Intestinal environment-improving agent

Abstract

To find out a useful living body-regulating function in a treated product of Abelmoschus manihot and to provide a functional use thereof.SOLUTION: An intestinal environment-improving agent comprising a processed product of Abutilon striata as an active ingredient, wherein the intestinal environment-improving agent increases bacteria of the genus Lactobacillusmurinus and / or the genus Parabacteroides.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] The present invention relates to an agent for improving the intestinal environment, which contains a treated product of black soldier fly as an active ingredient. [Background technology]

[0002] The world's population is expected to exceed 8 billion in 2023 and reach 9.8 billion by 2050. This rapid population growth will make it difficult to ensure a stable food supply and will also increase the burden on the environment.

[0003] In recent years, insects such as crickets and mealworms have been attracting attention as food. These insects are not only rich in protein and other nutrients, but also have a shorter life cycle than livestock and can be raised on food waste, making them a promising solution to the above-mentioned problems. Among these, black soldier flies are expected to be used as food or feed because of their shorter life cycle compared to other insects and their high protein and vitamin content. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2014 / 126982 [Non-patent literature]

[0005] [Non-Patent Document 1] Pan, F., Zhang, L., Li, M. et al. Predominant gut Lactobacillus murinus strain mediates anti-inflammaging effects in calorie-restricted mice. Microbiome 6, 54 (2018). https: / / doi.org / 10.1186 / s40168-018-0440-5 [Non-patent document 2] Yang X, Wu M, Qiu S, Chen Y, Yan Z, Luo S, Zhao J, Liu W, Li C, Liu KX. Lactobacillus murinus alleviates intestinal ischemia / reperfusion injury through promoting the release of interleukin-10 from M2 macrophages via Toll-like receptor 2 signaling. Microbiome. 2022 Mar 3;10(1):38. doi: 10.1186 / s40168-022-01227-w. PMID: 35241180; PMCID: PMC8896269.

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[0006] An object of the present invention is to discover useful bioregulatory functions of black soldier fly treated materials and to provide functional uses thereof.

[0007] As a result of extensive research into solving the above-mentioned problems, the inventors discovered that ingesting treated black soldier fly products improves the intestinal environment by promoting the growth of specific beneficial bacteria in the intestinal flora, and thus completed the present invention. [Means for solving the problem]

[0008] The present invention includes the following aspects. [1] An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, An agent for improving the intestinal environment, which improves the intestinal environment by increasing the number of bacteria of the genus Lactobacillus murinus and / or Parabacteroides. [2] An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, The intestinal environment improving agent improves the intestinal environment by increasing metabolites produced by intestinal bacteria, and the metabolites are one or more selected from the group consisting of uric acid, lauric acid, octanoic acid, and hydroxylamine. [3] An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, The intestinal environment improving agent reduces metabolites produced by intestinal flora, and the metabolites are one or more selected from the group consisting of fumaric acid, nonanoic acid, glutaric acid, 4-hydroxyproline, 1,6-anhydro-β-d-glucose, arabitol, ribitol, and 5-methoxytryptamine. [4] The intestinal environment improving agent according to [1] to [3], wherein the processed material is a dried processed material of black soldier fly. [5] The intestinal environment improving agent according to [1] to [4], wherein the black soldier fly is an American black soldier fly. [6] The intestinal environment improving agent according to any one of [1] to [5], which is a food or drink. [7] The intestinal environment improving agent according to any one of [1] to [5], which is a pharmaceutical product. [8] A preventive and ameliorative agent for inflammatory diseases containing treated black soldier fly material as an active ingredient. [9] An agent for preventing and improving allergic diseases containing treated black soldier fly material as an active ingredient. [Effects of the Invention]

[0009] The intestinal environment improving agent of the present invention increases bacteria of the genus Lactobacillus murinus and / or Parabacteroides, and can effectively prevent and improve inflammatory diseases, allergic diseases, multiple sclerosis, etc., which are associated with these bacteria. Furthermore, the intestinal environment improving agent of the present invention regulates the production of metabolites by intestinal bacteria, and can achieve preventive and improving effects against diseases associated with these metabolites. [Brief explanation of the drawings]

[0010] [Figure 1]Figure 1 shows the results of the alpha diversity analysis of the intestinal microbiota. Figure 1(A) shows the ACE index. Figure 1(B) shows the Chao1 index. Statistical analysis was performed using an unpaired t test. "**" indicates a significant difference (p<0.01). [Figure 2] Figure 2 shows the results of β-diversity (Unifrac distance) analysis of the intestinal microbiota. [Figure 3] Figure 3 shows the occupancy rate of Parabacteriodes bacteria. Statistical analysis was performed using the Mann-Whitney test. "**" indicates a significant difference (p<0.01). [Figure 4] FIG. 4 shows the blood metabolite levels in the non-administration group relative to the blood metabolite levels in the administration group in a volcano plot. [Figure 5] Figure 5 shows the relative ratios of blood metabolite levels that were significantly increased in the treated group compared to the untreated group. Statistical analysis was performed using an unpaired t test. * and ** indicate significant differences (*: p<0.05, **: p<0.01). [Figure 6] Figure 6 shows the relative ratios of blood metabolite levels that were significantly reduced in the treated group compared to the untreated group. Statistical analysis was performed using an unpaired t test. * and ** indicate significant differences (*: p<0.05, **: p<0.01). DETAILED DESCRIPTION OF THE INVENTION

[0011] The present invention will be described in detail below. The following description of the present invention may be based on preferred embodiments of the present invention, but the present invention is not limited to such embodiments. In this specification, the use of "to" to indicate a range of values ​​means that the values ​​before and after it are included as the lower and upper limits.

[0012] The intestinal environment improving agent of the present invention contains a treated black soldier fly as an active ingredient. The black soldier fly is not particularly limited as long as it is an insect belonging to the family Stratiomyidae of the order Diptera, and examples thereof include the American soldier fly, the American black soldier fly, the yellow-legged black soldier fly, the black soldier fly, the black soldier fly, and the small black soldier fly. One or more of these can be used, but the American black soldier fly is preferred. The form of the black soldier fly is also not particularly limited, and examples include larvae, pupae, and adults, but it is preferred to use larvae.

[0013] The method for obtaining a treated black soldier fly product is not particularly limited, and examples thereof include freezing, boiling, heating, drying, extraction, crushing, and squeezing, and these treatments may be combined. Among these, a dried black soldier fly product obtained by drying is preferred, and black soldier fly powder obtained by combining drying and crushing is more preferred.

[0014] The drying treatment is not particularly limited, and known methods can be used, such as hot air drying, low-temperature drying, vacuum freeze drying, pressure drying, etc., and one or more of these can be used. The pulverization treatment is not particularly limited, and can be carried out using known crushing devices, such as a cutter mill, roll mill, hammer mill, ball mill, bead mill, etc., and one or more of these can be used.

[0015] By ingesting the intestinal environment improving agent of the present invention, the intestinal environment of animals, including humans, can be improved.

[0016] The intestinal environment improving agent of the present invention has the effect of increasing the proliferation of Lactobacillus murinus, a type of Lactobacillus bacteria.

[0017] Lactobacillus murinus promotes the differentiation and induction of regulatory T cells, and increasing their presence in the intestinal flora can prevent and ameliorate inflammatory bowel disease (Patent Document 1). Furthermore, Lactobacillus murinus is involved in reducing intestinal permeability, reducing endotoxin, reducing the production of the inflammatory cytokine IL-8, and promoting the production of the anti-inflammatory cytokine IL-10 via TLR2. Therefore, increasing their presence in the intestinal flora can prevent and ameliorate inflammatory diseases (Non-Patent Documents 1 and 2). Furthermore, Lactobacillus murinus promotes the production of the anti-inflammatory cytokine IL-12 and suppresses the expression of OX40 ligand by intestinal cell CD11c, thereby shifting the Th1 / Th2 cell balance to a state in which Th1 cells are dominant, thereby potentially reducing allergic reactions such as food allergies (Non-Patent Document 3). In the present invention, intake of black soldier fly processed material increases the occupancy rate of Lactobacillus murinus, which has the effect of preventing and improving inflammatory diseases such as inflammatory bowel disease (ulcerative colitis and Crohn's disease) and irritable bowel syndrome, and allergic diseases such as food allergies.

[0018] The intestinal environment-improving agent of the present invention can improve the intestinal environment by promoting the growth of bacteria of the genus Parabacteroides. It has been reported that the presence of Parabacteroides bacteria is reduced in patients with multiple sclerosis compared to healthy individuals (Non-Patent Document 4). In the present invention, by ingesting black soldier fly-treated material, the proportion of Parabacteroides bacteria can be increased, thereby achieving preventive and ameliorative effects against multiple sclerosis.

[0019] The intestinal environment improving agent of the present invention can improve the intestinal environment by increasing or decreasing metabolites produced by intestinal bacteria, and for example, promotes the production of uric acid, lauric acid, octanoic acid, and hydroxylamine, and suppresses the production of fumaric acid, nonanoic acid, glutaric acid, 4-hydroxyproline, 1,6-anhydro-β-d-glucose, arabitol, ribitol, and 5-methoxytryptamine.

[0020] Uric acid has antioxidant and neuroprotective effects (Non-Patent Document 5), lauric acid has antioxidant and blood pressure lowering effects (Non-Patent Document 6), and appetite suppressant effects (Non-Patent Document 7). Octanoic acid has GABA synthesis promoting and antioxidant effects, as well as reducing intestinal permeability (Non-Patent Documents 8 and 9). Hydroxyamine has antioxidant and blood pressure lowering effects (Non-Patent Documents 10 and 11). In the present invention, by ingesting black soldier fly processed products, the production of uric acid, lauric acid, octanoic acid, and hydroxyamine is promoted, and effects such as antioxidant, blood pressure lowering, appetite suppressant, GABA synthesis promoting, intestinal permeability suppression, and blood pressure lowering can be obtained.

[0021] Glutaric acid is a neurotoxic substance that causes declines in cognitive and motor function. Furthermore, glutaric acid accumulates in large amounts in the bodies of patients with glutaric acidemia type 1, causing damage to the central nervous system (particularly the caudate nucleus and putamen of the striatum). 5-Methoxytryptamine is a substance that induces hyperglycemia (Non-Patent Document 12), and prolonged hyperglycemia can lead to the development of diabetes. In the present invention, the ingestion of black soldier fly processed products suppresses the production of glutaric acid and 5-methoxytryptamine, thereby preventing and improving declines in cognitive and motor function, glutaric acidemia type 1, hyperglycemia, and diabetes.

[0022] The intestinal environment improving agent of the present invention can be formulated by combining the above-mentioned active ingredients with a pharmaceutically acceptable carrier as needed. Examples of pharmaceutically acceptable carriers include glucose, lactose, starch, mannitol, dextrin, fatty acid glycerides, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, amino acids, gelatin, albumin, water, and physiological saline. Furthermore, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonicity agents, and excipients can be added as needed. The dosage form is not particularly limited, and examples include liquids, powders, granules, capsules, and tablets, and can be prepared according to conventional methods.

[0023] The intestinal environment-improving agent of the present invention can be applied to mammals including humans, such as humans, dogs, cats, cows, pigs, chickens, sheep, and horses, with humans being preferred. The dosage is not particularly limited, but for example, the daily dosage for an adult is about 0.5 to 10 g per kg of body weight, preferably about 1 to 5 g. For dogs, cats, and the like, the dosage is about 0.5 to 20 g per kg of body weight, preferably about 1.5 to 10 g. The dosage may be, for example, once a day or in multiple divided doses. The dosage interval may be, for example, every day or every few days.

[0024] The intestinal environment-improving agent of the present invention can also be in the form of a food or drink, and is prepared by blending the above-mentioned active ingredient with known food additives and / or food materials according to a conventional method, and the form is not particularly limited, and examples include starch-based foods such as bread, biscuits, pancakes, noodles, and tablet candy, confectioneries such as gum, candy, and Japanese sweets, meat foods such as ham and sausage, fish foods such as chikuwa and kamaboko, seafood foods, seasonings such as dressing, soy sauce, jam, and furikake, beverages such as tea, juice, soft drinks, and alcoholic beverages, etc. Foods and drinks include foods for specified health uses and foods with functional claims that have been approved or notified for their use in improving the intestinal environment.

[0025] The intestinal environment improving agent of the present invention can be in the form of feed or pet food, and is prepared according to a conventional method by appropriately blending the above-mentioned active ingredient with known feed ingredients, gelling agents, shape-retaining agents, pH adjusters, seasonings, preservatives, nutritional supplements, etc. The form is not particularly limited, and any form such as pellets, flakes, or mash can be exemplified, and the agent can be used, for example, as feed for livestock such as cows, pigs, chickens, sheep, and horses, or as pet food for dogs, cats, etc. [Example]

[0026] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to these examples.

[0027] [Materials and rearing conditions] Eleven 7-week-old C57BL / 6 mice weighing 20–25 g were purchased from Japan SLC Co., Ltd. One mouse per cage was housed at 24°C ± 2°C under a 12-hour light / dark cycle with free access to tap water and feed (AIN-93G; CLEA Japan, Inc.).

[0028] [Administration schedule] After one week of preliminary breeding under the above conditions, 8-week-old mice were randomly divided into a non-administered group (5 mice) and a treated group (6 mice). During the four-week test period, the non-administered group continued to receive the above feed, while the treated group received the above feed mixed with 5% black soldier fly powder by weight.

[0029] At the end of the study period, blood was collected from each group, and then the mice were dissected and the cecal contents were collected and stored at -80°C.

[0030] [Intestinal microbiota analysis] For intestinal microbiota analysis, DNA was extracted from cecal contents using the following method. Frozen cecal contents were thawed on ice, and 100 mg of each sample was suspended in 4 M guanidinium thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA. The suspension was disrupted with zirconia beads using a FastPrep FP100A (MP Biomedicals). DNA was extracted from the bead-treated suspension using a Magtration System 12GC and a GC series MagDEA DNA 200.

[0031] The resulting DNA was used as a template for PCR amplification of the 16S rRNA V3-V4 region using primers (341F and 785R). The resulting PCR products were analyzed using the Illumina MiSeq platform and MiSeq Reagent Kit V3 (both manufactured by Illumina) by paired-end PCR. The resulting sequences were then assigned to a taxonomic group using the EzBioCloud (https: / / www.ezbiocloud.net / ) database (Ver. 20230823). Alpha diversity analysis (Figure 1), beta diversity analysis (Figure 2), and intergroup comparison analysis (LEfSe analysis) (Table 1) confirmed that the proportion of Parabacteriodes bacteria was significantly higher in the treated group than in the untreated group (Figure 3).

[0032] (alpha diversity analysis) Alpha diversity analysis can analyze the diversity of the intestinal microbiota within each group. In this example, the ACE Index and Chao1 Index were used as indices. The higher the index, the higher the diversity of the intestinal microbiota. As shown in Figures 1(A) and 1(B), it was confirmed that the species diversity in the treated group was significantly increased compared to the untreated group.

[0033] (beta diversity analysis) Beta diversity analysis can analyze differences in the diversity of the intestinal microbiota between groups. In this example, the analysis was performed using a combination of UniFrac principal coordinate analysis and nonparametric multivariate analysis of variance. The greater the distance between the plots, the more different the composition of the intestinal microbiota. As shown in Figure 2, it was confirmed that the distance between the plots of the non-treated group and the treated group was significantly different.

[0034] (Intergroup comparison analysis (LEfSe analysis)) Characteristic bacterial species were identified in each group using intergroup comparison analysis (|LDA score|>3.5, mean relative abundance>1%). As shown in Table 1 below, the occupancy rate of Lactobacillus murinus was confirmed to be higher in the treatment group than in the non-treatment group.

[0035] [Table 1]

[0036] [Metabolome analysis] Metabolomic analysis was performed using blood samples from the untreated and treated groups. As shown in Figure 4, four metabolites (uric acid, lauric acid, caprylic acid, and hydroxylamine) were significantly increased in the treated group compared to the untreated group, and eight metabolites (fumaric acid, nonanoic acid, glutaric acid, 4-hydroxyproline, 1,6-anhydro-β-d-glucose, arabitol, ribitol, and 5-methoxytryptamine) were significantly decreased. Figure 5 shows the relative ratios of the blood metabolite levels in the treated group to those in the untreated group for the 12 metabolites that were significantly increased or decreased.

Claims

1. An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, The intestinal environment improving agent, wherein the intestinal environment improving agent increases the proliferation of bacteria of the genus Lactobacillus murinus and / or Parabacteroides.

2. An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, The intestinal environment improving agent, wherein the improvement of the intestinal environment is achieved by increasing metabolites produced by intestinal bacteria, and the metabolites are one or more selected from the group consisting of uric acid, lauric acid, octanoic acid, and hydroxylamine.

3. An intestinal environment improving agent containing a treated black soldier fly as an active ingredient, The intestinal environment improving agent, wherein the improvement of intestinal bacteria is achieved by reducing metabolites produced by intestinal bacterial flora, and the metabolites are one or more selected from the group consisting of fumaric acid, nonanoic acid, glutaric acid, 4-hydroxyproline, 1,6-anhydro-β-d-glucose, arabitol, ribitol, and 5-methoxytryptamine.

4. The intestinal environment improving agent according to claim 1 or 2, wherein the processed material is a dried processed material of black soldier fly.

5. The intestinal environment improving agent according to claim 1 or 2, wherein the black soldier fly is an American black soldier fly.

6. The intestinal environment improving agent according to claim 1 or 2, which is a food or drink.

7. The intestinal environment improving agent according to claim 1 or 2, which is a pharmaceutical product.

8. An agent for preventing and improving inflammatory diseases, containing treated black soldier fly material as an active ingredient.

9. An agent for preventing and improving allergic diseases, containing treated black soldier fly material as an active ingredient.

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