Composition and method for treating chronic endometritis

Abstract

To create a model animal for selecting therapeutic agents having superior effects in preventing and treating chronic endometritis, and to provide therapeutic agents and compositions having superior effects in preventing and treating chronic endometritis.SOLUTION: A model animal of chronic endometritis was produced by administering LPS into the uterus of a non-human animal, particularly by administering LPS into the uterus of a non-human animal having a deficiency in the SREBP1 gene. By using this model animal, superior effects of omega-3 fatty acids such as EPA or omega-3 fatty acid esters in the prevention and treatment of chronic endometritis were demonstrated, thereby solving the above problem.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a treatment method, a pharmaceutical composition for preventing and / or improving chronic endometritis, a method for manufacturing such a medicine, and a treatment method. The present invention also provides a food composition and a dietary composition. The composition of the present invention contains, as an active ingredient, an ω3 fatty acid, for example, eicosapentaenoic acid (EPA).

Background Art

[0002] In modern times troubled by a declining birthrate, recurrent implantation failure and recurrent miscarriage that do not respond to normal infertility treatment are issues to be overcome. As a typical cause, chronic endometritis (CE) has become a problem, but a treatment method has not been established.

[0003] Assisted Reproductive Technology (ART) has become the mainstream of infertility treatment, and infertile couples who were previously considered unable to conceive have been able to have children. However, in the current state of the advancement of ART technology, about 20% of cases of recurrent implantation failure where pregnancy does not occur even after multiple transfers of morphologically good embryos and recurrent miscarriage cases where miscarriage occurs repeatedly after pregnancy establishment remain (Non-Patent Document 1).

[0004] In recent years, chronic endometritis (CE), a persistent and mild inflammatory disease characterized by infiltration of plasma cells into the endometrial stromal region, has been attracting attention for its association with idiopathic infertility and habitual miscarriage and its association with a decline in ART outcomes.

[0005] In fact, it has been reported that CE is detected in about 20% of infertile women, and recurrent implantation failure is recognized in about 40% of them, and CE is detected in about 1% of recurrent miscarriage cases (Non-Patent Document 2), strongly suggesting that CE is a contributing factor to recurrent implantation failure and recurrent miscarriage.

[0006] On the one hand, the pathological condition of CE is not fully understood. In particular, in the dynamic endometrial tissue that repeats exfoliation (menstruation), proliferation, and differentiation every menstrual cycle, the reason why chronic inflammation persists is not clear.

[0007] ω3 polyunsaturated fatty acids such as EPA have anti-inflammatory activity, and it has been reported that a decrease in unsaturated fatty acids chronicizes inflammation (Non-Patent Document 3). In addition, knockout mice of GPR120, which is a receptor for ω3PUFA, develop miscarriages due to incomplete decidualization of the endometrium (Non-Patent Document 4).

[0008] As the first choice for CE treatment, treatment with antibacterial drugs having a broad spectrum including doxycycline is performed. However, it has been reported that 25% of cases have residual inflammation even after treatment with repeated administration of up to three types of antibacterial drugs (Non-Patent Document 5), and treatment-resistant CE is a major clinical problem. In addition, the emergence of resistant bacteria due to the use of multiple antibacterial drugs for a long period of time is also a concern.

[0009] Currently, no therapeutic agent having excellent effects in the prevention and treatment of chronic endometritis has been reported.

Prior Art Documents

Non-Patent Documents

[0010]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Summary of the Invention

Problems to be Solved by the Invention

[0011] An object of the present invention is to provide a treatment method, a pharmaceutical composition for preventing and / or improving chronic endometritis, a method for producing such a medicine, and a treatment method. The present invention also provides a food and drink composition and a diet composition.

Means for Solving the Problems

[0012] The inventors of the present invention prepared a non-human animal serving as a CE model by using LPS on non-human animals (preferably, REBP1-deficient animals and / or Srebf1-deficient). Furthermore, the inventors of the present invention completed the present invention by providing a glyceride-type omega-3 fatty acid (for example, EPA) as an active ingredient of a prophylactic or therapeutic agent for chronic endometritis.

[0013] The present invention provides, for example, the following. (Item 1) A pharmaceutical composition for preventing and / or treating chronic endometritis, which comprises a substance selected from the group consisting of omega-3 fatty acids and omega-3 fatty acid esters, a food and drink composition, or a diet composition. (Item 2) The pharmaceutical composition, food and drink composition, or diet composition according to Item 1, wherein the omega-3 fatty acid is selected from the group consisting of α-linolenic acid (ALA), stearidonic acid (SDA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA). (Item 3) The pharmaceutical composition, food and drink composition, or diet composition according to Item 2, wherein the omega-3 fatty acid is eicosapentaenoic acid (EPA). (Item 4) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid is an omega-3 fatty acid salt. (Item 5) The pharmaceutical composition, food or drink composition, or diet composition according to item 4, wherein the omega-3 fatty acid salt is a calcium salt of an omega-3 fatty acid. (Item 6) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid ester is a substance selected from the group consisting of omega-3 fatty acid glycerides, alcohol esters of omega-3 fatty acids, and phospholipids containing omega-3 fatty acids. (Item 7) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid ester is an omega-3 fatty acid triglyceride. (Item 8) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid ester is an alcohol ester of an omega-3 fatty acid. (Item 9) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid ester is an ethyl ester of an omega-3 fatty acid. (Item 10) The pharmaceutical composition, food or drink composition, or diet composition according to item 1, wherein the omega-3 fatty acid ester is an ethyl ester of eicosapentaenoic acid. (Item 11) Oral ingestion The pharmaceutical composition, food or drink composition, or diet composition according to item 1, characterized by oral ingestion. (Item 12) Non-human model animals for chronic endometritis. (Item 13) The non-human model animal according to item 12, which is a rodent. (Item 14) The non-human model animal according to item 12, which is a mouse. (Item 15) The non-human model animal according to item 12, prepared by administering LPS into the uterus. (Item 16) The non-human model animal according to item 12, having a deletion in the SREBP1 gene. (Item 17) The non-human model animal according to item 16, having a homozygous deletion in the SREBP1 gene. (Item 18) A method for preparing a non-human model animal of chronic endometritis, comprising the step of administering LPS into the uterus of a non-human animal. (Item 19) A method for preparing a non-human model animal of chronic endometritis, comprising the step of administering LPS into the uterus of a non-human animal having a deletion in the SREBP1 gene. [Effect of the Invention]

[0014] The present invention provides a model animal of chronic endometritis. Further, the present invention provides a treatment method, a pharmaceutical composition for preventing and / or improving chronic endometritis, a method for manufacturing such a medicine, and a treatment method. The present invention also provides a food composition and a diet composition. [Brief Description of the Drawings]

[0015]

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Mode for Carrying Out the Invention

[0016] The present invention will be described below. Throughout this specification, it should be understood that the singular expressions include the concepts of their plurals unless otherwise specified. Also, the terms used in this specification are to be understood as being used in the ordinary meanings commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meanings as commonly understood by those skilled in the art to which the present invention pertains. In case of conflict, this specification (including definitions) shall prevail. Also, in this specification, "wt%" is used interchangeably with "mass percent concentration".

[0017] (Definition of Terms) The definitions of the terms particularly used in this specification are listed below.

[0018] As used herein, the term "omega-3 fatty acid" refers to an unsaturated fatty acid having a carbon-carbon double bond at the ω3 position. Examples of omega-3 fatty acids include, but are not limited to, α-linolenic acid (ALA), stearidonic acid (SDA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA).

[0019] The omega-3 fatty acid used in the present invention may be, for example, a free fatty acid or in an ester form.

[0020] As used herein, the term "omega-3 fatty acid ester" refers to an ester of an omega-3 fatty acid, and typically includes, but is not limited to, an alcohol ester of an omega-3 fatty acid, an omega-3 fatty acid glyceride, and an omega-3 fatty acid-containing phospholipid.

[0021] As used herein, the term "alcohol ester of a fatty acid" refers to an ester of a fatty acid and an alcohol. Examples of the alcohol used for the production of the alcohol ester of a fatty acid include, but are not limited to, methanol, ethanol, propanol, isopropanol, butanol and its isomers, pentanol and its isomers, hexanol and its isomers, heptanol and its isomers, octanol and its isomers, or alcohols with a larger molecular weight and their isomers.

[0022] As used herein, the term "omega-3 fatty acid glyceride" includes, but is not limited to, triglycerides, diglycerides, and monoglycerides of omega-3 fatty acids, as well as various positional isomers thereof. Preferably, the "omega-3 fatty acid glyceride" of the present invention is a triglyceride.

[0023] As used herein, the term "omega-3 fatty acid-containing phospholipid" refers to an omega-3 fatty acid that forms an ester with a phospholipid. Examples of the phospholipid moiety forming the ester include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, their lyso forms, and their positional isomers.

[0024] As used herein, the term "pathology of chronic endometritis" means that (1) a certain number or more of CD138-positive plasma cells are observed in the endometrial stromal region, and typically, one or more CD138-positive plasma cells are recognized in two or more of the 10 observed fields, but is not limited thereto.

[0025] In CE, since the appearance of plasma cells in the lesion is significantly different from that of general inflammation, the appearance of plasma cells can be cited as "the pathological condition of chronic endometritis". In this case, the suppression of the appearance of plasma cells can be used as an index for the preventive effect and / or therapeutic effect of CE. Plasma cells that can be used as an index for the preventive effect and / or therapeutic effect of CE typically include, but are not limited to, CD138-positive plasma cells.

[0026] Examples of omega-3 fatty acids used in the present invention include, but are not limited to, α-linolenic acid (ALA), stearidonic acid (SDA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA). The omega-3 fatty acid used in the present invention may be a free fatty acid or a salt. Examples of omega-3 fatty acid salts include, but are not limited to, calcium salt, sodium salt, potassium salt, and magnesium salt.

[0027] The "model animal for chronic endometritis" used in the present invention is a model animal that exhibits at least one of the "pathological conditions of chronic endometritis". Pathologically, the "pathological conditions of chronic endometritis" include endometrial edematous changes, high stromal cell density, and infiltration of plasma cells into the endometrial stroma. Clinically, it is associated with repeated implantation failure and repeated miscarriage. The main diagnostic markers of chronic endometritis include, but are not limited to, the detection of CD138-positive plasma cells in the endometrial stroma.

[0028] Model animals for chronic endometritis are, for example, rodents, and typically include, but are not limited to, mice and rats. Model animals for chronic endometritis typically have a deficiency in the SREBP1 gene. The gene deficiency may be a heterozygous mutation or a homozygous mutation. Preferably, the gene deficiency is a homozygous deficiency.

[0029] (Pharmaceutical composition) The composition of the present invention is in the form of an orally deliverable dosage form or dosage unit. Non-limiting examples of suitable dosage forms include tablets (e.g., suspension tablets, chewable suspension tablets, rapidly dispersing tablets, chewable tablets, etc.), caplets, capsules (e.g., soft or hard gelatin capsules, or HPMC capsules), medicinal lozenges, sachets, cachets, troches, pellets, suspensions, elixirs, syrups, or any other solid dosage form reasonably adapted for oral administration. As used herein, the terms "oral delivery" and "oral administration" include any form of delivery in which the agent or composition is placed in the mouth of the subject under treatment, whether swallowed or not. Thus, this includes oral and sublingual administration, as well as esophageal administration. Alternatively, the compositions of the present invention may also be formulated for rectal, topical, or parenteral (e.g., subcutaneous, intramuscular, intravenous, and intradermal or infusion) delivery. (Composition for oral delivery) When the composition of the present invention is used as an oral delivery composition, it is preferably a composition that enables the effective utilization of the omega-3 fatty acid and / or omega-3 fatty acid ester as the active ingredient. In particular, when the omega-3 fatty acid and / or omega-3 fatty acid ester as the active ingredient is administered to livestock, the unsaturated fatty acids contained in feed and roughage are rapidly decomposed into fatty acids and glycerol. Unsaturated fatty acids are rapidly hydrogenated by bacteria and protozoa in the rumen and converted into saturated fatty acids. Fatty acids flow into the lower digestive tract and are digested and absorbed. A part of the unsaturated fatty acids passes through the rumen without being hydrogenated and is absorbed in the small intestine. Therefore, in order to effectively utilize the omega-3 fatty acid and / or omega-3 fatty acid ester by oral ingestion, it is necessary to suppress the hydrogenation and decomposition of the orally ingested unsaturated fatty acids. When unsaturated fatty acids are administered to livestock such as dairy cows, the reduction of omega-3 fatty acids and / or omega-3 fatty acid esters caused by the decomposition action (rumen fermentation) of rumen microorganisms becomes a particular problem. As a means of minimizing the influence of rumen microorganisms, for example, the use of "bypass fats and oils" has been proposed (Osamu Furukawa, Forage and Horticulture, Vol. 53, No. 4, 2005). As bypass fats and oils, (1) calcium fatty acid salts and (2) hydrogenated fatty acids (including partial hydrogenation) have been proposed. Also, the fatty acid source is mainly palm oil. In addition, calcium fatty acid salts and triglyceride-type bypass fat and oil preparations can also be used. (1. Action of calcium fatty acid salt) When a fatty acid is combined with calcium, it has the property of remaining in a bound state in an alkaline environment and separating (dissociating) in an acidic environment. That is, in the rumen with a pH of about 6 to 7, it is separated and decomposed, and in the abomasum with a strong acidity of pH 2 to 3, it is separated and decomposed. As a result, the reduction of omega-3 fatty acids and / or omega-3 fatty acid esters caused by the decomposition action (rumen fermentation) of rumen microorganisms is suppressed. (2. Use of calcium fatty acid salt in feed) Even when using calcium fatty acid salts for the purpose of minimizing the impact of rumen fermentation in the first stomach, since high levels of oil and fat addition pose risks such as a decrease in dry matter intake, it is recommended to keep it within the level of 6 - 7% in this dry matter. Usually, when combining roughage and concentrated feed, the fat content is generally expected to be about 3%, so it can be expected that the remaining 3 - 4% can be added with an oil and fat source. At that time, when using cottonseed, heated soybeans, etc. and they have an adverse effect on milk fat percentage and dry matter intake, part of their use can be replaced with bypass oil and fat. In this case, about 450 g per head per day is considered the upper limit as the amount of fatty acids. (Food or dietary composition) The composition of the present invention can be used as a food or dietary composition. The pharmaceutical composition or food or dietary composition containing omega-3 fatty acids and / or omega-3 fatty acids as active ingredients can be directly used as liquid, gel or solid foods, such as juice, soft drink, coffee, black tea, Japanese green tea, oolong tea, vegetable juice, natural fruit juice, milk beverage, milk, soy milk, sports drink, near-water beverage, nutritional supplement drink, coffee drink, cocoa, soup, dressing, mousse, jelly, yogurt, pudding, furikake, powdered milk for infants, processed milk, sports drink, nutritional drink, cake mix, bread, pizza, pie, cracker, biscuit, cake, cookie, spaghetti, macaroni, pasta, udon, soba, ramen, candy, soft candy, gum, chocolate, okaki, potato chips, snack, ice cream, sherbet, cream, cheese, powdered milk, condensed milk, milk beverage and other powdery or liquid dairy products, steamed bun, uiro, mochi, ohagi, soy sauce, tare, noodle soup, sauce, dashi stock, stew base, soup base, compound seasoning, curry base, mayonnaise, ketchup, retort curry, retort stew, retort soup, retort donburi, canned food, ham, hamburger, meatball, croquette, dumpling, pilaf, onigiri, frozen food and refrigerated food, chikuwa, kamaboko, rice in bento, sushi, milk for infants, baby food, baby food, sports food, nutritional supplement food, supplement, health food, etc., or can be processed into pellets, tablets, granules, etc. together with excipients such as dextrin, lactose, starch, flavors, pigments, etc. as needed, or can be coated with gelatin, etc. and formed into capsules for use as health foods, nutritional supplement foods, etc. The blending amount of omega-3 fatty acids and / or omega-3 fatty acids in these food or food composition is not limited, but for example, it is 1% (w / w) to 60% (w / w), 3% (w / w) to 50% (w / w), 5% (w / w) to 40% (w / w), 10% (w / w) to 30% (w / w), or 15% (w / w) to 25% (w / w). The blending amount of DHA in these food or food composition is not limited, but for example, it is 1% (w / w) to 80% (w / w), 10% (w / w) to 70% (w / w), 20% (w / w) to 60% (w / w), or 30% (w / w) to 55% (w / w). When used in supplements, etc., the blending amount of the oxide of unsaturated fatty acids is typically 0.1% to 10% or more. When using the composition of the present invention as a supplement, the active ingredient may be used as it is, or it may be formulated into soft capsules, tablets, powders, granules, etc. and administered. When using the composition of the present invention in foods and drinks, it may be added to the raw materials of foods and drinks and administered. (Use as an infertility treatment agent) Preventing and / or treating chronic endometritis leads to the improvement of the state of the mother's uterus, and therefore, it is predicted to have the effect of enhancing the implantation efficiency. Also for this reason, preventing and / or treating chronic endometritis is considered to lead to infertility treatment. In addition, enhancing the implantation efficiency in livestock (Japanese black cattle and racehorses) leads to a reduction in the burden on livestock farmers.

Examples

[0030] The present invention will be described in detail below with reference to examples, etc., but the present invention is not limited thereto.

[0031] SREBP1 (sterol regulatory element-binding protein 1) is a transcription factor that controls lipid synthesis and fatty acid metabolism. SREBP1-deficient mice (Srebf1 - / -) In this case, the unsaturation and elongation of fatty acids are suppressed, and the content of omega-3 unsaturated fatty acids such as EPA in each tissue is decreased. In addition, compared with wild-type mice, SREBP1-deficient mice (Srebf1- / -) show a delay in systemic inflammation caused by sepsis, but the intake of a diet rich in EPA (5% EPA diet) can partially avoid the delay of inflammation (Oishi et al., Cell Metab. 7; 25(2): 412-427, 2017).

[0032] As a mechanism by which EPA ingested as a diet exhibits an anti-inflammatory effect, the inventors found that in addition to the anti-inflammatory effect of EPA itself, dietary-derived EPA is incorporated into the phospholipids of cell membranes and changes the composition, thereby regulating cell functions.

[0033] Based on these results, we hypothesized that a decrease in ω3 unsaturated fatty acids might be present in the background of the pathogenesis of recurrent implantation failure and recurrent miscarriage due to CE. For hypothesis verification, first, we established a CE model independently. Next, we applied the established model to SREBP1-deficient mice (Srebf1 - / - ) that exhibit chronic inflammation and confirmed that miscarriages increased. Furthermore, we found that when a preparation of EPA, one of the ω3 unsaturated fatty acids, was supplemented, the pathological condition of CE was significantly improved. These results indicate that the EPA preparation is effective as a new therapeutic agent for CE.

[0034] (Example 1: Preparation of CE model mice) After wild-type mice (WT) were fed a diet without fish meal for 7 days, they were laparotomized under anesthesia. Since fish meal contains unsaturated fatty acids such as EPA, a fish meal-free diet was used to align the baseline. Needles were inserted into both uterine horns, and after injecting physiological saline (NS) or 10 μg / 100 μL of LPS (lipopolysaccharide), which is a bacterial membrane component, the abdomen was closed (Figure 1).

[0035] On the 7th day after surgery, the uterus was removed and CD138 immunostaining was performed. As a result, the number of plasma cells in the endometrial stroma per section increased significantly by administering LPS into the uterus (NS (n = 9) vs LPS (n = 9): 0.7 ± 0.6 cells vs 3.4 ± 1.9 cells (p < 0.05)) (Figure 2).

[0036] (Example 2: Srebf1 - / - (Analysis of mice) In Srebf1-deficient mice that develop chronic inflammation, the endometrial stroma was evaluated under non-LPS administration. The number of plasma cells in the endometrial stroma per section was significantly increased compared to wild-type mice (WT (n = 9) vs knockout mice (KO) (n = 5): 0.7 ± 0.6 cells vs 5.0 ± 4.9 cells (p < 0.05)). That is, in SREBP1-deficient mice, chronic inflammation was considered to occur in the uterus even without LPS administration.

[0037] The CE model established in Example 1 was applied to and analyzed in SREBP1-deficient mice (Srebf1 - / - ). As a result, when LPS was administered, the number of plasma cells in the endometrial stroma per section increased significantly compared to WT (WT (n = 9) vs KO (n = 5): 3.4 ± 1.9 cells vs 10.7 ± 3.0 cells (p < 0.05)) (Figure 3).

[0038] Next, the CE model established in Example 1 was created in SREBP1-deficient mice (Srebf1 - / - ) and wild-type mice, and they were co-housed with male mice of the same species after the 7th day after surgery. The morning when ovulation and copulation were observed in female mice was set as days post coitum (dpc) 0.5, and the pregnant uterus at dpc 16.5 was observed. In mice in which CE was induced in SREBP1-deficient mice (Srebf1 - / - ), findings of abortion accompanied by hematoma formation were observed significantly more frequently compared to wild-type mice (WT (n = 5) vs KO (n = 3): 0.8 ± 0.7 individuals vs 4.0 ± 2.2 individuals (p < 0.05)) (Figure 4).

[0039] This result indicates that induction of CE in SREBP1-deficient mice with chronic inflammation frequently causes miscarriage.

[0040] (Example 3: Improvement effect of EPA supplementation on CE pathology) Wild-type mice and Srebf1-deficient mice were fed a fish meal-free diet or a fish meal-free diet supplemented with 5% EPA (v / v) for 1 week, and then the CE model established in Example 1 was created.

[0041] After 7 days, the plasma cells observed in the endometrial stroma per section were WT: fish meal-free (n = 9) vs. 5% EPA diet (n = 8): 3.4 ± 1.9 cells vs 0.3 ± 0.4 cells (p < 0.05), Srebf1-deficient: fish meal-free (n = 5) vs 5% EPA diet n = 4): 10.7 ± 3.0 cells vs 3.0 ± 0.5 cells (p < 0.05) (Figure 5). That is, by supplementing with EPA (EPA ethyl ester) as a diet, the appearance of the number of plasma cells reflecting the disease state of CE could be significantly reduced.

[0042] (Example 4: General anti-inflammatory effect of EPA) The anti-inflammatory effect of EPA preparations is well-known. Non-Patent Document 3 (Serhan et al., Nature. 5; 510(7503):92-101, 2014) discloses the anti-inflammatory action of EPA preparations and its mechanism. Also, JP-T-2005-532394 (Patent Document 1) and JP-A-2016-128482 (Patent Document 2) describe the improvement effect of EPA preparations on the inflammatory symptoms in endometritis, respectively. These evaluated general inflammatory markers (TNFα, IL-1b in Non-Patent Document 3, CRP, WBC in Patent Document 1, IFNγ, TNF-α, IL-12 in Patent Document 2).

[0043] In contrast, the inflammation of chronic endometritis (CE) is a "chronic inflammation" with a different nature from the "general inflammation" discussed in the above literature. In particular, in CE, the appearance of plasma cells in the lesion is significantly different from general inflammation. Therefore, from the above literature that only evaluates general inflammation markers, the anti-inflammatory effect of EPA preparations on the inflammation of CE is unpredictable. In other words, the improvement effect of EPA on CE found this time far exceeds the level predicted from the prior art.

[0044] Therefore, first, the following experiment showed that EPA reduces the expression of IL-1β after LPS administration in mouse uterine tissue and has an inhibitory effect on general inflammation.

[0045] Wild-type mice (WT) and Srebf1-deficient mice were fed a fish meal-free diet or a fish meal-free diet supplemented with 5% EPA (v / v) for 1 week. Then, in the same manner as the CE model described in the patent application, after injecting 10 mg / 100 mL of LPS into the uterus, the abdomen was closed. Six hours after administration, the uterus was removed, and the expression of IL-1β in the uterine tissue was evaluated by real-time PCR. As a result, the relative expression level of IL-1β tended to decrease with the addition of EPA in both WT and KO (Figure 6).

[0046] This result shows the general anti-inflammatory effect of EPA preparations in our experimental system and is predictable from the prior literature mentioned at the beginning.

[0047] (Example 5: Effect of EPA on CE) If the inflammation seen in CE is "general inflammation", ibuprofen, which is widely used as an anti-inflammatory drug, should also improve the pathological condition of CE. To verify this hypothesis, following Patent Document 2, the CE model was used to compare the anti-inflammatory effects of ibuprofen and EPA.

[0048] (1) Conventional anti-inflammatory agent (ibuprofen) does not suppress the appearance of plasma cells Wild-type mice were bred for one week with a fish meal-free diet and normal drinking water (EPA− / ibuprofen−), or a fish meal-free diet and drinking water containing ibuprofen (0.4 mg / mL) (EPA− / ibuprofen+), or a diet with 5% EPA (v / v) added to a fish meal-free diet and normal drinking water (EPA+ / ibuprofen−), and then CE model mice were created in the same manner as described in the patent application. Seven days later, the number of plasma cells observed in the endometrial stroma per section was as follows: EPA− / ibuprofen− (n = 9) vs EPA− / ibuprofen+ (n = 9) vs EPA+ / ibuprofen− (n = 8): 3.4 ± 1.9 cells vs 3.7 ± 2.7 cells vs 0.3 ± 0.4 cells (Figure 7).

[0049] From this, it was revealed that ibuprofen, a common anti-inflammatory agent, has no effect on suppressing the appearance of plasma cells in CE, while EPA has an effect on suppressing the appearance of plasma cells.

[0050] CE is diagnosed by the appearance of plasma cells in the endometrial stroma. Plasma cells are known to differentiate from B cells in response to the invasion of antigens such as viruses and bacteria and are mainly involved in humoral immunity, but the mechanism by which plasma cells appear in CE is unknown. Plasma cells are immune memory cells with properties different from general inflammatory markers, and CE characterized by their appearance has a pathological condition different from general inflammatory diseases.

[0051] (2) Miscarriage in the CE model was avoided by EPA intake Srebf1-deficient mice were bred for one week with a fish meal-free diet or a diet with 5% EPA (v / v) added to a fish meal-free diet, and then CE model mice were created in the same manner as described in the patent application. After the operation, they were co-housed with male mice of the same species, and the pregnant uterus at dpc16.5 was observed. The number of miscarriages observed in Srebf1-deficient mice fed a fish meal-free diet tended to decrease when fed a diet with 5% EPA added (EPA− (n = 3) vs EPA+ (n = 2): number of miscarriages 4.0 ± 2.2 vs 1.0 ± 0.5) (see Figure 8).

[0052] The above results shown in Figure 8 indicate that the EPA formulation improved miscarriages caused as a result of CE. Note that many cases of CE are asymptomatic and do not present symptoms such as general inflammatory conditions like fever or pain. Rather, implantation failure and miscarriage caused as a result of CE are the most clinically significant problems.

[0053] From the above, it became clear that the EPA formulation not only has the effect of reducing the number of plasma cells and suppressing chronic inflammation, but also suppressing miscarriages caused as a result of CE, not only for normal inflammatory markers (such as IL-1b). These results show excellent effects that cannot be predicted from the general anti-inflammatory effects of EPA formulations.

[0054] As described above, the present invention has been illustrated using preferred embodiments of the present invention, but the present invention should not be construed as being limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. Those skilled in the art will understand that equivalent ranges can be implemented based on the description of the present invention and common general knowledge from the description of the specific preferred embodiments of the present invention. It is understood that patents, patent applications, and documents cited in this specification should be incorporated by reference in their entirety as if the content itself were specifically described in this specification.

Industrial Applicability

[0055] The present invention provides non-human animals serving as models of chronic endometritis, as well as treatment methods, pharmaceutical compositions, and methods for manufacturing such medicaments for preventing and / or improving chronic endometritis. The present invention also provides food and dietary compositions.

Claims

1. A pharmaceutical composition for preventing and / or treating chronic endometritis, the pharmaceutical composition, food composition, or dietary composition comprising a substance selected from the group consisting of omega-3 fatty acids and omega-3 fatty acid esters.

2. 2. The pharmaceutical composition, food composition, or dietary composition of claim 1, wherein the omega-3 fatty acid is selected from the group consisting of alpha-linolenic acid (ALA), stearidonic acid (SDA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA).

3. The pharmaceutical composition, food composition, or dietary composition according to claim 2 , wherein the omega-3 fatty acid is eicosapentaenoic acid (EPA).

4. The pharmaceutical composition, food composition, or dietary composition according to claim 1 , wherein the omega-3 fatty acid is an omega-3 fatty acid salt.

5. The pharmaceutical composition, food composition, or dietary composition according to claim 4 , wherein the omega-3 fatty acid salt is an omega-3 fatty acid calcium salt.

6. The pharmaceutical composition, food composition, or dietary composition described in claim 1, wherein the omega-3 fatty acid ester is a substance selected from the group consisting of omega-3 fatty acid glycerides, alcohol esters of omega-3 fatty acids, and phospholipids containing omega-3 fatty acids.

7. The pharmaceutical composition, food composition, or dietary composition according to claim 1 , wherein the omega-3 fatty acid ester is an omega-3 fatty acid triglyceride.

8. The pharmaceutical composition, food composition, or dietary composition according to claim 1 , wherein the omega-3 fatty acid ester is an alcohol ester of an omega-3 fatty acid.

9. 2. The pharmaceutical composition, food composition, or dietary composition according to claim 1, wherein the omega-3 fatty acid ester is an ethyl ester of an omega-3 fatty acid.

10. 2. The pharmaceutical, beverage or feed composition according to claim 1, wherein the omega-3 fatty acid ester is an ethyl ester of eicosapentaenoic acid.

11. 2. The pharmaceutical composition, food composition, or dietary composition according to claim 1, which is taken orally.

12. A non-human animal model of chronic endometritis.

13. The non-human model animal of claim 12, which is a rodent.

14. The non-human model animal of claim 12, which is a mouse.

15. The non-human model animal of claim 12 , which is produced by administering LPS in utero.

16. The non-human model animal of claim 12 , which has a defect in the SREBP1 gene.

17. The non-human model animal according to claim 16 , which has a homozygous deficiency in the SREBP1 gene.

18. A method for producing a non-human animal model of chronic endometritis, comprising administering LPS into the uterus of the non-human animal.

19. A method for producing a non-human animal model of chronic endometritis, comprising administering LPS into the uterus of a non-human animal having a defect in the SREBP1 gene.

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