Phyllanthus emblica extract and its use with probiotics

Abstract

The application belongs to the technical field of food and medicine, and particularly relates to an emblica extract and application of the emblica extract and probiotics. The application discloses, for the first time, application of active ingredients in controlling fat in the body, maintaining or restoring a healthy level of blood fat, maintaining or restoring a healthy level of blood sugar, auxiliary protection or recovery of chemical liver injury and / or regulation of intestinal flora; wherein the active ingredients include a first active ingredient and / or a second active ingredient, the first active ingredient is an emblica extract, and the second active ingredient is probiotics, a fermentation culture thereof, a dried product thereof or a mixture thereof.

Description

Technical Field

[0001] This invention relates to the fields of food and pharmaceutical technology, specifically to Phyllanthus emblica extract and its application with probiotics. Background Technology

[0002] With societal development and the expansion of e-commerce, people have become accustomed to convenient lifestyles, leading to reduced travel and a gradual decline in bodily functions. Simultaneously, prolonged bed rest and sitting, increased work pressure, irregular sleep patterns, and high-fat, high-sugar diets can all cause abnormal blood sugar and lipid levels, damage and dysfunction of the liver and pancreatic β-cells, resulting in digestive and metabolic diseases such as fatty liver, hyperglycemia, and pancreatitis. Pancreatic β-cells, located in the central part of the islets of Langerhans, account for approximately 60%-80% of the total islet cells and are the core cells of the pancreatic endocrine system. The core function of pancreatic β-cells is to secrete insulin, and the liver is a key target organ for insulin's metabolic regulation. Blood lipids (lipid metabolism) are directly regulated by insulin; conversely, abnormal liver function or dyslipidemia can directly damage pancreatic β-cell function through metabolic feedback, forming a two-way "regulation-feedback" relationship. When pancreatic β-cell function declines and insulin secretion is insufficient, the above regulation fails, directly leading to dyslipidemia, the typical hyperlipidemia manifestation of "metabolic syndrome." Abnormalities in the liver (such as fatty liver) and blood lipids can, in turn, damage pancreatic β cells, forming a vicious cycle of "decreased function → organ damage → further decline in function".

[0003] Therefore, regulating / assisting in the regulation of abnormal blood sugar, lowering hyperglycemia, reducing the damage of hyperglycemia to organs such as the liver and pancreas, reducing visceral fat accumulation, regulating inflammatory states, and regulating intestinal metabolism have important practical significance and application value. Summary of the Invention

[0004] The first aspect of this invention aims to provide the application of the active ingredient.

[0005] A second aspect of the present invention is to provide a combination of active ingredients.

[0006] The third aspect of this invention is to provide a product.

[0007] The fourth aspect of this invention aims to provide a method.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A first aspect of the present invention provides the use of an active ingredient in the preparation of a product; The active ingredients include a first active ingredient, which is an extract of Phyllanthus emblica; The product is food, health supplement, or medicine; The food and health products mentioned above have one or more of the following functions: b1) Control body fat; b2) Maintain or restore healthy levels of blood lipids (e.g., cholesterol, triglycerides); b3) Maintain or restore healthy blood sugar levels; b4) It has an auxiliary protective or restorative effect on chemically induced liver injury; b5) Regulates gut microbiota; The drug has one or more of the following functions: a1) Lowering lipids; a2) Prevention or treatment of mitochondrial function-related diseases; a3) Prevention or treatment of liver diseases; a4) Prevention or treatment of pancreatic diseases; a5) Prevention or treatment of inflammatory diseases.

[0009] In this invention, the amla extract is an amla water extract.

[0010] In this invention, the amla extract is an alcohol extract.

[0011] In this invention, the amla extract can be commercially available or obtained by extracting amla using conventional plant extraction methods in the art.

[0012] In this invention, the preparation method of the amla extract includes the following steps: mixing amla with water or ethanol, extracting, and separating solid and liquid to obtain amla extract.

[0013] In this invention, the extraction conditions are: extraction at 60-120℃ for 10-120 min; further, extraction at 80-105℃ for 30-60 min.

[0014] In this invention, the mass ratio of Phyllanthus emblica to solvent (water or ethanol) is 1:(5-15); more specifically, it is 1:(10-12).

[0015] In this invention, the solid-liquid separation method is filtration; preferably, it is filtration using a 100nm ceramic membrane.

[0016] In this invention, the solid-liquid separation is followed by concentration.

[0017] In this invention, the concentration conditions are 50-70°C and a vacuum degree of 0.07-0.09 MPa.

[0018] In this invention, the concentration process further includes drying.

[0019] In this invention, the drying process involves drying to a constant weight.

[0020] In this invention, the drying is spray drying.

[0021] In this invention, the drying conditions are an inlet air temperature of 180-205°C and an outlet air temperature of 80-105°C.

[0022] For example, the present invention provides a method for preparing Phyllanthus emblica extract, comprising the following steps: Phyllanthus emblica extract, pulverization, hot water or hot ethanol extraction (greater than 60 degrees Celsius, 10-12 times the mass of water, extraction for 30-60 minutes), filtration (100nm ceramic membrane filtration), concentration (50-70 degrees Celsius, vacuum degree 0.07-0.09 MPa), spray drying (inlet air temperature 180-205 degrees Celsius, outlet air temperature 80-105 degrees Celsius), and sieving.

[0023] In this invention, the active ingredient further includes a second active ingredient, which is a probiotic, its fermentation culture, its dried product, or a mixture thereof.

[0024] In this invention, the ratio of the first active ingredient to the second active ingredient by mass to colony count is (5-20) mg: 5 x 10 mg. 9 CFU; further to (10-14) mg: 5 x 10 9 CFU, for example, 5mg: 5 x 10 9 CFU; 10mg: 5 x 10 9 CFU; 12mg: 5 x 10 9 CFU; 14mg: 5 x 10 9 CFU; 18mg: 5 x 10 9 CFU; 20mg: 5 x 10 9 CFU.

[0025] In this invention, the probiotics include Akkermansia myxophilus (… Akkermansia muciniphila ).

[0026] In this invention, the first active ingredient and the second active ingredient exist independently or in combination.

[0027] In this invention, the first active ingredient and the second active ingredient are applied simultaneously, separately, or sequentially.

[0028] In this invention, the dosage form of the product includes: solid dosage form, semi-solid dosage form, or liquid dosage form.

[0029] In this invention, the food or health product also includes excipients acceptable for use in food or health products.

[0030] In this invention, the excipients acceptable for use in food or health products include at least one of the following: wetting agents, emulsifiers, preservatives, antioxidants, buffers, diluents, lubricants, solvents, suspending agents, solubilizers, thickeners, stabilizers, sweeteners, and flavorings.

[0031] In this invention, the dosage form of the food or health product is a dosage form suitable for children or an adult.

[0032] In this invention, the dosage form of the food or health product is selected from oral dosage forms or non-oral dosage forms.

[0033] In this invention, the drug also includes pharmaceutically acceptable excipients.

[0034] In this invention, the pharmaceutically acceptable excipients include at least one of diluents, excipients, binders, humectants, surfactants, lubricants, and disintegrants.

[0035] In this invention, the dosage form of the drug is either a dosage form suitable for children or a dosage form suitable for adults.

[0036] In this invention, the dosage form of the drug is selected from gastrointestinal dosage forms or non-gastrointestinal dosage forms.

[0037] In this invention, the gastrointestinal dosage form includes at least one of the following: powder, tablet, granule, capsule, sustained-release, solution, dry suspension, effervescent tablet, emulsion, suspension, syrup, drops, and chewable tablet.

[0038] In this invention, the non-gastrointestinal drug delivery dosage forms include at least one of the following: injectable dosage forms (e.g., injections, including various injections such as intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, and intracavitary injections); respiratory dosage forms (e.g., sprays, aerosols, powder inhalers, etc.); skin dosage forms (e.g., topical solutions, lotions, liniments, ointments, plasters, pastes, patches, etc.); mucosal dosage forms (e.g., eye drops, nasal drops, ophthalmic ointments, mouthwashes, sublingual tablets, adhesive tablets, films, etc.); and cavity dosage forms (e.g., suppositories, aerosols, effervescent tablets, drops, pills, etc., for use in the rectum, vagina, urethra, nasal cavity, ear canal, etc.).

[0039] In this invention, the product is applied to animals.

[0040] In this invention, the animal is a mammal; further selected from humans, non-human primates (e.g., orangutans, apes), rodents (e.g., rats, mice, guinea pigs), pets (e.g., cats, dogs), livestock (e.g., horses, cattle, sheep, pigs, rabbits); and further selected from humans.

[0041] In this invention, the lipid-lowering method is selected from reducing visceral fat and / or local fat in obese individuals.

[0042] In this invention, the mitochondrial function-related diseases include mitochondrial function-related diseases caused by diabetes.

[0043] In this invention, the mitochondrial function-related diseases include those caused by a high-fat diet and diabetes.

[0044] In this invention, the mitochondrial function-related diseases include, but are not limited to, at least one of metabolic diseases, nervous system diseases, aging, and muscle diseases.

[0045] In this invention, the metabolic disease is a metabolic disorder, metabolic disturbance, or a disease caused by a metabolic disturbance, including but not limited to: obesity and obesity-related diseases, diabetes, dyslipidemia, glucose intolerance, impaired glucose tolerance, hyperglycemia, hyperinsulinemia, hyperlipoproteinemia, hypertriglyceridemia, uremia, ketoacidosis, hypoglycemia, thrombotic diseases, non-alcoholic fatty liver disease (NAFLD), and non-alcoholic steatohepatitis (NASH).

[0046] In this invention, the causes of the metabolic diseases include, but are not limited to, at least one of the following: diabetes, high-fat diet, high-cholesterol diet, high-sugar diet, high blood lipids, high blood sugar, or high cholesterol.

[0047] In this invention, obesity and obesity-related diseases include, but are not limited to, at least one of the following: overweight, obesity, metabolic syndrome, cardiovascular and cerebrovascular diseases, hyperlipidemia, hypercholesterolemia, hypertension, insulin resistance syndrome, obesity-related gastroesophageal reflux disease, and steatohepatitis.

[0048] In this invention, obesity and obesity-related diseases include, but are not limited to: obesity caused by a high-fat diet, obesity caused by high cholesterol, obesity caused by a high-sugar diet, obesity caused by high fat and high cholesterol, obesity caused by high fat and high sugar, obesity caused by high fat, high cholesterol and high sugar, or obesity in patients with NAFLD or NASH.

[0049] In this invention, the obesity is peripheral obesity and / or central obesity. In this invention, the obesity is diet-induced obesity and / or metabolic obesity. In particular, the obesity is abdominal obesity or apple-shaped obesity, such as excessive visceral fat.

[0050] In this invention, the diabetes includes, but is not limited to: type 1 diabetes, type 2 diabetes, gestational diabetes, HDAC activity-mediated diabetes, diabetic neuropathy, diabetic retinopathy, diabetic foot, diabetes caused by pancreatic β-cell damage, diabetes caused by insulin resistance, and diabetes caused by obesity.

[0051] In this invention, the causes of diabetes include, but are not limited to: pancreatic islet cell dysfunction, decreased insulin secretion, increased insulin resistance, high-fat diet, high-sugar diet, high cholesterol, high blood lipids, high blood sugar, NAFLD or NASH.

[0052] In this invention, the cardiovascular disease or cerebrovascular disease includes, but is not limited to: atherosclerosis, coronary heart disease, hypertension, cardiovascular disease in patients with NAFLD or NASH, cerebrovascular disease in patients with NAFLD or NASH, and high cholesterol disease.

[0053] In this invention, the metabolic disease is selected from obesity, diabetes, hyperglycemia, hypercholesterolemia, hyperlipidemia, metabolic syndrome caused by a high-fat diet, glucose intolerance caused by a high-fat diet, lipid metabolism abnormalities caused by a high-fat diet, and dyslipidemia caused by a high-fat diet.

[0054] In this invention, the metabolic diseases are selected from obesity, diabetes, hyperglycemia, hypercholesterolemia, hyperlipidemia, high-fat diet and metabolic syndrome caused by diabetes, high-fat diet and glucose intolerance caused by diabetes, high-fat diet and lipid metabolism abnormalities caused by diabetes, and high-fat diet and dyslipidemia caused by diabetes.

[0055] In this invention, the neurological diseases include, but are not limited to, at least one of the following: Friedreich ataxia, amyotrophic lateral sclerosis (ALS), adrenoleukodystrophy, Alexander disease, Alpert disease, familial fatal insomnia, Huntington's disease, Kennedy's disease, Crabbe's disease, Lyme disease, Machado-Joseph disease, multiple sclerosis, multiple system atrophy, Niemann-Pick disease, Pick's disease, primary lateral sclerosis, progressive supranuclear palsy, Rafe-Syme disease, Sandhoff's disease, diffuse demyelinating sclerosis, spinocerebellar ataxia, subacute mixed spinal cord degeneration, toxic encephalopathy, disseminated spongiform encephalopathy, and ataxia-telangiectasia.

[0056] In this invention, the muscle diseases include, but are not limited to, sarcopenia and / or Duchenne muscular dystrophy.

[0057] In this invention, the liver disease includes liver disease caused by diabetes.

[0058] In this invention, the liver disease includes liver disease caused by a high-fat diet and diabetes.

[0059] In this invention, the liver diseases include, but are not limited to, at least one of: fatty liver, NAFLD (non-alcoholic fatty liver disease), NASH (non-alcoholic steatohepatitis), abnormal liver function, extrahepatic cholestasis, hepatitis, liver injury, intrahepatic cholestasis, liver fibrosis, cirrhosis, and liver cell damage.

[0060] In this invention, the liver injury can be caused by prolonged strenuous work; liver injury caused by long-term alcohol consumption; fatty liver, drug-induced liver disease, or liver injury caused by genetic metabolic factors.

[0061] In this invention, the liver disease is selected from non-alcoholic steatohepatitis (NASH) caused by a high-fat diet and diabetes, non-alcoholic fatty liver disease (NAFLD) caused by a high-fat diet and diabetes, hepatitis caused by a high-fat diet and diabetes, abnormal liver function caused by a high-fat diet and diabetes, liver fibrosis caused by a high-fat diet and diabetes, and liver damage (e.g., hepatocyte damage) caused by a high-fat diet and diabetes.

[0062] In this invention, the pancreatic disease includes pancreatic diseases caused by diabetes.

[0063] In this invention, the pancreatic diseases include those caused by a high-fat diet and those caused by diabetes.

[0064] In this invention, the pancreatic diseases include, but are not limited to, at least one of: pancreatitis (e.g., acute pancreatitis, chronic pancreatitis), pancreatic fibrosis, and pancreatic cancer.

[0065] In this invention, the causes of pancreatic diseases include, but are not limited to, at least one of the following: high-fat diet, high-cholesterol diet, high-sugar diet, high blood lipids, high blood sugar or high cholesterol, and diabetes.

[0066] In this invention, the inflammatory disease includes inflammatory diseases caused by diabetes.

[0067] In this invention, the inflammatory diseases include those caused by a high-fat diet and diabetes.

[0068] In this invention, the inflammatory disease is selected from inflammation related to metabolic diseases, such as obesity-related inflammation; diseases related to liver inflammation, such as hepatitis (e.g., alcoholic hepatitis, fatty liver disease, autoimmune hepatitis, drug-induced hepatitis); diseases related to pancreatic inflammation, such as pancreatitis (e.g., biliary pancreatitis, hyperlipidemic pancreatitis, alcoholic pancreatitis, autoimmune pancreatitis, drug-induced pancreatitis, traumatic pancreatitis, hereditary / idiopathic pancreatitis); diseases related to bronchial inflammation, such as bronchitis; and diseases related to cervical inflammation, such as... Cervicitis; conjunctivitis; esophageal inflammation; myocarditis; proctitis; scleritis; gum inflammation; bone inflammation; lung inflammation (alveolitis); respiratory tract inflammation (e.g., bronchial asthma); acute respiratory distress syndrome (ARDS); inflammatory skin diseases (e.g., contact allergy, atopic dermatitis); fibrotic diseases (e.g., pulmonary fibrosis); encephalitis.

[0069] A second aspect of the present invention provides an active ingredient combination comprising the first active ingredient and the second active ingredient of the first aspect of the present invention.

[0070] In this invention, the ratio of the first active ingredient to the second active ingredient by mass to colony count is (5-20) mg: 5 x 10 mg. 9 CFU; further to (10-14) mg: 5 x 10 9 CFU, for example, 5mg: 5 x 10 9 CFU; 10mg: 5 x 10 9 CFU; 12mg: 5 x 10 9 CFU; 14mg: 5 x 10 9 CFU; 18mg: 5 x 10 9 CFU; 20mg: 5 x 10 9 CFU.

[0071] For example, a combination of active ingredients or a product comprising 12 mg of Phyllanthus emblica extract and 5 x 10 9 CFU (Ackermansia muciniphila) Akkermansia muciniphila ).

[0072] A third aspect of the invention provides a product comprising a combination of active ingredients from the second aspect of the invention.

[0073] In some embodiments, the product is food, health supplement, or medicine.

[0074] In some embodiments, the product further includes food or health food-grade or pharmaceutically acceptable excipients.

[0075] A fourth aspect of the invention provides a method for administering to a subject an effective amount of the product of the first aspect of the invention, or the product of the third aspect of the invention, or a combination of active ingredients of the second aspect of the invention; The method has one or more of the following functions: b1) Control body fat; b2) Maintain or restore healthy levels of blood lipids (e.g., cholesterol, triglycerides); b3) Maintain or restore healthy blood sugar levels; b4) It has an auxiliary protective or restorative effect on chemically induced liver injury; b5) Regulating gut microbiota; or The method has one or more of the following functions: a1) Lowering lipids; a2) Prevention or treatment of mitochondrial function-related diseases; a3) Prevention or treatment of liver diseases; a4) Prevention or treatment of pancreatic diseases; a5) Prevention or treatment of inflammatory diseases.

[0076] In some implementations, the method does not involve the diagnosis or treatment of the disease.

[0077] In some implementations, the subject may include mammals, such as humans or non-human mammals.

[0078] In some embodiments, the non-human mammals may include, but are not limited to, non-human primates (e.g., monkeys, orangutans), mice, rats, hamsters, gerbils, cats, dogs, guinea pigs, rabbits, horses, sheep, cattle, pigs, etc.

[0079] The beneficial effects of this invention are: This invention discloses for the first time the application of active ingredients in controlling body fat, maintaining or restoring healthy blood lipid levels, maintaining or restoring healthy blood sugar levels, assisting in the protection or recovery of chemically induced liver damage, regulating intestinal flora, lowering lipids, and preventing or treating mitochondrial function-related diseases, liver diseases, pancreatic diseases, and inflammatory diseases; wherein, the active ingredients include a first active ingredient, which is Phyllanthus emblica extract.

[0080] Furthermore, the active ingredient further includes a second active ingredient, which is a probiotic, its fermentation culture, its dried product, or a mixture thereof; wherein, for the first time, it has been discovered that the first and second active ingredients have a synergistic effect in controlling body fat, maintaining or restoring healthy blood lipid levels, maintaining or restoring healthy blood glucose levels, assisting in the protection or recovery of chemically induced liver damage, regulating intestinal flora, lowering lipids, and preventing or treating mitochondrial function-related diseases, liver diseases, pancreatic diseases, and inflammatory diseases. Attached Figure Description

[0081] Figure 1 The diagram shows the construction and experimental procedure of a type 2 diabetes mouse model.

[0082] Figure 2 The body weights of mice under different treatments are shown.

[0083] Figure 3 The weight of visceral fat in mice under different treatments is shown.

[0084] Figure 4 TEM results of pancreatic β cells from mice under different treatments are shown.

[0085] Figure 5 The results of mitochondrial-associated functional protein (MFN2) expression detection in pancreatic tissues of mice under different treatments are shown: A shows the results of MFN2 expression detection in pancreatic tissues of mice under different treatments; B shows the results of MFN1 expression detection in pancreatic tissues of mice under different treatments; C shows the results of MFN2 expression detection in pancreatic tissues of mice under different treatments.

[0086] Figure 6 The concentrations of fasting blood glucose (FBG) in mice under different treatments are shown.

[0087] Figure 7 The concentrations of glycated serum protein (GSP) in mice under different treatments are shown.

[0088] Figure 8 The concentrations of triglycerides (TG) in mice under different treatments are shown.

[0089] Figure 9 The concentrations of total cholesterol (TC) in mice under different treatments are shown.

[0090] Figure 10 The concentrations of low-density lipoprotein cholesterol (LDLC) in mice under different treatments are shown.

[0091] Figure 11The concentrations of high-density lipoprotein cholesterol (HDLC) in mice under different treatments are shown.

[0092] Figure 12 The blood glucose levels and their corresponding areas under the curves for mice under different treatments at 0, 15, 30, 60, and 120 min after glucose injection are shown.

[0093] Figure 13 The blood glucose levels and their corresponding areas under the curves for mice under different treatments at 0, 15, 30, 60, and 120 min after insulin injection are shown.

[0094] Figure 14 The concentrations of the pro-inflammatory factor tumor necrosis factor-α in mice under different treatments are shown.

[0095] Figure 15 The concentrations of interleukin-6, a pro-inflammatory factor, in mice under different treatments are shown.

[0096] Figure 16 The concentrations of the anti-inflammatory factor interleukin-10 in mice under different treatments are shown.

[0097] Figure 17 The concentrations of interleukin-4, an anti-inflammatory factor, are shown in mice under different treatments.

[0098] Figure 18 The specific activity of alanine aminotransferase (ALT) in mice under different treatments is shown.

[0099] Figure 19 The specific activity of aspartate aminotransferase (AST) in mice under different treatments is shown.

[0100] Figure 20 The liver weight index of mice under different treatments is shown.

[0101] Figure 21 The concentrations of triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDLC) in the livers of mice under different treatments are shown: A shows the concentration of triglycerides (TG) in the livers of mice under different treatments; B shows the concentration of total cholesterol (TC) in the livers of mice under different treatments; and C shows the concentration of low-density lipoprotein cholesterol (LDLC) in the livers of mice under different treatments.

[0102] Figure 22 The pancreatic weight index of mice under different treatments is shown.

[0103] Figure 23 HE staining results of pancreatic tissue from mice under different treatments are shown.

[0104] Figure 24 The results of TUNEL staining of pancreatic tissue from mice under different treatments are shown.

[0105] Figure 25 The results of gut microbiota analysis in mice under different treatments are shown. Detailed Implementation

[0106] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention in any way. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concept of the invention. Such structures and techniques have also been described in many publications.

[0107] definition Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0108] Unless the context clearly indicates otherwise, the terms “a” and “an” as used herein include plural references. For example, reference to “a cell” includes multiple such cells and equivalents known to those skilled in the art, etc.

[0109] As used herein, the term "about" indicates a range of ±20% of the following value. In some embodiments, the term "about" indicates a range of ±10% of the following value. In some embodiments, the term "about" indicates a range of ±5% of the following value.

[0110] Those skilled in the art will understand that when a numerical range is listed herein, it is intended to indicate that any intermediate value or range of values ​​within the listed numerical range is also within the scope of this invention.

[0111] As used herein, the terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations thereof, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0112] The term "maintain" as used in this article refers to the continuous stabilization of various physiological indicators such as blood lipids, blood sugar, gut microbiota, liver status, and body fat metabolism within the healthy and normal range. This method aims to prevent abnormal increases or decreases in these indicators and to prevent metabolic disorders, liver damage, gut microbiota imbalance, and excessive fat accumulation.

[0113] The term "recovery" used in this article refers to the process by which this method reverses various indicators that deviate from healthy standards when the body has already experienced abnormal states such as high blood lipids, abnormal blood sugar, chemical liver damage, intestinal flora imbalance, and excessive accumulation of body fat, thereby restoring damaged metabolism, liver status, and flora structure to normal healthy levels.

[0114] In this paper, the ratio of the first active ingredient to the second active ingredient is expressed as the ratio of the mass of the first active ingredient to the colony count of the second active ingredient as (5–20) mg: 5 × 10⁻⁶ mg. 9 CFU, this ratio is a proportional relationship. For example, if the colony count of the second active ingredient is 5 × 10⁻⁶... 8 CFU or 5×10 10 CFU corresponds to a mass of 0.5mg–2mg or 50mg–200mg of the first active ingredient, and the above proportions all fall within the protection scope of this invention. For example, an active ingredient combination or product includes 50mg of Phyllanthus emblica extract and 5 x 10 10 CFU (Ackermansia muciniphila) Akkermansia muciniphila ).

[0115] In this document, the food, health product / method / application / product / combination described has one or more of the following functions: b1) Helps control body fat; b2) Helps maintain healthy blood lipid levels (such as cholesterol and triglycerides); b3) Helps maintain healthy blood sugar levels; b4) It has an auxiliary protective effect against chemical liver injury; b5) Regulate the gut microbiota.

[0116] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise specified, the materials and reagents used in these examples are commercially available. For reagents whose manufacturers are listed, similar products from other manufacturers are substituted.

[0117] The experimental methods involved in the embodiments of this invention are as follows: I. Preparation of Phyllanthus emblica extract The preparation method of Phyllanthus emblica extract includes the following steps: Phyllanthus emblica extract, pulverization, hot water extraction (95 degrees Celsius, 10 times the mass of water, extraction for 60 min), filtration (100 nm ceramic membrane filtration), concentration (60 degrees Celsius, vacuum degree 0.07 MPa), spray drying (inlet air temperature 200 degrees Celsius, outlet air temperature 100 degrees Celsius), and sieving.

[0118] II. Verification of the effects of Phyllanthus emblica extract and its interaction with probiotics The experimental methods involved in Examples 1-7 are as follows: 1. Laboratory animals Sixty 5-week-old male SPF-grade C57BL / 6J mice, weighing 16-20 g, were used. After 7 days of acclimatization feeding, the mice were randomly divided into a normal control group (n=10) and a disease model group (n=50) for subsequent modeling and feeding experiments.

[0119] 1.1. Construction of a type 2 diabetic mouse model Modeling process as follows Figure 1 As shown, male C57BL / 6J mice were grouped and acclimatized for one week under a strict 12-hour light-12-hour dark environment, constant temperature of 20-22 ℃, and relative humidity of 65%-70% before the experiment began. Type 2 diabetes model mice were induced by a combination of a high-fat diet and streptozotocin (STZ). The disease model group was fed a 60% high-fat diet (Jiangsu Xietong Biotechnology, batch number XTHF60), while the normal control group was fed a normal maintenance diet (Jiangsu Xietong Biotechnology, batch number XTCON50J). Six weeks later (week 8), mice in each group were fasted but allowed free access to water for 12 hours. Mice in the model group were intraperitoneally injected with STZ (40 mg / kg, dissolved in 0.1 mM sodium citrate buffer, pH=4.4, protected from light and kept on ice, prepared fresh and used within 30 minutes) for 3 consecutive days. Mice in the normal control group were intraperitoneally injected with the same volume of 0.1 mM sodium citrate buffer. One week later (week 9, based on the first STZ injection), fasting blood glucose (FBG) values ​​were measured using a blood glucose meter by tail reduction. Mice with a fasting blood glucose value ≥11.1 mmol / L were considered to have successfully modeled type 2 diabetes.

[0120] 1.2. Experimental Grouping and Processing (1) Blank control group (CON group, n=10 animals, ig (gavage administration). Equal volume of distilled water): fed with ordinary maintenance feed and free access to water.

[0121] (2) Type 2 diabetes model group (DM group, n=10, ig equal volume of distilled water): fed with 60% high-fat diet and free access to water.

[0122] (3) Metformin positive control group (MET group, n=10, ig 300 mg / kg): fed with 60% high-fat diet and free access to water.

[0123] (4) Phyllanthus emblica extract group (PEL group, n=10 animals, ig 400 mg / kg, Phyllanthus emblica extract obtained from “I. Preparation of Phyllanthus emblica extract”): fed with 60% high-fat diet and free access to water.

[0124] (5) Probiotic group A (PA group, n=10, ig 5 x 10) 9 CFU AKK): Feed with 60% high-fat feed and free access to water.

[0125] (6) Phyllanthus emblica extract + probiotic A group (PEL+PA group or PELP group, n=10, ig 400 mg / kg + 5 x 10 9 CFU AKK): Feed with 60% high-fat feed and free access to water.

[0126] Note: 1) After the diabetic mouse model was successfully established (week 9), all groups were fed for 10 weeks (i.e., until week 18).

[0127] 2) The gavage dosage for all groups should be adjusted according to the dynamic changes in body weight.

[0128] 3). In groups (5) and (6), each mouse was given 100 mg of bacterial powder / 20 g by gavage daily (of which, the bacterial powder of probiotic A was AKK live bacterial powder, purchased from Guangdong Qingyunshan Traditional Chinese Medicine Innovation Research and Development Co., Ltd.), and 100 mg / 100 uL bacterial powder solution was prepared and given 100 uL / 20 g by gavage. The bacterial solution was prepared and used immediately, and the dosage was dynamically adjusted according to the body weight.

[0129] 2. Mouse weight measurement (once a week after the experiment officially begins (week 9)).

[0130] 3. At the end of the experiment (at the end of week 18), the weight of the mouse liver, pancreas, and visceral adipose tissue were measured.

[0131] 4. Measure peripheral glucose and lipid metabolism indicators, liver indicators, and inflammatory markers. (1). Fasting blood glucose (FBG): After the experiment officially started (week 9), blood was collected from the tail tip of the mice and fasting blood glucose was measured once a week.

[0132] (2). Glycated serum protein (GSP): After the experiment (at the end of week 18), blood was collected from the eyeballs to measure the level of glycated serum protein in the mouse serum.

[0133] (3). Four lipid parameters: After the experiment (at the end of week 18), blood was collected from the eyeballs to measure the levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), and low-density lipoprotein cholesterol (LDLC) in the mouse serum.

[0134] (4) Liver function indicators: After the experiment (at the end of week 18), blood was collected from the eyeballs to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the mouse serum.

[0135] (5) Inflammatory markers: After the experiment (at the end of week 18), blood was collected from the eyeballs to measure the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-4 (IL-4), and interleukin-10 (IL-10) in the serum of mice.

[0136] 5. Assay for pancreatic β-cell function (1) Intraperitoneal glucose tolerance test (IPGTT): After ten consecutive weeks of administration (i.e., at the end of week 18), the intraperitoneal glucose tolerance test (IPGTT) was performed. All mice were fasted for 12 h before the test but allowed free water. After intraperitoneal injection of 2 g / kg glucose, blood glucose levels were monitored at 0, 15, 30, 60, and 120 min, and the area under the curve was recorded and calculated.

[0137] (2). Insulin Tolerance Test (ITT): After ten weeks of continuous administration (i.e., at the end of week 18), the insulin tolerance test (ITT) was performed. Mice were fasted for 6 hours, and fasting blood glucose was measured first, followed by insulin injection (0.75 U / kg). Blood glucose values ​​were measured at 0, 15, 30, 60, and 120 min after injection. The area under the curve was calculated by plotting the curve of blood glucose concentration over time.

[0138] 6. Immunohistochemical staining of pancreatic tissue HE staining: Immunohistochemical staining of pancreatic tissue was performed at the end of the experiment (at the end of week 18, i.e., week 19). Fresh mouse pancreatic tissue was removed and fixed in 4% paraformaldehyde for at least 24 hours. After removal, the tissue was placed in a fume hood, trimmed with a scalpel, labeled accordingly, and dehydrated. The tissue was then immersed in different concentrations of alcohol and anhydrous ethanol. After the paraffin was melted, the paraffin-impregnated pancreatic tissue was placed in a specialized embedding machine for cooling and trimming. Finally, it was cut to a thickness of approximately 4 μm. The tissue sections were placed in 40°C warm water to float and flatten them. The sections were removed using a glass slide, air-dried, and placed at room temperature. The pancreatic tissue sections were dewaxed with xylene, anhydrous ethanol, and alcohol. Hippocampal samples were dehydrated, embedded in paraffin, frozen, and dewaxed. The sections were then immersed in anhydrous ethanol, 95% alcohol, and 85% alcohol for 30 seconds, 2 minutes, and 2 minutes respectively. Stain with hematoxylin for 4 minutes, wash with tap water, then differentiate, re-blue, and rinse. Finally, dehydrate in 85% and 95% alcohol solutions, stain with eosin for 5 minutes, mount, examine under a microscope, and finally acquire images for analysis.

[0139] 7. TUNEL apoptosis detection of pancreatic β cells TUNEL staining: At the end of the experiment (at the end of week 18, i.e., week 19), TUNEL apoptosis detection of pancreatic β-cells was performed. Mouse pancreatic tissue samples were fixed and dehydrated with 4% paraformaldehyde, then embedded in paraffin and sectioned. Next, after dewaxing, hydration, and enzymatic digestion, the samples were labeled with terminal transferase-mediated dUTP using a TUNEL assay kit and incubated at 37 °C. Finally, the labeling signal was observed under a microscope to identify apoptotic cells.

[0140] 8. Ultrastructural observation of pancreatic β cells (observation of mitochondrial cristae in mouse pancreatic β cells) At the end of the experiment (at the end of week 18, i.e. week 19), ultrastructural observation of pancreatic β cells was performed. After pancreatic tissue dissection, a 1 mm sample of pancreatic tail tissue was taken. 3 The sample was fixed overnight in 2.5% glutaraldehyde, and the PBS buffer was changed the next day. It was then fixed with osmium tetroxide, followed by gradient dehydration, infiltration embedding, trimming, sectioning, and staining. The sample was then observed and images were acquired using a transmission electron microscope.

[0141] 9. Detection of expression of mitochondrial-related functional proteins in pancreatic tissue (1). Western blot: mitochondrial fusion protein MFN2.

[0142] (2). Western blot: mitochondrial fission proteins Drp1 and MFF.

[0143] (3). Western blot: mitochondrial internal reference COXIV.

[0144] The relative expression levels of Drp1 / COXIV, MFF / COXIV, and MFN2 / COXIV were calculated.

[0145] At the end of the experiment (at the end of week 18, i.e., week 19), the expression of mitochondrial-related functional proteins in pancreatic tissue was detected. Proteins were extracted from the pancreas and quantified. After electrophoresis on an SDS-PAGE gel, the samples were transferred to PVDF or nitrocellulose membranes. The membranes were then blocked with TBST buffer containing 5% nonfat milk powder or BSA to reduce nonspecific binding, and incubated overnight with primary antibodies against Drp1, MFN2, MFF, and COXIV. After washing away unbound primary antibodies, the membranes were incubated with HRP-labeled secondary antibodies and developed using chemiluminescent substrates. Finally, the density of the target protein bands relative to the density of the internal control COXIV band was recorded and analyzed using an imaging system to assess the expression levels of Drp1, MFN2, and MFF.

[0146] 10. Gut microbiota analysis After ten weeks of continuous administration (i.e., at the end of week 18), fecal samples were collected from animals, and bacterial genomic DNA was extracted from the fecal samples using the QIAamp® DNAStool Mini Kit (Qiagen, Germany) according to the manufacturer's instructions. DNA quality, including purity and integrity, was assessed by agarose gel electrophoresis and a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Acceptable DNA samples were sonicated to an average length of approximately 350 bp for sequencing library preparation. Library quality and concentration were assessed by Qubit fluorescence assay and RT-qPCR, followed by sequencing on an IlluminaNovaSeq 6000 platform by Novogene Technology Co., Ltd. (Beijing, China). Raw sequencing data were denoised and amplicon sequence variants (ASVs) were generated using the DADA2 plugin in QIIME2 software (version 2022.02), followed by classification and annotation in QIIME2. At the genus level, the relative abundance levels of beneficial and harmful bacteria in each group of samples were analyzed.

[0147] 11. Synergistic Efficiency Calculation Model Based on the experimental results, the highest single-drug effect model was used to calculate the indicators of synergistic effect.

[0148] (1) Calculation of synergistic effects of reduced indicators: For indicators such as FBG, GSP, IPGTT AUC, ITT AUC, TG, TC, LDL-C, TNF-α, IL-6, ALT, and AST, where a decrease in value indicates improvement, the following formula is used: ΔHSA=min(PEL, PA)-PELP Wherein, PEL represents the mean index of the group treated with Phyllanthus emblica alone, PA represents the mean index of the group treated with AKK live bacteria alone, and PELP represents the mean index of the group treated with combined intervention.

[0149] When ΔHSA>0, it indicates that PELP is lower than the best single-drug group among the two single-drug groups, suggesting that the combined intervention is superior to either single intervention and has a combined enhancement effect or synergistic effect.

[0150] (2) Calculation of synergistic effect of rising indicators For indicators such as HDL-C, IL-10, and IL-4, where "increased values ​​represent improvement," the following formula is used: ΔHSA = PELP - max(PEL, PA) When ΔHSA>0, it indicates that PELP is higher than the best single-drug group among the two single-drug groups, suggesting that the combined intervention is superior to either single intervention.

[0151] Example 1: Phyllanthus emblica extract and its effect on reducing visceral fat with probiotics At the end of the experiment (at the end of week 18), the weight of visceral adipose tissue in mice under different treatments was as follows: Figure 3 As shown: Compared with the blank control group, the visceral fat weight of the model group was significantly increased, indicating that the high-fat diet successfully induced obesity and visceral fat accumulation in rats; while the treatment with amla extract or its combination with probiotics significantly reduced visceral fat weight, improved fat accumulation induced by the high-fat diet, and thus improved obesity, and its effect was comparable to that of the metformin positive control group; and the visceral fat weight after treatment with amla extract and probiotics was lower than that after treatment with amla extract or probiotics alone, indicating that amla extract and probiotics have a synergistic effect in reducing visceral fat.

[0152] Example 2: The mitochondrial repair effect of Phyllanthus emblica extract and its combination with probiotics (1). Observation of the ultrastructure of pancreatic β cells (observation of mitochondrial cristae in mouse pancreatic β cells) At the end of the experiment (at the end of week 18), the TEM results of pancreatic β cells in mice under different treatments were as follows: Figure 4As shown: In the blank control group, the mitochondrial cristae were neatly and densely arranged, with regular orientation, perpendicular or parallel to the long axis of the mitochondria, and the matrix electron density was uniform; in the model group, the mitochondria were significantly larger and rounder, the matrix was lighter (electron density decreased), the mitochondrial cristae were disordered, shorter, reduced or even disappeared, and vacuolization occurred; while the use of Phyllanthus emblica extract or its combination with probiotics can improve the morphology and cristae of mitochondria and repair mitochondria.

[0153] (2) Detection of expression of mitochondrial-related functional proteins and proteins related to mitochondrial dynamics signaling pathways in pancreatic tissue. The expression results of mitochondrial fission proteins and proteins related to mitochondrial dynamics signaling pathways in pancreatic tissue are as follows: Figure 5 As shown, compared with the blank control group (CON), the expression of MFN2 protein in the pancreatic tissue of diabetic model mice (DM) was significantly decreased, while the expression of DRP1 and MFF proteins was significantly increased, suggesting that the mitochondrial dynamics balance in pancreatic tissue is disrupted in type 2 diabetes, manifested as inhibited mitochondrial fusion and enhanced mitochondrial division. Metformin (MET) significantly reversed the above abnormal changes, showing increased MFN2 expression and decreased DRP1 and MFF expression. PEL intervention alone significantly increased MFN2 expression and decreased DRP1 and MFF levels, suggesting that PEL has a certain role in improving mitochondrial dynamics disorders. PA intervention alone can also upregulate MFN2 and downregulate DRP1 and MFF to some extent, but its overall effect is weaker than that of PEL.

[0154] Although the PELP group did not significantly restore MFN2 expression, suggesting its limited effect in promoting mitochondrial fusion, its inhibitory effect on mitochondrial division-related proteins was more pronounced. Compared with the PEL and PA groups alone, DRP1 expression in the PELP group was further reduced, and statistical grouping showed differences compared to the PEL and PA groups, indicating that the combined intervention of PELP was more effective than single-component intervention in inhibiting DRP1-mediated mitochondrial division, suggesting that Phyllanthus emblica extract and probiotics have a synergistic effect in inhibiting DRP1-mediated mitochondrial division. Meanwhile, the MFF expression level in the PELP group was also lower than that in the PEL and PA groups, and closer to that in the MET group and the normal control group, indicating that PELP also showed a superior regulatory trend compared to PEL and PA alone in inhibiting MFF-related mitochondrial division, suggesting that Phyllanthus emblica extract and probiotics have a synergistic effect in inhibiting MFF-mediated mitochondrial division.

[0155] In summary, these results indicate that type 2 diabetes can induce abnormal mitochondrial dynamics in mouse pancreatic tissue, primarily manifested as downregulation of MFN2 and upregulation of DRP1 / MFF. The advantage of PELP combined intervention lies mainly in its stronger inhibitory effect on DRP1 / MFF-mediated excessive mitochondrial division, rather than primarily through promoting MFN2-mediated mitochondrial fusion. Compared with PEL or PA alone, PELP can more effectively reduce the expression of DRP1 and MFF proteins in pancreatic tissue under diabetic conditions, suggesting that PELP may exert a protective effect against diabetes-related pancreatic damage by inhibiting abnormal mitochondrial division, reducing mitochondrial fragmentation, and maintaining mitochondrial homeostasis in pancreatic β-cells.

[0156] Example 3: The hypoglycemic and hypolipidemic effects of Phyllanthus emblica extract and its combination with probiotics (1) Peripheral blood glucose and lipid metabolism indicators 1) Fasting blood glucose Ten weeks after administration, the concentrations of fasting blood glucose in mice under different treatments were as follows: Figure 6 As shown: Compared with the blank control group, the fasting blood glucose in the model group was significantly increased, indicating that the mice had persistent hyperglycemia; PEL+PA can significantly reduce fasting blood glucose in the hyperglycemic model (P<0.01), and its effect is more obvious than that of PEL alone (P<0.05), PA or metformin alone (P<0.05); indicating that Phyllanthus emblica extract and probiotics have a synergistic effect in reducing fasting blood glucose.

[0157] 2) Glycated serum protein At the end of the experiment (at the end of week 18), the concentrations of glycated serum protein (GSP) in mice under different treatments were as follows: Figure 7 As shown, compared with the blank control group, the glycated serum protein in the model group was significantly increased, indicating that the mice had persistent enhanced glycation stress. All interventions improved GSP abnormalities to varying degrees, with PELP showing the most stable and significant effect (P<0.0001). The improvement effect of PELP was more significant than that of PEL (P<0.001), PA (P<0.01), and metformin alone (P<0.05).

[0158] Glycemic index (GSP) was used as a long-term glycemic control indicator for evaluation. Results showed that the mean GSP in the model group was 29.50 mmol / L, in the Phyllanthus emblica extract group it was 14.14 mmol / L, in the probiotic A group it was 15.56 mmol / L, and in the combination group of Phyllanthus emblica extract and probiotic A it was 6.74 mmol / L. For GSP reduction, a leading indicator of improvement, the highest single-drug effect model was used. The combination group showed a further reduction of 7.40 mmol / L compared to the optimal single-drug group, indicating that the combination group was superior to either single-drug group. This demonstrates that the combined use of Phyllanthus emblica extract and probiotics has a synergistic effect in reducing GSP.

[0159] 3) Four blood lipid tests At the end of the experiment (at the end of week 18), high-fat diet / streptozotocin (HFD / STZ) induced diabetic mice showed significant abnormalities in lipid metabolism. Compared with the blank control group (CON), the serum triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDLC) levels in the diabetic model group (DM) were significantly increased. Figure 8 , Figure 9 , Figure 10 ), while high-density lipoprotein cholesterol (HDLC) levels were significantly reduced ( Figure 11 This indicates that lipid homeostasis was disrupted. Drug and dietary interventions improved these changes to varying degrees. Among the intervention groups, PELP showed the most comprehensive lipid-regulating effect, manifested by a decrease in TG, TC, and LDLC levels, while simultaneously restoring HDLC. TG levels in the PELP group were significantly lower than in the PEL or PA groups, suggesting that the combined use of PEL and AKK is superior to either intervention alone in triglyceride regulation. In summary, these data indicate that PELP can effectively alleviate type 2 diabetes-related dyslipidemia and is significantly superior to PEL or PA alone in regulating serum TG levels.

[0160] Serum triglycerides (TG) were used as an indicator of lipid metabolism improvement. Since a decrease in TG represents improvement, the highest single-drug effect model was used for analysis. Results showed that the mean TG level in the Phyllanthus emblica extract group was 2.17 mmol / L, the mean TG level in the probiotic A group was 2.15 mmol / L, and the mean TG level in the Phyllanthus emblica extract and probiotic A combination group was 0.89 mmol / L. The TG level in the PEL+PA group was lower than the best single-drug group, with a HSA difference of 1.26 mmol / L, indicating that the combined use of Phyllanthus emblica extract and probiotics is superior to either component alone in reducing TG and improving lipid metabolism, demonstrating a synergistic effect.

[0161] Serum total cholesterol (TC) was used as an indicator of lipid metabolism improvement. Since a decrease in TC represents improvement, the highest single-drug effect model was used for analysis. Results showed that the mean TC in the Phyllanthus emblica extract group was 2.58 mmol / L, the mean TC in the probiotic A group was 2.66 mmol / L, and the mean TC in the Phyllanthus emblica extract and probiotic combination group was 1.67 mmol / L. The TC level in the PEL+PA group was lower than the best single-drug group, with a HSA difference of 0.91 mmol / L, indicating that the combined use of Phyllanthus emblica extract and probiotics is superior to either component alone in reducing TC and improving lipid metabolism, demonstrating a synergistic effect.

[0162] (2) Assay of pancreatic β-cell function 1) Intraperitoneal glucose tolerance test (IPGTT) At the end of week 18 of the experiment, the blood glucose levels and corresponding areas under the curves of mice under different treatments at 0, 15, 30, 60, and 120 min after glucose injection are shown in the figure below. Figure 12 As shown: Compared with the blank control group (CON), the area under the curve (AUC) of the IPGTT in diabetic model (DM) mice increased, indicating persistent glucose intolerance. All interventions improved this metabolic abnormality to varying degrees, with PELP showing the most stable and significant effect. During the IPGTT, PELP shifted the overall blood glucose response curve downward and significantly reduced the corresponding AUC, with a stronger effect than PA or PEL alone; indicating that the combined use of Phyllanthus emblica extract and probiotics has a synergistic effect in reducing IPGTT AUC. Overall, these results suggest that PELP has a superior effect on improving glucose tolerance in T2DM mice compared with single-drug treatment.

[0163] 2) Insulin Tolerance Test (ITT) At the end of week 18 of the experiment, the blood glucose levels and corresponding areas under the curve for mice under different treatments at 30, 60, 90, and 120 minutes after insulin injection are shown in the figure below. Figure 13As shown, compared with the blank control group (CON), the area under the ITT curve (AUC) of diabetic model mice (DM) increased, indicating a persistent decrease in insulin sensitivity. All interventions improved this metabolic abnormality to varying degrees, with PELP showing the most stable and significant effect. During the ITT, PELP shifted the overall response curve downward and significantly reduced the corresponding AUC, exhibiting a stronger effect than PA or PEL alone; indicating that the combined use of Phyllanthus emblica extract and probiotics has a synergistic effect in reducing ITT AUC. Overall, these results suggest that PELP has a superior effect on improving insulin sensitivity in T2DM mice compared to single-drug treatment.

[0164] (3) Body weight index like Figure 2 As shown, after 10 weeks of drug administration, the weight gain of mice in the DM group was significantly lower than that in the CON group (P<0.05), while MET, PEL, PA and PELP could partially restore weight, with PELP showing relatively stable growth in the later stage.

[0165] Example 4: Phyllanthus emblica extract and its anti-inflammatory effects with probiotics At the end of the experiment (at the end of week 18), the high-fat diet / streptozotocin (HFD / STZ) induced diabetic mice showed significant abnormalities in their inflammatory state. Compared with the blank control group (CON), the diabetic model mice exhibited significant pro-inflammatory features, with elevated levels of TNF-α and IL-6. Figure 14 , Figure 15 ), while IL-10 and IL-4 levels decreased ( Figure 16 , Figure 17 PEL and PELP interventions reduced the elevation of pro-inflammatory cytokines and restored the levels of anti-inflammatory cytokines, suggesting improved inflammatory homeostasis. In summary, these data indicate that PEL and PELP can effectively alleviate type 2 diabetes-related inflammatory dysregulation.

[0166] Serum interleukin-6 (IL-6) was used as a pro-inflammatory marker for evaluation. Since a decrease in IL-6 indicates improved inflammation, a maximum single-drug effect model was used for analysis. Results showed that the mean IL-6 level in the Phyllanthus emblica extract group was 23.73 pg / mL, the mean IL-6 level in the probiotic A group was 34.58 pg / mL, and the mean IL-6 level in the combination group of Phyllanthus emblica extract and probiotic A was 18.50 pg / mL. The IL-6 level in the combination group was lower than that of the best single-drug group, with a difference in HSA of 5.23 pg / mL, indicating that the combination of Phyllanthus emblica extract and probiotics is superior to either component alone in reducing IL-6 and alleviating inflammation, demonstrating a synergistic effect.

[0167] Serum interleukin-4 (IL-4) was used as an anti-inflammatory marker for evaluation. Since elevated IL-4 levels indicate improved inflammation, a maximum single-drug effect model was employed for analysis. Results showed that the mean IL-4 level in the Phyllanthus emblica extract group was 19.44 pg / mL, in the probiotic A group it was 24.19 pg / mL, and in the combined Phyllanthus emblica extract and probiotic A group it was 32.18 pg / mL. The combined group's IL-4 level was higher than the best single-drug group, with a HSA difference of 7.99 pg / mL, indicating that the combined use of Phyllanthus emblica extract and probiotics is superior to either component alone in increasing IL-4 and enhancing anti-inflammatory effects, demonstrating a synergistic effect.

[0168] Example 5: Phyllanthus emblica extract and its effect on liver function in combination with probiotics (1) Liver function indicators At the end of the experiment (at the end of week 18), the specific activity of alanine aminotransferase (ALT) in mice under different treatments was as follows: Figure 18 As shown: Compared with the blank control group, the alanine aminotransferase level in the model group was significantly increased; however, treatment with Phyllanthus emblica extract or its combination with probiotics significantly reduced alanine aminotransferase levels, with effects comparable to the metformin positive control group; the specific activity of aspartate aminotransferase (AST) in mice treated with different methods was as follows: Figure 19 As shown, compared with the blank control group, the aspartate aminotransferase in the model group was significantly increased; while the use of Phyllanthus emblica extract or its combination with probiotics can significantly reduce aspartate aminotransferase, and its effect is comparable to that of the metformin positive control group.

[0169] (2). Liver weight index At the end of the experiment (at the end of week 18), the liver weight index (liver weight / body weight, Liver Index) of mice under different treatments was as follows: Figure 20 As shown, compared with the blank control group, the liver weight index of the model group was significantly increased, suggesting diabetes-related hepatomegaly or increased metabolic burden; while the treatment with Phyllanthus emblica extract or its combination with probiotics significantly reduced the liver weight index, and its effect was comparable to that of the metformin positive control group; moreover, the liver weight index after treatment with Phyllanthus emblica extract and probiotics was lower than that after treatment with Phyllanthus emblica extract or probiotics alone, indicating that Phyllanthus emblica extract and probiotics have a synergistic effect in reducing liver weight index.

[0170] (3) Hepatic lipid metabolism At the end of the experiment (at the end of week 18), compared with the blank control group (CON), the levels of triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDLC) in the liver of diabetic model mice (DM) were significantly increased, indicating that liver lipid homeostasis was significantly disrupted in diabetic patients. Figure 21 PELP treatment significantly reduced these lipid indicators, suggesting that combined intervention with PEL and AKK can effectively reduce hepatic lipid accumulation in T2DM mice.

[0171] Example 6: Phyllanthus emblica extract and its effect on improving pancreatic function in combination with probiotics (1) Pancreatic weight index At the end of the experiment (at the end of week 18), the pancreatic index (pancreatic weight / body weight) of mice under different treatments was as follows: Figure 22 As shown, compared with the blank control group, the pancreatic weight index of the model group was significantly reduced; however, the treatment with Phyllanthus emblica extract or its combination with probiotics significantly increased the pancreatic weight index, suggesting that it can improve pancreatic tissue damage or pancreatic atrophy in diabetic model mice, and its effect was comparable to that of the metformin positive control group; moreover, the pancreatic weight index after treatment with Phyllanthus emblica extract and probiotics was higher than that after treatment with Phyllanthus emblica extract or probiotics alone, indicating that Phyllanthus emblica extract and probiotics have a synergistic effect in increasing the pancreatic weight index.

[0172] (2) Immunohistochemical staining of pancreatic tissue At the end of the experiment (at the end of week 18), the HE staining results of pancreatic tissue from mice under different treatments are as follows: Figure 23 As shown: in the blank control group, the pancreatic tissue cells of mice were tightly arranged, while the cells of the model group mice showed obvious vacuolization, tissue disorder, loose cell arrangement and local structural damage. The vacuolization was significantly improved by using Phyllanthus emblica extract or its combination with probiotics.

[0173] (3). TUNEL apoptosis detection of pancreatic β cells TUNEL labeling can detect apoptosis-related DNA breaks and is commonly used to assess β-cell death, a key factor in β-cell dysfunction during the progression of type 2 diabetes mellitus (T2DM). At the end of the experiment (at the end of week 18), the TUNEL staining results of pancreatic tissues from mice under different treatments are as follows: Figure 24 As shown, compared with the blank control group, the pancreatic tissue of the model group mice showed a large number of green fluorescent positive signals, indicating that pancreatic β cells underwent significant apoptosis. The apoptosis was significantly improved by using Phyllanthus emblica extract or by treating it with probiotics.

[0174] Example 7: Phyllanthus emblica extract and its effect on regulating intestinal flora with probiotics At the end of the experiment (at the end of week 18), the relative abundance of gut microbiota at the genus level in each group is shown in the figure. Figure 25 Based on the relative abundance results at the genus level, the DM group showed a significant imbalance in bacterial community compared to the CON group. unclassified_Muribaculaceae, Bacteroides, Dubosiella, Thomasclavelia When probiotic / metabolism-related bacteria decrease, at the same time Enterococcus, Helicobacter and unclassified_Desulfovibrionaceae An increase in potentially conditionally pathogenic or inflammation-related bacteria suggests impaired gut microbiota homeostasis in patients with type 2 diabetes mellitus (T2DM).

[0175] In terms of single-drug intervention, the Phyllanthus emblica extract (PEL) group showed a better effect on improving gut microbiota. Compared with the DM group, PEL significantly improved gut microbiota. unclassified_Muribaculaceae The relative abundance increased from 12.23% to 19.23%; Dubosiella It rose from 0.22% to 8.01%. Bacteroides The percentage also increased from 4.02% to 4.42%. Meanwhile, PEL significantly reduced the potential for harmful bacteria. Enterococcus It decreased from 4.44% to 0.63%. Helicobacter The percentage decreased from 3.40% to 1.99%. This indicates that the intervention of Phyllanthus emblica extract alone mainly regulates the intestinal flora by restoring the production of short-chain fatty acids and metabolic regulation-related flora, and inhibiting some inflammation-related bacteria.

[0176] The probiotic A (PA) group exhibited a different type of regulatory characteristic. The PA group significantly improved... Bacteroides The relative abundance of [a substance] increased from 4.02% in the DM group to 10.35%, approaching or exceeding the level in the CON group; meanwhile, Helicobacter The percentage decreased significantly, from 3.40% to 1.04%, suggesting that probiotics have an improving effect on some beneficial metabolic bacteria and inflammation-related bacteria. However, in the PA group... Enterococcus It rose to 11.63%. unclassified_Desulfovibrionaceae It was also slightly higher than the DM group, indicating that probiotics are not comprehensive in inhibiting certain potentially harmful bacteria, and their flora regulation has a certain degree of selectivity.

[0177] The combined intervention group (PELP) showed a certain complementary / synergistic regulatory trend. Compared with the DM group, PELP significantly improved... unclassified_Muribaculaceae It rose from 12.23% to 19.44%; Bacteroides It rose from 4.02% to 9.65%, approaching the 8.22% of the CON group; Dubosiella It also rose from 0.22% to 1.62%. Meanwhile, Helicobacter The percentage decreased from 3.40% to 1.86%, lower than the DM group and close to the PEL group. This indicates that the combined use of Phyllanthus emblica and AKK live bacteria can simultaneously retain the benefits of Phyllanthus emblica and improve overall health. Muribaculaceae The advantages of AKK live bacteria promotion Bacteroides The restorative effect is characterized by a complementary remodeling effect on probiotic / metabolism-related microbiota. The combined intervention of Phyllanthus emblica and AKK live bacteria shows complementary and partially synergistic trends in enhancing some beneficial bacteria and restoring the microbiota structure, particularly evident in… Muribaculaceae and Bacteroides On the recovery.

[0178] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. Application of active ingredient combinations in product preparation; The combination of active ingredients includes a first active ingredient and a second active ingredient. The first active ingredient is Phyllanthus emblica extract, and the second active ingredient is probiotics, their fermentation culture, their dried product, or a mixture thereof; The product is food, health supplement, or medicine; The food and health products mentioned above have one or more of the following functions: b1) Control body fat; b2) Maintain or restore healthy levels of blood lipids (e.g., cholesterol, triglycerides); b3) Maintain or restore healthy blood sugar levels; b4) It has an auxiliary protective or restorative effect on chemically induced liver injury; b5) Regulates gut microbiota; The drug has one or more of the following functions: a1) Lowering lipids; a2) Prevention or treatment of mitochondrial function-related diseases; a3) Prevention or treatment of liver diseases; a4) Prevention or treatment of pancreatic diseases; a5) Prevention or treatment of inflammatory diseases.

2. Application of active ingredients in product preparation; The active ingredients include a first active ingredient, which is an extract of Phyllanthus emblica; The product has one or more of the following functions: b1) Control body fat; b2) Maintain or restore healthy levels of blood lipids (e.g., cholesterol, triglycerides); b3) Maintain or restore healthy blood sugar levels; b4) It has an auxiliary protective or restorative effect on chemically induced liver injury; b5) Regulates gut microbiota; The products are preferably food or health products.

3. The application according to any one of claims 1 to 2, characterized in that, The dosage forms of the product include: solid dosage forms, semi-solid dosage forms, or liquid dosage forms; Preferably, the food or health product further includes excipients acceptable for use in food or health products; Preferably, the excipients acceptable for the food or health product include at least one of the following: wetting agents, emulsifiers, preservatives, antioxidants, buffers, diluents, lubricants, solvents, suspending agents, solubilizers, thickeners, stabilizers, sweeteners, and flavorings; Preferably, the dosage form of the food or health product is a dosage form suitable for children or adults; Preferably, the dosage form of the food or health product is selected from oral dosage forms or non-oral dosage forms.

4. The application according to claim 1, characterized in that, The probiotics include Akkermansia mycotoxin-loving bacteria (Akermansia spp.) Akkermansia muciniphila ); Preferably, the first active ingredient and the second active ingredient exist independently or in combination; Preferably, the first active ingredient and the second active ingredient are applied simultaneously, separately, or sequentially; Preferably, the drug further includes pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include at least one of diluents, excipients, binders, humectants, surfactants, lubricants, and disintegrants; Preferably, the dosage form of the drug is a dosage form suitable for children or an adult; Preferably, the dosage form of the drug is selected from gastrointestinal dosage forms or non-gastrointestinal dosage forms.

5. The application according to claim 1 or 4, characterized in that, The mitochondrial function-related diseases include at least one of metabolic diseases, nervous system diseases, aging, and muscle diseases; Preferably, the metabolic disease is a metabolic disorder, metabolic disturbance, or a disease caused by a metabolic disturbance; preferably including at least one of the following: obesity and obesity-related diseases, diabetes, dyslipidemia, glucose intolerance, impaired glucose tolerance, hyperglycemia, hyperinsulinemia, hyperlipoproteinemia, hypertriglyceridemia, uremia, ketoacidosis, hypoglycemia, and thrombotic diseases. Preferably, the obesity and obesity-related diseases include at least one of the following: overweight, obesity, metabolic syndrome, cardiovascular and cerebrovascular diseases, hyperlipidemia, hypercholesterolemia, hypertension, insulin resistance syndrome, obesity-related gastroesophageal reflux disease, and steatohepatitis. Preferably, the neurological disease includes at least one of Friedreich's ataxia, amyotrophic lateral sclerosis (ALS), adrenoleukodystrophy, Alexander disease, Alpert disease, familial fatal insomnia, Huntington's disease, Kennedy's disease, Crabbe's disease, Lyme disease, Machado-Joseph disease, multiple sclerosis, multiple system atrophy, Niemann-Pick disease, Pick's disease, primary lateral sclerosis, progressive supranuclear palsy, Rafe-Syme disease, Sandhoff's disease, diffuse demyelinating sclerosis, spinocerebellar ataxia, subacute mixed spinal cord degeneration, toxic encephalopathy, disseminated spongiform encephalopathy, and ataxia-telangiectasia. Preferably, the muscle disease includes sarcopenia and / or Duchenne muscular dystrophy; Preferably, the liver disease includes at least one of the following: fatty liver, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), abnormal liver function, extrahepatic cholestasis, hepatitis, liver injury, intrahepatic cholestasis, liver fibrosis, cirrhosis, and hepatocyte damage. Preferably, the pancreatic disease includes at least one of pancreatitis, pancreatic fibrosis, and pancreatic cancer; Preferably, the inflammatory disease is selected from inflammation related to metabolic diseases; diseases related to liver inflammation; diseases related to pancreatic inflammation; diseases related to bronchial inflammation; diseases related to cervical inflammation; diseases related to conjunctival inflammation; diseases related to esophageal inflammation; diseases related to myocardial inflammation; diseases related to rectal inflammation; diseases related to scleral inflammation; diseases related to gum inflammation; diseases related to bone inflammation; lung inflammation; respiratory tract inflammation; acute respiratory distress syndrome; inflammatory skin diseases; fibrotic diseases; and encephalitis.

6. A combination of active ingredients, comprising a first active ingredient and a second active ingredient; The first active ingredient is Phyllanthus emblica extract; the second active ingredient is probiotics, their fermentation cultures, their dried products, or mixtures thereof.

7. The active ingredient combination according to claim 6, characterized in that, The probiotics include Akkermansia mycotoxin-loving bacteria (Akermansia spp.) Akkermansia muciniphila ); Preferably, the first active ingredient and the second active ingredient exist independently or in combination; Preferably, the first active ingredient and the second active ingredient are applied simultaneously, separately, or sequentially.

8. A product comprising the combination of active ingredients as described in any one of claims 6 to 7.

9. The product according to claim 8, characterized in that, The product is food, health supplement, or medicine; Preferably, the dosage form of the product includes: solid dosage form, semi-solid dosage form, or liquid dosage form; Preferably, the product further includes food or health product-acceptable excipients or pharmaceutically acceptable excipients.

10. The product according to claim 9, characterized in that, The food or health product also includes excipients acceptable for use in food or health products; Preferably, the excipients acceptable for the food or health product include at least one of the following: wetting agents, emulsifiers, preservatives, antioxidants, buffers, diluents, lubricants, solvents, suspending agents, solubilizers, thickeners, stabilizers, sweeteners, and flavorings; Preferably, the dosage form of the food or health product is a dosage form suitable for children or adults; Preferably, the dosage form of the food or health product is selected from oral dosage forms or non-oral dosage forms; or, The drug also includes pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include at least one of diluents, excipients, binders, humectants, surfactants, lubricants, and disintegrants; Preferably, the dosage form of the drug is a dosage form suitable for children or an adult; Preferably, the dosage form of the drug is selected from gastrointestinal dosage forms or non-gastrointestinal dosage forms.

11. A method of administering to a subject an effective amount of the product according to any one of claims 1 to 5, the combination of active ingredients according to claims 6 to 7, or the product according to claims 8 to 10; The method has one or more of the following functions: b1) Control body fat; b2) Maintain or restore healthy levels of blood lipids (e.g., cholesterol, triglycerides); b3) Maintain or restore healthy blood sugar levels; b4) It has an auxiliary protective or restorative effect on chemically induced liver injury; b5) Regulate the gut microbiota.

Images