Use of hemerocallis citrina baroni in dispelling effects of alcohol and protecting liver

Abstract

A use of Hemerocallis citrina Baroni or an extract thereof as an active ingredient in preparing a composition for dispelling effects of an alcohol. In addition, the composition can also be used for auxiliary protection against alcoholic liver damage.

Description

Daylily's use in sobering up and protecting the liver Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a new use of daylily for sobering up and protecting the liver. Background Art

[0002] Wine culture has a long and rich history in Chinese civilization. Wine has permeated the entire 5,000-year history of Chinese civilization, occupying a vital place in every aspect of Chinese life, from literary and artistic creation, cultural entertainment, to culinary delights. However, alcohol is a damp, hot, and toxic substance. Excessive alcohol consumption, either short-term or long-term, can severely damage the functions of various organs. Research has linked alcohol to over 200 diseases and contributes to approximately 3 million deaths worldwide each year. For example, alcohol can irritate the brain, causing short-term mental states such as excitement, agitation, and irritability. Long-term alcohol consumption can also lead to alcoholic encephalopathy. Chronic ethanol intoxication can cause hand tremors and impaired ability to hold objects, which can be life-threatening in severe cases. Alcohol irritates the gastric mucosa, making patients more susceptible to symptoms such as stomach pain, stomach cramps, diarrhea, vomiting, and indigestion. It can also lead to gastritis, gastric ulcers, gastric bleeding, and gastric mucosal erosions. Alcohol accelerates vasoconstriction and dilation, and the brain and heart are particularly important blood supply sites. Excessive alcohol consumption can cause palpitations, chest tightness, and shortness of breath. Long-term drinking can lead to coronary heart disease, myocardial infarction, and cerebral hemorrhage. Alcohol is particularly damaging to the liver. As the primary organ for ethanol metabolism and detoxification, excessive and long-term drinking can increase the metabolic burden on the liver, leading to pathological liver damage and the accumulation of fatty liver. Alcohol can also cause alcoholic hepatitis, which, if not treated and controlled promptly, can lead to cirrhosis and liver cancer. In order to alleviate the damage caused to the body by single or multiple long-term drinking and improve the quality of public health, the research and development of alcohol detoxification and liver protection products has become a research hotspot.

[0003] Therefore, there is an urgent need to develop a product that effectively relieves alcohol and protects the liver.

[0004] Summary of the Invention

[0005] The main purpose of the present invention is to provide a product that can effectively sober up and protect the liver.

[0006] In a first aspect, the present invention provides use of Hemerocallis citrina Baroni or an extract thereof as an active ingredient in preparing a composition for sobering up.

[0007] In another preferred embodiment, the hangover relief comprises the step of accelerating ethanol metabolism, more specifically, increasing the activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) to reduce the levels of ethanol and acetaldehyde in the body.

[0008] In another preferred embodiment, the hangover remedy comprises one or more selected from the following groups:

[0009] Reduce the rate of intoxication, delay the latency period of intoxication, shorten the duration of intoxication, improve behavioral changes after intoxication, restore motor ability after intoxication, relieve alcohol intoxication, and / or reduce the mortality rate of acute alcohol intoxication.

[0010] In another preferred embodiment, the extract is selected from the following group: a water extract of daylily, an organic solvent extract of daylily, or an active component separated from the daylily extract (such as daylily polysaccharides, daylily flavonoids, daylily alkaloids, daylily nucleosides or a combination thereof), or a combination thereof.

[0011] In another preferred embodiment, the extraction solvent used for the extract is selected from: water, C1-C5 alcohol, petroleum ether, ethyl acetate, dichloromethane, dichloroethane, acetone, or a combination thereof.

[0012] In another preferred embodiment, the extraction solvent used for the extract is a C1-C5 alcohol selected from the following group: methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol or tert-butanol.

[0013] In another preferred embodiment, the extraction solvent used for the extract is a 100-0% C1-C5 alcohol-water (v / v) mixed solvent, preferably, 20-90% C1-C5 alcohol, more preferably 90-40% C1-C5 alcohol, more preferably 80-60% C1-C5 alcohol, such as 95% C1-C5 alcohol, 90% C1-C5 alcohol, 85% C1-C5 alcohol, 80% C1-C5 alcohol, 75% C1-C5 alcohol, 70% C1-C5 alcohol, 65% C1-C5 alcohol, 60% C1-C5 alcohol, 50% C1-C5 alcohol, 40% C1-C5 alcohol, 30% C1-C5 alcohol, 20% C1-C5 alcohol, 10% C1-C5 alcohol or 5% C1-C5 alcohol, wherein 20%-30% C1-C5 alcohol, or 75%-95% C1-C5 alcohol-water extract is a preferred extract.

[0014] In another preferred embodiment, the extraction solvent used for the extract is a 100-0% ethanol-water (v / v) mixed solvent, preferably, 20-90% ethanol, more preferably 90-40% ethanol, more preferably 80-60% ethanol, such as 95% ethanol, 90% ethanol, 85% ethanol, 80% ethanol, 75% ethanol, 70% ethanol, 65% ethanol, 60% ethanol, 50% ethanol, 40% ethanol, 30% ethanol, 20% ethanol, 10% ethanol or 5% ethanol, among which 20%-30% ethanol and 75%-95% ethanol-water extracts are preferred extracts.

[0015] In another preferred embodiment, the extract is prepared by the following method: extracting the daylily in an extraction solvent at 80-120° C. for 1-5 times, combining the extracts and removing the solvent to obtain the daylily extract.

[0016] In another preferred embodiment, the extraction time for each extraction is independently 30 min-2 h, preferably 45 min-1.5 h or 1 h.

[0017] In another preferred embodiment, the weight ratio of the daylily to the extraction solvent on a dry weight basis is 1:1-50, preferably 1:2-40, such as 1:5, 1:6, 1:7, 1:7.5, 1:8, 1:10, 1:12, 1:15, 1:20, 1:25, 1:30, 1:30 or 1:40.

[0018] In another preferred embodiment, the extraction times are 1, 2, 3, 4 or 5 times.

[0019] In another preferred embodiment, the solvent is removed by the following methods: concentration under reduced pressure, spray drying, freeze drying, or a combination thereof.

[0020] In another preferred embodiment, the daylily is the whole plant, flower buds, roots, stems, leaves, or a combination thereof, preferably, the daylily is the flower buds.

[0021] In another preferred embodiment, the composition is also used for auxiliary protection against alcoholic liver damage (ie, for protecting the liver).

[0022] In another preferred embodiment, the auxiliary protection against alcoholic liver damage includes one or more uses selected from the following group:

[0023] (a) Alleviate acute or chronic liver cell and liver damage caused by alcohol and protect the liver, such as reducing alcohol-induced increases in transaminases, triglycerides, and cholesterol, restoring albumin levels, and improving alcohol-induced hypoglycemia;

[0024] (b) reducing oxidative stress damage, improving inflammatory damage, and preventing hepatitis; and / or

[0025] (c) Improve alcoholic liver disease, alleviate fatty degeneration or fat accumulation in liver tissue caused by alcohol, and reduce the content of lipid droplets, triglycerides, cholesterol, etc.;

[0026] (d) Improve liver aging caused by long-term drinking, improve changes in liver aging-related markers, and restore liver health.

[0027] In another preferred embodiment, the composition is a food, a special medical food, a health product or a pharmaceutical composition.

[0028] In another preferred embodiment, the composition further comprises a physiologically acceptable carrier.

[0029] In another preferred embodiment, the composition is in the form of a solid preparation or a liquid preparation.

[0030] The second aspect of the present invention provides a composition comprising:

[0031] (a) daylily or a daylily extract as an active ingredient; and

[0032] (b) a physiologically acceptable carrier.

[0033] In another preferred embodiment, the daylily or daylily extract is as described above.

[0034] In another preferred embodiment, the composition is in the form of a solid preparation or a liquid preparation. In another preferred embodiment, the composition is a food, a special medical food, a health product or a pharmaceutical composition.

[0035] The third aspect of the present invention provides use of the composition described in the second aspect of the present invention in preparing a food, a special medical food, a health product or a pharmaceutical composition for sobering up.

[0036] In another preferred embodiment, the food, special medical food, health product or pharmaceutical composition is also used to assist in protecting alcoholic liver damage.

[0037] The fourth aspect of the present invention provides the use of the composition described in the second aspect of the present invention in the preparation of food, special medical food, health product or pharmaceutical composition for auxiliary protection against alcoholic liver damage (i.e., for liver protection).

[0038] In a fifth aspect, the present invention provides a method for sobering up, comprising the steps of administering the active ingredient of the present invention, or administering the composition of the second aspect of the present invention, to a subject in need thereof, thereby sobering up.

[0039] In another preferred embodiment, the active ingredient or composition can be administered before, during or after drinking alcohol.

[0040] In another preferred embodiment, the method is also a method for assisting in protecting alcoholic liver damage.

[0041] In another preferred embodiment, the subject is a human or non-human mammal, such as a rat, mouse, or monkey.

[0042] 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 described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 shows the results of the ethanol metabolism rate test at the cellular level of daylily

[0044] Figure 2 shows the results of the antioxidant capacity test at the cellular level of daylily

[0045] Figure 3 shows the results of the test on the metabolic kinetics of serum ethanol in drunken mice by daylily.

[0046] Figure 4 shows the results of behavioral (rotarod) test on drunk mice by daylily

[0047] Figure 5 shows the results of behavioral (pole climbing) tests on drunk mice using daylily.

[0048] Figure 6 shows the results of behavioral (righting reflex) test of daylily on drunk mice

[0049] Figure 7 shows the results of the cell survival rate test of Daylily in the alcohol-induced liver cell injury model

[0050] Figure 8 Results of the test on the mortality rate of mice with acute alcohol poisoning by daylily

[0051] Figure 9 shows the test results of the protective ability of daylily on the liver of mice with acute alcoholic liver injury

[0052] Figure 10 shows the test results of regulating lipid droplet content at the cellular level of daylily

[0053] Figure 11 shows the effect of daylily on the liver status and liver-to-body ratio of mice with chronic alcoholic liver disease

[0054] Figure 12 shows the test results of daylily on liver function indicators (AST, ALT) of mice with chronic alcoholic liver disease

[0055] Figure 13 shows the results of the daylily test on albumin and blood sugar in mice with chronic alcoholic liver disease

[0056] Figure 14 shows the test results of daylily on inflammatory indicators (interleukin-6 and TNF-α) in mice with chronic alcoholic liver disease

[0057] Figure 15 shows the test results of daylily on key enzymes of ethanol metabolism (alcohol dehydrogenase, acetaldehyde dehydrogenase, cytochrome P450 2E1) in mice with chronic alcoholic liver disease

[0058] Figure 16 shows the results of the test on the antioxidant index superoxide dismutase activity of daylily in mice with chronic alcoholic liver disease

[0059] Figure 17 shows the test results of daylily on serum and liver tissue triglyceride and cholesterol in mice with chronic alcoholic liver disease

[0060] Figure 18 shows the results of H&E staining test on liver tissue pathology sections of mice with chronic alcoholic liver disease.

[0061] Figure 19 shows the results of the Oil Red O staining test on liver tissue sections of mice with chronic alcoholic liver disease.

[0062] FIG20 shows the test results of Daylily on serum and liver tissue aging indicators (CXCL-1, MMP-1, etc.) of mice with chronic alcoholic liver disease;

[0063] FIG21 shows the results of the test of SA-β-gal positive cell rate, an indicator of aging, in liver tissue sections of mice with chronic alcoholic liver disease using Daylily;

[0064] FIG22 shows the test results of daylily on the aging indicators p53 and γH2AX in the liver tissue of mice with chronic alcoholic liver disease; FIG23 shows the safety test results of daylily on mice. DETAILED DESCRIPTION

[0065] After extensive and in-depth research, the inventors discovered for the first time that daylily and its extract can effectively sober up and / or protect the liver, thus providing a new, safe and effective sober up and liver protection product. Based on this, the present invention was completed.

[0066] the term

[0067] Active ingredient

[0068] As used herein, the active ingredient of the present invention comprises daylily or a daylily extract.

[0069] Daylily (Hemerocallis citrina Baroni, Daylily), also known as daylily, forget-me-not, daylily, daylily flower, brain-strengthening vegetable, tranquilizing vegetable, green onion flower, lemon daylily, daylily flower, etc., is an angiosperm of the genus Hemerocallis of the family Asparagaceae, Asparagaceae, Magnoliaceae, with stomachic, diuretic, and detumescent effects. It has certain medicinal value and is also an edible vegetable with a long history of consumption. Through extensive screening of Chinese herbal medicine libraries, the inventors discovered for the first time that daylily (especially its alcohol-water extract) is an edible Chinese herbal medicine with excellent alcohol-relieving and liver-protecting activities and high safety, thereby providing a drug, food, or health product composition with daylily or its extract as an active ingredient for alcohol-relieving and liver-protecting.

[0070] The raw material of the daylily flower that can be used in the present invention is usually dry daylily flower or its chopped or crushed form. The daylily flower can be the whole plant, flower bud, root, stem, leaf, or a combination thereof, preferably, the flower bud or root of the daylily flower.

[0071] More preferably, the present invention also provides a daylily extract having excellent activity and being very suitable for preparing a composition product; or active components (daylily polysaccharides, daylily flavonoids, daylily alkaloids, daylily nucleosides or combinations thereof) separated from the daylily extract.

[0072] Preferably, the extraction solvent used in the daylily extract of the present invention is selected from: water, C1-C5 alcohol, petroleum ether, ethyl acetate, dichloromethane, dichloroethane, acetone, or a combination thereof. More preferably, the extraction solvent used in the extract is a 100-0% ethanol-water (v / v) mixed solvent, preferably, 90-40% ethanol, more preferably 80-60% ethanol, such as 95% ethanol, 90% ethanol, 85% ethanol, 80% ethanol, 75% ethanol, 70% ethanol, 65% ethanol, 60% ethanol, 50% ethanol, 40% ethanol, 30% ethanol, 20% ethanol, 10% ethanol, or 5% ethanol, wherein the 20-30% ethanol or 75%-95% ethanol-water extract is a preferred extract.

[0073] In another preferred embodiment, the extract is prepared by the following method: extracting the daylily in an extraction solvent at 80-120° C. for 1-5 times, combining the extracts and removing the solvent to obtain the daylily extract.

[0074] Hangover and liver protection

[0075] As used herein, hangover relief refers to when a person or animal, after consuming alcohol, has less alcohol content, lower concentration and / or shorter existence time in the subject's body, compared to not using the active ingredient or composition of the present invention, thereby reducing the physical and mental effects or damage of alcohol. The hangover relief of the present invention includes preventing drunkenness by accelerating ethanol metabolism, rapidly decomposing ethanol into acetaldehyde, and ultimately decomposing into carbon dioxide and water, thereby reducing or delaying alcohol's inhibition of the central nervous system, reducing the possibility of the subject reaching a drunken state (alcohol poisoning), alleviating alcohol-induced behavioral disorders, or prolonging the incubation period of drunkenness. In addition, the hangover relief of the present invention also includes "sobering up", which allows the subject to wake up from a drunken state, restore consciousness or restore self-control. It usually does not refer to natural sobriety, but rather to taking hangover relief substances to accelerate ethanol metabolism, which serves the purpose of accelerating sobriety and reducing the harm of ethanol to the body.

[0076] The metabolism of alcohol in the body primarily relies on two enzymes in the liver: alcohol dehydrogenase and acetaldehyde dehydrogenase. Alcohol dehydrogenase removes two hydrogen atoms from alcohol molecules, breaking down ethanol into acetaldehyde. Acetaldehyde dehydrogenase, on the other hand, removes two hydrogen atoms from acetaldehyde, converting it into acetic acid, which ultimately decomposes into carbon dioxide and water. Alcohol and acetaldehyde are the most harmful to the body. A deficiency in acetaldehyde dehydrogenase prevents acetaldehyde from being completely broken down into acetic acid, instead allowing it to remain in the body as acetaldehyde. This leads to symptoms of intoxication such as nausea, loss of balance, slurred speech, impaired judgment, and unconsciousness. Experiments have demonstrated that the active ingredients or compositions of the present invention can increase the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, accelerating alcohol metabolism and thus achieving hangover relief.

[0077] As used herein, "liver protection" refers to protecting the liver and its auxiliary protective function against alcoholic liver damage. The liver is the primary organ for the breakdown and metabolism of alcohol. Drinking alcohol can lead to liver damage, even to alcoholic fatty liver disease, alcoholic cirrhosis, and alcoholic liver failure. The higher the alcohol level in the body and the longer the drinking time, the more severe the liver damage tends to be. Experimental studies have shown that the active ingredients or compositions of the present invention can alleviate alcohol-induced liver cell damage, improve alcohol-induced liver function impairment, reduce liver cell fat accumulation, enhance liver cell antioxidant and anti-inflammatory functions, promote liver rejuvenation, and / or slow the progression of alcoholic liver disease.

[0078] The "prevention," "treatment," "improvement," and "relief" described herein include delaying and stopping the progression of a disease or symptom, or eliminating the disease, and do not require 100% inhibition, elimination, or reversal. In some embodiments, the composition of the present invention can alleviate, shorten, or reverse one or more symptoms or indicators of drunkenness or alcoholic liver damage, for example, by 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 composition of the present invention.

[0079] Some representative hangover indicators (but not limited to): drunkenness rate, righting reflex disappearance rate of drunken mice, righting reflex disappearance time and recovery time, pole climbing time, pole climbing score, rod rotation time, grip strength, hanging endurance, mouse serum ethanol concentration, mouse liver ADH and ALDH activity and expression, and CYP2E1 expression.

[0080] Some representative liver protection indicators (but not limited to): mortality rate of mice with acute alcohol poisoning, liver-to-body ratio, ALT, AST, ALB, Glu, TNF-α, IL-6, TG, CHOL, lipid droplet content, pathological morphology, aging indicators SA-β-gal, CXCL-1, CXCL-3, MMP-1, MMP-7, IL-1β, p16, p53, γH2AX.

[0081] Compositions and methods of administration

[0082] The present invention also provides compositions comprising the active ingredients of the present invention and a physiologically acceptable carrier.

[0083] The composition can be used to sober up. Experiments have shown that the active ingredient of the present invention can accelerate ethanol metabolism, thereby sobering up. More specifically, it can reduce ethanol and acetaldehyde levels in the body by increasing the activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), thereby delaying the onset of intoxication, shortening the duration of intoxication, improving behavioral symptoms after intoxication, restoring motor ability after intoxication, alleviating alcohol poisoning, and / or reducing the mortality rate of acute alcohol poisoning.

[0084] Furthermore, the compositions of the present invention can also be used to protect the liver, particularly by reducing acute or chronic alcohol-induced liver cell and liver damage. More specifically, the active ingredients of the present invention can reduce alcohol-induced increases in transaminases, triglycerides, and cholesterol, restore albumin levels, and alleviate alcohol-induced hypoglycemia; reduce oxidative stress, alleviate inflammatory damage, and prevent hepatitis; and / or improve alcoholic liver disease, alleviate alcohol-induced fatty degeneration or fat accumulation in liver tissue, and reduce lipid droplet, triglyceride, and cholesterol levels.

[0085] The compositions of the present invention include, but are not limited to, foods, special medical foods, health products and medicines.

[0086] In the composition, the active ingredient of the present invention can be used alone as the sole active ingredient, or can be used in combination with other active ingredients.

[0087] Typically, the composition of the present invention contains an effective amount of the active ingredient. The term "effective amount" or "effective dose" refers to an amount that can produce a function or activity (i.e., alcohol sobering, liver protection) on humans and / or animals and is acceptable to humans and / or animals.

[0088] As used herein, the term "physiologically acceptable" ingredients are substances that are suitable for use in humans and / or mammals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., substances with a reasonable benefit / risk ratio. The term "physiologically acceptable carrier" refers to a carrier for administering a therapeutic agent, including various excipients and diluents.

[0089] The compositions of the present invention contain a safe and effective amount of the active ingredient of the present invention and a physiologically acceptable carrier, which can be in solid or liquid form. The carriers include (but are not limited to): water, NaCl, physiological saline solution, lactated Ringer's solution, regular sucrose, regular glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, and pigments. Such preparations can be sterilized and, if necessary, mixed with adjuvants that do not adversely react with or interfere with the activity of the compounds provided herein, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, colorants, and / or aromatic substances. Those of ordinary skill in the art will recognize that other carriers and excipients are suitable for the disclosed compounds.

[0090] The compositions of the present invention can be administered to a subject by any suitable route, including orally, subcutaneously, intravenously, intramuscularly, transdermally, by inhalation spray, topically, rectally, nasally, or buccally. Preferably, the active compound is administered orally, subcutaneously, intravenously, intramuscularly, or transdermally.

[0091] Compositions of the invention suitable for administration will typically be discrete units in solid form, for example, in the form of tablets, capsules, powders, granules, pills, patches, gels, ointments, suppositories, or in liquid form, for example, oral liquids, injectable or infusible solutions or suspensions, or lyophilized powders.

[0092] The composition is preferably prepared under sterile conditions. The choice of carrier should match the mode of administration, which is well known to those skilled in the art.

[0093] The effective amount of the active ingredient of the present invention may vary with the mode of administration and the amount of alcohol consumed by the subject, the degree of intoxication, the severity of the intoxicated behavior, etc. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease or symptoms to be treated or improved by the subject, the patient's weight, the patient's immune status, the route of administration, etc. Typically, the active ingredient of the present invention can be administered daily at a dose of about 0.1 mg / kg-10 g / kg, 1 mg / kg-1 g / kg, or 10 mg / kg-1000 mg / kg of animal body weight. For example, depending on the urgency of the subject's condition, several divided doses may be administered daily, or the dose may be reduced proportionally.

[0094] The present invention also provides a method for sobering up, comprising the steps of administering an active ingredient of the present invention or a composition of the present invention to a subject in need thereof, thereby sobering up. The active ingredient or composition of the present invention can be administered before, during, or after drinking. In particular, the active ingredient or composition can be administered from 6 hours before drinking to the start of drinking, such as 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 0.5 hours, 10 minutes, 5 minutes, or 1 minute before drinking, at any time during drinking, or within 6 hours after drinking, such as 1 minute, 5 minutes, 10 minutes, 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, or 5 hours after drinking.

[0095] In another preferred embodiment, the subject is a human or non-human mammal, such as a rat, mouse, or monkey.

[0096] The main advantages of the present invention include:

[0097] A) The present invention is the first to discover the use of daylily in sobering up and accelerating ethanol metabolism.

[0098] B) The present invention is the first to discover that daylily has the ability to alleviate acute alcohol poisoning and resist alcoholic liver disease.

[0099] C) The active ingredients of the present invention are derived from edible Chinese medicines / vegetables and are safe and effective.

[0100] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which no specific conditions are specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0101] Example 1 Preparation of Daylily Samples

[0102] The present invention uses daylily samples from Anhui, Fujian, Heilongjiang, Shanxi, Hunan, Sichuan, and Gansu. The daylily (dried flower buds) are crushed and decocted three times using water, 10% ethanol, 20% ethanol, 30% ethanol, 40% ethanol, 50% ethanol, 75% ethanol, 95% ethanol, petroleum ether, ethyl acetate, dichloromethane, n-butanol, and other organic solvents at a certain material-liquid ratio. The crude extracts are then concentrated under reduced pressure and freeze-dried to obtain daylily crude extracts from different production areas and extracted using different methods, and the yields are calculated for each extract.

[0103] Weigh 20 g of dry material of daylily (originating from Anhui, Fujian, Heilongjiang, Shanxi, Hunan, Sichuan, and Gansu), crush it in a grinder, and boil it three times with 75% ethanol at 100°C with a material-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried. 1.52 g was obtained from Anhui, with a yield of 7.6%; 1.78 g was obtained from Fujian, with a yield of 8.9%; 1.82 g was obtained from Heilongjiang, with a yield of 9.1%; 1.54 g was obtained from Shanxi, with a yield of 7.7%; 1.62 g was obtained from Hunan, with a yield of 8.1%; 1.49 g was obtained from Sichuan, with a yield of 7.45%; and 1.92 g was obtained from Gansu, with a yield of 9.6%. These are the daylilies (75% ethanol extracts) from different origins.

[0104] Weigh 10 g of dried daylily buds, grind them into powder, and boil them three times with 95% ethanol at 100°C with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.394 g with a yield of 13.94%, which is daylily bud (95% ethanol extract).

[0105] Weigh 5 g of dried daylily, grind it into powder, and boil it with 60% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.029 g with a yield of 20.58%, which is daylily (60% ethanol extract).

[0106] Weigh 5 g of dried daylily buds, grind them into powder, and boil them three times with 50% ethanol at 100°C with a material-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.378 g with a yield of 27.56%, which is daylily bud (50% ethanol extract).

[0107] Weigh 5 g of dried daylily, grind it into powder, and boil it with 40% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.362 g with a yield of 27.24%, which is daylily (40% ethanol extract).

[0108] Weigh 5 g of dried daylily, grind it into powder, and boil it with 30% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.453 g with a yield of 29.06%, which is daylily (30% ethanol extract).

[0109] Weigh 5 g of dried daylily buds, grind them into powder, and boil them three times with 20% ethanol at 100°C with a material-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.665 g with a yield of 33.33%, which is daylily buds (20% ethanol extract).

[0110] Weigh 5 g of dried daylily, grind it into powder, and boil it with 10% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.362 g with a yield of 27.24%, which is daylily (10% ethanol extract).

[0111] Weigh 5 g of dried daylily, grind it with a grinder, and boil it with deionized water at 100°C for 3 times with a material-liquid ratio of 1:40, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1.136 g with a yield of 22.76%, which is the daylily (water extract).

[0112] Weigh 15 g of dried daylily, grind it with a grinder, and boil it with petroleum ether at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts are combined, concentrated under reduced pressure, and then freeze-dried to obtain 0.1 g with a yield of 0.67%, which is the daylily (petroleum ether extract).

[0113] Weigh 15 g of dried daylily, grind it with a grinder, and boil it with ethyl acetate at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 0.15 g with a yield of 1%, which is daylily (ethyl acetate extract).

[0114] Weigh 15 g of dried daylily buds, grind them into powder, and boil them in dichloromethane at 100°C for 3 times with a solid-liquid ratio of 1:7.5, each time for 60 min. The extracts were combined, concentrated under reduced pressure, and then freeze-dried to obtain 0.22 g with a yield of 1.47%, which is the daylily bud (dichloromethane extract).

[0115] Weigh 15 g of dried daylily buds, grind them into powder, and boil them with n-butanol at 100°C for three times with a solid-liquid ratio of 1:7.5, each time for 60 min. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 0.5 g with a yield of 3.33%, which is daylily bud (n-butanol extract).

[0116] Example 2 Daylily promotes ethanol metabolism in hepatocytes

[0117] This experiment measures the ethanol content in the culture medium to determine the efficacy of drugs that promote ethanol metabolism. HepG2 cells were divided into a normal control group, a model group, a positive drug control group (75% ethanol extract of Hovenia dulcis, 200 μg / mL and dihydromyricetin, 100 μM), and a daylily group. The cells were cultured normally for 12 hours. The positive drug and different daylily stock solutions were diluted in culture medium to the test concentration (200 μg / mL), and the corresponding test solution was added to each well. After incubation with the cells for 6 hours, alcohol modeling was performed. Except for the cells in the normal control group, the culture medium was supplemented with 300 mM ethanol solution and incubated for 4 hours. After modeling, the supernatant was collected and the culture medium ethanol concentration was measured using the Megazyme-Alcohol Kit. The data were summarized, a bar graph was plotted using Graph Pad, and significant differences were calculated (t-test).

[0118] The data from the hepatocyte ethanol metabolism experiment are shown in Figure 1. Compared with the model group, daylily extracts extracted with different solvents had different abilities to promote hepatocyte ethanol metabolism. The 20%, 30%, 50%, 75%, and 95% ethanol extracts of daylily were the most active. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. This indicates that daylily can promote ethanol metabolism at the cellular level.

[0119] Example 3 Daylily reduces alcohol-induced oxidative stress

[0120] Alcohol can induce liver cell damage, promote high expression of reactive oxygen species, and lead to oxidative stress. This experiment investigated the antioxidant capacity of drugs by testing the content of reactive oxygen species in liver cells. HepG2 cells were divided into normal control group, model group, and daylily group. The cells were cultured normally for 12 hours, and the positive drug and daylily mother solution were diluted with culture medium to the test concentration (75% ethanol extract, 200μg / mL), and the corresponding test solution was added to each well. After incubation with the cells for 6 hours, alcohol modeling was performed. Except for the cells in the normal control group, the culture medium contained 300mM ethanol solution and incubated for 16 hours. After the modeling was completed, the ROS expression content was detected using a commercially available DCFH-DA probe. The data were summarized, fluorescence quantification was performed using Image J, a bar graph was drawn using Graph Pad, and significant differences were calculated (t-test).

[0121] The experimental results are shown in Figure 2. Compared with the control group, the fluorescence intensity of daylily was significantly reduced after administration, indicating that daylily reduced the increase in the active oxygen content of cells caused by alcohol. * indicates P < 0.05, which indicates that daylily can play an antioxidant role at the cellular level.

[0122] Example 4 Daylily promotes ethanol metabolism in drunken mice

[0123] This study examined the effects of drugs on ethanol metabolism in mice by measuring blood alcohol levels in intoxicated mice. Eight-week-old ICR male mice were divided equally according to body weight into an alcohol group, an alcohol-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg), and an alcohol-positive drug group (75% ethanol extract of Hovenia dulcis, 0.5, 1, and 2 g / kg). Thirty minutes before alcohol administration, the alcohol-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg) and the alcohol-positive drug group (0.5, 1, and 2 g / kg) were administered via oral gavage. The alcohol group received an equivalent volume of vehicle. Subsequently, all groups were given alcohol by gavage at a dose of 7 g / kg. Blood was collected from the mice by orbital sampling 0.5, 1, 2, 4, 8, and 12 hours after alcohol consumption. Serum was collected by centrifugation after standing at room temperature. Serum ethanol concentration was measured using a Megazyme-Alcohol kit, and serum acetaldehyde concentration was measured by HPLC derivatization. The data were summarized, curves were fitted using Graph Pad, and significant differences were calculated (t-test).

[0124] The experimental results, shown in Figure 3(A), show that serum ethanol levels in mice increased after drinking alcohol, reaching a peak approximately 1-3 hours later. Compared with the alcohol group, the alcohol + daylily (0.5, 1, and 2 g / kg) groups reduced serum alcohol concentrations at all time points and restored serum ethanol concentrations to normal levels more quickly, indicating that daylily can effectively promote ethanol metabolism in intoxicated mice. Concurrently, the alcohol + daylily (1 g / kg) group reduced serum acetaldehyde concentrations at all time points and restored serum acetaldehyde concentrations to normal levels more quickly.

[0125] Eight-week-old ICR male mice were divided equally into alcohol group, alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 0.8 g / kg) group, alcohol + daylily (water extract, 0.8 g / kg) group, alcohol + daylily (10% ethanol extract, 0.8 g / kg) group, alcohol + daylily (20% ethanol extract, 0.8 g / kg) group, alcohol + daylily (30% ethanol extract, 0.8 g / kg) group, alcohol + daylily (40% ethanol extract, 0.8 g / kg) group, alcohol + daylily (50% ethanol extract, 0.8 g / kg) group and alcohol + daylily (75% ethanol extract, 0.8 g / kg) group according to their body weight. Thirty minutes before alcohol administration, the alcohol-positive drug group (75% ethanol extract of Hovenia dulcis (Huangzi) fruit, 0.8 g / kg) and the alcohol-Daphne lily (Daphne dasyphylla) group were administered by oral gavage. The alcohol group received an equivalent volume of vehicle. All groups were then gavaged with alcohol at a dose of 7 g / kg. Blood was collected from the mice via orbital sampling at 0.5, 1, 2, 4, 8, and 12 hours after alcohol consumption. Serum was collected by centrifugation at room temperature and assayed for serum ethanol concentration using the Megazyme-Alcohol Kit. Data were compiled, curves were fitted using GraphPad, and significant differences were calculated using t-tests.

[0126] The experimental results are shown in Figure 3(B). The serum ethanol content of mice increased after drinking alcohol, and the serum ethanol content peaked about 1-3 hours later. Compared with the alcohol group, all alcohol + daylily groups reduced the serum alcohol concentration 0-6 hours after drinking and restored the serum ethanol concentration to normal levels more quickly. ** indicates P < 0.01, *** indicates P < 0.001, which indicates that daylily can effectively promote ethanol metabolism in drunken mice.

[0127] Example 5 Daylily improves drunken behavior in mice (rotarod)

[0128] This study examined the effects of drugs on coordination, balance, and endurance in mice after alcohol intoxication by testing their behavioral rotarod. Eight-week-old ICR male mice were divided equally according to body weight into a control group, an alcohol group, an alcohol-plus-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg), and an alcohol-plus-Daily Lily (75% ethanol extract, 0.5, 1, and 2 g / kg) groups. All groups underwent three days of rotarod training prior to the experiment. Thirty minutes before alcohol administration, the alcohol-plus-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg) and the alcohol-plus-Daily Lily (0.5, 1, and 2 g / kg) were administered via oral gavage. The control and alcohol groups were then gavaged with an equal volume of vehicle. Subsequently, all groups except the control group were gavaged with alcohol at a dose of 4.5 g / kg. Ten minutes after alcohol administration, the mice underwent rotarod behavioral testing, and the time spent on the rod was recorded. The data were summarized, histograms were drawn using Graph Pad, and significant differences were calculated (t-test).

[0129] The experimental results are shown in Figure 4. Compared with the blank group, the alcohol group significantly reduced the time mice spent on the rod (*** indicates P < 0.001). Compared with the alcohol group, the alcohol + daylily (0.5, 1, and 2 g / kg) groups significantly increased the time mice spent on the rod (** indicates P < 0.01, *** indicates P < 0.001). This suggests that daylily can effectively improve the motor function of mice after intoxication and help sober them up.

[0130] Example 6 Daylily improves drunken behavior (pole climbing) in mice

[0131] This study examined the effects of drugs on coordination, balance, and endurance in mice after alcohol intoxication by measuring behavioral pole climbing time and scores. Eight-week-old ICR male mice were divided equally according to body weight into a control group, an alcohol group, an alcohol-plus-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg), and an alcohol-plus-Daily Lily (75% ethanol extract, 0.5, 1, and 2 g / kg) groups. All groups underwent three days of pole climbing training prior to the experiment. Thirty minutes before alcohol administration, the alcohol-plus-positive drug group (75% ethanol extract of Hovenia dulcis, 1 g / kg) and the alcohol-plus-Daily Lily (0.5, 1, and 2 g / kg) were administered via oral gavage. The control and alcohol groups were gavaged with an equal volume of vehicle. Subsequently, all groups except the control group were gavaged with alcohol at a dose of 4.5 g / kg. Ten minutes after drinking, mice were tested for pole climbing behavior, their climbing performance was scored, and their climbing time was recorded. Data were summarized, plotted in a histogram using Graph Pad, and significant differences were calculated (t-test).

[0132] The experimental results are shown in Figure 5. Compared with the blank group, the alcohol group significantly reduced the time and improved the climbing score of the mice. *** indicates P < 0.001. Compared with the alcohol group, the alcohol + daylily (0.5, 1, and 2 g / kg) groups significantly increased the climbing time and reduced the climbing score of the mice. ** indicates P < 0.01. This indicates that daylily can effectively improve the motor ability of mice after intoxication and play a role in sobering up.

[0133] Example 7 Daylily improves drunken behavior (righting reflex) in mice

[0134] This study investigated the effects of drugs on the latency to intoxication and sobriety, as well as endurance and gripping strength, by measuring the loss and recovery time of the righting reflex after alcohol intoxication in mice, as well as their gripping and hanging endurance after righting reflex recovery. Eight-week-old ICR male mice were divided equally according to body weight into an alcohol group, an alcohol-infused Hovenia dulcis (75% ethanol extract, 1 g / kg), and an alcohol-infused Daylily (75% ethanol extract, 0.5, 1, and 2 g / kg) group. Thirty minutes before alcohol administration, the alcohol-infused Hovenia dulcis (1 g / kg) and alcohol-infused Daylily (0.5, 1, and 2 g / kg) groups were administered via oral gavage. The alcohol group received an equivalent volume of vehicle. Subsequently, all groups were gavaged with alcohol at a dose of 7 g / kg. The loss and recovery time of the righting reflex after alcohol intoxication were recorded, and the intoxication rate, latency to intoxication, and sobriety time were calculated. Gripping strength and hanging endurance were also measured after sobering. The data were summarized, histograms were drawn using Graph Pad, and significant differences were calculated (t-test).

[0135] The experimental results are shown in Figure 6 (AC). Compared with the alcohol group (drunkenness rate 100%), alcohol + daylily (0.5, 1, 2g / kg) reduced the drunkenness rate of mice (75%, 25%, 16.67%), and alcohol + daylily (2g / kg) significantly increased the incubation period of drunkenness in mice, *** indicates P < 0.001, and alcohol + daylily (1, 2g / kg) significantly shortened the sobering time of drunken mice, * indicates P < 0.05, ** indicates P < 0.01. This shows that daylily can reduce the drunkenness rate of mice, play a role in preventing drunkenness and accelerating sobering. As shown in Figure 6 (DE), compared with the alcohol group, alcohol + daylily (1, 2g / kg) restored the grip and hanging endurance of mice after sobering up, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. This shows that daylily can promote sobering up in mice and restore the behavioral ability of drunken mice.

[0136] Example 8 Daylily hepatocyte protection experiment

[0137] This study examined the protective effects of drugs against alcohol-induced hepatocellular injury by testing cell viability after modeling and drug administration, using a CCK-8 assay. Cells were divided into a normal control group, a model group, a positive drug control group (silybin, 100 μM), and a daylily group. Cells were cultured normally for 12 hours. The positive drug and different daylily stock solutions were diluted in culture medium to the test concentrations (50 μg / mL and 200 μg / mL), and the corresponding test substance solution was added to each well. After a 6-hour incubation, the alcohol model was established. Except for cells in the normal control group, the culture medium was supplemented with 350 mM ethanol and incubated for 16 hours. After modeling, the supernatant was discarded, and cell viability was determined using the Cell Counting Kit-8 (CCK8) assay. Data were summarized, plotted as bar graphs using Graph Pad, and significant differences were calculated (t-test).

[0138] The data of the liver cell protection experiment are shown in Figure 7. Compared with the model group, daylily (petroleum ether, ethyl acetate, dichloromethane, n-butanol extract) has no effect on protecting liver cells. Daylily (20%, 30%, 50%, 75%, 95% ethanol extract) and daylily extracts from different origins all have liver cell protection effects. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. This shows that daylily extracted with different ethanol contents and daylily from different origins can play a role in protecting liver cells in the alcohol-induced liver cell damage model. Example 9 Daylily reduces the mortality rate of acute alcoholic liver poisoning

[0139] This study established an acute alcoholic liver intoxication mouse model and investigated the effects of drugs on mortality in this condition by measuring mouse mortality. Eight-week-old ICR male mice were divided equally according to body weight into a control group, an alcohol group, an alcohol + Hovenia dulcis (1 g / kg) group, an alcohol + Caulis dasyphylla (75% ethanol extract, 1 g / kg), and an alcohol + Caulis dasyphylla (75% ethanol extract, 2 g / kg) groups. Thirty minutes before alcohol administration, the alcohol + Hovenia dulcis (1 g / kg), alcohol + Caulis dasyphylla (1 g / kg), and alcohol + Caulis dasyphylla (2 g / kg) groups were administered via oral gavage. The control and alcohol groups were then gavaged with an equal volume of vehicle. Subsequently, all groups except the control group were gavaged with alcohol at a dose of 9 g / kg. Body weight changes and mortality rates were recorded over three days. Data were summarized, and bar graphs and survival curves were plotted using GraphPad. Statistical significance was calculated using a t-test.

[0140] The experimental results are shown in Figure 8. Compared with the blank group, the survival rate of the alcohol group (6.66%) was significantly reduced (*** indicates P < 0.001); the survival rate of the alcohol + daylily (1g / kg) group (60%) was significantly increased (** indicates P < 0.01); and the survival rate of the alcohol + daylily (2g / kg) group (80%) was significantly increased (*** indicates P < 0.001). This indicates that daylily can improve the survival rate of mice with acute alcohol intoxication in a dose-dependent manner.

[0141] Example 10 Daylily reduces acute alcoholic liver damage

[0142] This study established a mouse model of acute alcoholic liver injury and evaluated the effects of drugs on acute alcoholic liver injury by examining liver-to-body ratio (LBR) and pathological sections. Eight-week-old ICR male mice were divided equally according to body weight into a control group, an alcohol group, an alcohol-Houyuzi (1 g / kg) group, and an alcohol-Daihuali (75% ethanol extract, 1 g / kg) group. Thirty minutes before alcohol administration, the alcohol-Houyuzi (1 g / kg) and alcohol-Daihuali (1 g / kg) groups were administered via oral gavage. The control and alcohol groups were gavaged with an equal volume of vehicle. Subsequently, all groups except the control group were gavaged with alcohol at a dose of 8 g / kg. Nine hours after alcohol administration, mice were sacrificed and dissected. Liver images were taken, liver weights were measured, and LBR was calculated. The livers were fixed, paraffin-sectioned, and stained with H&E. Data were summarized, bar graphs were plotted using GraphPad, and significant differences were calculated using a t-test.

[0143] The experimental results are shown in Figure 9. Compared with the blank group, the livers in the alcohol group showed a significant whitening and yellowing, a significantly reduced liver-to-body ratio, and more lipid droplets and vacuoles were observed in the pathological sections (* indicates P < 0.05). Compared with the livers in the alcohol group, the livers treated with alcohol plus daylily (1g / kg) showed some recovery in color, a significantly increased liver-to-body ratio, and a return to normal levels. No significant changes in fat were observed in the pathological sections (* indicates P < 0.05). This suggests that daylily can effectively combat acute alcoholic liver damage and protect the liver.

[0144] Example 11 Daylily attenuates alcohol-induced lipid droplet accumulation

[0145] This experiment established an alcohol-induced lipid droplet accumulation cell model and investigated the effects of drugs on lipid droplet accumulation using Nile red staining. HepG2 cells were divided into a normal control group, a model group, and a daylily group. The cells were cultured normally for 12 hours. The positive drug and daylily mother solution were diluted with culture medium to the test concentration (75% ethanol extract, 200 μg / mL), and the corresponding test solution was added to each well. After incubation with the cells for 6 hours, alcohol modeling was performed. Except for the cells in the normal control group, the culture medium was supplemented with 300mM ethanol solution and incubated for 16 hours. After modeling, the lipid droplet expression content was detected using Nile red staining. The data were summarized, fluorescence quantification was performed using Image J, bar graphs were drawn using Graph Pad, and significant differences were calculated (t-test).

[0146] The experimental results are shown in FIG10 . Compared with the control group, the Nile red fluorescence intensity of the daylily (200 μg / mL) was significantly weakened compared with the model group, P=0.087, which indicates that the daylily reduced the lipid droplet accumulation in hepatocytes caused by alcohol.

[0147] Example 12 Treatment of Chronic Alcoholic Liver Disease with Daylily (75% Ethanol Extract)

[0148] This study established a chronic alcoholic liver disease mouse model to investigate the effects of drugs on liver function, pathological parameters, inflammatory markers, and lipid accumulation, thereby assessing their potential impact on chronic alcoholic liver disease. Eight-week-old C57BL / 6J male mice were divided equally according to body weight into three groups: a control group, an alcohol group, an alcohol plus silybin (100 mg / kg) group, an alcohol plus Hovenia dulcis (1 g / kg) group, an alcohol plus daylily (0.5 g / kg), an alcohol plus daylily (1 g / kg), and an alcohol plus daylily (2 g / kg). Prior to drug administration, the mice were acclimated to a Lieber-DeCarli liquid diet (1%-4% alcohol) for 5 days. On the sixth day, drug administration was initiated. All groups, except the control and model groups, were administered the different drugs by gavage. The control and alcohol groups received an equal volume of vehicle. Fresh Lieber-DeCarli liquid diet (4% alcohol) was replaced daily in the afternoon. Drug administration lasted for 4 weeks. On the last day of the experimental period, the final alcohol gavage was administered at 9:00 AM. Orbital blood was collected from the mice at 6:00 PM. The mice were then sacrificed, dissected, and their livers removed. The livers were photographed, weighed, and the liver-to-body ratio was calculated. Liver tissue was obtained and the corresponding experiments were performed. Data were summarized, plotted as bar graphs using Graph Pad, and significant differences were calculated (t-test).

[0149] The experimental results are shown in Figure 11. Compared with the blank group, the liver of the alcohol group turned significantly whiter and yellower, and the liver-to-body ratio increased. *** indicates P < 0.001. Compared with the liver of the alcohol group, the liver color of the alcohol + daylily (2g / kg) group recovered, and the liver-to-body ratio decreased significantly and returned to normal levels, which indicates that daylily can improve liver morphology. ** indicates P < 0.01.

[0150] As shown in Figure 12, compared with the blank group, the serum ALT and AST levels in the alcohol group were significantly increased, indicating that the model successfully caused liver dysfunction in mice, *** indicates P < 0.001; compared with the livers of the alcohol group, the serum ALT and AST levels in the alcohol + daylily (0.5, 1, 2 g / kg) groups were significantly decreased, indicating that daylily can effectively alleviate the damage to liver function caused by this model, *** indicates P < 0.001.

[0151] As shown in Figure 13, compared with the blank group, the serum ALB content in the alcohol group was significantly reduced, indicating that the model successfully caused liver dysfunction in mice, resulting in reduced albumin production. ** indicates P < 0.01. Compared with the alcohol group, the serum albumin content in the alcohol + daylily (0.5, 1, 2 g / kg) groups was significantly increased, indicating that daylily can effectively alleviate the damage to liver function caused by this model. ** indicates P < 0.01, *** indicates P < 0.001. Compared with the blank group, the blood glucose content in the alcohol group was significantly reduced, indicating that the model was successful. *** indicates P < 0.001. Compared with the alcohol group, the blood glucose content in the alcohol + daylily (2 g / kg) group was significantly increased, indicating that daylily can effectively alleviate the effect of this model on blood glucose. ** indicates P < 0.01. In addition, compared with the model group, the alcohol + daylily (2 g / kg) group effectively reduced serum triglyceride and cholesterol levels, indicating that daylily has a lipid-lowering effect.

[0152] As shown in Figure 14, compared with the blank group, the TNF-α and IL-6 levels in the serum and tissues of the alcohol group were significantly increased, indicating that the model successfully caused the accumulation of inflammatory factors, *** indicates P < 0.001; compared with the alcohol group, the serum TNF-α and IL-6 levels in the alcohol + daylily (0.5, 1, 2 g / kg) groups were significantly decreased, indicating that daylily can effectively play an anti-inflammatory role, *** indicates P < 0.001.

[0153] As shown in Figure 15, compared with the blank group, the ADH enzyme activity in the liver of the alcohol group was significantly decreased, and the expression level was increased, indicating that the model successfully caused insufficient alcohol metabolic enzyme activity. * indicates P < 0.05, ** indicates P < 0.01. Compared with the alcohol group, the ADH enzyme activity in the liver of the alcohol + daylily (0.5, 1, 2 g / kg) group was significantly increased, and the expression level was decreased, indicating that daylily can accelerate ethanol metabolism by activating ADH. * indicates P < 0.05, *** indicates P < 0.001. As shown in Figure 15, compared with the blank group, the ALDH enzyme activity in the liver of the alcohol group was decreased, and the expression level was increased, indicating that the model successfully caused insufficient alcohol metabolic enzyme activity. Compared with the alcohol group, the ALDH enzyme activity in the liver of the alcohol + daylily (2 g / kg) group was significantly increased, indicating that daylily can accelerate ethanol metabolism by activating ALDH. * indicates P < 0.05. As shown in Figure 16, compared with the blank group, the SOD content in the liver of the alcohol group was decreased, indicating that the model successfully caused oxidative stress in the liver; compared with the alcohol group, the SOD content in the liver of the alcohol + daylily (2 g / kg) group was significantly increased, indicating that daylily can play an antioxidant role, * indicates P < 0.05; compared with the blank group, the expression level of CYP2E1 enzyme in the liver of the alcohol group was increased, indicating that the model successfully caused insufficient activity of alcohol metabolism enzymes; compared with the alcohol group, the expression level of CYP2E1 enzyme in the liver of the alcohol + daylily (0.5, 1, 2 g / kg) group was decreased, indicating that daylily can regulate alcohol metabolism-related enzymes, *** indicates P < 0.001.

[0154] As shown in Figure 17, compared with the blank group, the triglyceride and cholesterol levels in the liver and serum of mice in the alcohol group were significantly increased, indicating that the model was successful, *** indicates P < 0.001; compared with the alcohol group, the triglyceride and cholesterol levels in the alcohol + daylily (0.5, 1, 2 g / kg) group were significantly reduced, indicating that daylily can effectively alleviate the effect of the model on fat accumulation, *** indicates P < 0.001.

[0155] The experimental results are shown in Figure 18. Compared with the blank group, more lipid droplet vacuoles appeared in the liver pathological sections of the alcohol group, which indicated that the model successfully caused the formation and accumulation of lipid droplets. Compared with the liver of the alcohol group, the lipid droplet vacuoles in the pathological sections of the alcohol + daylily (0.5, 1, 2 g / kg) group were significantly less and smaller, which indicated that daylily can reduce the accumulation of lipid droplets.

[0156] The experimental results are shown in Figure 19. Compared with the blank group, the results of Oil Red O staining of the liver sections of the alcohol group showed that the model successfully caused fat accumulation and alcoholic fatty liver; compared with the liver of the alcohol group, the red fat in the Oil Red O staining sections of the alcohol + daylily (0.5, 1, 2 g / kg) was significantly less and smaller, which indicates that daylily can reduce the formation and accumulation of fat in liver tissue.

[0157] Example 13 Daylily improves liver aging caused by drinking

[0158] This study established a chronic alcoholic liver disease mouse model and examined the effects of drugs on aging markers (such as SASP and SA-β-gal) to assess their potential impact on chronic alcoholic liver disease. Eight-week-old C57BL / 6J male mice were divided equally according to body weight into three groups: a control group, an alcohol group, an alcohol plus silybin (100 mg / kg) group, an alcohol plus Hovenia dulcis (1 g / kg) group, an alcohol plus daylily (0.5 g / kg), an alcohol plus daylily (1 g / kg), and an alcohol plus daylily (2 g / kg). Prior to drug administration, the mice were acclimated to a Lieber-DeCarli liquid diet (1%-4% alcohol) for 5 days. On the sixth day, all groups, except the control and model groups, were administered the various drugs by gavage. The control and alcohol groups received an equal volume of vehicle. Fresh Lieber-DeCarli liquid diet (4% alcohol) was replaced daily in the afternoon. Drug administration lasted for 4 weeks. On the last day of the experimental period, the final alcohol gavage was administered at 9:00 AM. Orbital blood was collected from the mice at 6:00 PM. The mice were then sacrificed, dissected, and their livers removed. The livers were photographed, weighed, and the liver-to-body ratio was calculated. Liver tissue was obtained and the corresponding experiments were performed. Data were summarized, plotted as bar graphs using Graph Pad, and significant differences were calculated (t-test).

[0159] The experimental results are shown in Figure 20. Compared with the blank group, the alcohol group showed significantly increased levels of serum CXCL1, MMP-1, and p16, and tissue CXCL-1, CXCL-3, IL-1β, MMP-1, MMP-7, and p16. This indicates that long-term alcohol consumption in mice significantly increases aging-related indicators and that long-term alcohol consumption is associated with liver aging. Compared with the serum and liver of the alcohol group, alcohol + daylily (0.5, 1, and 2 g / kg) significantly decreased serum CXCL1, MMP-1, and p16, and tissue CXCL-1, CXCL-3, IL-1β, MMP-1, MMP-7, and p16. This suggests that daylily can reduce the accumulation of aging indicators in liver tissue and play an anti-liver aging role. *** indicates P < 0.001.

[0160] The experimental results are shown in Figure 21. Compared with the blank group, the alcohol group showed a significant increase in the rate of senescence-associated β-galactosidase (SA-β-gal)-positive cells in tissue sections, indicating that long-term alcohol consumption in mice will lead to a significant increase in senescence-related indicators, accompanied by liver aging. Compared with the alcohol group, alcohol + daylily (0.5, 1, and 2 g / kg) significantly reduced the rate of senescence-associated β-galactosidase (SA-β-gal)-positive cells in liver sections, suggesting that daylily can reduce the accumulation of senescence indicators in liver tissue and play an anti-liver aging role. *** indicates P < 0.001.

[0161] The experimental results are shown in Figure 22. Compared with the blank group, the alcohol group showed significantly increased levels of p53 and γH2AX in tissues, indicating that long-term alcohol consumption in mice significantly increases aging-related markers and that long-term alcohol consumption is associated with liver aging. Compared with liver sections from the alcohol group, alcohol plus daylily (0.5, 1, and 2 g / kg) significantly decreased p53 and γh2AX levels, suggesting that daylily can reduce the accumulation of aging markers in liver tissue and play an anti-aging role. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0162] Example 14 Safety Experiment of Daylily

[0163] This experiment uses 14 days of continuous high-dose administration to examine the effects of the drug on the body weight ratio of important organs, serum indicators, and pathological sections to determine the safety of the drug. Eight-week-old C57BL / 6J male mice were divided into a blank group, a daylily (5g / kg) group, and a daylily (10g / kg) group according to their body weight. The mice were gavage-administered for 14 days, with each group receiving different doses of daylily crude extract, and the blank group receiving the same volume of vehicle. The weight changes of the mice were recorded daily. On the 15th day, blood was collected from the mouse orbits and dissected, the mouse organs were weighed, and the body weight ratio was calculated. The liver was fixed, paraffin-sectioned, and then stained with H&E. The data were summarized, a bar graph was drawn using Graph Pad, and significant differences (t-test) were calculated.

[0164] The experimental results are shown in Figure 23. Compared with the blank group, the daylily (5g / kg) group and the daylily (10g / kg) group had no toxicity to the important organs of mice, which shows that daylily has good safety.

[0165] In summary, this study experimentally demonstrated that daylily or its extract has significant alcohol detoxification and liver protection effects at the cellular and animal levels. On the one hand, it promotes the activation of alcohol metabolism enzymes ADH and ALDH, accelerating ethanol and acetaldehyde metabolism; improves the state of intoxication in subjects, including delaying the onset of intoxication, shortening the duration of intoxication, improving behavioral changes, restoring exercise capacity, and reducing mortality in mice with acute alcohol poisoning. On the other hand, it significantly improves liver morphology, liver-to-body ratio, and pathology in acute alcoholic liver injury; reduces alcohol-induced hepatocellular and liver damage, exerts a liver-protective effect, and improves chronic alcoholic liver injury. This includes reducing alcohol-induced elevations in transaminases, triglycerides, and cholesterol, restoring albumin levels, and ameliorating alcohol-induced hypoglycemia; reducing the upregulation of inflammatory markers (TNF-α and IL-6) in alcoholic liver disease; and reducing cellular reactive oxygen species. It also reduces alcohol-induced lipid droplet content and fat accumulation in liver tissue, and restores liver aging induced by long-term alcohol consumption. This suggests that daylily or its extract has broad application prospects in alcohol detoxification and the prevention or treatment of alcoholic liver disease.

[0166] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. Use of Hemerocallis citrina Baroni or an extract thereof as an active ingredient in preparing a composition for sobering up.

2. The use according to claim 1, characterized in that The hangover relieving method comprises one or more selected from the following groups: Reduce the rate of intoxication, delay the latency period of intoxication, shorten the duration of intoxication, improve behavior after intoxication, restore motor ability after intoxication, relieve alcohol intoxication, and / or reduce the mortality rate of acute alcohol intoxication.

3. The use according to claim 1, characterized in that The extract is selected from the following group: water extract of daylily, organic solvent extract of daylily, or active components separated from daylily extract (such as daylily polysaccharides, daylily flavonoids, daylily alkaloids, daylily nucleosides, or combinations thereof), or a combination thereof.

4. The use according to claim 1, characterized in that The extraction solvent used for the extract is a 100-0% C1-C5 alcohol-water (v / v) mixed solvent.

5. The use according to claim 1, characterized in that The extraction solvent used for the extract is a 100-0% ethanol-water (v / v) mixed solvent, preferably, 20-90% ethanol, more preferably 90-40% ethanol, more preferably 80-60% ethanol, such as 95% ethanol, 90% ethanol, 85% ethanol, 80% ethanol, 75% ethanol, 70% ethanol, 65% ethanol, 60% ethanol, 50% ethanol, 40% ethanol, 30% ethanol, 20% ethanol, 10% ethanol or 5% ethanol.

6. The use according to claim 1, characterized in that The extraction solvent used for the extract is 20%-30% ethanol or 75%-95% ethanol-water extract.

7. The use according to claim 1, characterized in that The daylily is the whole plant, flower buds, roots, stems, leaves, or a combination thereof, preferably, the daylily is a flower bud.

8. The use according to claim 1, characterized in that The extract is prepared by the following method: extracting the daylily in an extraction solvent at 80-120° C. for 1-5 times, combining the extracts and removing the solvent, thereby obtaining the daylily extract.

9. The use according to claim 1, characterized in that The composition is also used for auxiliary protection against alcoholic liver damage (ie, for protecting the liver).

10. The use according to claim 1, characterized in that The auxiliary protection against alcoholic liver damage includes one or more uses selected from the following group: (a) Alleviate acute or chronic liver cell and liver damage caused by alcohol and protect the liver, such as reducing the increase of transaminase, triglyceride, cholesterol, etc. caused by alcohol, restoring albumin level, and improving the decrease of blood sugar caused by alcohol; (b) reducing oxidative stress damage, improving inflammatory damage, and preventing hepatitis; and / or (c) Improve alcoholic liver disease, alleviate fatty degeneration or fat accumulation of liver tissue caused by alcohol, and reduce the content of lipid droplets, triglycerides, cholesterol, etc.; (d) Improve liver aging caused by long-term drinking, improve changes in liver aging-related markers, and restore liver health. Kang.

11. The use according to claim 1, characterized in that The composition is a food, a special medical food, a health product or a pharmaceutical composition.

12. A composition, characterized in that The composition contains: (a) daylily or a daylily extract as an active ingredient; and (b) a physiologically acceptable carrier.

13. The composition according to claim 12, characterized in that The active ingredient is a 100-0% C1-C5 alcohol-water (v / v) mixed solvent extract of day lily.

14. The composition according to claim 12, characterized in that The active ingredient is a 20%-30% ethanol or 75%-95% ethanol-water extract of day lily.

15. Use of the composition according to any one of claims 12 to 14 in the preparation of food, special medical food or health care product for auxiliary protection against alcoholic liver damage.

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