Application of lily and lotus seed composition in improvement of obesity and type 2 diabetes mellitus

Abstract

The invention provides application of a lily and lotus seed composition in improvement of obesity and type 2 diabetes mellitus. Specifically, the invention provides application of a lily and lotus seed composition in preparation of drugs for intervening or treating glucose metabolism disorder diseases. The lily and lotus seed composition can effectively (a) reduce body weight; (b) reducing fatty liver; (c) reducing fasting blood glucose; and / or (d) cholesterol is reduced, and the composition disclosed by the invention has a remarkable synergistic effect when being used in a combined manner.

Description

Technical Field

[0001] The present invention belongs to the field of chemical medicine, and particularly relates to the application of a lily and lotus seed composition in improving obesity and type 2 diabetes. Background Art

[0002] Diabetes is one of the most common chronic diseases at present. Patients may simultaneously develop complications such as obesity, cardiovascular diseases, retinopathy, nephropathy, and nervous system damage, and it may reduce the life expectancy of patients. Obesity is a chronic multi-system disease, and obese people may suffer from diseases such as insulin resistance, type 2 diabetes, dyslipidemia, and hypertension.

[0003] Therefore, finding drugs or foods that can effectively prevent and control blood sugar elevation and effectively regulate the body's glucose and lipid metabolism not only helps to prevent the onset of diabetes in humans, control diabetes, and prevent the further expansion of the obese population, but also helps to relieve and treat the diseases of diabetes and obese people themselves, which has important practical significance for the prevention and treatment of diabetes and obesity.

[0004] Therefore, there is an urgent need in this field to develop a new food, dietary supplement, health product, or drug that can effectively prevent and / or treat obesity and type 2 diabetes. Summary of the Invention

[0005] The present invention provides a new food, dietary supplement, health product, or drug that can effectively prevent and / or treat obesity and type 2 diabetes.

[0006] In the first aspect of the present invention, there is provided the use of a lily-lotus seed composition for preparing a preparation or composition, and the preparation or composition is used for:

[0007] (a) alleviating fatty liver;

[0008] (b) reducing blood sugar;

[0009] (c) reducing cholesterol; and / or

[0010] (d) reducing body weight;

[0011] Wherein, the content ratio (wt:wt) of the lily and lotus seed is: 4:1 - 2:1.

[0012] In another preferred example, the content of the lily and lotus seed is based on the dry weight of the lily and lotus seed.

[0013] In another preferred example, the content ratio (wt:wt) of the lily and lotus seed is: 3:1.

[0014] In another preferred example, the reducing blood sugar includes reducing fasting blood sugar.

[0015] In another preferred embodiment, the preparation or composition is also used to reduce the body fat percentage.

[0016] In another preferred embodiment, the preparation or composition is for an object on a high-fat diet.

[0017] In another preferred embodiment, the object is a human or non-human mammal (such as a rodent).

[0018] In another preferred embodiment, the object is a human.

[0019] In another preferred embodiment, the fatty liver is a fatty liver caused by a high-fat diet.

[0020] In another preferred embodiment, the composition is a pharmaceutical composition, a dietary supplement, a food composition, a feed composition, or a health product composition.

[0021] In another preferred embodiment, the preparation or composition is administered to humans and non-human mammals.

[0022] In another preferred embodiment, the composition is a pharmaceutical composition.

[0023] In another preferred embodiment, the pharmaceutical composition is used to treat disorders of glycolipid metabolism.

[0024] In another preferred embodiment, the pharmaceutical composition is used to treat diseases including those selected from the group consisting of: obesity, type 2 diabetes, fatty liver, hyperlipidemia, or a combination thereof.

[0025] In another preferred embodiment, the pharmaceutical composition is used to treat diseases including those selected from the group consisting of: obesity, type 2 diabetes, or a combination thereof.

[0026] In another preferred embodiment, the pharmaceutical composition contains a lily and lotus seed composition and a pharmaceutically acceptable carrier.

[0027] In another preferred embodiment, the pharmaceutical composition is a solid preparation, a liquid preparation, or a semi-solid preparation.

[0028] In another preferred embodiment, the dosage form of the pharmaceutical composition includes: oral preparations, injections, and lyophilized products.

[0029] In another preferred embodiment, the pharmaceutical composition is administered by the following routes: oral, intravenous, intramuscular, subcutaneous, and inhalation.

[0030] In another preferred embodiment, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

[0031] In another preferred embodiment, the dosage form of the pharmaceutical composition is an oral preparation, preferably: tablets, pills, capsules, granules, or oral liquids.

[0032] In a second aspect of the present invention, there is provided an active ingredient combination, which comprises:

[0033] (Z1) A first active ingredient, which is lily or its extract; and

[0034] (Z2) A second active ingredient, which is lotus seed or its extract;

[0035] Wherein, the content ratio (wt:wt) of the first active ingredient to the second active ingredient is: 4:1 - 2:1.

[0036] In another preferred embodiment, the contents of the lily and the lotus seed are based on their dry weights.

[0037] In another preferred embodiment, the extract is an extract converted to the corresponding weight ratio of lily or lotus seed.

[0038] In another preferred embodiment, the lily or the lily extract contains the following active ingredients: lily polysaccharide, saponin, alkaloid, flavonoid.

[0039] In another preferred embodiment, the lotus seed or the lotus seed extract contains the following active ingredients: neferine, polysaccharide, flavonoid.

[0040] In another preferred embodiment, the total content of the first active ingredient and the second active ingredient is 70 - 100%, preferably 80 - 100 wt%, more preferably 90 - 100 wt%, based on the total weight of the active ingredients, and the weight is the dry weight.

[0041] In another preferred embodiment, the ratio R 1 (wt:wt) of the content of the first active ingredient to the content of the second active ingredient is 4:1 - 2:1.

[0042] In another preferred embodiment, the ratio R 1 (wt:wt) of the content of the first active ingredient to the content of the second active ingredient is 3:1.

[0043] In another preferred embodiment, the effective content of the first active ingredient is calculated based on the content or weight of the lily polysaccharide contained therein; while the effective content of the second active ingredient is calculated based on the content or weight of the neferine contained therein,

[0044] And the ratio R 2 (wt:wt) of the effective contents of the first active ingredient and the second active ingredient is: (12 - 4):1.

[0045] In another preferred embodiment, the ratio R 2 (wt:wt) of the effective amounts of the first active ingredient and the second active ingredient is: (8 - 7):1.

[0046] In another preferred embodiment, in the combination, the first active ingredient and the second active ingredient are independent of each other or are mixed with each other.

[0047] In a third aspect of the present invention, there is provided a pharmaceutical composition comprising:

[0048] (Z1) A first active ingredient, which is lily or an extract thereof;

[0049] (Z2) A second active ingredient, which is lotus seed or an extract thereof; and

[0050] (Z3) A pharmaceutically acceptable carrier;

[0051] Wherein, the content ratio (wt:wt) of the first active ingredient to the second active ingredient is: 4:1 - 2:1.

[0052] In another preferred embodiment, the contents of the lily and the lotus seed are based on the dry weights of the lily and the lotus seed.

[0053] In another preferred embodiment, the extract is an extract that has been converted to the corresponding weight ratio of lily and lotus seed.

[0054] In another preferred embodiment, the content ratio (wt:wt) of the lily to the lotus seed is: 3:1.

[0055] In another preferred embodiment, the pharmaceutical composition is administered to subjects on a high-fat diet and subjects not on a high-fat diet.

[0056] In another preferred embodiment, the pharmaceutical composition is administered to subjects on a high-fat diet.

[0057] In another preferred embodiment, the lily or the lily extract contains the following active ingredients: lily polysaccharide, saponin, alkaloid, flavonoid.

[0058] In another preferred embodiment, the lotus seed or the lotus seed extract contains the following active ingredients: neferine, polysaccharide, flavonoid.

[0059] In another preferred embodiment, the total content of the first active ingredient and the second active ingredient is 0.01 - 99%, preferably 0.1 - 90 wt%, more preferably 1 - 80 wt%, based on the total weight of the active ingredients.

[0060] In another preferred embodiment, the ratio R of the content of the first active ingredient to the content of the second active ingredient 1 (wt:wt) is 4:1 - 2:1.

[0061] In another preferred embodiment, the ratio R of the content of the first active ingredient to the content of the second active ingredient 1 (wt:wt) is 3:1.

[0062] In another preferred embodiment, the dosage forms of the pharmaceutical composition include tablets, granules, capsules, pills, injections, or oral liquids.

[0063] In another preferred embodiment, in the combination, the first active ingredient and the second active ingredient are independent of each other or are mixed with each other.

[0064] In the fourth aspect of the present invention, there is provided a feed composition, which contains:

[0065] (Z1) a first active ingredient, and the first active ingredient is lily or an extract thereof;

[0066] (Z2) a second active ingredient, and the second active ingredient is lotus seed or an extract thereof; and

[0067] (Z3) a food-acceptable carrier

[0068] Wherein, the content ratio (wt:wt) of the first active ingredient and the second active ingredient is: 4:1 - 2:1, and the total content of the first active ingredient and the second active ingredient is 10wt% - 30wt%.

[0069] In another preferred embodiment, the content ratio (wt:wt) of the lily and the lotus seed is: 3:1.

[0070] In another preferred embodiment, the total content of the first active ingredient and the second active ingredient is 15% - 20%.

[0071] In another preferred embodiment, the total content of the first active ingredient and the second active ingredient is 16%, and the content ratio (wt:wt) of the first active ingredient to the second active ingredient is: 3:1.

[0072] In another preferred embodiment, the pharmaceutical composition is administered to a high-fat diet subject and a non-high-fat diet subject.

[0073] In another preferred embodiment, the pharmaceutical composition is administered to a high-fat diet subject.

[0074] In another preferred embodiment, the subject is a mammal (such as a rodent).

[0075] In the fifth aspect of the present invention, there is provided a method for treating obesity or type 2 diabetes, which includes the step of: administering to a subject in need the combination of active ingredients described in the second aspect of the present invention, or the pharmaceutical composition described in the third aspect of the present invention.

[0076] In another preferred example, the objects include humans and non-human mammals (such as rodents).

[0077] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 Shows that the lily lotus seed composition inhibits high-fat diet-induced obesity in mice: (A) Weight changes and percentage of weight changes of mice fed different types of rodent diets for 6 weeks (B); (C) Body weight and percentage of weight changes of mice after 6 weeks of feeding different types of rodent diets (D); (E) Food intake changes of mice fed different types of rodent diets for 6 weeks (g / day); (F) Food intake changes of mice fed different types of rodent diets for 6 weeks (kcal / Body Weight / day); (G) Effects of experimental treatments on the livers of mice, and their livers were weighed and calculated; (H) Effects of experimental treatments on the epididymal fat of mice, and their epididymal fat was weighed and calculated; Data are expressed as mean ± SEM, and the number of mice in the normal diet, high-fat diet, lotus seed, lily, and lily lotus seed groups was n = 8, 8, 7, 8, 8, 8, 8), *P < 0.05, **P < 0.01, ***P < 0.001, representing significant differences compared with the normal diet group, #P < 0.05, ##P < 0.01, P < 0.001 representing significant differences compared with the high-fat diet group.

[0079] Figure 2 Shows that the lily lotus seed composition controls the increase in blood glucose in high-fat diet-induced obese mice: (A) Detection of fasting blood glucose in mice after 3 weeks of experimental treatment; (B and C) Detection of glucose tolerance in mice after 5 weeks of experimental treatment, and the blood glucose changes of mice in different groups 2 h after fasting glucose injection; (D) Area under the blood glucose change curve of mice in different groups 2 h after fasting glucose injection; Data are expressed as mean ± SEM, *P < 0.05, ***P < 0.001, representing significant differences compared with the normal diet group; #P < 0.05, P < 0.001 representing significant differences compared with the high-fat diet group.

[0080] Figure 3 Shows that the lily lotus seed composition controls the increase in total cholesterol levels in the serum of high-fat diet-induced obese mice: The measurement results of total cholesterol in the serum of mice; Data are expressed as mean ± SEM, *P < 0.05, ***P < 0.001, representing significant differences compared with the normal diet group; #P < 0.05, P < 0.001 representing significant differences compared with the high-fat diet group. Detailed implementation mode

[0081] Through extensive and in-depth research and a large number of screenings, the present inventor unexpectedly discovered for the first time that a combination of lily and lotus seeds in a specific ratio (4:1 - 2:1, preferably 3:1) can effectively improve obesity and type 2 diabetes. Research shows that the combination of lily and lotus seeds can effectively (a) reduce body weight; (b) reduce fatty liver; (c) reduce fasting blood glucose; and / or (d) reduce cholesterol, and the combination of lily and lotus seeds in a specific ratio has a very significant synergistic effect, which can significantly reduce the body fat percentage. Thus, it can effectively improve or treat sugar metabolism disorder diseases including obesity and type 2 diabetes. Based on this, the present invention was completed.

[0082] Term

[0083] As used herein, the term "comprising" can be open-ended, semi-closed, or closed. In other words, this term includes the following limitations: "containing", "substantially consisting of...", "consisting of...".

[0084] As used herein, the term "combination of active ingredients" refers to a combination composed of a first active ingredient and a second active ingredient. Among them, the combination of active ingredients can be a mixture formed by blending the first active ingredient and the second active ingredient (including mixtures in solid phase, liquid phase, or other states), or it can be a combination composed of the first active ingredient and the second active ingredient without blending. For example, place the first active ingredient in a first container, place the second active ingredient in a second container, and place the above-mentioned first container and second container in the same medicine box. Such a medicine box is also covered by this term.

[0085] As used herein, "pharmaceutically acceptable salt" refers to a salt formed by the compound of the active ingredient of the present invention with an acid or a base, which is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is the salts formed by the compound of the active ingredient of the present invention with an acid. Acids suitable for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzene sulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid. A preferred class of salts is the salts formed by the compound of the active ingredient of the present invention with a base. Bases suitable for forming salts include, but are not limited to: inorganic bases such as sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, sodium phosphate, etc., organic bases such as ammonia water, triethylamine, diethylamine, etc. Another preferred class of salts is the salts formed by the active ingredient of the present invention with metal ions, including but not limited to magnesium salts, sodium salts, calcium salts, potassium salts, etc.

[0086] As used herein, "prevention" and "treatment" include delaying and halting the progression of a disease, or eliminating the disease, and do not require 100% inhibition, eradication, and reversal. In some embodiments, the compositions or pharmaceutical compositions of the present invention prevent, mitigate, inhibit, and / or reverse ischemic reperfusion injury by, for example, at least about 10%, at least about 30%, at least about 50%, or at least about 80% compared to the levels observed in the absence of the compositions or pharmaceutical compositions of the present invention.

[0087] Lily

[0088] Lily is the dried fleshy scale leaf of Lilium lancifolium Thunb., Lilium brownii F.E.Brown var. viridulum Baker., or Lilium pumilum DC. of the Liliaceae family. According to the Chinese Pharmacopoeia, its main effects are to nourish yin and moisten the lungs, and to clear the heart and calm the mind.

[0089] The lily bulb is rich in various medicinal active ingredients such as polysaccharides, alkaloids, steroidal saponins, and flavonoids. Among them, flavonoids and polysaccharides have antioxidant effects.

[0090] The saponins and alkaloids in lily have anti-tumor effects. Saponins can induce cell cycle arrest of tumor cells and reduce the toxicity induced by chemotherapy or radiotherapy.

[0091] Lily polysaccharide in lily promotes insulin secretion and thus has a certain hypoglycemic effect by enhancing the activity of antioxidant enzymes in the body, improving the body's antioxidant function, and reducing the damage of oxygen free radicals to pancreatic islet β cells.

[0092] In addition, the anti-inflammatory, antibacterial, antidepressant, and immune system regulatory effects of the active ingredients in lily have also been discovered and confirmed.

[0093] Lotus Seed

[0094] Lotus seed is the seed of Nelumbo nucifera Gaertn. of the Nymphaeaceae family. According to the Chinese Pharmacopoeia, its main effects are to invigorate the spleen and stop diarrhea, and to nourish the heart and calm the mind.

[0095] Lotus seeds contain flavonoids, polysaccharides, and various alkaloids, and the proportion of amino acids in lotus seeds is relatively balanced, so it is considered a high-quality protein source.

[0096] The research on the efficacy and mechanism of the active substances in lotus seeds is very extensive: N-methyl liensinine, liensinine, and isoliensinine in lotus seeds induce autophagy and apoptosis by activating ROS and MAP kinases, and thus have a certain anti-tumor effect.

[0097] Isoliensinine can improve the body's sensitivity to insulin by activating adenosine monophosphate-activated protein kinase (AMPK) in skeletal muscle, white adipose tissue, and the liver, as well as glucose transporter GLUT4 in adipose tissue and muscle, thereby achieving the effect of lowering blood sugar.

[0098] Lotus seed polysaccharide enhanced the immune response of H22 tumor-bearing mice by improving the spleen and thymus indices and increasing serum cytokines.

[0099] In addition, the anti-inflammatory, antioxidant, and beneficial effects on the digestive system of the functional components in lotus seeds have also been gradually confirmed.

[0100] Composition and Application

[0101] Based on the unexpected findings of the present invention, the inventors of the present invention provide various different applications of the active ingredients of the present invention.

[0102] Typically, the active ingredient of the present invention can be used to prepare a preparation or composition for:

[0103] (a) Alleviating fatty liver;

[0104] (b) Lowering blood sugar;

[0105] (c) Lowering cholesterol; and / or

[0106] (d) Lowering body weight;

[0107] Wherein, the content ratio (wt:wt) of lily to lotus seed is: 4:1 - 2:1.

[0108] The present invention also provides various different compositions containing the active ingredient of the present invention, and the compositions include (but are not limited to): pharmaceutical compositions, health product compositions, feed compositions.

[0109] Taking the pharmaceutical composition as an example, as used herein, the term "active ingredient" refers to memantine or a pharmaceutically acceptable salt thereof that can be used in the present invention.

[0110] As used herein, the term "effective amount" or "effective dose" refers to an amount that can produce a function or activity in humans and / or animals and is acceptable to humans and / or animals.

[0111] As used herein, the components of the term "pharmaceutically acceptable" are suitable for humans and / or mammals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier used for administering a therapeutic agent, including various excipients and diluents.

[0112] The pharmaceutical composition of the present invention contains a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier. Such carriers include (but are not limited to): saline, buffer solution, glucose, water, glycerol, ethanol, and combinations thereof. Generally, the pharmaceutical preparation should be matched with the administration method, and the dosage forms of the pharmaceutical composition of the present invention are injection, oral preparation (tablet, capsule, oral liquid), transdermal agent, sustained-release agent. For example, it is prepared by conventional methods with physiological saline or an aqueous solution containing glucose and other adjuvants. The said pharmaceutical composition should be manufactured under aseptic conditions.

[0113] The effective amount of the active ingredient described in the present invention may vary with the mode of administration, the severity of the disease to be treated, etc. The selection of the preferred effective amount can be determined by those of ordinary skill in the art according to various factors (such as through clinical trials). The said factors include but are not limited to: the pharmacokinetic parameters of the said active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated in the patient, the patient's body weight, the patient's immune status, the route of administration, etc. Generally, when the active ingredient of the present invention is administered at a dose of about 0.00001 mg - 50 mg / kg of animal body weight per day (preferably 0.0001 mg - 10 mg / kg of animal body weight), satisfactory effects can be obtained. For example, due to the urgent requirements of the treatment condition, several separate doses may be administered per day, or the dose may be proportionally reduced.

[0114] The pharmaceutically acceptable carriers described in the present invention include (but are not limited to): water, saline, liposome, lipid, protein, protein-antibody conjugate, peptide substance, cellulose, nanogel, or combinations thereof. The selection of the carrier should be matched with the administration method, which are well-known to those of ordinary skill in the art.

[0115] The main advantages of the present invention:

[0116] 1. The lily and lotus seed composition of the present invention can significantly control weight gain.

[0117] 2. The lily and lotus seed composition of the present invention can effectively control blood glucose elevation, including fasting blood glucose, postprandial blood glucose, etc.

[0118] 3. The lily and lotus seed composition of the present invention can control the elevation of total cholesterol level in serum.

[0119] 4. The lily and lotus seed composition of the present invention in a specific ratio has excellent synergistic effects in controlling weight, controlling blood glucose and reducing cholesterol, etc.

[0120] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight percentage and weight parts.

[0121] Materials and Methods

[0122] Experimental Animals

[0123] Eight-week-old male C57BL / 6 mice were purchased from the Shanghai branch of Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0124] Lily and lotus seed raw materials

[0125] Dried lily slices were purchased from Hongyun Planting Professional Cooperative in Lianhua County; skinned lotus seeds with cores were purchased from Fuxing Food Co., Ltd. in Guangchang County.

[0126] Main Reagents, Instruments and Equipment

[0127] Neferine reference substance (Dalian Meilun Biotechnology Co., Ltd.); Anthrone (Sinopharm Chemical Reagent Co., Ltd.); Triglyceride detection kit (Nanjing Jiancheng Bioengineering Institute), Total cholesterol detection kit (Nanjing Jiancheng Bioengineering Institute); Blood glucose meter Freestyle and blood glucose test strips (Abbott); Glucose (Sinopharm Chemical Reagent Co., Ltd.).

[0128] High-speed multi-functional pulverizer (2500Y) was purchased from Yongkang Bo'ou Hardware Products Co., Ltd.; Multi-functional microplate reader (Various Flash) was purchased from Thermo Fisher Scientific Co., Ltd.; Ultrasonic instrument (SB25-12DT) was purchased from Ningbo Xinzhi Biotechnology Co., Ltd.; High-performance liquid chromatograph (Ultimate3000) was purchased from Thermo Fisher Scientific Co., Ltd.; High-speed refrigerated centrifuge (Centrifuge 5424) was purchased from Eppendorf (China) Co., Ltd.

[0129] Experimental Methods

[0130] Determination of the content of core components in lily and lotus seeds

[0131] Determination of lily polysaccharide

[0132] The anthrone-sulfuric acid method was used to determine the content of polysaccharides in lilies. 1.0 g of lily powder was accurately weighed, water was added, and ultrasonic extraction was carried out (the ratio of material to liquid was 1:20, the extraction time was 20 min, and the extraction temperature was 75 °C), and it was shaken well. 1.5 ml of the supernatant was accurately measured, 7.5 ml of ethanol was added, and it was shaken well, centrifuged (3000 rpm, 15 min), the precipitate was taken, dissolved in water, and fixed to 50 ml, and it was shaken well. 1.0 ml was accurately measured, 4.0 ml of 0.2% anthrone-sulfuric acid solution was added (0.2 g of anthrone was taken, dissolved and diluted to 100 mL with concentrated sulfuric acid), mixed well, quickly cooled in an ice-water bath, and then 200 μl of the test solution was taken and placed in a 96-well plate. Using an enzyme-linked immunosorbent assay reader, according to the ultraviolet-visible spectrophotometry method, the absorbance of the test sample was measured at a wavelength of 620 nm, and then the weight of anhydrous glucose contained in the test solution was obtained according to the standard curve.

[0133] Determination of methyl liensinine

[0134] High performance liquid chromatography was used to determine the methyl liensinine in lotus seeds. Chromatographic conditions: C13 was used as the filler; methanol-85% formic acid-aqueous solution (1:1000) was used as the mobile phase; the detection wavelength was 282 nm. The number of theoretical plates calculated by the methyl liensinine peak should be not less than 5000. About 0.5 g of lotus seed powder was accurately weighed, 25 ml of 2% hydrochloric acid-methanol solution was added, ultrasonic treatment and degassing were carried out (power 250 W, frequency 40 kHz) for 30 min, shaken well, centrifuged (3000 rpm, 10 min), 5 ml of the filtrate was accurately measured, and the filtrate was blown to near dryness with a nitrogen blower. The residue was dissolved in methanol, transferred to a 10 ml volumetric flask, diluted to the scale, shaken well, filtered, and the filtrate was taken as the test solution. 3 μl of each test solution was accurately pipetted and injected into the high performance liquid chromatograph, and the absorption peak area at 282 nm was detected. The content of methyl liensinine in the test solution was obtained according to the standard curve.

[0135] Example 1

[0136] 1.1 Experimental procedures

[0137] 1.1.1 Preparation of feed containing lilies and lotus seeds

[0138] Dried lilies and dried lotus seeds were respectively ground into powder with a high-speed multi-functional grinder, and lily and lotus seed powders were added to the high-fat diet for mice in different proportions.

[0139] The compositions of each feed under different addition ratios are shown in Table 1-2, and the addition ratios are shown in Table 3. The energy provided by the fat in the feed is about 60%.

[0140] Table 1. Raw material compositions of normal feed, 60% fat function, 16% lily group and 16% lotus seed group

[0141]

[0143] Table 2 Feed composition of 12% lily + 4% lotus seed group, 8% lily + 8% lotus seed group, and 4% lily + 12% lotus seed group

[0144]

[0145] 1.1.2 Animal grouping and model construction

[0146] After 1 week of adaptive feeding, the mice were randomly divided into 7 groups (Table 1). Mice in different groups were fed different diets for 6 weeks. Mice in group 1 were fed a normal diet, mice in group 2 were fed a 60% high-fat diet as the high-fat control group, and mice in groups 3 - 7 were fed a 60% high-fat diet containing different proportions of lily and lotus seed respectively.

[0147] Table 3: Mouse grouping and intervention feed composition

[0148] Group number Grouping Dietary intervention Energy kcal Quantity 1 Normal diet group Normal diet 2032.2 8 rats 2 High-fat diet group High-fat diet 4057.2 8 rats 3 16% lotus seed High-fat diet + 16% lotus seed 3957.1 7 rats 4 16% lily High-fat diet + 16% lily 3967.6 8 rats 5 12% lily + 4% lotus seed High-fat diet + 12% lily + 4% lotus seed 3967.609 8 rats 6 8% lily + 8% lotus seed High-fat diet + 8% lily + 8% lotus seed 3962.454 8 rats 7 4% lily + 12% lotus seed High-fat diet + 4% lily + 12% lotus seed 3959.799 8 rats

[0149] 1.1.3 Index monitoring

[0150] (1) Body weight and food intake: The body weight and food intake of the mice were measured every week, and the data were recorded.

[0151] (2) Fasting blood glucose: After 3 weeks of dietary intervention, the fasting blood glucose of the mice was measured after a 5-hour fasting period.

[0152] (3) Glucose tolerance test: After 5 weeks of dietary intervention, the mice were fasted for 5 hours, and then intraperitoneally injected with 1.0 g / kg BW of glucose. The blood glucose of the mice was measured at 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min after glucose injection.

[0153] (4) Sample collection: After 6 weeks of dietary intervention, the mice were anesthetized with an overdose of isoflurane, and then blood was collected from the apex of the heart into an anticoagulation tube. The blood was centrifuged at 3000 r / min for 15 min at 4°C, and the supernatant was taken and stored at -80°C. The liver and epididymal fat of the mice were taken and weighed, and then stored at -80°C.

[0154] (5) Serum index detection: The total cholesterol content in the serum of the mice was measured using a total cholesterol detection kit.

[0155] 1.2 Experimental results

[0156] 1.2.1 Content of core components of lily and lotus seed

[0157] The content of lily polysaccharide in lily was measured to be approximately 12.18% by the anthrone-sulfuric acid method, and the content of neferine in lotus seed was measured to be approximately 4.81% by high performance liquid chromatography.

[0158] Table 4: Determination Results of Core Component Contents in Lily and Lotus Seed

[0159] Core ingredient Lily polysaccharide Neferine Content (%) 12.18±0.08 4.81±0.22

[0160] 1.2.2 Effects of Lily and Lotus Seed Composition on Mice Induced by High-Fat Diet

[0161] The effects of the lily and lotus seed composition on various indexes of mice induced by high-fat diet are shown in Table 5

[0162] Table 5

[0163]

[0164] Note: Data are expressed as Mean±SEM, *P<0.05, **P<0.01, ***P<0.001, indicating significant differences compared with the low-fat control group; #P<0.05, indicating significant differences compared with the high-fat control group

[0165] The following conclusions can be drawn from Table 5

[0166] (1) The lily and lotus seed composition slows down the weight gain of mice induced by high-fat diet

[0167] As Figure 1 shown in A-D, during the 6-week intervention with different diets, the body weight of the high-fat diet group was higher than that of the normal diet group of mice. The body weight and body weight change of the lotus seed group alone and the 4% lily + 12% lotus seed group were significantly higher than those of other groups during the whole intervention process; the body weight and body weight change of the 12% lily + 4% lotus seed group and the 8% lily + 8% lotus seed group were close to those of the high-fat diet group during the whole intervention process, being at a medium level; while the overall body weight and body weight change of the 12% lily + 4% lotus seed group were lower than those of the model group and close to those of the normal diet group during the whole intervention process. Thus, among the lily and lotus seed groups, the 12% lily + 4% lotus seed group was closest to the normal diet group and had an extremely excellent effect on controlling the body weight gain of mice. And this effect could not be achieved when using lily or lotus seed group alone or in equal proportion. Moreover, other lily and lotus seed groups such as the 4% lily + 12% lotus seed group not only could not achieve the effect of reducing body weight, but might even increase the body weight

[0168] Therefore, the research of the present invention shows that a lily and lotus seed composition with a specific ratio (such as the 12% lily + 4% lotus seed group) has a synergistic effect in controlling high-fat-induced weight gain, and its weight-reducing effect is better than using any one alone or using the two (lily, lotus seed) in equal proportion

[0169] In order to further verify whether the treatment with the lily and lotus seed composition has an impact on the food intake of mice, the food intake of mice was measured and calculated weekly during the experiment

[0170] The results are asFigure 1 As shown in Figures E - F, the lily group, lotus seed group, and lily - lotus seed group had no significant effect on the food intake of mice, indicating that the differences in the body weights of the experimental group mice were not caused by food intake.

[0171] After 6 weeks of treatment, the mice were sacrificed for organ index analysis. The results are as Figure 1 shown in Figure G. The liver weight and liver index (the percentage of liver weight to body weight) of the high - fat diet group were significantly higher than those of the normal diet group; compared with the high - fat diet group, the lily group, lotus seed group, and lily - lotus seed group could all control the increase in mouse liver weight to varying degrees. Among them, the liver weights of the 12% lily + 4% lotus seed group and 4% lily + 12% lotus seed group had significant differences compared with the high - fat diet group. Among them, the 12% lily + 4% lotus seed group had an extremely excellent effect on reducing liver weight, and the reduction was better than that of the single lily group and single lotus seed group.

[0172] Meanwhile, the epididymal fat weight of the high - fat diet group was significantly higher than that of the normal diet group; as Figure 1 shown in Figure H, among the lily - lotus seed groups with different ratios, the 12% lily + 4% lotus seed group had an extremely significant effect on reducing the epididymal fat weight in mice. And the reduced weight was more than the sum of the weights reduced by using lily alone or lotus seed alone. This reflects its synergistic effect in reducing fat weight.

[0173] (2) The lily - lotus seed composition can control the increase in blood glucose of obese mice induced by high - fat diet

[0174] After 3 weeks of different feed interventions, fasting blood glucose was measured. The results are as Figure 2 shown in Figure A. The fasting blood glucose of the lotus seed group and high - fat diet group mice was significantly higher than that of the normal diet group, and the fasting blood glucose of the 16% lily group and 12% lily + 4% lotus seed group mice was significantly lower than that of the high - fat diet group. It can be inferred that lily can better control blood glucose and maintain relatively stable blood glucose compared with lotus seed.

[0175] After 5 weeks of different feed interventions, an oral glucose tolerance test was conducted. The results are as Figure 2 shown in Figures B - D. During the 2 hours after glucose administration, the blood glucose of the high - fat diet group mice at each detection time point was significantly higher than that of the normal diet group, and the blood glucose of the 12% lily + 4% lotus seed group mice at each detection time point was significantly lower than that of the high - fat diet group. Moreover, the area under the blood glucose curve of the 12% lily + 4% lotus seed group mice was significantly lower than that of the high - fat diet group and had no significant difference compared with the normal diet group, while the areas under the blood glucose curves of the lily - lotus seed groups with other ratios and the single groups had no significant differences compared with the high - fat group. This shows that among the lily - lotus seed groups with different ratios, the 12% lily + 4% lotus seed group has an extremely excellent effect on controlling glucose tolerance abnormalities, and the control of blood glucose is better than the sum of the single lily group and single lotus seed group. This reflects its synergistic effect in controlling the increase in blood glucose induced by high - fat diet.

[0176] (3) The lily lotus seed composition can control the increase in the total cholesterol level in the serum of obese mice induced by a high-fat diet.

[0177] After 5 weeks of intervention with different feeds, the serum of the mice was collected. The results are as Figure 3 shown. The results of the total cholesterol measurement showed that the total cholesterol levels in the serum of the mice in the lotus seed group and the lily lotus seed group were reduced to varying degrees compared with those of the mice in the high-fat feed group. Among them, in the lily lotus seed groups with different proportions, the group with 12% lily + 4% lotus seed had an extremely excellent effect on controlling the increase in the serum total cholesterol level, and the control of total cholesterol was better than the sum of the lily group and the lotus seed group alone. This reflects its synergistic effect in controlling the increase in total cholesterol induced by a high-fat diet.

[0178] Discussion

[0179] In this study, the contents of two substances that may affect the body's blood glucose regulation ability in lily and lotus seed, namely lily polysaccharide and methyl liensinine, were determined. And the lily and lotus seed were replaced into the high-fat mouse diet according to a certain proportion, and the mice were fed to construct a sugar / lipid metabolism disorder model to explore the efficacy of the lily lotus seed composition in the core formula of the biscuit. Five groups of mice, namely the normal feed group, the high-fat feed group, the lily group, the lotus seed group, and the lily lotus seed group, were set up for preventive intervention, and the body weight, food intake, organ weight, fasting blood glucose, glucose tolerance, total cholesterol, and triglyceride in the serum of each group of mice were measured and analyzed. The analysis found that the lily lotus seed composition can slow down the increase in the body weight, liver and epididymal fat weight of the diet-induced mice, reduce blood glucose and blood lipids, and improve the body's sugar metabolism ability, but has no obvious effect on its food intake.

[0180] In summary, the lily lotus seed composition is superior to the oral liquid in assisting in reducing blood glucose and maintaining the body's sugar metabolism ability, and both have the effect of reducing total cholesterol.

[0181] This study first discovered the role of a lily lotus seed composition in a specific proportion in controlling the abnormal glucose and lipid metabolism of diet-induced obese mice. Specifically, it shows that (1) slowing down the weight gain and the accumulation of body fat in obese mice; (2) controlling the increase in blood glucose and glucose tolerance abnormalities; (3) controlling the increase in serum total cholesterol; (4) controlling liver enlargement. The lily lotus seed composition has a better effect than using lily and lotus seed alone, especially in controlling body weight, body fat, liver enlargement, and blood lipids.

[0182] The study first compared the effects of lily and lotus seed in regulating glycolipid metabolism in vivo. The study found that (1) in terms of controlling body weight, body fat and blood glucose levels, the effect of using lily alone was stronger than that of using lotus seed alone. The rich dietary fiber, more than 30 steroidal saponins and up to 12% lily polysaccharide in lily are important active ingredients for controlling body weight and blood glucose. (2) In terms of controlling hepatomegaly and serum cholesterol, using lotus seed alone was stronger than lily, and liensinine, polyphenols and flavonoids in lotus seed played an important role.

[0183] The present invention can be used for the control and improvement of glycolipid metabolism disorders such as obesity, type 2 diabetes, fatty liver, hyperlipidemia, etc.

[0184] The formula of the present invention can be applied to the development of products such as food, feed, health food, special dietary food, special medical food, medicine, etc.

[0185] All the documents mentioned in the present invention are cited in this application as references, as if each document was cited separately as a reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. Use of a combination of lily and lotus seed, characterized in that, for preparing a preparation or composition, and the preparation or composition is used for: (a) Alleviating fatty liver; (b) Lowering blood sugar; (c) Lowering cholesterol; and / or (d) Reducing body weight; wherein, the content ratio (wt:wt) of the lily and the lotus seed is: 4:1 - 2:

1.

2. The use according to claim 1, characterized in that, the fatty liver is the fatty liver caused by a high-fat diet.

3. The use according to claim 1, characterized in that, the composition is a pharmaceutical composition, a dietary supplement, a food composition, a feed composition or a health care product composition.

4. The use according to claim 1, characterized in that, the pharmaceutical composition is used for treating diseases including those selected from the group consisting of: obesity, type 2 diabetes, fatty liver, hyperlipidemia, or a combination thereof.

5. The use according to claim 1, characterized in that, the dosage form of the pharmaceutical composition includes: oral preparations, injections, lyophilized products.

6. An active ingredient combination, characterized in that, the active ingredient combination includes: (Z1) A first active ingredient, and the first active ingredient is lily or an extract thereof; and (Z2) A second active ingredient, and the second active ingredient is lotus seed or an extract thereof; wherein, the content ratio (wt:wt) of the first active ingredient to the second active ingredient is: 4:1 - 2:

1.

7. The active ingredient combination according to claim 6, characterized in that, the total content of the first active ingredient and the second active ingredient is 70 - 100 wt%, preferably 80 - 100 wt%, more preferably 90 - 100 wt%, based on the total weight of the active ingredient combination, and the weight is dry weight.

8. The active ingredient combination according to claim 6, characterized in that, in the combination, the first active ingredient and the second active ingredient are independent of each other or are mixed with each other.

9. A pharmaceutical composition, characterized in that, the pharmaceutical composition contains: (Z1) A first active ingredient, and the first active ingredient is lily or an extract thereof; (Z2) A second active ingredient, and the second active ingredient is lotus seed or an extract thereof; and (Z3) A pharmaceutically acceptable carrier; wherein, the content ratio (wt:wt) of the first active ingredient and the second active ingredient is: 4:1 - 2:

1.

10. A feed composition, characterized in that, the feed composition contains: (Z1) A first active ingredient, and the first active ingredient is lily or an extract thereof; (Z2) A second active ingredient, and the second active ingredient is lotus seed or an extract thereof; and (Z3) A food-acceptable carrier wherein, the content ratio (wt:wt) of the first active ingredient and the second active ingredient is: 4:1 - 2:1, and the total content of the first active ingredient and the second active ingredient is 10 wt% - 30 wt%.

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