses) as defined in the Enforcement Regulations of the Health Promotion Act (Ministry of Health, Labor and Welfare Ordinance No. 86 of April 30, 2003), and similar labeling, etc.

[0043] According to another aspect of the present invention, the composition for lowering heart rate of the present invention can be provided as a pharmaceutical composition. The pharmaceutical composition of the present invention may be used for promoting heart rate reduction, suppressing an increase in heart rate, maintaining a normal heart rate, or preventing and / or treating diseases such as cardiovascular diseases (e.g., heart failure, myocardial infarction, angina pectoris, arteriosclerosis, etc.), coronary diseases, and hypertensive complications (hypertensive cardiac hypertrophy, congestive heart failure, cerebral hemorrhage, cerebral infarction, and hypertensive nephropathy). The pharmaceutical composition of the present invention can be produced according to standard manufacturing procedures for such foods, except for the inclusion of lactic acid bacteria. Here, the pharmaceutical composition refers to the composition of the present invention prepared as an oral or parenteral formulation according to standard methods. The formulation may be carried out using additives acceptable for formulation. Examples of additives acceptable for formulation include excipients, stabilizers, preservatives, humectants, emulsifiers, lubricants, sweeteners, colorants, flavorings, buffers, antioxidants, and pH adjusters. When the pharmaceutical composition is an oral preparation, it can take the form of a solid preparation such as a tablet, powder, fine granules, granules, capsules, pills, or sustained-release preparation, or a liquid preparation such as a solution, suspension, or emulsion. When the pharmaceutical composition is a parenteral preparation, it can take the form of an injection, suppository, or the like. From the viewpoint of ease of ingestion (administration) to a subject, oral preparations are preferred for pharmaceutical compositions.

[0044] The intake amount of the composition of the present invention is not particularly limited as long as the effects of the present invention are achieved, and can be appropriately adjusted depending on the age, health condition, body weight, etc. of the subject. Typically, it is 0.01 to 10,000 mg / kg body weight per day, preferably 0.1 to 1,000 mg / kg body weight, more preferably 0.5 to 300 mg / kg body weight, and even more preferably 1 to 100 mg / kg body weight. Furthermore, the dry mass of the lactic acid bacteria is preferably 0.001 to 1,000 mg / kg body weight, more preferably 0.01 to 100 mg / kg body weight, more preferably 0.05 to 30 mg / kg body weight, and even more preferably 0.1 to 10 mg / kg body weight. Furthermore, the number of lactic acid bacteria is preferably 10 4 ~10 12 per kg body weight, preferably 10 5~10 11 per kg body weight, more preferably 10 6 ~10 10 pcs / kg body weight, particularly preferably 10 6 ~10 8 Pieces / kg body weight.

[0045] To maximize the effectiveness of the composition of the present invention, it is preferable to continuously ingest (administer) it for a long period of time, specifically, for at least three days, and more preferably for at least one week. Examples of the administration period include 1 to 6 weeks, 1 to 12 weeks, 2 to 10 weeks, 4 to 10 weeks, and 4 to 12 weeks. As used herein, "continuously" means that a predetermined amount of the composition of the present invention is continuously ingested every day.

[0046] According to another aspect of the present invention, there is provided a method for reducing heart rate in a subject, comprising administering to the subject an effective amount of lactic acid bacteria belonging to the genus Lactobacillus and / or a composition containing lactic acid bacteria belonging to the genus Lactobacillus.

[0047] The subjects who take the lactic acid bacteria and / or composition in the above-mentioned method are not particularly limited as long as they are subjects who need to lower their heart rate, for example, subjects who have symptoms of tachyarrhythmia or subjects whose heart rate tends to increase.

[0048] In the method for lowering heart rate, the intake amount and intake period of the lactic acid bacteria and / or composition are not particularly limited as long as the effects of the present invention are achieved, and can be appropriately adjusted depending on the age, health condition, weight, etc. of the subject. Typically, in the method of the present invention, the intake amount and intake period of the lactic acid bacteria and / or composition are the same as the intake amount and intake period of the lactic acid bacteria in the composition of the present invention.

[0049] According to another aspect of the present invention, there is provided a method for treating and / or preventing a disease and / or disorder caused by an elevated heart rate in a subject, the method comprising administering to the subject an effective amount of lactic acid bacteria belonging to the genus Lactobacillus and / or a composition containing lactic acid bacteria belonging to the genus Lactobacillus.

[0050] Diseases and disorders caused by an increase in heart rate include, but are not limited to, cardiovascular diseases (e.g., heart failure, myocardial infarction, angina pectoris, arteriosclerosis, etc.), coronary artery disease, hypertensive complications (hypertensive cardiac hypertrophy, congestive heart failure, cerebral hemorrhage, cerebral infarction, hypertensive nephropathy), etc.

[0051] According to another aspect of the present invention, there is provided use of lactic acid bacteria belonging to the genus Lactobacillus for lowering heart rate.

[0052] According to another aspect of the present invention, there is provided use of a lactic acid bacterium belonging to the genus Lactobacillus for the manufacture of a composition for lowering heart rate.

[0053] According to another aspect of the present invention, there is provided a lactic acid bacterium belonging to the genus Lactobacillus for reducing heart rate.

[0054] As the lactic acid bacteria belonging to the genus Lactobacillus used in the above-mentioned other embodiment, the lactic acid bacteria belonging to the genus Lactobacillus used in the composition of the present invention described above can be used. [Example]

[0055] The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples.

[0056] Example 1: Confirmation of the heart rate lowering effect of a composition containing lactic acid bacteria (1) Subject selection (screening) Subjects for the confirmation test of the heart rate lowering effect were selected based on the following inclusion criteria A and exclusion criteria B. A: Selection criteria Healthy men and women aged 20 to 64 -Those with fasting blood glucose levels of 100-125 mg / dL at the time of screening -Those with a blood hemoglobin A1c (HbA1c) level of 5.6 to 6.4 at the time of screening According to the "Report of the Committee on the Classification and Diagnostic Criteria of Diabetes (International Standardization Version)" by Kiyono et al., Diabetes, Vol. 55, No. 7 (2012), subjects who meet the fasting blood glucose and HbA1c values ​​shown in selection criterion A are diagnosed as either a group in which suspicion of diabetes cannot be ruled out, or a group in which the subject does not currently have diabetes but is at high risk of developing diabetes in the future. In other words, subjects who meet selection criterion A are thought to be experiencing a decline in the metabolic function of sugar and lipids due to chronic inflammation of adipose tissue associated with aging and unhealthy lifestyles. Specifically, increased levels of inflammatory cytokines, such as monocyte chemotactic protein-1 (MCP-1) and IL-6, in adipose tissue promote macrophage infiltration into adipose tissue and reduce the production of hormones necessary for normal glucose and lipid metabolism (Xu H et al. "Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance," J Clin Invest, 112: 1821-1830, 2003; Weisberg SP et al. "CCR2 modulates inflammatory and metabolic effects of high-fat feeding," J Clin Invest, 116: 115-124, 2006). Therefore, subjects who meet the fasting blood glucose and HbA1c levels listed in inclusion criterion A are likely to experience an increase in blood IL-6 levels over time if they do not undergo lifestyle modification or other interventions. Furthermore, because this study was conducted between August and November, a time when people tend to gain weight, the amount of IL-6 produced by fat cells increased, which in turn tended to increase the amount of IL-6 in the blood. B: Exclusion criteria -People taking medication that affects blood sugar levels - Subjects who have taken supplements or health foods that affect blood sugar levels within one month prior to the screening test - Subjects who had the habit of consuming fermented milk or lactic acid bacteria drinks at least three times a week for the three months prior to the screening test -People who have an allergy to milk -Those who have been diagnosed with diabetes as a result of screening tests and require drug treatment -Those suffering from severe systemic diseases -People with chronic illnesses that require daily medication - Subjects who are deemed unsuitable as subjects based on clinical examinations during screening tests -Those who have received a blood transfusion of more than 200 mL within the past month or more than 400 mL within the past three months Heavy alcohol drinkers Alcoholics or drug addicts -Those who have participated in other clinical trials within one month prior to obtaining consent to participate in this study Pregnant or breastfeeding women -Other individuals who are deemed inappropriate to be subjects by the investigator

[0057] From 1,593 volunteers, 126 subjects were selected based on the above criteria. The age, sex, weight, and BMI of the subjects in the control and test groups are shown in Table 1. [Table 1]

[0058] (2) Preparation of the composition Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus were added as lactic acid bacteria starters to a mixture containing raw milk, skim milk powder, cream, sugar, and stevia, and yogurt was prepared by fermenting at a temperature of 43°C for approximately 3 hours. Heat-treated Lactobacillus plantarum OLL2712 strain (accession number: FERM BP-11262) was added to the prepared yogurt to obtain 112 g of a composition (test composition) to be ingested by the test group. The bacterial cell mass of the lactic acid bacteria in the test composition was 5 x 10 9 The average number of lactic acid bacteria consumed was more than 112g per 112g. On the other hand, 112g of the same yogurt as used in the test compositions (yogurt not containing the heat-treated lactic acid bacteria Lactobacillus plantarum OLL2712 strain (Accession Number: FERM BP-11262)) was used as a control composition (control composition). Each composition was stored refrigerated until consumed by the subjects in each group. The heat treatment of the lactic acid bacteria was performed by concentrating the bacterial cells to the above concentration and then heating at 60°C for 10 minutes.

[0059] (3) Effect on blood levels of IL-6 The effect of the composition of the present invention on the blood level of IL-6 was evaluated according to the following procedure. First, each of the subjects (62 individuals) in the test group selected in (1) above was instructed to take 112 g of the test composition daily for 12 weeks. Additionally, each of the subjects (64 individuals) in the control group selected in (1) above was instructed to take 112 g of the control composition daily for 12 weeks. Blood samples were taken from each group at 4, 8, and 12 weeks after the start of intake, and the serum IL-6 levels (concentrations) were measured using a Bio-Plex multiplex system.

[0060] The IL-6 measurement values ​​at each time point were evaluated using the Mann-Whitney U test. The results are shown in Table 2. All values ​​in the table are in pg / mL. [Table 2]

[0061] As shown in Table 2, in the test group that ingested the test composition, the blood IL-6 level did not significantly increase after initiating ingestion of the test composition compared to before initiation of ingestion, and actually decreased slightly. On the other hand, in the control group that ingested the control composition, the blood IL-6 level tended to increase over 12 weeks after initiating ingestion of the control composition, and a significant increase was observed 12 weeks after initiation of ingestion. As described in "A: Selection Criteria" in (1) above, given that this Example targeted subjects with a tendency for increased blood IL-6 levels, it was shown that the increase in blood IL-6 levels was suppressed in the test group (i.e., the blood IL-6 level was reduced), while the increase in blood IL-6 levels was not suppressed in the control group (i.e., the blood IL-6 level was not reduced).

[0062] (4) Effects on interleukin-10 production in bone marrow-derived dendritic cells and peritoneal macrophages The effects of Lactobacillus plantarum OLL2712 on interleukin-10 (IL-10) production in bone marrow-derived dendritic cells and peritoneal macrophages were evaluated according to the following procedure.

[0063] The amount of IL-10 produced by lactic acid bacteria in bone marrow-derived dendritic cells was measured as follows. Bone marrow fluid extracted from the femur of a BALB / c mouse (manufactured by Japan SLC Co., Ltd.) was passed through a 70 μm cell strainer and then hemolyzed. Rabbit IgG was added to prevent nonspecific adsorption. Next, biotin-labeled anti-CD4 antibody, anti-CD8 antibody, and IA antibody were added. d(MHCII marker) antibody was added and the mixture was left on ice for 30 minutes. Streptavidin magnetic beads and anti-B220 antibody magnetic beads were then added. After passing the mixture through a 40 μm cell strainer again, the negative fraction was collected using an auto MACS DEPLETE. This procedure removed T cells, B cells, and antigen-presenting cells from the bone marrow fluid, allowing the isolation of only immature dendritic cells.

[0064] The resulting immature dendritic cells were cultured in 10 ml of RPMI-10 containing 10% GM-CSF, and after 3 days, 5 ml of RPMI-10 containing GM-CSF was added. After an additional 5 days, floating cells were collected and used as bone marrow-derived dendritic cells.

[0065] The obtained immature dendritic cells were cultured at 10 5 The cells were seeded onto a 96-well plate at 10 μg / mL of heat-killed Lactobacillus plantarum OLL2712 cells per well, and 10 μg / mL of heat-killed Lactobacillus plantarum OLL2712 cells was added. After 24 hours, the culture supernatant was collected, and the amount (concentration) of IL-10 in the collected culture supernatant was measured using a mouse ELISA kit, thereby measuring the amount of IL-10 produced by bone marrow-derived dendritic cells. The antibodies and ELISA kit used were all purchased from Becton Dickinson.

[0066] When the amount of IL-10 produced by bone marrow-derived dendritic cells was measured using the above procedure, the amount of IL-10 produced by bone marrow-derived dendritic cells when Lactobacillus plantarum OLL2712 was added at a concentration of 10 μg / mL was 1319 ± 62 pg / mL (mean ± standard deviation).On the other hand, the amount of IL-10 produced by bone marrow-derived dendritic cells when Lactobacillus plantarum OLL2712 was not added was 0 pg / mL.

[0067] These results indicate that Lactobacillus plantarum OLL2712 has the ability to induce IL-10 production in bone marrow-derived dendritic cells.

[0068] The amount of IL-10 produced by lactic acid bacteria in peritoneal macrophages was measured as follows: BALB / c mice (manufactured by Japan SLC Co., Ltd.) were intraperitoneally administered 2 ml of 4% thioglycollate medium (Becton Dickinson) and housed for 4 days. Five ml of PBS (lavage fluid) was injected intraperitoneally, and the lavage fluid was then collected to obtain peritoneal macrophages.

[0069] The obtained peritoneal macrophages were cultured at 10 5 Cells were seeded at 1000 cells / well in a 96-well plate, and 10 μg / mL of heat-killed Lactobacillus plantarum OLL2712 cells were added. After 24 hours, the culture supernatant was collected, and the amount (concentration) of IL-10 in the collected culture supernatant was measured using a mouse ELISA kit, and the amount of IL-10 produced by peritoneal macrophages was measured. The antibodies and ELISA kit used were all purchased from Becton Dickinson.

[0070] When the amount of IL-10 produced in peritoneal macrophages was measured using the above procedure, When Lactobacillus plantarum OLL2712 was added at a concentration of 10 μg / mL, the amount of IL-10 produced by peritoneal macrophages was 1178 ± 298 pg / mL (mean ± standard deviation).On the other hand, when Lactobacillus plantarum OLL2712 was not added, the amount of IL-10 produced by peritoneal macrophages was 0 pg / mL.

[0071] These results indicated that Lactobacillus plantarum OLL2712 has the activity of inducing IL-10 production in peritoneal macrophages.

[0072] (5) Evaluation of heart rate reduction effect The effect of the compositions of the present invention on heart rate was evaluated according to the following procedure. First, the heart rate of each subject (62 subjects) in the test group was measured immediately before initiating the ingestion of the test composition. Then, each subject in the test group was instructed to ingest 112 g of the test composition daily for 12 weeks. Meanwhile, the heart rate of each subject (64 subjects) in the control group was measured immediately before initiating the ingestion of the control composition. Then, each subject in the control group was instructed to ingest 112 g of the control composition daily for 12 weeks. During the study, subjects in each group were asked to maintain their normal diet and lifestyle habits (e.g., quantity and quality of exercise). After 12 weeks of initiating the ingestion of each composition, each subject's heart rate was measured on the upper arm after resting. The heart rate measurements of each subject at each time point were analyzed and evaluated using the Wilcoxon signed-rank test. The results are shown in Table 3.

[0073] [Table 3]

[0074] As shown in Table 3, in the test group, the heart rate 12 weeks after initiating ingestion of the test composition was significantly lower than that immediately before initiating ingestion. On the other hand, in the control group, no significant change was observed between the heart rate 12 weeks after initiating ingestion of the control composition and the heart rate immediately before initiating ingestion. These results demonstrate that the composition of the present invention significantly reduces the heart rate in subjects.

[0075] (6) Correlation between blood IL-6 levels and heart rate The correlation between the amount of IL-6 in the blood and the heart rate was investigated according to the following procedure. Specifically, the correlation between the amount of IL-6 in the blood of all subjects in the test group and the control group before starting to take each composition (week 0), as measured in (3) above, and the heart rate of all subjects in the test group and the control group before starting to take each composition (week 0), as measured in (5) above, was confirmed. Figure 1 shows a graph showing the correlation between the amount of IL-6 in the blood and the heart rate of all subjects in the test group and the control group before starting to take each composition (week 0).

[0076] The results in Figure 1 show that there is a significant correlation between the amount of IL-6 in the blood and heart rate (R=0.425, P<0.001).

[0077] As described above, the results in Table 2 showed that the test group that ingested the test composition inhibited an increase in blood IL-6 levels, and even tended to decrease. Furthermore, the results in Figure 1 showed that there was a high correlation between blood IL-6 levels and heart rate. These results suggested that the decrease in heart rate in the test group was due to the inhibition of an increase in the amount of IL-6 in the subjects' blood. [Industrial Applicability]

[0078] According to the present invention, the heart rate in a subject can be significantly lowered, and the risk of diseases caused by an elevated heart rate, such as cardiovascular disease, coronary disease, and hypertensive complications, can be reduced.

Claims

1. A composition for lowering heart rate containing lactic acid bacteria belonging to the genus Lactobacillus as an active ingredient. The composition, wherein the lactic acid bacterium is Lactobacillus plantarum OLL2712 strain deposited under accession number FERM BP-11262.

2. The composition of claim 1, for human use.

3. A composition described in claim 1 or 2, wherein the lactic acid bacteria include dead cells of lactic acid bacteria.

4. The composition according to any one of claims 1 to 3, wherein the lactic acid bacteria comprise heat-killed lactic acid bacteria.

5. The composition according to any one of claims 1 to 4, wherein the composition is an oral composition.

6. The composition according to any one of claims 1 to 5, wherein the composition is a food composition.

7. The composition according to any one of claims 1 to 5, wherein the composition is a pharmaceutical composition.

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