Application of day lily in dispelling effects of alcohol and protecting liver

Abstract

The invention provides a hangover alleviating and liver protecting application of day lily. More specifically, the invention provides the use of hemerocallis citrina Baroni or an extract thereof as an active ingredient in the preparation of a composition for dispelling the effects of alcohol. In addition, the composition can also be used for auxiliary protection of alcoholic liver injury (namely liver protection).

Description

Technical Field

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

[0002] Studies have shown that alcohol is associated with the onset of more than 200 diseases and causes about 3 million deaths worldwide each year. For example, alcohol can stimulate brain nerves, which can easily cause mental states such as excitement, agitation, and irritability in the short term. Long-term alcohol intake can also easily cause alcoholic encephalopathy. Chronic ethanol poisoning can cause hand tremors and reduce the ability to hold objects, which can be life-threatening in severe cases. Alcohol can irritate the gastric mucosa, making patients more likely to experience symptoms such as stomach pain, stomach cramps, diarrhea, vomiting, and indigestion. It can also easily cause gastritis, gastric ulcers, gastric bleeding, gastric mucosal erosion and other diseases. Alcohol can accelerate blood vessel contraction and expansion, and the brain and heart are particularly important blood supply sites. Drinking a lot of alcohol can easily induce palpitations, chest tightness, shortness of breath, etc. Long-term drinkers can induce coronary atherosclerotic heart disease, myocardial infarction, cerebral hemorrhage and other diseases. Among them, alcohol is particularly harmful to the liver. The liver is the most important organ for ethanol metabolism and detoxification. Excessive drinking and long-term drinking can easily increase the metabolic burden on the liver, leading to pathological liver damage, fatty degeneration and accumulation, and the formation of fatty liver. Under the stimulation of alcohol, it is also easy to cause alcoholic hepatitis. If it is not treated and controlled in time, it is easy to cause 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 is effective in sobering up and protecting the liver. Summary of the invention

[0004] The main purpose of the invention is to provide a product that can effectively sober up and protect the liver.

[0005] 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.

[0006] In another preferred embodiment, the hangover sobering method comprises accelerating ethanol metabolism, more specifically, reducing ethanol and acetaldehyde levels in the body by increasing the activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH).

[0007] In another preferred embodiment, the hangover remedy comprises one or more selected from the following groups:

[0008] 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.

[0009] 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.

[0010] 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.

[0011] 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.

[0012] 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.

[0013] 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.

[0014] In another preferred embodiment, the extract is prepared by the following method: extracting the day lily in an extraction solvent at 80-120° C. for 1-5 times, combining the extracts and removing the solvent, thereby obtaining the day lily extract.

[0015] 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.

[0016] In another preferred embodiment, the weight ratio of the day lily to the extraction solvent is 1:1-50 on a dry weight basis, 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.

[0017] In another preferred embodiment, the extraction times are 1, 2, 3, 4 or 5 times.

[0018] In another preferred embodiment, the solvent is removed by the following methods: concentration under reduced pressure, spray drying, freeze drying, or a combination thereof.

[0019] In another preferred embodiment, the daylily is the whole plant, flower buds, roots, stems, leaves, or a combination thereof, preferably, the daylily flower buds.

[0020] In another preferred embodiment, the composition is also used for auxiliary protection against alcoholic liver damage (ie, for protecting the liver).

[0021] In another preferred embodiment, the auxiliary protection against alcoholic liver damage includes one or more uses selected from the following group:

[0022] (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;

[0023] (b) reducing oxidative stress damage, improving inflammatory damage, and preventing hepatitis; and / or

[0024] (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.;

[0025] (d) Improve liver aging caused by long-term drinking, improve changes in liver aging-related markers, and restore liver health.

[0026] In another preferred embodiment, the composition is a food, a special medical food, a health product or a pharmaceutical composition.

[0027] In another preferred embodiment, the composition further comprises a physiologically acceptable carrier.

[0028] In another preferred embodiment, the dosage form of the composition is a solid preparation or a liquid preparation.

[0029] The second aspect of the present invention provides a composition comprising:

[0030] (a) daylily or a daylily extract as an active ingredient; and

[0031] (b) a physiologically acceptable carrier.

[0032] In another preferred embodiment, the daylily or daylily extract is as described above.

[0033] 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.

[0034] 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.

[0035] In another preferred embodiment, the food, special medical food, health product or pharmaceutical composition is also used to assist in protecting alcoholic liver damage.

[0036] 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 care product or pharmaceutical composition for auxiliary protection against alcoholic liver damage (i.e., for liver protection).

[0037] The fifth aspect of 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, for sobering up.

[0038] In another preferred embodiment, the active ingredient or composition can be administered before, during or after drinking alcohol.

[0039] In another preferred embodiment, the method is also a method for assisting protection against alcoholic liver damage.

[0040] In another preferred embodiment, the subject is a human or a non-human mammal, such as a rat, a mouse, or a monkey.

[0041] 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 embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 Showing the results of the ethanol metabolism rate test at the cellular level of daylily

[0043] Figure 2 Shows the results of the antioxidant capacity test at the cellular level of daylily

[0044] Figure 3 Shows the results of the test on the metabolic dynamics of serum ethanol in drunken mice by daylily

[0045] Figure 4 Shows the results of behavioral (rotarod) test on drunk mice by daylily

[0046] Figure 5 Shows the results of behavioral (pole climbing) tests on drunk mice by daylily

[0047] Figure 6 Shows the results of behavioral (righting reflex) tests of daylily on drunk mice

[0048] Figure 7 Shows the results of the cell survival test of daylily in the alcohol-induced hepatocyte injury model

[0049] Figure 8 Results: The results of the test on the mortality rate of acute alcohol poisoning mice with daylily

[0050] Fig. 9 The results of the test show that daylily has the ability to protect the liver of mice with acute alcoholic liver injury

[0051] Fig.10 Shows the test results of regulating lipid droplet content at the cellular level of daylily

[0052] Fig.11 The results of the liver-to-body ratio test showing the effect of daylily on the liver status and liver volume ratio of mice with chronic alcoholic liver disease

[0053] Fig.12 The results of the test of daylily on liver function indexes (AST, ALT) of mice with chronic alcoholic liver disease are shown.

[0054] Fig.13 The results of the test showing the effect of daylily on albumin and blood sugar in mice with chronic alcoholic liver disease

[0055] Fig.14 Shows the test results of daylily on inflammatory indicators (interleukin-6 and TNF-α) in mice with chronic alcoholic liver disease

[0056] Fig.15 The results of the test of daylily on key enzymes of ethanol metabolism (alcohol dehydrogenase, acetaldehyde dehydrogenase, cytochrome P450 2E1) in mice with chronic alcoholic liver disease are shown.

[0057] Fig.16 The results of the test on the antioxidant index superoxide dismutase activity of daylily in mice with chronic alcoholic liver disease are shown

[0058] Fig.17 The results of the test of daylily on serum and liver tissue triglyceride and cholesterol in mice with chronic alcoholic liver disease are shown

[0059] Fig.18Shows the results of H&E staining test on liver tissue pathology of mice with chronic alcoholic liver disease

[0060] Fig.19 The results of the Oil Red O staining test of liver tissue sections of mice with chronic alcoholic liver disease are shown.

[0061] Fig. 20 The results of the test of daylily on serum and liver tissue aging indicators (CXCL-1, MMP-1, etc.) of chronic alcoholic liver disease mice are shown;

[0062] Fig.21 The results of the test of SA-β-gal positive cell rate in liver tissue sections of chronic alcoholic liver mice using daylily are shown;

[0063] Fig. 22 The results of the test of daylily on p53 and γH2AX, the aging indicators of liver tissue in chronic alcoholic liver mice are shown; Fig.23 Shown are the results of safety testing of daylily on mice. DETAILED DESCRIPTION

[0064] After extensive and in-depth research, the inventors have 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. On this basis, the present invention was completed.

[0065] the term

[0066] Active ingredients

[0067] As used herein, the active ingredient of the present invention comprises daylily or a daylily extract.

[0068] Daylily (Hemerocallis citrina Baroni, Daylily), also known as daylily, forget-me-not, daylily, daylily flower, brain-boosting vegetable, tranquilizing vegetable, green onion flower, lemon daylily, daylily flower, etc., is an angiosperm of the genus Hemerocallis of the family Asparagaceae, Asparagaceae, and has the effects of stomachic, diuretic, and detumescence. It has certain medicinal value and is also an edible vegetable with a long history of consumption. The inventors have found for the first time through a large number of screenings of Chinese herbal medicine libraries that daylily (especially its alcohol-water extract) is an edible Chinese herbal medicine with excellent alcohol-resolving 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-resolving and liver-protecting.

[0069] The daylily raw material that can be used in the present invention is usually dry daylily or its chopped and crushed form. The daylily can be the whole plant, flower bud, root, stem, leaf, or a combination thereof, preferably, the daylily flower bud or root.

[0070] 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.

[0071] 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 20-30% ethanol or 75%-95% ethanol-water extract is a preferred extract.

[0072] In another preferred embodiment, the extract is prepared by the following method: extracting the day lily in an extraction solvent at 80-120° C. for 1-5 times, combining the extracts and removing the solvent, thereby obtaining the day lily extract.

[0073] Hangover and liver protection

[0074] As used herein, hangover relief refers to when a person or animal, after taking in alcohol, makes the alcohol content in the body of the object less, the concentration is lower and / or the existence time is shorter than that without using the active ingredient or composition of the present invention, so that the physical and mental effects or damage of alcohol are smaller. The hangover relief of the present invention includes preventing drunkenness, by accelerating ethanol metabolism, making ethanol quickly decompose into acetaldehyde, and finally decomposing into carbon dioxide and water, thereby reducing or delaying the inhibition of alcohol on the central nervous system, reducing the possibility of the subject reaching a drunken state (alcoholism), alleviating the action disorder caused by alcohol, or prolonging the incubation period of drunkenness. In addition, the hangover relief of the present invention also includes "sobering up", so that the subject wakes up from the state of drunkenness, restores consciousness or restores self-control. It usually does not refer to natural sobriety, but adopts the sobering up of taking alcohol-resolving substances, accelerates ethanol metabolism, and serves the purpose of accelerating sobriety and reducing the harm of ethanol to the body.

[0075] The decomposition and metabolism of alcohol in the body mainly depends on two enzymes in the enzyme system of the liver: one is alcohol dehydrogenase, and the other is acetaldehyde dehydrogenase. Alcohol dehydrogenase can remove two hydrogen atoms in alcohol molecules, so that ethanol is decomposed into acetaldehyde. Acetaldehyde dehydrogenase can remove two hydrogen atoms in acetaldehyde, so that acetaldehyde is converted into acetic acid, and finally decomposed into carbon dioxide and water. Ethanol and acetaldehyde are the most harmful to the body. Among them, the lack of acetaldehyde dehydrogenase prevents acetaldehyde from being completely decomposed into acetic acid, but acetaldehyde continues to remain in the body, resulting in nausea and vomiting, loss of balance, slurred speech, weakened judgment, coma and discomfort after drinking. Experiments have shown that the active ingredient or composition of the present invention can increase the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, accelerate the metabolism of alcohol, and thus achieve hangover relief.

[0076] As used herein, liver protection means protecting the liver, which refers to the auxiliary protective function against alcoholic liver damage. The liver is the main organ for the decomposition of alcohol. Drinking alcohol may cause liver damage, and even alcoholic fatty liver, alcoholic cirrhosis and alcoholic liver failure. The higher the level of alcohol in the body and the longer the drinking time, the more serious the liver damage tends to be. Experiments have shown that the active ingredient or composition of the present invention can alleviate alcohol damage to liver cells, improve alcohol's decline in liver function, reduce fat accumulation in liver cells, improve liver cell antioxidant and anti-inflammatory functions, promote liver rejuvenation, and / or delay the progression of alcoholic liver disease.

[0077] The "prevention", "treatment", "improvement" and "relief" described in the present invention include delaying and stopping the progression of the disease or symptoms, or eliminating the disease, and do not require 100% inhibition, elimination and reversal. In some embodiments, the composition of the present invention can reduce, shorten or reverse one or more symptoms or indicators of drunkenness or alcoholic liver damage, such as reducing, shortening and / or reversing 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.

[0078] Some representative hangover sobering 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, CYP2E1 expression.

[0079] 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.

[0080] Compositions and methods of administration

[0081] The present invention also provides compositions comprising the active ingredients of the present invention and a physiologically acceptable carrier.

[0082] The composition can be used to sober up. Experiments have shown that the active ingredient of the present invention can accelerate ethanol metabolism to sober up, and more specifically, can reduce the levels of ethanol and acetaldehyde in the body by increasing the activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), thereby delaying the time of drunkenness, shortening the duration of drunkenness, improving behavior after drunkenness, restoring exercise ability after drunkenness, alleviating alcohol poisoning, and / or reducing the mortality rate of acute alcohol poisoning.

[0083] In addition, the composition of the present invention can also be used to protect the liver. In particular, it can reduce acute or chronic liver cell and liver damage caused by alcohol. More specifically, the active ingredients of the present invention can reduce the increase of transaminase, triglycerides, cholesterol, etc. caused by alcohol, restore albumin levels, improve the hypoglycemia caused by alcohol; reduce oxidative stress damage, improve inflammatory damage, prevent hepatitis; and / or improve alcoholic liver disease, alleviate fatty degeneration or fat accumulation of liver tissue caused by alcohol, and reduce the content of fat droplets, triglycerides, cholesterol, etc.

[0084] The compositions of the present invention include, but are not limited to, foods, special medical foods, health products and medicines.

[0085] 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.

[0086] Generally, 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 function) on humans and / or animals and is acceptable to humans and / or animals.

[0087] As used herein, the term "physiologically acceptable" ingredients are suitable for use in humans and / or mammals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), i.e., materials with a reasonable benefit / risk ratio. The term "physiologically acceptable carrier" refers to a carrier for administration of a therapeutic agent, including various excipients and diluents.

[0088] The composition of the present invention contains 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 carrier includes (but is not limited to): water, NaCl, physiological saline solution, lactated Ringer's solution, conventional sucrose, conventional glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates, such as lactose, straight-chain starch or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine and pigments, etc. Such preparations can be sterilized and, if necessary, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, colorants and / or aromatic substances that do not react harmfully with the compounds provided herein or interfere with the activity of the compounds provided herein. Those of ordinary skill in the art will recognize that other carriers and excipients are suitable for the disclosed compounds.

[0089] The compositions of the present invention can be administered to the subject by any suitable route, including orally, subcutaneously, intravenously, intramuscularly, transdermally, by inhalation spray, topically, rectally, nasally, buccally, with the active compound being administered to the subject. Preferably, the active compound is administered orally, subcutaneously, intravenously, intramuscularly, transdermally.

[0090] Compositions of the invention suitable for administration will typically be discrete units in solid form, such as tablets, capsules, powders, granules, pills, patches, gels, ointments, suppositories, or in liquid form, such as oral liquids, injectable or infusible solutions or suspensions, or lyophilized powder injections.

[0091] 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.

[0092] 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 drunkenness, the severity of drunken 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 symptom to be treated or improved by the subject, the patient's body weight, the patient's immune status, the route of administration, etc. Generally, the active ingredient of the present invention can be administered 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 per day. For example, several divided doses may be administered per day, or the dose may be reduced proportionally, depending on the urgency of the subject's condition.

[0093] The present invention also provides a method for sobering up, comprising the steps of administering the active ingredient of the present invention to a subject in need, or administering the composition of the present invention, so as to sober up. The active ingredient or composition of the present invention can be administered before, during or after drinking. In particular, from 6 hours before drinking to the beginning of drinking, such as 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 0.5 hours, 10 minutes, 5 minutes, 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.

[0094] In another preferred embodiment, the subject is a human or a non-human mammal, such as a rat, a mouse, or a monkey.

[0095] The main advantages of the present invention include:

[0096] A) The present invention is the first to discover the use of daylily in sobering up and accelerating ethanol metabolism.

[0097] B) The present invention is the first to discover the ability of daylily to relieve acute alcohol poisoning and resist alcoholic liver disease.

[0098] C) The active ingredients of the present invention are derived from edible Chinese medicine / vegetables and are safe and effective.

[0099] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0100] Example 1 Preparation of Daylily Samples

[0101] The daylily samples used in the present invention come from Anhui, Fujian, Heilongjiang, Shanxi, Hunan, Sichuan and Gansu. After the daylily (dried flower buds) are crushed, 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 are used for decoction three times according to a certain material-liquid ratio, and the daylily crude extracts from different origins and different extraction methods are obtained by reduced pressure concentration and freeze drying, and the yields are calculated respectively.

[0102] Weigh 20 g of dry material of day lily (originating from Anhui, Fujian, Heilongjiang, Shanxi, Hunan, Sichuan, and Gansu), grind it into powder, boil it with 75% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, and 60 min each time. Combine the extracts, concentrate under reduced pressure, and then freeze-dry. 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%, which is the day lily (75% ethanol extract) from different origins.

[0103] Weigh 10 g of dried daylily, grind it with a grinder, boil it with 95% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, and boil it for 60 min each time. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.394 g with a yield of 13.94%, which is daylily (95% ethanol extract).

[0104] Weigh 5 g of dried daylily, grind it in a grinder, 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. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.029 g with a yield of 20.58%, which is daylily (60% ethanol extract).

[0105] Weigh 5 g of dried daylily, grind it in a grinder, and boil it with 50% ethanol 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 1.378 g with a yield of 27.56%, which is daylily (50% ethanol extract).

[0106] Weigh 5 g of dried daylily, grind it in a grinder, 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. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.362 g with a yield of 27.24%, which is daylily (40% ethanol extract).

[0107] Weigh 5 g of dried daylily, grind it in a grinder, and boil it three times with 30% ethanol at 100°C 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 1.453 g with a yield of 29.06%, which is daylily (30% ethanol extract).

[0108] Weigh 5 g of dried daylily, grind it with a grinder, boil it with 20% ethanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, and boil it for 60 min each time. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.665 g with a yield of 33.33%, which is daylily (20% ethanol extract).

[0109] Weigh 5 g of dried daylily, grind it in a grinder, 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. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.362 g with a yield of 27.24%, which is daylily (10% ethanol extract).

[0110] Weigh 5 g of dried daylily, grind it with a grinder, boil it with deionized water at 100°C for 3 times with a solid-liquid ratio of 1:40, and boil it for 60 min each time. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 1.136 g with a yield of 22.76%, which is daylily (water extract).

[0111] 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. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 0.1 g with a yield of 0.67%, which is daylily (petroleum ether extract).

[0112] 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).

[0113] Weigh 15 g of dried daylily, grind it with a grinder, boil it with dichloromethane at 100°C for 3 times with a solid-liquid ratio of 1:7.5, and boil it for 60 min each time. Combine the extracts, concentrate under reduced pressure, and then freeze-dry to obtain 0.22 g with a yield of 1.47%, which is the daylily (dichloromethane extract).

[0114] Weigh 15 g of dried daylily, grind it with a grinder, boil it with n-butanol at 100°C for 3 times with a solid-liquid ratio of 1:7.5, and boil it for 60 min each time. 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 (n-butanol extract).

[0115] Example 2 Daylily promotes ethanol metabolism in liver cells

[0116] This experiment determines the efficacy of drugs in promoting ethanol metabolism by testing the ethanol content in the culture medium. HepG2 cells were divided into a normal control group, a model group, a positive drug control group (75% ethanol extract of Hovenia dulcis, 200μg / mL and dihydromyricetin, 100μM) and a daylily group. The cells were cultured normally for 12 hours, and the positive drugs and different daylily mother solutions were diluted with the culture medium to the concentration to be tested (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 was incubated for 4 hours. After the modeling was completed, the supernatant was taken and the ethanol concentration of the culture medium was detected by the Megazyme-Alcohol kit. The data were summarized, a bar graph was drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0117] Experimental data on ethanol metabolism in hepatocytes Figure 1 As shown, compared with the model group, the extracts of daylily extracted by different solvents have different abilities to promote ethanol metabolism in hepatocytes, among which the 20%, 30%, 50%, or 75% or 95% ethanol extracts of daylily have the best activity, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001. This indicates that daylily can promote ethanol metabolism at the cellular level.

[0118] Example 3 Daylily reduces alcohol-induced oxidative stress

[0119] Alcohol can induce liver cell damage, promote high expression of reactive oxygen species, and lead to oxidative stress. This experiment examines 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 concentration to be tested (75% ethanol extract, 200μg / mL), and the corresponding test solution was added to each well. After incubation with 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 commercially available DCFH-DA probe was used to detect the expression content of ROS. The data were summarized, quantified by fluorescence using Image J, histograms were drawn using Graph Pad, and significant differences (t-test) were calculated.

[0120] The experimental results are as follows Figure 2 As shown, compared with the control group, the fluorescence intensity was significantly reduced after administration of daylily, which indicates that daylily reduced the increase in the content of active oxygen in cells caused by alcohol. * indicates P < 0.05, which suggests that daylily can play an antioxidant role at the cellular level.

[0121] Example 4 Daylily promotes ethanol metabolism in drunken mice

[0122] This experiment tested the blood ethanol content of drunk mice to investigate the effect of drugs on the metabolism of ethanol in mice. Eight-week-old ICR male mice were divided into alcohol group, alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg) group and alcohol + daylily (75% ethanol extract, 0.5, 1, 2g / kg) group according to their body weight. Thirty minutes before alcohol administration, alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg) group and alcohol + daylily (0.5, 1, 2g / kg) group were administered by oral gavage, and the alcohol group was gavaged with the same volume of solvent. After that, all groups were gavaged with alcohol, and the alcohol dose was 7g / kg. After drinking, blood was collected from the mice 0.5, 1, 2, 4, 8, and 12 hours after drinking by orbital blood sampling. Serum was centrifuged after standing at room temperature, and serum was detected by Megazyme-Alcohol kit and serum acetaldehyde concentration by HPLC derivatization. The data were summarized, curves were fitted using Graph Pad, and significant differences were calculated (t-test).

[0123] The experimental results are as follows Figure 3 As shown in (A), 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, the alcohol + daylily (0.5, 1, 2 g / kg) group reduced the serum alcohol concentration at all time points and restored the serum ethanol concentration to normal levels faster, indicating that daylily can effectively promote the ethanol metabolism of drunk mice. At the same time, the alcohol + daylily (1 g / kg) group reduced the serum acetaldehyde concentration at all time points and restored the serum acetaldehyde concentration to normal levels faster.

[0124] Eight-week-old ICR male mice were evenly divided 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. 30 minutes before alcohol administration, the alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 0.8 g / kg) and the alcohol + daylily group were administered by oral gavage, and the alcohol group was gavaged with the same volume of solvent. After that, all groups were gavaged with alcohol, and the alcohol dose was 7 g / kg. After drinking, the blood of mice was collected by orbital blood sampling at 0.5, 1, 2, 4, 8, and 12 hours after drinking. After standing at room temperature, the serum was centrifuged and the serum ethanol concentration was detected by Megazyme-Alcohol kit. The data were summarized, the curve was fitted with Graph Pad, and the significant difference (t-test) was calculated.

[0125] The experimental results are as follows Figure 3 As shown in (B), the serum ethanol content of mice increased after drinking, 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.

[0126] Example 5 Daylily improves drunken behavior in mice (rotarod)

[0127] This experiment tests the behavioral rotarod of drunk mice to investigate the effects of drugs on the coordination, balance and endurance of mice after drinking. Eight-week-old ICR male mice were divided into a blank group, an alcohol group, an alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg), and an alcohol + daylily (75% ethanol extract, 0.5, 1, 2g / kg) group according to their body weight. Before the experiment, all groups of mice were trained in the rotarod for three days. 30 minutes before alcohol administration, the alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg) and the alcohol + daylily (0.5, 1, 2g / kg) groups were administered by oral gavage, and the blank group and the alcohol group were gavaged with the same volume of solvent. Afterwards, all groups except the blank group were gavaged with alcohol, with an alcohol dose of 4.5g / kg. Ten minutes after drinking, the mouse rotarod behavior test was performed, and the time the mouse was on the rod was recorded. The data were summarized, histograms were drawn using Graph Pad, and significant differences were calculated (t-test).

[0128] The experimental results are as follows Figure 4 As shown, compared with the blank group, the alcohol group significantly reduced the time of mice on the rod, *** indicates P < 0.001; compared with the alcohol group, the alcohol + daylily (0.5, 1, 2g / kg) group significantly increased the time of mice on the rod, ** indicates P < 0.01, *** indicates P < 0.001. This shows that daylily can effectively improve the motor ability of mice after drunkenness and play a role in sobering up.

[0129] Example 6 Daylily improves drunken behavior of mice (pole climbing)

[0130] This experiment tests the behavioral climbing time and climbing score of mice after drinking to investigate the effect of drugs on the coordination, balance and endurance of mice after drinking. Eight-week-old ICR male mice were divided into a blank group, an alcohol group, an alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg), and an alcohol + daylily (75% ethanol extract, 0.5, 1, 2g / kg) group according to their body weight. Before the experiment, all groups of mice were trained to climb the pole for three days. Thirty minutes before giving alcohol, the alcohol + positive drug group (75% ethanol extract of Hovenia dulcis, 1g / kg) and the alcohol + daylily (0.5, 1, 2g / kg) groups were administered by oral gavage, and the blank group and the alcohol group were gavaged with the same volume of solvent. Afterwards, all groups except the blank group were gavaged with alcohol, and the alcohol dose was 4.5g / kg. Ten minutes after drinking, the mice were tested for pole climbing behavior, the pole climbing status of the mice was scored, and the pole climbing time of the mice was recorded. The data were summarized, a bar graph was drawn using Graph Pad, and significant differences were calculated (t-test).

[0131] The experimental results are as follows Figure 5As shown in the figure, compared with the blank group, the alcohol group significantly reduced the climbing time of mice and increased the climbing score of mice, *** indicates P < 0.001; compared with the alcohol group, the alcohol + daylily (0.5, 1, 2g / kg) group significantly increased the climbing time of mice and reduced the climbing score of mice, ** indicates P < 0.01. This shows that daylily can effectively improve the motor ability of mice after drunkenness and play a role in sobering up.

[0132] Example 7 Daylily improves drunken behavior (righting reflex) in mice

[0133] This experiment tested the behavioral righting reflex disappearance and recovery time of mice after drinking, as well as the hanging endurance and gripping strength after the righting reflex was recovered, so as to investigate the effects of drugs on the incubation period of drunkenness and sobering time of mice after drinking, as well as the endurance and gripping strength of drunken mice. Eight-week-old ICR male mice were divided into alcohol group, alcohol + Hovenia dulcis (75% ethanol extract, 1g / kg) and alcohol + Daylily (75% ethanol extract, 0.5, 1, 2g / kg) groups according to their body weight. 30 minutes before drinking, the alcohol + Hovenia dulcis (1g / kg) group and alcohol + Daylily (0.5, 1, 2g / kg) were administered by oral gavage, and the alcohol group was gavaged with the same volume of solvent. After that, all groups were gavaged with alcohol, with an alcohol dose of 7g / kg. The disappearance of the righting reflex and the recovery time of mice after drinking were recorded, the drunkenness rate, the incubation period of drunkenness and the sobering time were counted, and the gripping strength and hanging endurance of mice after sobering were tested. The data were summarized, histograms were drawn using Graph Pad, and significant differences were calculated (t-test).

[0134] The experimental results are as follows Figure 6 As shown in (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, alcohol + daylily (1, 2g / kg) significantly shortened the sobering time of drunk mice, * indicates P<0.05, ** indicates P<0.01. This shows that daylily can reduce the drunkenness rate of mice, prevent drunkenness and accelerate sobering. Figure 6 As shown in (DE), compared with the alcohol group, alcohol + daylily (1, 2 g / kg) restored the grasping strength and hanging endurance of mice after sobering up, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. This indicates that daylily can promote sobering up mice and restore the behavioral ability of drunk mice.

[0135] Example 8 Daylily hepatocyte protection experiment

[0136] This experiment tested the cell viability after modeling and drug administration, and used CCK-8 experiments to investigate the protective effect of drugs on alcohol-induced hepatocyte injury. The cells were divided into a normal control group, a model group, a positive drug control group (silybin, 100μM) and a daylily group. The cells were cultured normally for 12 hours, and the positive drugs and different daylily mother solutions were diluted with culture medium to the concentrations to be tested (50μg / mL and 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 350mM ethanol solution and incubated for 16 hours. After the modeling was completed, the supernatant was discarded, and the cell survival rate was detected by CellCounting Kit-8 (CCK8). The data were summarized, a bar graph was drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0137] Hepatocyte protection experimental data such as Figure 7 As shown, compared with the model group, daylily (petroleum ether, ethyl acetate, dichloromethane, n-butanol extracts) did not have a protective effect on hepatocytes, daylily (20%, 30%, 50%, 75%, 95% ethanol extracts) and daylily extracts from different origins all had a hepatocyte protective effect, * 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 hepatocytes in the alcohol-induced hepatocyte injury model. Example 9 Daylily reduces the mortality rate of acute alcoholic liver poisoning

[0138] This experiment established an acute alcoholic liver poisoning mouse model and investigated the effect of drugs on the mortality of acute alcoholic liver poisoning by measuring the mortality of mice. Eight-week-old ICR male mice were divided into a blank group, an alcohol group, an alcohol + Hovenia dulcis (1g / kg) group, an alcohol + Daylily (75% ethanol extract, 1g / kg) and an alcohol + Daylily (75% ethanol extract, 2g / kg) group according to their body weight. Thirty minutes before alcohol administration, the alcohol + Hovenia dulcis (1g / kg) group, the alcohol + Daylily (1g / kg) group and the alcohol + Daylily (2g / kg) group were administered by oral gavage, and the blank group and the alcohol group were gavaged with the same volume of solvent. Afterwards, all groups except the blank group were gavaged with alcohol, and the alcohol dose was 9g / kg. The weight changes and mortality of mice were recorded within three days. The data were summarized, and the bar graph and survival curve were drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0139] The experimental results are as follows Figure 8As shown, compared with the blank group, the survival rate of the alcohol group (6.66%) was significantly reduced, *** indicates P < 0.001: compared with the survival rate of the alcohol group, the survival rate of alcohol + daylily (1g / kg) (60%) was significantly increased, ** indicates P < 0.01; compared with the survival rate of the alcohol group, the survival rate of alcohol + daylily (2g / kg) (80%) was significantly increased, *** indicates P < 0.001. This shows that daylily can improve the survival rate of mice with acute alcohol poisoning, and it is dose-dependent.

[0140] Example 10 Daylily reduces acute alcoholic liver damage

[0141] In this experiment, an acute alcoholic liver injury mouse model was constructed, and the effect of drugs on acute alcoholic liver injury was determined by examining the liver-to-body ratio and pathological sections of mice. Eight-week-old ICR male mice were divided into a blank group, an alcohol group, an alcohol + Hovenia dulcis (1 g / kg) group, and an alcohol + Daylily (75% ethanol extract, 1 g / kg) according to their body weight. Thirty minutes before alcohol administration, the alcohol + Hovenia dulcis (1 g / kg) group and the alcohol + Daylily (1 g / kg) group were administered by oral gavage, and the blank group and the alcohol group were gavaged with the same volume of solvent. Afterwards, all groups except the blank group were gavaged with alcohol, and the alcohol dose was 8 g / kg. Nine hours after alcohol gavage, the mice were killed for dissection, the appearance of the mouse liver was photographed, the liver weight was weighed, and the liver-to-body ratio was calculated. The liver was fixed, paraffin sectioned, and H&E stained. The data were summarized, a bar graph was drawn using Graph Pad, and significant differences (t-test) were calculated.

[0142] The experimental results are as follows Fig. 9 As shown, compared with the blank group, the liver of the alcohol group turned white and yellow, the liver-to-body ratio was significantly reduced, and more fat droplets and vacuoles appeared in the pathological sections, * indicates P < 0.05; compared with the liver of the alcohol group, the liver color of the alcohol + daylily (1g / kg) was restored, the liver-to-body ratio was significantly increased and returned to normal levels, and no obvious fat changes were found in the pathological sections, * indicates P < 0.05. This shows that daylily can effectively resist acute alcoholic liver damage and play a role in protecting the liver.

[0143] Example 11 Daylily attenuates alcohol-induced lipid droplet accumulation

[0144] In this experiment, an alcohol-induced lipid droplet accumulation cell model was constructed, and the effect of drugs on lipid droplet accumulation was investigated by 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, and the positive drug and the daylily mother solution were diluted with culture medium to the concentration to be tested (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 lipid droplet expression content was detected using Nile red dye. The data were summarized, and the fluorescence was quantified using Image J, and the bar graph was drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0145] The experimental results are as follows Fig.10 As shown, compared with the control group, the Nile red fluorescence intensity of the daylily (200 μg / mL) group 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.

[0146] Example 12 Treatment of chronic alcoholic liver disease with Daylily (75% ethanol extract)

[0147] This experiment constructs a mouse model of chronic alcoholic liver disease and investigates the effects of drugs on liver function, pathological indicators, inflammatory indicators, lipid accumulation and other indicators, so as to determine the effects of drugs on chronic alcoholic liver disease. Eight-week-old C57BL / 6J male mice were divided into a blank group, an alcohol group, an alcohol + silybin (100 mg / kg) group, an alcohol + Hovenia dulcis (1 g / kg) group, an alcohol + daylily (0.5 g / kg), an alcohol + daylily (1 g / kg) and an alcohol + daylily (2 g / kg) according to their body weight. Before formal administration, the mice will be adapted to Lieber-DeCarli liquid feed (alcohol content 1%-4%) for 5 days. On the sixth day, formal administration was carried out. Except for the blank group and the model group, the other groups were given different drugs by gavage. The blank group and the alcohol group were gavaged with the same volume of solvent. Freshly prepared Lieber-DeCarli liquid feed (alcohol content 4%) was replaced every afternoon. The drug was administered for a total of 4 weeks. On the last day of the experimental cycle, the last alcohol gavage was performed at 9:00 am, and the mice were blooded from their eye sockets at 18:00 pm. The mice were killed, dissected, and the liver was removed. The appearance of the mouse liver was photographed, the liver weight was weighed, and the liver-to-body ratio was calculated. The liver tissue was taken and the corresponding experiments were performed. The data were summarized, a bar graph was drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0148] The experimental results are as follows Fig.11As shown, 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 was restored, and the liver-to-body ratio was significantly reduced and returned to normal levels, which indicates that daylily can improve liver morphology, ** indicates P < 0.01.

[0149] like Fig.12 As shown, 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 liver of the alcohol group, the serum ALT and AST levels in the alcohol + daylily (0.5, 1, 2 g / kg) group were significantly decreased, indicating that daylily can effectively alleviate the damage to liver function in this model, *** indicates P < 0.001.

[0150] like Fig.13 As shown, compared with the blank group, the serum ALB content of the alcohol group was significantly reduced, indicating that the model successfully caused liver dysfunction in mice, thereby reducing albumin production, ** indicates P < 0.01; compared with the alcohol group, the serum albumin content of the alcohol + daylily (0.5, 1, 2g / kg) group 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 of 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 of the alcohol + daylily (2g / 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 (2g / kg) group effectively reduced serum triglyceride and cholesterol levels, indicating that daylily has a lipid-lowering effect.

[0151] like Fig.14 As shown, compared with the blank group, the levels of TNF-α and IL-6 in 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 levels of serum TNF-α and IL-6 in the alcohol + daylily (0.5, 1, 2 g / kg) group were significantly decreased, indicating that daylily can effectively play an anti-inflammatory role, *** indicates P < 0.001.

[0152] like Fig.15As shown in the figure, compared with the blank group, the ADH enzyme activity in the liver of the alcohol group was significantly reduced, and the expression level was increased, which indicated that the model successfully caused insufficient activity of alcohol metabolic enzymes, * 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, 2g / kg) group was significantly increased, and the expression level was reduced, which indicated that daylily can accelerate ethanol metabolism by activating ADH, * indicates P < 0.05, *** indicates P < 0.001. Fig.15 As shown in the figure, compared with the blank group, the ALDH enzyme activity in the liver of the alcohol group was reduced and the expression level was increased, which indicated that the model successfully caused insufficient activity of alcohol metabolic enzymes; compared with the alcohol group, the ALDH enzyme activity in the liver of the alcohol + daylily (2g / kg) group was significantly increased, which indicated that daylily can accelerate ethanol metabolism by activating ALDH, * indicates P<0.05. Fig.16 As shown, compared with the blank group, the SOD content in the liver of the alcohol group was decreased, which indicated 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 (2g / kg) group was significantly increased, which indicated that daylily can play an antioxidant role, * indicates P<0.05; compared with the blank group, the expression of CYP2E1 enzyme in the liver of the alcohol group was increased, which indicated that the model successfully caused insufficient activity of alcohol metabolic enzymes; compared with the alcohol group, the expression of CYP2E1 enzyme in the liver of the alcohol + daylily (0.5, 1, 2g / kg) group was decreased, which indicated that daylily can regulate alcohol metabolism-related enzymes, *** indicates P<0.001.

[0153] like Fig.17 As shown, 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.

[0154] The experimental results are as follows Fig.18 As shown, 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 induced the formation and accumulation of lipid droplets; compared with the liver of the alcohol group, the lipid droplet vacuoles in the pathological sections of alcohol + daylily (0.5, 1, 2 g / kg) were significantly reduced and smaller, which indicated that daylily can play a role in reducing the accumulation of lipid droplets.

[0155] The experimental results are as follows Fig.19As shown, compared with the blank group, the results of Oil Red O staining of liver sections in the alcohol group showed that the model successfully caused fat accumulation and alcoholic fatty liver occurred; compared with the liver in the alcohol group, the red fat in the Oil Red O staining sections of alcohol + daylily (0.5, 1, 2g / kg) was significantly less and smaller, indicating that daylily can reduce the formation and accumulation of fat in liver tissue.

[0156] Example 13 Daylily improves liver aging caused by drinking

[0157] This experiment constructs a chronic alcoholic liver mouse model to investigate the effects of drugs on aging indicators (such as SASP, SA-β-gal, etc.), so as to determine the effects of drugs on chronic alcoholic liver aging. Eight-week-old C57BL / 6J male mice were divided into a blank group, an alcohol group, an alcohol + silybin (100 mg / kg) group, an alcohol + Hovenia dulcis (1 g / kg) group, an alcohol + daylily (0.5 g / kg), an alcohol + daylily (1 g / kg), and an alcohol + daylily (2 g / kg) according to their body weight. Before formal administration, the mice will be adapted to Lieber-DeCarli liquid feed (alcohol content 1%-4%) for 5 days. On the sixth day, formal administration was carried out. Except for the blank group and the model group, the other groups were given different drugs by gavage. The blank group and the alcohol group were gavaged with the same volume of solvent. Freshly prepared Lieber-DeCarli liquid feed (alcohol content 4%) was replaced every afternoon. The drug was administered for a total of 4 weeks. On the last day of the experimental cycle, the last alcohol gavage was performed at 9:00 am, and the mice were blooded from their eye sockets at 18:00 pm. The mice were killed, dissected, and the liver was removed. The appearance of the mouse liver was photographed, the liver weight was weighed, and the liver-to-body ratio was calculated. The liver tissue was taken and the corresponding experiments were performed. The data were summarized, a bar graph was drawn using Graph Pad, and the significant difference (t-test) was calculated.

[0158] The experimental results are as follows Fig. 20 As shown, compared with the blank group, the alcohol group had significantly increased levels of serum CXCL1, MMP-1, p16, and tissue CXCL-1, CXCL-3, IL-1β, MMP-1, MMP-7, and p16, indicating that long-term drinking mice will have a significant increase in aging-related indicators, and long-term drinking is accompanied by liver aging. Compared with the serum and liver of the alcohol group, alcohol + daylily (0.5, 1, 2g / kg), serum CXCL1, MMP-1, p16, tissue CXCL-1, CXCL-3, IL-1β, MMP-1, MMP-7, p16 were significantly reduced, indicating that daylily can reduce the accumulation of aging indicators in liver tissue and play an anti-liver aging role, *** indicates P < 0.001.

[0159] The experimental results are as follows Fig.21As shown, compared with the blank group, the rate of senescence-associated β-galactosidase (SA-β-gal) positive cells in the alcohol group tissue slices increased significantly, indicating that long-term drinking mice will have a significant increase in aging-related indicators, accompanied by liver aging. Compared with the liver slices of the alcohol group, the rate of senescence-associated β-galactosidase (SA-β-gal) positive cells in alcohol + daylily (0.5, 1, 2g / kg) was significantly reduced, indicating 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 as follows Fig. 22 As shown, compared with the blank group, the levels of p53 and γH2AX in the alcohol group tissues were significantly increased, indicating that long-term drinking mice will have a significant increase in aging-related indicators, and long-term drinking is accompanied by liver aging. Compared with the liver sections of the alcohol group, alcohol + daylily (0.5, 1, 2g / kg), p53, γh2AX content was significantly reduced, indicating that daylily can reduce the accumulation of aging indicators in liver tissue and play an anti-liver aging role, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

[0161] Example 14 Safety Experiment of Daylily

[0162] This experiment uses 14 days of continuous high-dose administration to examine the effects of the drug on the body-to-viscera 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 will be gavaged for 14 days, and each administration group will be given different doses of daylily crude extract, and the blank group will be given the same volume of solvent. The weight changes of the mice will be recorded daily. On the 15th day, blood was collected from the mouse orbits and dissected, the mouse organs were weighed, and the body-to-viscera ratio was calculated. The liver was fixed, paraffin sectioned, and then H&E stained. The data were summarized, a bar graph was drawn using Graph Pad, and significant differences (t-test) were calculated.

[0163] The experimental results are as follows Fig.23 As shown, 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 indicates that daylily has good safety.

[0164] In summary, this study proved through experiments that daylily or its extract can significantly play a role in sobering up and protecting the liver at the cellular and animal levels. On the one hand, it can promote the activation of alcohol metabolism-related enzymes ADH and ALDH, accelerate the metabolism of ethanol and acetaldehyde; improve the state of subjects after being drunk, including delaying the time of drunkenness, shortening the duration of drunkenness, improving the behavior after being drunk, restoring the motor ability after being drunk, and reducing the mortality rate of mice with acute alcohol poisoning. On the other hand, it can significantly improve the morphology, liver-to-body ratio and pathology of the liver in acute alcoholic liver injury; reduce alcohol-induced liver cell and liver damage, play a role in protecting the liver, and improve chronic alcoholic liver injury, including reducing the increase of blood transaminases, triglycerides and cholesterol caused by alcohol, restoring albumin levels, and improving the decrease of blood sugar caused by alcohol; reducing the upregulation of inflammatory indicators (TNF-α, IL-6) caused by alcoholic liver; and reducing the level of cellular reactive oxygen. Reduce the lipid droplet content and fat accumulation of liver tissue caused by alcohol; restore liver aging caused by long-term drinking. This shows that daylily or its extract has broad application prospects in the fields of sobering up and preventing or treating alcoholic liver disease.

[0165] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. 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 claims attached to this application.

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.

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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