CN106573024A

CN106573024A - Composition for improving liver function, containing extract of dendropanax morbifera

Info

Publication number: CN106573024A
Application number: CN201580040390.8A
Authority: CN
Inventors: 朴贤济; 朴明用; 李晧振; 申章禹; 金昇熏; 金成奎; 金相虎
Original assignee: Yuhan Corp
Current assignee: Yuhan Corp
Priority date: 2014-05-29
Filing date: 2015-05-28
Publication date: 2017-04-19
Also published as: WO2015183027A1; KR20150138068A; KR102413705B1

Abstract

The present invention provides an extract of Dendropanax morbifera for improving liver function, containing a chlorogenic acid and/or a polyphenol as a marker component. The extract according to the present invention is excellent in antioxidant activity and hepatocyte protective efficacy, thereby rapidly recovering the liver function damaged by hepatotoxic substances and thus improving liver function, and promotes the degradation of alcohol and acetaldehyde, thereby relieving hangovers and protecting the hepatocyte and thus improving liver function.

Description

For improving liver function, the extract containing yellow paint wood compositionss

Technical field

It relates to a kind of compositionss for improving liver function, it includes natural plant extracts or detached from its Compound.

Background technology

Liver is the vitals of human body, and it is related to several functions, including：Blood storage and circulation, metabolism such as lipid metabolism, Bile secretion, the storage of various nutrition, blood flow are adjusted and detoxified.However, because Human body package is to various pollutant and has Noxious material, over-burden for liver, and also by major injuries such as stress, excessive drinking, smokings.

Liver be prevent by the intake of exterior materials caused by damage significant defense organ.For this reason, liver is worked as During function diminuendo, serious disease may be caused by viral or various medicines etc..This is likely to result in the exception in immune system, its Cause other diseases.Especially, due to a variety of causes, infect including the Excess free enthalpy containing fatty food and ethanol, virus, The for example various medicines of noxious substance, nutritional disorder etc., acute or chronic disease may occur, cause fatty liver, hepatitis, jaundice, Liver cirrhosis, hepatocarcinoma etc..Additionally, causing fatty liver, wherein lipid accumulation by food Excess free enthalpy fat or Excess free enthalpy ethanol In hepatic tissue, and in this case, in serum GOT (aspartate transaminase, AST), GPT (alanine aminotransferase, ALT), γ-GPT (γ-. glutamyl transferase) etc. level raise.

According to cause, hepatopathy is categorized as the mediated hepatopathy of viral hepatopathy, alcoholic hepatic disease, medicine, fat Liver, autoimmunity hepatopathy, metabolic hepatopathy and other hepatopathys.Because liver is the organ with huge buffer capacity, The disease of liver in many cases the starting stage it is subjective be not noticeable, and simply after sizable progress just by It was found that.For this reason, not only in Korea, and in the world, hepatopathy is all main causes of death.

As described above, liver function loss of energy is not discovered, and cause the defense function and function of detoxification of human body Loss, so as to cause human body in many problems.Therefore, for development has Liver protection effect and prepares as safe preparation Medicine and functional health food, there are needs.

Meanwhile, yellow paint wood (Dendropanax morbifera) is the warm temperature that one kind belongs to Araliaceae (Araliaceae) Band subtropical evergreen broad-leaved, its growth up to 15m height, and be distributed in Korea's Southern Coast region, Japan the torrid zone and Subtropical area and Taiwan.The botanical term of yellow paint wood is Dendropanax morbifera Nakai, from yellow paint wood The resin collected of bark contain Benzoinum, and therefore paid close attention to as the trees with outstanding pharmacological effect, and It was used as natural coating in the past.Nearest research is disclosed, yellow paint wood in anticancer therapy, treating diabetes, sclerous tissueses' cell again The aspect such as raw is effective.

However, for yellow paint wood extract pharmacological effect disclosed in those be very not enough, and depend on The pharmacological activity of the content of the key component of these extracts or the component or optimal pharmacological effect are unknown.Including Yellow paint is wooden in interior many natural materials, according to the difference of the content of component or the component contained in these natural materials It is different, huge difference may be shown in terms of their effect.Therefore, in order to obtain the effect wanted, not only for right These natural materials itself and composition and its content to showing optimum efficiency, have the further need to studying Will.

Disclosure

Technical problem

The disclosure is intended to provide a kind of yellow paint wood extract for improving liver function, and it contains chlorogenic acid.

The disclosure is also intended to provide a kind of compositionss for improving liver function, and it includes the yellow paint wood containing chlorogenic acid and carries Thing is taken as active component.

The disclosure is also intended to provide a kind of method for improving liver function, and what it included applying effective dose contains chlorogenic acid Yellow paint wood extract.

The disclosure be also intended to provide containing chlorogenic acid yellow paint wood extract prepare for improve liver function medicine or Purposes in food.

The disclosure is also intended to provide a kind of yellow paint wood extract for improving liver function, and it contains polyphenol.

The disclosure is also intended to provide a kind of compositionss for improving liver function, and it includes the wood of the yellow paint containing polyphenol and extracts Thing is used as active component.

The disclosure is also intended to provide a kind of method for improving liver function, it include applying effective dose containing polyphenol Yellow paint wood extract.

The disclosure is also intended to yellow paint of the offer containing polyphenol wood extract and is preparing medicine or food for improving liver function Purposes in product.

Technical scheme

The present inventor has made intensive studies, and is found that displaying pharmacological effect (including passing through in yellow paint wood It is still drank after a night the Hepatic function improvement that mitigation (hangover relief) and hepatocyte protection (hepatocyte protection) reach) Component, and from yellow paint wood extract separated index component (index component) first, and identify separation Component structure, and it was found that, yellow paint containing chlorogenic acid and/or polyphenol wood extract is thin for by mitigation regulating liver-QI of being still drank after a night Born of the same parents' Hepatic function improvement for reaching of protection has an outstanding effect, and have also discovered the extract for showing optimum activity component and Their content, so as to complete the disclosure.

The disclosure provides a kind of yellow paint wood extract for improving liver function, and it contains chlorogenic acid.

The disclosure also provides a kind of compositionss for improving liver function, and it includes the wood extract of the yellow paint containing chlorogenic acid As active component.

The disclosure also provides a kind of food comprising above-mentioned composition.

The disclosure also provides a kind of functional food comprising the food.

The disclosure also provides a kind of yellow paint wood extract for improving liver function, and it contains polyphenol.

The disclosure also provides a kind of compositionss for improving liver function, and it includes the wood extract of the yellow paint containing polyphenol and makees For active component.

The disclosure also provides a kind of functional food comprising the food.

In the disclosure, chlorogenic acid has the structure represented by following formula 1：

Formula 1

The yellow paint wood extract for improving liver function according to the disclosure containing chlorogenic acid is because it contains chlorogenic acid And there is remarkable result to Hepatic function improvement.

As it is used herein, statement " Hepatic function improvement " refers to the reparation and treatment of the liver function for dying down, and anticipate Including, for example, not exclusively improve (including mitigation of being still drank after a night, is caused by Hepatoxic substance (hepatotoxic substance) Hepatocyte injury (hepatocyte damage), alcoholic liver (alcoholic liver) and hepatic injury (liver damage) Treatment, or removing toxic substances (detoxification)), and be the prevention and treatment of hepatic injury.Additionally, the statement is intended to include liver The various functions for the following in the improvement of all or some：Such as drink for living habit, high fat diet or The continuous bile secretion of fatigue, synthesis, excretion and metabolism.Especially, state " Hepatic function improvement " be intended to include by AST and (it is the damage to biliary system for ALT levels (it is the biomarker that liver parenchyma sexual cell is damaged), γ-GT and ALP levels Biomarker) and bilirubin level (it is the biomarker of the damage to bile secretion function) return to normal water It is flat.

As the example of Hepatic function improvement, be still drank after a night mitigation (alleviation) or mitigate suppress or reduce after drinking Headache, diarrhoea, inappetence, Nausea and vomiting, shiver with cold, cold sweat of appearance etc., and it is by body is from cognitive competence reduction and transports Kinetic force reduces recovering, and maintains normal blood and hormonal status.Needing the colony of such Hepatic function improvement includes, example Such as, the common people for preventing hepatopathy and lifting liver health are needed；Need to lift liver function or prevention hepatopathy and with liver disease The prognosis of disease or the patient with unconscious initial stage hepatopathy；Or need that the liver of liver removing toxic substances is treated or lifted for hepatopathy Function improves and with the patient of disease such as fatty liver, hepatitis, jaundice, liver cirrhosis, hepatocarcinoma etc., and any needs liver function The people of improvement.

The wood extract of the yellow paint containing chlorogenic acid according to the disclosure is to use the natural material yellow paint of safety wooden standby, And the single compound unlike, side effect is not caused when human body is applied to.Additionally, being had according to the extract of the disclosure There is outstanding antioxidant activity regulating liver-QI cell protection activity, and therefore outstanding effect is shown to Hepatic function improvement.

In the disclosure, as yellow paint wood, especially restriction can be not added with using branch, leaf, root, the fruit etc. of trees.It is excellent Selection of land, using the branch (including xylem) and leaf of trees.Yellow paint wood used in the disclosure is widely distributed in Korea, Japan, platform The ground such as gulf, with low cost acquisition, and can buy or directly collect.

The extracting method that can be used in the disclosure includes that hot water extraction, Soakage extraction, reflux cooling are extracted and ultrasound Extract.Extraction is preferably carried out 1 to 5 time.The Extraction solvent for using in the disclosure is water, alcohol or their mixture.It is preferred that Ground, Extraction solvent is selected from water, C₁To C₄The solvent of lower alcohol and their mixture.It is highly preferred that Extraction solvent is water, first Alcohol-water solution or ethanol water, but not limited to this.The amount of the Extraction solvent for using is the 5- of the weight of the branch of yellow paint wood or leaf 15 times.For the hot water extract of yellow paint wood, extraction preferably carries out 3-48 hours at 90 to 120 DEG C.For yellow paint wood For methanol and ethanol extraction, extraction preferably carries out 3-48 hours at 60 to 90 DEG C.

The disclosure also provides a kind of yellow paint wood extract fraction by the way that yellow paint wood extract classification is obtained.Unless in addition Point out, in the description of the disclosure, it is believed that yellow paint wood extract includes yellow paint wood extract fraction.Using known in the art Separation method, carries out the classification of yellow paint wood extract.Preferably, can be by the way that yellow paint wood extract be suspended in water, subsequently Extracted to obtain the soluble extract of yellow paint wood with solvent such as hexane, chloroform, ethyl acetate, butanol etc..Specifically, can be with By adding hexane to obtain the fraction and water-soluble fraction that dissolve in hexane to yellow paint wood extract, and chlorine can be used The imitative fraction for extracting water-soluble fraction to obtain water-soluble fraction He dissolve in chloroform.Furthermore, it is possible to dissolving in The fraction addition ethyl acetate of water, to obtain the fraction and water-soluble fraction of ethyl acetate are dissolved in, and finally, can be with With the water-soluble fraction of butyl alcohol extraction, to obtain the fraction and water-soluble fraction that dissolve in butanol.Preferably, Ke Yiyi Secondary use normal hexane, ethyl acetate and n-butyl alcohol are used to be classified.By this stage division, it is possible to obtain preferably contain containing being in The yellow paint wood extract fraction of the chlorogenic acid of amount scope.

Furthermore, it is possible to by carrying out silica gel column chromatography (ethyl acetate fraction is used into hexane/ethyl acetate/methanol mixed Thing is used as eluting solvent) 5-11 kind fraction is separated into, so as to obtain chlorogenic acid.In order to be further purified, can be by being made Standby type high performance liquid chromatography (HPLC), using 20-70% as mobile phase, and will merge from the eluate of chromatographic column eluting and Concentration, so as to obtain compound, the chlorogenic acid purification that will be obtained.Except said method, it is also possible to by conventional for taking The method of synthesis and the classification of Dai Ji synthesizes the compound from yellow paint wood for mitigating activity with being still drank after a night according to the disclosure.

The content of the effective biological active substanceies in raw material herbal material be likely to be dependent on the place of production, acquisition time and Storage period and condition of storage and great variety, and the effective physiology in the raw material herbal material with identical weight lives The content of property material may change up to 100 times.For those reasons, it is very difficult to by raw material herbal material standardization.In this public affairs In opening, in order to overcome this problem and obtain show needed for maximized pharmacological effect yellow paint wood extract, various Chlorogenic acid is selected in preparation method as important biological active substanceies, and determines optimal pharmacological activity.Especially, because Content for yellow paint wood Content of Chlorogenic Acid depends on the place of production, acquisition time, Storage period and condition of storage and great variety, and because The great variety depending on its content such as hepatocyte protection activity, antioxidant activity for chlorogenic acid, determines that chlorogenic acid is optimal Content range so as to by the Hepatic function improvement effect of extract maximize.

Preferably, the wood extract of the yellow paint containing chlorogenic acid contains at least 0.10 weight based on the gross weight of yellow paint wood extract Amount %, preferred 0.10-6.5 weight %, more preferably 0.30-6.5 weight %, even more preferably the amount of 1.0-6.5 weight % is green Ortho acid.

In the range of the chlorogenic acid content, compared with those with other content ranges, extract shows outstanding place Liquor-saturated mitigation regulating liver-QI cytoprotective effect, and outstanding effect is displayed that compared with single Chlorogenic acid compound.Particularly, it Advantage be that it can represent the maximized protection of liver poisoning for being caused by hepatotoxic compound such as carbon tetrachloride and imitate Really.

The yellow paint wood extract for improving liver function containing chlorogenic acid can also containing the new of the structure with following formula 2 4-O- caffeoyl guinic acids (4-dicaffeoylquinic acid (the cryptochlorogenic of chlorogenic acid and/or the structure with following formula 3 acid))：

Formula 2

Formula 3

When the compound and/or formula 3 according to the yellow paint wood extract for improving liver function of the disclosure also containing formula 2 During compound, it has more preferable antioxidant activity and more preferable hepatocyte protection and liver function improvement.In the disclosure In, can be by said extracted method and stage division, by the compound of the formula 2 also containing the constitutional isomer for being chlorogenic acid And/or the yellow paint wood extract of the compound of formula 3 is incorporated in extract and its fraction.Compound also containing formula 2 and/or The yellow paint wood extract of the compound of formula 3 can contain neochlorogenic acid, and gross weight of its amount based on yellow paint wood extract is at least 0.13 weight %, more preferably preferred 0.13-1.6 weight %, 0.30-1.6 weight %, even more preferably 0.40-1.6 weight %. Additionally, it can contain 4-O- caffeoyl guinic acids (4-dicaffeoylquinic acid), gross weight of its amount based on yellow paint wood extract is at least 0.11 weight %, more preferably preferred 0.11-1.2 weight %, 0.12-1.2 weight %, even more preferably 0.25-1.2 weight %.

Additionally, the wood extract of the yellow paint containing chlorogenic acid and its constitutional isomer can contain total chlorogenic acid (chlorogenic acid With the summation of its constitutional isomer), its amount be at least 0.34 weight %, preferred based on the gross weight of yellow paint wood extract 0.34-9.3 weight %, more preferably 1.0-9.3 weight %, even more preferably 1.5-9.3 weight %.

The disclosure provides a kind of compositionss for improving liver function, and it includes the wood extract of the yellow paint containing chlorogenic acid and makees For active component.

The compositionss for improving liver function according to the disclosure can be Pharmaceutical composition, food according to its desired purposes Product compositionss, functional food or functional health food.

The compositionss of the disclosure can in a variety of manners be prepared according to its desired purposes by conventional method, including oral Preparation, such as powder, granule, tablet, capsule, suspension, pill, extract, emulsion, syrup or aerosol, and injectable is molten Liquid such as sterile injectable solution, and can be administered via various approach, including it is oral, intravenous, endoperitoneal, subcutaneous , intrarectal and local approach.

The example of the pharmaceutical carrier in this compositions, excipient and diluent can be included to be included：Lactose, glucose, Sucrose, Sorbitol, mannitol, xylitol, erithritol, maltose alcohol, starch, arabic gum, alginate, gelatin, phosphoric acid Calcium, calcium silicates, cellulose, methylcellulose, amorphous cellulose, crystal fibre element, carboxymethyl cellulose, carboxymethyl cellulose Calcium, sodium carboxymethyl cellulose, Explotab, cyclodextrin, rudimentary substituted hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxyl Propyl cellulose, xanthan gum, guar gum, agar, citric acid, sodium citrate, sodium alginate, polyvinylpyrrolidone, water, hydroxyl It is essence of Niobe, nipasol, Talcum, magnesium stearate, sucrose fatty acid ester, shell calcium (shell calcium), hard Fatty acid magnesium, Talcum and mineral oil.

The compositionss of the disclosure can also include as the vitamin B of additive, vitamin C, Vitamin E, β for adding- Carotene, Ca, Mg, Zn, lecithin, alanine, taurine (taurin), maltol, Fructose, oligosaccharide, Ganoderma (Ganoderma lucidum), glutamate, Glu, shitosan, aspartic acid, Cordyceps (Cordyceps sinensis), turn Fructus Jujubae (Hovenia dulcis) extract, Japanese alder (Alnus japonica) extract, Milk Thistle (Milk thistle) Deng.

The compositionss of the disclosure can also include filler, anticoagulant, lubricant, wetting agent, spice, emulsifying agent, anti-corrosion Agent etc..Include tablet, pill, powder, granule, capsule etc., and such solid system for Orally administered solid preparation Agent is outside compositionss also comprising at least one excipient, for example, starch, Calcium Carbonate, sucrose, Lactose or gelatin.Except simple Excipient, it is also possible to using lubricant such as magnesium stearate or Talcum.Include suspension, molten for Orally administered liquid preparation Liquid, emulsion and syrup, and various excipient can be contained, for example, except commonly used simple diluent water and liquid Wetting agent, aromatic, aromatic substance and preservative outside body paraffin.

When the compositionss of the disclosure are pharmaceutical compositions, it is administered with pharmacy effective dose.As it is used herein, art Language " pharmacy effective dose " refers to be enough to the available rational benefit/risk for any medical therapy than treatment disease Amount.Can determine the effective dose level of compositionss depending on various factors, including the species of the disease of patient, disease are tight Severe, the activity of medicine, the sensitivity to medicine, administration time, route of administration, discharge rate, the persistent period for the treatment of and group Medicine and other factors known to medical field that compound is applied in combination.Can by the compositionss of the disclosure be administered alone or with Combination with other therapeutic agents is administered, and can sequentially or simultaneously be administered with conventional therapeutic agent.Can be by compositionss with single or multiple Dosage forms for administration.In view of all above-mentioned factors, it is important that so that maximum can be shown in the case where side effect is not caused The minimum of effect is administered compositionss, and those skilled in the art can readily determine that the amount.Specifically, the disclosure The effective dose of the active component in compositionss can depend on age, sex and the body weight of patient and change, and active component Dosage can be 10-20000mg, preferred 100-5000mg.However, the dosage is not intended to limit the disclosure by any way Scope, because it can depend on severity, the age of patient, sex and body weight of disease etc. changing.

The disclosure provides a kind of food for improving liver function or a kind of functional food for improving liver function, its Comprising compositionss, the compositionss contain the wood extract of the yellow paint containing chlorogenic acid as active component.

As it is used herein, term " food " refers to the natural material or finished with more than one nutrients Food.

As it is used herein, term " functional food " is referred in addition to the function of supply nutrient with beneficial The food of the function of health (including biophylaxises, disease prevention and recovery, biorhythm regulation etc.).In the disclosure, function Property food there is beneficial effect to Hepatic function improvement.

According to the disclosure wood of the yellow paint containing chlorogenic acid extract can be used alone or with other food or food group Subassembly is used, and can be suitably used according to any conventional method.

The food of extract can be added to be included, for example, various food, beverage, chewing gum, tea, confection, vitamin Complex, functional health food, powder, granule, tablet, capsule, fruit jelly or beverage, but to it, there is no particular limitation.

Preferably, can in order to mitigated by being still drank after a night and hepatocyte protection and the purpose that improves liver function adds extract To in Foods or drinkses.In this case, the amount of the extract in Foods or drinkses can be based on the gross weight of food For 0.01-15 weight %, and for beverage composition for treating dental erosion, can be with every 100mL beverage composition for treating dental erosion 0.01-20g, preferred 0.1-5g Amount addition extract.Except it contains extract using the amount that indicates as in addition to solvent, the food of the disclosure is for it He does not have special restriction at component.The food of the disclosure can be containing various aromatic or natural carbohydrate as addition Agent, seems the situation of conventional beverage.The example of natural carbohydrate include saccharide for example glucose, Fructose, maltose, sucrose, Dextrin, cyclodextrin etc., and sugar alcohol such as xylitol, Sorbitol, erithritol etc..Aromatic includes natural aromatic agent (thaumatin (thaumatin), Flos Chrysanthemi extract (for example, Lay bud enlightening glycosides A (rebaudioside A), glycyrrhizin etc.)) and synthesis fragrance Agent (saccharin, Aspartame etc.).Additionally, the food of the disclosure can also contain various nutrients, vitamin, mineral (electrolysis Matter), synthesis and natural aromatic, coloring agent, filler, pectic acid and its salt, alginic acid, citric acid, sodium citrate and Its salt, protecting colloid thickening agent, pH adjusting agent, stabilizer, preservative, glycerol, alcohol and the carbonating for carbonated beverages Agent.Such additive is generally used with the amount of the extract of 0.01 to the about 50 weight portions/weight portion disclosure.

As it is used herein, term " polyphenol " refers to the compound that each molecule contains more than one phenolic group group.

The yellow paint wood extract for improving liver function according to the disclosure containing polyphenol is to use safe natural material Yellow paint is wooden standby, and when it is used for human body, compared with other polyphenolic substances, causes considerably less side effect.When this When disclosed extract contains polyphenol, it has significant antioxidant activity regulating liver-QI cytoprotective effect, and therefore shows The outstanding effect to Hepatic function improvement.

The yellow paint wood extract for improving liver function containing polyphenol contains At least 1.2 weight %, more preferably preferred 1.2-10.8 weight %, 2.2-10.8 weight %, even more preferably 4.4-10.8 weights The total polyphenols of the amount of amount %.

In the range of above-mentioned polyphenol content, extract is still drank after a night compared with those with other content ranges with outstanding Mitigate regulating liver-QI cytoprotective effect, and outstanding effect is displayed that compared with polyphenolic substance.Particularly, it is for by liver poison Property the liver poisoning that causes of material such as carbon tetrachloride show maximum protection effect.

The detailed description of the yellow paint wood extract for improving liver function containing polyphenol with above for containing chlorogenic acid For improve liver function yellow paint wood extract described in as, difference is above-mentioned polyphenol.

The disclosure also provides a kind of compositionss for improving liver function, what it contained as active component containing polyphenol Yellow paint wood extract.The compositionss for improving liver function containing the wood extract of the yellow paint containing polyphenol as active component Detailed description with above for containing the wood of the yellow paint containing the chlorogenic acid extract as active component for improving liver function As described in the compositionss of energy, difference is above-mentioned polyphenol.

The disclosure also provides a kind of food for improving liver function or the functional food for improving liver function, and it contains There is the wood extract of the yellow paint containing polyphenol as active component.

The food for improving liver function containing the wood extract of the yellow paint containing polyphenol as active component is used for Improve the detailed description of the functional food of liver function and above for containing the yellow paint containing chlorogenic acid as active component The food for improving liver function of wooden extract or as improving described in the functional food of liver function, difference It is above-mentioned polyphenol.

The disclosure also provides a kind of method for improving liver function, and it includes applying the Huang containing chlorogenic acid of effective dose The wooden extract of paint.

As it is used herein, term " effective dose " refers to effectively relax, prevent or treat for liver function show The amount of the wood extract of the yellow paint containing chlorogenic acid of effect.

Included the wood extract administration of the yellow paint containing chlorogenic acid according to the method for improving liver function of the disclosure, from And liver function recovery is provided and the symptom of deterioration of liver function is treated and suppress or avoid.In the management of Hepatic function improvement, relax, The active component of prevention or therapeutic dose is by depending on the person's character and severity of hepatopathy or the liver function patient's condition and depending on applying Changed with the approach of active component.The dosage and frequency of administration is by depending on the age of individual subjects, body weight and reaction Change.Those skilled in the art are considered that such factor is readily selected suitable dosage.Additionally, according to the disclosure Method for improving liver function can also include by effective dose auxiliary Hepatic function improvement additional active agents with contain it is green The yellow paint wood extract of ortho acid is administered together.Here, additional active agents can show extract with the wood of the yellow paint containing chlorogenic acid The synergy or supplementary result of thing.

Preferably, it is the method for treating hepatopathy according to the method for improving liver function of the disclosure, it includes By the wood extract administration of the yellow paint containing chlorogenic acid of the effective dose in treatment.

The disclosure also provides the wood extract of the yellow paint containing chlorogenic acid and is preparing medicine or food for improving liver function In purposes.Can be by the wooden extract of the yellow paint containing chlorogenic acid for preparing medicine or food and pharmaceutical excipient, dilution Agent or carrier etc. mix, and can combine it with other active agents so that active component can have synergy.

The disclosure also provides a kind of method for improving liver function, and it is included the wood of the yellow paint containing polyphenol of effective dose Extract is administered.Preferably, it is the method for treating hepatopathy according to the method for improving liver function of the disclosure, its bag Include the wood extract administration of the yellow paint containing polyphenol of the upper effective dose for the treatment of.

The disclosure also provides the wood extract of the yellow paint containing polyphenol in the medicine or food for improving liver function is prepared Purposes.

The detailed description of above-mentioned ameliorative way and purposes and being used for above for the yellow paint wood extract including effective dose Improve the method or the yellow paint wood extract containing chlorogenic acid of liver function for improving the medicine of liver function or the preparation of food In purposes described in as.

The content mentioned in purposes of the invention, compositionss and ameliorative way is used in an identical manner, unless they It is conflicting.

【Beneficial effect】

Had for the wood extract of the yellow paint containing chlorogenic acid that improves liver function according to the disclosure very outstanding anti- Anti-oxidant active and mitigation regulating liver-QI cytoprotective effect of being significantly still drank after a night, and therefore make the hepatic injury caused by Hepatoxic substance Recover to normal condition, and mitigated by being still drank after a night and for the hepatocyte protection of hepatic injury caused by ethanol and noxious substance With the remarkable result to Hepatic function improvement.

Additionally, being had according to the wood extract of the yellow paint containing polyphenol for improving liver function of the disclosure very outstanding Antioxidant activity and mitigation regulating liver-QI cytoprotective effect of being significantly still drank after a night, and therefore damage the liver caused by Hepatoxic substance Wound is recovered to normal condition, and mitigated by being still drank after a night and hepatic injury for being caused by ethanol and noxious substance hepatocyte protection And there is the remarkable result to Hepatic function improvement.

Description of Drawings

Fig. 1 shows the change of the DPPH antioxidant activities of the function as yellow paint wood extract concentrations.

Fig. 2 shows the HepG2 cell protection activities of the yellow paint wood extract of the function as concentration.

Fig. 3 shows the change of the time dependence of the BAC by being caused with yellow paint wood extract for treating.

Fig. 4 shows the change of the time dependence of the Blood Acetaldehyde concentration by being caused with yellow paint wood extract for treating.

Fig. 5 is shown in acute liver poisoning model by with the yellow paint serum AST that causes of wood extract for treating and ALT water Flat change.

Fig. 6 shows the change of the hepatic tissue in acute liver poisoning model by being caused with yellow paint wood extract for treating.

Invention pattern

The advantages and features of the disclosure and obtain their mode and will be apparent from reference to following illustrative examples. However, the disclosure is not limited to illustrative examples disclosed below, and can implement in many different forms；More precisely, These examples are provided so that open will be thoroughly and completely, and by the model of those skilled in the art's totally tansitive disclosure Enclose.The scope of the present disclosure will be limited by appended claims.

Preparation example：The preparation of yellow paint wood extract or fraction

For prepare yellow paint wood hot water extract, the alcohol extracting thing of yellow paint wood or yellow paint wood extract fraction method such as Under, and describe to use different extraction units in embodiment 1 to 10, collect season and the yellow paint wood or yellow of extracting method preparation The instantiation of the wooden extract fraction of paint.

(1) preparation of the hot water extract of yellow paint wood

The branch and/or leaf portion of the yellow paint wood that will be collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture in the annual and moon (season) Use as former state or use after being dried.The branch and/or leaf portion of the drying of the yellow paint wood of the collection of 100g are placed on by adhesive-bonded fabric Made by bag for extracting, and subsequently with the water of 1,000mL (10 times of volumes) in the extractor equipped with reflux condenser 90 To 100 DEG C of extraction 12-48 hours.Extract is cooled to into room temperature (1 to 30 DEG C) subsequently to filter.By such extraction and filtration behaviour It is repeated twice, and filtrate is merged.Brix (Brix) concentration (%) of the filtrate of merging is measured before concentration, and is subsequently passed through The Brix concentration of 15-25% is concentrated the filtrate to using rotary evaporator (Heidolph).Using spray dryer by concentrate It is dried, is powder so as to prepare the hot water extract of the yellow paint wood of 15-25g (15-25% yields).

(2) preparation of the alcohol extracting thing of yellow paint wood

The branch and/or leaf portion of the drying of the yellow paint wood of the collection of 100g are placed on into the bag for extracting by made by adhesive-bonded fabric In, and subsequently with the methanol of 1,000mL (10 times of volumes), the ethanol of 1,000mL (10 times of volumes), 1,000mL (10 times of volumes) Water/carbinol mixture or 1,000mL (10 times of volumes) water/alcohol mixture in the extractor equipped with reflux condenser 3-48 hours are extracted at 60 to 90 DEG C.Extract is cooled to into room temperature (1 to 30 DEG C) subsequently to filter.By such extraction and filtration Operation is repeated twice, and filtrate is merged.Measure the Brix concentration (%) of the filtrate of merging before concentration, and subsequently by using Rotary evaporator (Heidolph) concentrates the filtrate to the Brix concentration of 15-30%.Concentrate is dried using spray dryer, It is powder so as to prepare the alcohol extracting thing of the yellow paint wood of 9-15g (9-15% yields).

(3) preparation of the fraction of yellow paint wood extract

The alcohol extracting thing of the yellow paint wood prepared in preparation example (2) is dispersed in water, and appropriate amount is added then to it Hexane, ethyl acetate and butanol, so as to subsequently obtain the soluble fraction of these solvents.Each in these fraction is existed The lower concentration of decompression is simultaneously freezed, subsequent lyophilization at -30 DEG C, so as to prepare the hexane level of the yellow paint wood extract according to the disclosure Point, ethyl acetate fraction and butanol fraction.

Embodiment 1：The preparation of the hot water extract of yellow paint wood

According to the method for above-mentioned preparation example (1), the Huang that will be collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture in summer The branch and/or the leaf portion water of 10 times of volumes of the drying of paint wood is in the extractor equipped with reflux condenser at 90 to 100 DEG C Extract 24 hours.Extract is filtered, concentration and subsequent spray are dried, so as to prepare dry extract.

Embodiment 2：The preparation of the hot water extract of yellow paint wood

According to the method for above-mentioned preparation example (1), will be in spring, fall and winter on Jeollanam-Do, Korea Changxing Jun Heguan islands Prefecture collect yellow paint wood drying branch and/or the leaf portion water of 10 times of volumes in the extractor equipped with reflux condenser 90 to 100 DEG C are extracted 24 hours.Extract is filtered, concentration and subsequent spray are dried, so as to prepare dry extract.

Embodiment 3：The preparation of the methanolic extract of yellow paint wood

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood It is little that dry branch and/or the leaf portion methanol of 10 times of volumes extracts 24 in the extractor equipped with reflux condenser at 60 to 70 DEG C When.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 4：The preparation of the ethanol extraction of yellow paint wood

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood It is little that dry branch and/or the leaf portion ethanol of 10 times of volumes extracts 24 in the extractor equipped with reflux condenser at 70 to 90 DEG C When.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 5：Water/the alcohol mixture (7 of yellow paint wood:3 v/v) extract preparation

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood The dry branch and/or leaf portion water/alcohol mixture (7 of 10 times of volumes:3v/v) in the extractor equipped with reflux condenser Extract 24 hours at 70 to 90 DEG C.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 6：Water/the alcohol mixture (3 of yellow paint wood:7 v/v) extract preparation

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood The dry branch and/or leaf portion water/alcohol mixture (3 of 10 times of volumes:7v/v) in the extractor equipped with reflux condenser Extract 24 hours at 70 to 90 DEG C.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 7：Water/the alcohol mixture (15 of yellow paint wood:85 v/v) extract preparation

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood The dry branch and/or leaf portion water/alcohol mixture (15 of 10 times of volumes:85v/v) in the extraction equipped with reflux condenser Device is extracted 24 hours at 70 to 90 DEG C.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 8：Water/the alcohol mixture (3 of yellow paint wood leaf:7 v/v) extract preparation

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood The dry leaf portion water/alcohol mixture (3 of 10 times of volumes:7v/v) in the extractor equipped with reflux condenser 70 to 90 DEG C are extracted 6 hours.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 9：Water/the alcohol mixture (3 of yellow paint wooden branch:7 v/v) extract preparation

According to the method for above-mentioned preparation example (2), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood Dry branch portion (including the xylem) water/alcohol mixture (3 of 10 times of volumes:7v/v) in carrying equipped with reflux condenser Take in device and extracted 6 hours at 70 to 90 DEG C.Extract is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extraction Thing.

Embodiment 10：The preparation of the ethyl acetate fraction of yellow paint wood extract

According to the method for above-mentioned preparation example (3), by the yellow paint collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture wood The dry branch and/or leaf portion water/alcohol mixture (3 of 10 times of volumes:7v/v) in the extractor equipped with reflux condenser Extract 24 hours at 70 to 90 DEG C.Extract is filtered, concentrated, is dispersed in water and subsequently with hexane classification, acetic acid is used thereafter Ethyl ester is classified remaining water layer.Ethyl acetate is filtered, is concentrated and subsequently lyophilization, so as to prepare dry extract.

Embodiment 11：Extracting and developing and identification of the chlorogenic acid from yellow paint wood

(1) extract and separate

By yellow paint wood (the Dendropanax morbifera directly collected Jeollanam-Do, Korea Changxing Jun Heguan islands prefecture Lev. branch and/or leaf portion) is used as former state or used after being dried.The material that 100g the is dried water-second as solvent of 2L Alcohol mixture (3:7v/v) extract and be dried.Solvent is evaporated in vacuo to obtain extract.Extract is dissolved in into water (1L) in, and subsequently successively with the classification of normal hexane (5x1L), ethyl acetate (5x1L) and n-butyl alcohol (3x1L).By ethyl acetate level Point with n-hexane-ethyl acetate (from 100% normal hexane to 100% ethyl acetate gradient) and acetate-methanol (from 100% ethyl acetate is to 100% methanol elution gradient) eluting, and the silicagel column to being obtained as mobile phase using mixture Chromatograph fraction carries out using 20-70% methanol as the preparative liquid chromatography of mobile phase, so as to obtain chlorogenic acid.

(2) identification of compound

Respectively on the UV/VIS spectrophotometers of Agilent 8453 and JASCO FT/IR-680 plus spectrophotometers Record UV (ultraviolet)-Vis absorption spectras and infrared (IR) absorption spectra.Bruker is marked on using tetramethylsilane (TMS) as interior ID and 2D nuclear magnetic resonance, NMR (NMR) experiment is carried out on Avance 400MHz FT-NMR spectrometers.In Waters Q-TOF high-resolution matter During spectrum counts (High-Resolution Mass Spectrometer) (HRMS), electro-spray ionization-mass spectrography (ESI- is obtained MS) compose.Using silica gel 60 (70-230 mesh, Merck, Darmstadt, Germany), column chromatography is carried out.In the 0.25mm of coating The F of thick Kieselgel 60₂₅₄On (Merck, Germany) plate, using in UV (254 and 365nm) visualizations and 10% (v/v) sulfur Acid spraying is subsequently heated (reach 5min at 120 DEG C), carries out thin layer chromatography (Thin Layer Chromatography, TLC) point Analysis.

(3) identification of chlorogenic acid

In high resolution mass spec method (HRMS), detached compound chlorogenic acid shows the [M-H of m/z 353.0927⁺] point Daughter ion peak, it corresponds to C₁₆H₁₈O₉Molecular formula.UV-Vis absorption spectras are displayed in the maximum absorption at 326nm.Compound Nuclear magnetic resonance, NMR¹H spectrums show signals below：δ2.02-1.74(4H,m,H-2,6),3.54(1H,m,H-3),3.92(1H,m,H- 4),5.07(1H,m,H-5),6.98(1H,dd,H-2’),6.77(1H,d,H-3’),7.03(1H,d,H-6’),7.43(1H,d, H-7 '), and 6.16 (1H, d, H-8 ').Nuclear magnetic resonance, NMR¹³C spectrums show δ 73.96 (C-1), 36.76 (C-2), 70.74 (C-3), 68.49(C-4),71.35(C-5),37.64(C-6),126.04(C-1’),121.79(C-2’),116.17(C-3’), 148.75(C-4’),145.98(C-5’),115.21(C-6’),145.37(C-7’),114.75(C-8’),166.20(C- , and 175.42 (COOH) 9 ').Chlorogenic acid has the structure represented by following formula 1：

Formula 1

Embodiment 12：Neochlorogenic acid and 4-O- caffeoyl guinic acids (constitutional isomer of chlorogenic acid) carrying from yellow paint wood Take, separate and identify

With with identical mode in embodiment 11, by with water/alcohol mixture solvent (3:7v/v) extract yellow paint wood, base Classification and silica gel column chromatography in gradient elution, obtains neochlorogenic acid and 4-O- caffeoyl guinic acids.By embodiment 11 Authentication method, identifies the neochlorogenic acid and 4-O- caffeoyl guinic acids for obtaining.

The structural analyses of detached compound show m/z353.0927 [M-H in high resolution mass spec method (HRMS)⁺] Molecular ion peak, it corresponds to C₁₆H₁₈O₉Molecular formula.UV-Vis absorption spectras are displayed in the maximum absorption at 326nm.This is indicated The presence of aromatics conjugated system.Indicated by yellow paint wood extract and fraction institute using high performance liquid chromatography (HPLC) result of the comparison The retention time at the peak for obtaining is as the commercial criterion of neochlorogenic acid and 4-O- caffeoyl guinic acids.Neochlorogenic acid has by following formula 2 structures for representing, and 4-O- caffeoyl guinic acids have the structure represented by following formula 3：

Formula 2

Formula 3

Embodiment 13：The analysis of the content of chlorogenic acid and constitutional isomer in yellow paint wood extract and fraction

It is green using label component in each in high performance liquid chromatography (HPLC) measurement yellow paint wood extract and fraction The content of ortho acid.For the analysis of content, using Atlantis T3 posts (4.6x250mm, 5 μm).As mobile phase A, using containing There is the water/acetonitrile mixture (95 of 0.1% formic acid:5 v/v), and as Mobile phase B, prepare the water/second containing 0.1% formic acid Mixture of nitriles (5:95 v/v), filtered by 0.45 μm of filter paper, and deaerated 10 minutes with helium.As for measurement markers thing group Divide the detector of the HPLC of content, measured using ultraviolet (UV) detector, and the UV wavelength in 326nm.Standard is molten Liquid and each 20 μ L injections of sample solution, for measuring.By using the external standard method of commercial criterion, by the content amount of label component Change, and in order to prepare standard solution, using commercial criterion (including chlorogenic acid (Sigma-Aldrich, Lot SLBF3987V) With constitutional isomer neochlorogenic acid (Sigma-Aldrich, Lot BCBK8245V) and 4-O- caffeoyl guinic acid (Sigma- Aldrich, Lot BCBK6135V)) calculate yellow paint wood extract and fraction in chlorogenic acid and constitutional isomer content.Make With water/carbinol mixture (5:5v/v) standard solution is prepared, and also use water/carbinol mixture (5:5v/v) prepare yellow paint wood Extract and fraction.

Gradient elution is carried out as shown in Table 1 below.

Table 1：Gradient elution

Time (min)	Mobile phase A (%)	Mobile phase B (%)
			0	98	2
25	75	25
			30	10	90
40	10	90
			41	98	2
45	98	2

The yellow paint wood extract and fraction prepared according to embodiment 1 to 10 is tested according to said method, to measure yellow paint wood The content of extract and fraction Content of Chlorogenic Acid and constitutional isomer.The result of measurement is shown in table 2 below.

Table 2：The content range (weight %) of yellow paint wood extract and fraction Content of Chlorogenic Acid and constitutional isomer

As seen in upper table 2, the content of chlorogenic acid and its constitutional isomer depends on various conditions and changes, including The part of medical herbs, collect season and extracting method.

Embodiment 14：The measurement of the Determination of Polyphenols of yellow paint wood extract and fraction

Determination of Polyphenols is measured using Folin-Denis methods.By yellow paint wood extract and fraction each be dissolved in it is dense In spending the methanol for 2mg/mL.Subsequently, by the 10% forint-Qiao Ka ladder Austria (folin- being dissolved in water of 0.15mL Ciocalteu) reagent is added in the sample solution of 30 μ L, and is allowed being stored at room temperature 10 minutes, and to its addition The 7.5%Na of 0.12mL₂CO₃Saturated solution, and be sufficiently stirred for.Then, it is allowed to mixture solution being stored at room temperature 40 minutes, and Subsequently use absorbance of the spectrophotometer measurement at 760nm.As standard material, prepare in the concentration of 0-200 μ g/mL and do not have Gallate-based, and to analyze with sample identical mode, to produce standard calibration curve.Calculate by mg/g (gallic acid equivalant) The content of phenolic compound.

The yellow paint wood extract prepared in embodiment 1 to 10 and the Determination of Polyphenols of fraction are shown in table 3 below.

Table 3：The Determination of Polyphenols of yellow paint wood extract and fraction

	Determination of Polyphenols (mg/g, GAE¹⁾)
		Control	-0.40
Embodiment 1	12.03
		Embodiment 2	22.27
Embodiment 3	44.13
		Embodiment 4	34.72
Embodiment 5	59.98
		Embodiment 6	84.24
Embodiment 7	56.00
		Embodiment 8	108.2
Embodiment 9	80.19
		Embodiment 10	75.68

¹⁾GAE：Gallic acid equivalant.

Such as visible in upper table 3, polyphenol content depends on various conditions and changes, including part, the collection season of medical herbs And extracting method.

Test example 1：The measurement of the antioxidant activity of yellow paint wood extract and fraction

Change of the measurement as the antioxidant activity of the yellow paint wood extract of chlorogenic acid or the function of polyphenol content.In order to DPPH (2,2- diphenyl -1- picrylhydrazyls) antioxidant activity is evaluated, the energy for removing (scavenge) DPPH free radicals is checked Power.The yellow paint wood extract and fraction of 100 μ L of various concentration are added to into the 300 μM of DPPH for dissolving in ethanol of 100 μ L Reagent, and be sufficiently mixed, and mixture is incubated in the dark 30 minutes, and subsequently measure the absorbance at 517nm.

Fig. 1 shows that the antioxidant of the yellow paint wood extract prepared in embodiment 5 to 8 of the function as concentration is lived Property.It is such as visible wherein, in the concentration of 1mg/mL, antioxidant activity highest, and it is maintained at constant level.Therefore, exist The concentration of 0.25mg/mL is carried out for comparing DPPH free radical SC₅₀(scavenging capacity (scavenging activity) 50%) Test.

Table 4 below shows the yellow paint wood extract prepared in embodiment 1 to 10 and fraction of the concentration of 0.25mg/mL DPPH antioxidant activities.

Table 4：The DPPH antioxidant activities of yellow paint wood extract and fraction

	DPPH antioxidant activities (%)
		Control	0.00
Embodiment 1	37.51
		Embodiment 2	38.09
Embodiment 3	57.47
		Embodiment 4	51.22
Embodiment 5	60.55
		Embodiment 6	71.22
Embodiment 7	60.13
		Embodiment 8	86.48
Embodiment 9	68.57
		Embodiment 10	65.43

Such as visible in upper table 4, there is the wood extract of the yellow paint containing chlorogenic acid and/or polyphenol outstanding antioxidant to live Property.

Test example 2：The measurement of the hepatocyte protection effect of yellow paint wood extract and fraction

As chlorogenic acid or the function of polyphenol concentration, the hepatocyte protection effect of yellow paint wood extract is measured.In order to measure The hepatocyte protection effect of yellow paint wood extract and fraction, using human hepatoma HepG2 cell.HepG2 cells is thin with 50,000 The density kind in born of the same parents/hole in 96 orifice plates, and containing 10% hyclone (FBS) and antibiotic (penicillin and streptomycin) In RPMI culture medium, cultivate 24 hours under conditions of 37 DEG C and 5% carbon dioxide.Then, culture medium is washed with physiology salt Wash, and subsequently with each in the yellow paint wood extract of embodiment 1 to 10 and the chlorogenic acid in the RPMI culture medium without FBS Process 30 minutes.Then, by cell tert-butyl hydroperoxide (t-BHP) the reagent culture 24 hours of 500uM, to induce liver damage Wound (that is, cell death).As positive control, using LDH Cell lysis buffers cell death is induced.After the completion of culture, The amount of the LDH that kits discharge from hepatocyte is determined using LDH.

Fig. 2 shows the hepatocyte of yellow paint wood extract (being obtained in embodiment 5 to 8) of the function as concentration (HepG2) protection activity.Such as visible wherein, in the concentration of 200ug/mL, extract is directed to by tert-butyl hydroperoxide (t- BHP) hepatocyte injury that (its be Strong oxdiative stress) be caused has outstanding protection activity.Additionally, in the concentration of 200ug/mL The hepatocyte protection activity of the yellow paint wood extract prepared in embodiment 1 to 10 and fraction is compared, and in table 5 below In result of the comparison is shown.

Table 5：The hepatocyte protection activity of yellow paint wood extract and fraction

	Hepatocyte protection activity (%)
		Control (feminine gender)	0.00
Embodiment 1	9.51
		Embodiment 2	14.5
Embodiment 3	21.7
		Embodiment 4	16.3
Embodiment 5	33.3
		Embodiment 6	40.7
Embodiment 7	32.4
		Embodiment 8	50.1
Embodiment 9	38.8
		Embodiment 10	37.1

In addition show, compared with the independent chlorogenic acid of same concentrations, there is yellow paint wood extract outstanding hepatocyte to protect Shield effect.

Test example 3：Being still drank after a night in vivo for yellow paint wood extract mitigates the measurement of effect

Checked the mitigation effect of being still drank after a night of the yellow paint wood extract of the disclosure.

(1) laboratory animal and rearing conditions

As animal, using the male Sparague-Dawley rats (average weight of 5 week old：180g ± 5g), and often Group is made up of 10 rats.By rat stable breeding under the following conditions：22 DEG C of temperature；45 ± 10% relative humidity；15-20 leads to Wind；12-hr is bright/12-hr dark cycles；The intensity of illumination of 150-250Lux；With the noise level of below 50db.In order that big Mus adapt to surrounding, temperature, humidity, feeding changed during testing etc., adapt to rat within 1 week before experiment starts.

(2) experimental technique

After the adaptation of 1 week, by the male SD rat fasting (fast) of 5 week old, and subsequently it is divided into 6 groups, 10 per group Rat.To negative control group vehicle (vehicle) oral administration, to the test group yellow paint wood for preparing in example 2 Extract oral is administered, and to positive controls with the Yeomyeong 808 of 1500mg/kg body weight (Grami Co., Ltd.s, Korea) oral administration.After 30 minutes, 40% ethanol, and 1,3 and 5 hours after application are applied to Oral Administration in Rats, to blood Liquid is sampled.Blood is centrifuged 15 minutes at -4 DEG C and 3000rpm, to separate serum.

(3) measurement of alcohol levels in blood samples concentration

Using test kit (r-biopharm, Cat.No.10 176 290 035), measure 1 hour after ethanol is applied, 3 hours and the alcohol concentration of the serum for obtaining from blood sample for 5 hours.NAD reactions to contained 3.0mL in test kit are molten In liquid, for matched group, the distilled water of 0.1mL is added and the mixing in cuvette, and for test group, by 0.1mL's Serum in distilled water 1:50 diluents are added and mixed in cuvette.After the reaction of 3 minutes, measure at 340nm Absorbance.Then, the enzyme ADH (alcohol dehydrogenase) of 0.05mL contained in test kit is added in it, and is reacted 5 minutes, And the subsequent measurement absorbance at 340nm.

Alcohol concentration (μ g/mL) is calculated using the equation shown in table 6 below.

Table 6：The calculation equation of alcohol concentration

(4) measurement of Blood Acetaldehyde concentration

Using test kit, (035) r-biopharm, Cat.No.10 668 613, is measured 1 hour after ethanol is applied, 3 The acetaldehyde concentration of hour and the serum for obtaining from blood sample for 5 hours.NAD reactions to contained 3.0mL in test kit are molten In liquid, for matched group, the distilled water of 0.2mL is added and is mixed in test tube, and for test group, by 0.2mL not The serum of dilution is added and mixed in test tube.After the reaction of 3 minutes, the absorbance at 340nm is measured.Then, Xiang Qi The enzyme ALDH (aldehyde dehydrogenase) of 0.05mL contained in test kit is added in, and is reacted 5 minutes, and the subsequently survey at 340nm Amount absorbance.

Acetaldehyde concentration (μ g/mL) is calculated using the equation shown in table 7 below.

Table 7：The calculation equation of acetaldehyde concentration

Acetaldehyde concentration (μ g/mL)=(V × M.W.)/(ε × d × v × 2 × 1000) × △ A
	The amount (mL) of V=final mixtures
The amount (mL) of v=processed sample
	The molecular weight (44.05g/mol) of M.W.=acetaldehyde
The length (1cm) of the path that d=light passes through cuvette
	ε=6.3 (L × mmol^-1×cm^-1) (in the wavelength of 340nm)
△ A (change of absorbance)=[(A2-A1) test group-(A2-A1) matched group]

(4) experimental result

Table 8 below shows the result of measurement alcohol levels in blood samples concentration.It is such as visible in table 8, when by with preparing in example 2 When the group of yellow paint wood extract for treating compares with negative control group, it can be seen that dense with the alcohol levels in blood samples in negative control group Degree is compared, with yellow paint wood extract for treating group in alcohol levels in blood samples concentration by time dependence and concentration dependent in the way of Substantially reduce.

Additionally, as seen in Figure 3, the alcohol degradation effect that yellow paint wood extract shows is similar to commercially available product Alcohol degradation effect.

Table 8：The change of the alcohol levels in blood samples concentration by being caused with yellow paint wood extract for treating

Table 9 below and Fig. 4 show the result of measurement Blood Acetaldehyde concentration.

It is such as visible in table 9, when by with prepare in example 2 yellow paint wood extract for treating group and negative control group When relatively, it can be seen that compared with the Blood Acetaldehyde concentration in negative control group, in the group with yellow paint wood extract for treating Blood Acetaldehyde concentration by time dependence and concentration dependent in the way of substantially reduce.

Additionally, as seen in Figure 4, alcohol of the acetaldehyde degradation effect that yellow paint wood extract shows better than commercially available product Degradation effect.

Table 9：The change of the Blood Acetaldehyde concentration by being caused with yellow paint wood extract for treating

Test example 4：The measurement of the internal hepatocyte protection activity of yellow paint wood extract

(1) laboratory animal and rearing conditions

Used as animal, male ICR rats (the Institute of Cancer Research) mice using 5 week old is (flat Equal weight：30.06 ± 0.72g to 30.13 ± 0.83g), and be made up of 8 mices per group.Mice is enclosed under the following conditions Support：22 DEG C of temperature；45 ± 10% relative humidity；15-20 divulges information；12-hr is bright/12-hr dark cycles；150-250Lux Intensity of illumination；With the noise of below 50db.In order that mice adapt to during testing change surrounding, temperature, humidity, Feeding etc., adapts to rat in 1 week before experiment starts.

(2) experimental technique

It is by the ICR mice groups of 5 week old and subsequently 48 little before being administered with carbon tetrachloride after the adaptation of 1 week When, test specimen (the there is 0.5%CMC) oral administration being dissolved in distilled water of 24 hours and 2 hours.With carbon tetrachloride 48 hours after administration, by mouse anesthesia, and to blood sampling.By carbon tetrachloride with 1:During 499 (v/v) are diluted in olive oil, And Intraperitoneal medication, until the final dose of 20 μ L/kg.Matched group (normal group) (is had into 0.5% with solvent medium CMC) oral administration, and subsequently only use olive oil Intraperitoneal medication.Induction group vehicle (having 0.5%CMC) is orally given Medicine, and subsequently use carbon tetrachloride Intraperitoneal medication.The fasting in 8 hours from before postmortem by laboratory animal.By test group with 50 Hes The yellow paint wood extract administration prepared in embodiment 6 of 200mg/kg, and by positive controls with 50mg/kg's Silymarin (Sigma, Lot BCBJ0393V) oral administration.In order to observe the change of the body weight between experiment periods, open in experiment At (the 2nd day) on date before the date (the 0th day) of beginning and fasting measured the body weight of animal.Experiment ending, mice fasting 8 is little When, while only being raised with drinking-water, and measure the fasting body weight of each mice before postmortem.18 is little after being administered with carbon tetrachloride When, with the abdominal part that each mice is opened under diethyl etherization, and blood sampling is carried out from ventral aorta.Allow the blood of sampling Being stored at room temperature 30 minutes, and subsequently in non-heparinized (non-heparinized) centrifuge tube (SST pipes) 4 DEG C with 3000rpm is centrifuged 10 minutes, to separate serum, subsequently in -70 DEG C of storages.Albumen sampling (EP is carried out to the liver that a part is extracted Pipe), and the liver of remainder is fixed in neutral buffered formalin.

(3) measurement of blood ALT and AST levels

18 hours after being administered with carbon tetrachloride, with the abdominal part that each mice is opened under diethyl etherization, and from abdomen master Tremulous pulse carries out blood sampling.Using automatic biochemistry analyzer (HITACHI 7080), turn from the blood measuring aspartic acid of sampling The level of ammonia enzyme (AST) and alanine aminotransferase (ALT), they are blood biochemical markers things.

From impaired liver in blood discharge enzyme activity for measurement liver toxicity for be highly useful method.With Due to hepatic necrosis regulating liver-QI disorganization caused by liver poisoning process, the activity of the enzyme in serum such as AST and ALT due to Transaminase improves to the release in blood.It is such as visible in table 10 below and Fig. 5, due to being administered with carbon tetrachloride, compared to just Those often organized, serum AST and ALT levels substantially increase, and specify by the hepatic injury of Carbon Tetrachloride Induced.In addition show, In the test group with yellow paint wood extract for treating and in positive controls (Silymarin), by being led with carbon tetrachloride administration The increase of the AST and ALT activity of cause is substantially suppressed in the way of concentration dependent.It is found especially that, the yellow paint of 200mg/kg Wooden extract is similar with positive control Silymarin, has outstanding effect to Liver protection.

Table 10：In the acute hepatic injury model by Carbon Tetrachloride Induced by with yellow paint wood extract for treating caused by The change of AST and ALT levels

(4) liver histological analysis

To be fixed on for the remainder for measuring the hepatic tissue of the extraction in addition to hepatic tissue part of protein concentration In the formalin of 10% neutral buffered.Taken off with alcohol by fixed tissue water displacement wash and while concentration is gradually increased Water, is afterwards processed it with toluene, and is subsequently embedded in paraffin.With microtome (Lipshow model-45, Diversified Equipment Com., Lockport, USA) by the tissue slice of embedding into 4 μm of thickness, and subsequently Dyeed with hematoxylin and eosin, subsequently observed with optical microscope (Olympus JP/BX51, Tokyo, Japan).

As shown in FIG. 6, in normal group (control), there is the blood vessel of same shape with strand with radial direction hepatic cords Pattern is arranged towards the edge of lobules of liver, while hepatocyte keeps the normal lobules structure relative to central vein, and by four Chlorination carbon (CCl₄) inducing the hepatic tissue of liver poisoning, the normal structure of lobules of liver is unclear, and sinus hepaticus shape structure is also moved back Change.Additionally, coagulation necrosiss and the inflammatory cell of serious hepatic tissue is observed around the central vein of lobules of liver, and Observe that there is the hepatocellular swelling for losing core (nuclear loss) and change is damaged in the clear-cut margin ground of slough.With In the case of the test group of yellow paint wood extract administration in accordance with an embodiment of the present disclosure, in some parts it was observed that faint is swollen It is swollen, but it will be seen that the necrosis of the liver parenchyma tissue around central vein is significantly reduced, and various Histological research's results It is normal.Particularly, in the group of the yellow paint wood extract administration with 200mg/kg, hepatocyte is with radial direction strand pattern row Cloth, while normal lobules structure is formed relative to central vein, similar to the knot of the group of the Silymarin administrations with 50mg/kg Really, and total result of study is normal, while cell volume does not have big change.Such result and blood ALT and AST The measurement result of level is consistent with the measurement result of antioxidant enzymatic activity with the lipid peroxide concentration of hepatic tissue, and secretly Show that yellow paint wood extract has for the acute liver poisoning by Carbon Tetrachloride Induced and protect liver tonifying and protecting effects.

(5) measurement of the lipid peroxide concentration of hepatic tissue and antioxidant enzymatic activity

By using OxiSelect^TMTBARS determines test kit (MDA is quantitative) (CELL BIOLAB, USA) measurement the third two The concentration of aldehyde (MDA) uses OxiSelect determining the LPO levels of hepatic tissue^TM Total Glutathione (GSSG/GSH) Assay Kit (CELL BIOLAB, USA) and OxiSelect^TMTotal antioxidant capacity (TAC) Determine test kit (CELL BIOLAB, USA) measurement glutathion (GSH) concentration and total antioxidant capacity (total antioxidant capacity,TAC).Be associated ground with the antioxidant activity analysis of tissue, is determined using SOD try respectively Agent box-WST (Dokindo, Japan) and OxiSelect^TMCatalase activity determines test kit (CELL BIOLAB, USA) Superoxide dismutase (SOD) activity and catalase activity in measurement tissue.According to the scheme recommended by manufacturer, enter The active measurement of row projects and quantization program.

By various biochemical reactions, during aerobic metabolism, superoxide anion, and the hydroxyl produced from it are produced Base free radical destruction tissue and macromole, so that their losses of function.SOD is the enzyme defence system in internal antioxidant defense mechanisms System, is typical scavenger enzyme in vivo, in being primarily present in mitochondrion, and superoxide radical is reduced and peroxidating is produced Hydrogen (H₂O₂), so as to protect live body.As shown in table 11 below, significantly reduced by the liver poisoning that induction is administered with carbon tetrachloride SOD is active, and in the test group with yellow paint wood extract for treating and in positive controls (Silymarin), and with four The group of chlorination carbon administration is compared, and SOD activity is dramatically increased in the way of concentration dependent.Especially, the yellow paint wood of 200mg/kg Extract shows the increase of significant SOD activity, similar to positive control (Silymarin), implies that it has to Liver protection Color effect.

As shown in table 11 below, in total antioxidant capacity (TAC), shown by the liver poisoning that induction is administered with carbon tetrachloride It is active that work reduces TAC.In the test group with yellow paint wood extract for treating and in positive controls (Silymarin), with The group being administered with carbon tetrachloride is compared, and TAC activity is dramatically increased in the way of concentration dependent.Such result show similar to Pattern to the result of SOD activity measurements.Especially, the yellow paint wood extract of 200mg/kg shows the increasing of significant TAC activity Plus, similar to positive control (Silymarin), imply that it has outstanding effect to Liver protection.

It is, by the enzyme of hydrogen-peroxide reduction Cheng Shui, and internal antioxidation to be formed together with SOD that catalase is function The basis of agent enzyme system.It is generally known that when by stress (stress) promote various metabolism to produce reactive oxygen, and therefore When SOD activity increases, catalase activity also increases.Therefore, the change of catalase activity shows and SOD activity changes The similar pattern of pattern.As shown in table 11 below, in the pattern similar to SOD activity changes, the yellow paint of 200mg/kg is wooden Extract shows dramatically increasing for catalase activity, similar to positive control (Silymarin), implies that it eliminates reaction Property oxygen, with protect biological tissue not by metabolic process in vivo produce poisonous peroxide affected, so as to recover to receive The function of the hepatic tissue of damage.

When the oxidative stress in cell increases the generation of free radical and reduces the antioxidant activity of cell, fat is produced Matter peroxide.This lipid peroxide is by the main cause of the hepatic injury of Carbon Tetrachloride Induced.As shown in table 11 below, The concentration of lipid peroxide (malonaldehyde (MDA)) is significantly higher than in normal group in carbon tetrachloride administration group, and uses yellow paint The test group that wooden extract is treated together with positive control (Silymarin) is shown in the way of concentration dependent to lipid peroxy The effect for significantly inhibiting of the formation of compound.

Learn from the research of the liver poisoning mechanism to Carbon Tetrachloride Induced, because GSH is played directly remove reactive oxygen Effect or the effect of the cofactor as glutathion peroxidase, and therefore show that the height for reactive oxygen protects effect Really, so it plays sizable work during the highly reactive poisonous metabolite by Carbon Tetrachloride Induced is removed With.Increase by cell death caused by oxidative stress is caused by the reduction of glutathion (GSH) concentration.Therefore, such as exist Shown in table 11 below, in carbon tetrachloride administration group, GSH concentration is substantially less than in normal group.Additionally, with yellow paint wood extract In the test group treated together with positive controls (Silymarin), GSH concentration is dramatically increased in the way of concentration dependent.

Table 11：In the acute hepatic injury model by Carbon Tetrachloride Induced by with yellow paint wood extract for treating caused by fat The change of matter peroxide and antioxidant enzymatic activity

Preparation example 1：The preparation of tablet

The yellow paint wood extract prepared in embodiment 1 to 10 according to sizing and/or fraction, and and crystalline fibre Element, Lactose, starch etc. uniformly mix.Compound particles are granulated, and are subsequently mixed with magnesium stearate, sucrose fatty acid ester etc., Subsequently compacting, so as to prepare tablet.

The component for using in tablets and the amount of the component are shown in table 12 below.

Table 12：The component of tablet

Preparation example 2：The preparation of capsule

The yellow paint wood extract prepared in embodiment 1 to 10 according to sizing and/or fraction, and with shell calcium, crystalline substance State cellulose etc. uniformly mixes.Mixture is filled in gelatine capsule, so as to prepare capsule.

The amount of component and component used in the preparation of capsule is shown in table 13 below.

Table 13：The component of capsule

The yellow paint wood extract prepared in embodiment 1 to 10 and/or fraction：250mg
	Shell calcium (KP)：125mg
Crystalline cellulose：(KP)65mg

Preparation example 3：The preparation of liquid preparation

Prepare a kind of compositionss, the yellow paint wood that in embodiment 1 to 10 prepares of the compositionss comprising 0.15 weight % Extract and/or fraction, the liquid fructose of 10 weight %, the Mel of 2 weight %, the concentrated Succus Mali pumilae (60bx) of 2 weight %, Guarana (Guarana) extract of 0.5 weight %, the citric acid monohydrate of 0.5 weight %, the sodium citrate of 0.1 weight %, With the taurine of 0.1 weight %, and to said composition add purification water, so as to prepare liquid preparation.

Preparation example 4：The preparation of healthy beverage

The yellow paint wood extract prepared in embodiment 1 to 10 according to sizing and/or fraction, and with citric acid, Oligosaccharide, Fructus Chaenomeliss (Chaenomeles sinensis) fruit concentrate, Korea's Fructus Pruni salicinae (Korean plum) concentrate, cattle sulphur The uniform mixing such as sour (Taurin).Then, the water of purification is added to mixture, is subsequently stirred at 85 DEG C and is heated about 1 hour.Will The solution of gained is filtered, in being placed on aseptic container, sealing, and sterilization, and subsequently cold preservation, so as to prepare beverage.

The amount of component and the component used in the preparation of healthy beverage is shown in table 14 below.

Table 14：The component of healthy beverage

Claims

1. it is a kind of for improve liver function yellow paint wood (Dendropanax morbifera) extract, it contains chlorogenic acid.

2. the wooden extract of yellow paint described in claim 1, wherein the improvement of the liver function is any in the following One or more：Be still drank after a night mitigation, hepatocyte protection, be protected from alcoholic liver injury and be protected from Hepatoxic substance and cause Hepatic injury.

3. the wooden extract of yellow paint described in claim 1, wherein the improvement of the liver function is appointing in selected from the following Hepatic function improvement in what one or more hepatopathy：Fatty liver, hepatitis, jaundice, liver cirrhosis and hepatocarcinoma.

4. the wooden extract of yellow paint described in claim 1, the yellow paint wood extract is also containing neochlorogenic acid and 4-O- caffeoyls Quinic acid.

5. the wooden extract of yellow paint described in claim 4, wherein yellow paint wood extract is obtained in the following manner：Will Yellow paint wooden branch, leaf or both are added to selected from water, C₁To C₄In any solvent of lower alcohol and their mixture；Subsequently 3-48 hours are extracted at 60 to 120 DEG C.

6. the wooden extract of yellow paint described in claim 5, wherein the gross weight based on yellow paint wood extract is with 0.10-6.5 The amount of weight % contains the chlorogenic acid.

7. the wooden extract of yellow paint described in claim 6, wherein the gross weight based on yellow paint wood extract is with 0.13-1.6 The amount of weight % contains neochlorogenic acid, and with the amount of 0.11-1.2 weight % 4-O- caffeoyl guinic acids are contained, and with 0.34- The amount of 9.3 weight % contains total chlorogenic acid.

8. a kind of compositionss for improving liver function, the compositionss for improving liver function are included according to claim 1 To the yellow paint wood extract any one of 7 as active component.

9. compositionss described in claim 8, the compositionss are also comprising one or more in the following：Vitamin B, vitamin C, Vitamin E, beta-carotene, Ca, Mg, Zn, lecithin, alanine, taurine (taurin), maltol, fruit Sugar, oligosaccharide, Ganoderma (Ganoderma lucidum), glutamate, Glu, shitosan, aspartic acid, Cordyceps (Cordyceps Sinensis), Semen Hoveniae (Fructus Hoveniae) (Hovenia dulcis) extract, Japanese alder (Alnus japonica) extract, Milk Thistle and Their mixture.

10. compositionss described in claim 9, wherein the compositionss are any form in the following：Liquid Body, suspension, powder, granule, tablet, capsule, pill and extract.

11. it is a kind of for Hepatic function improvement and prevention functional food, the functional food include according to claim 8 institute The compositionss stated.

A kind of 12. food for improving liver function, the food includes compositionss according to claim 8.

The food for improving liver function described in 13. claim 12, wherein the food be selected from tea, fruit jelly, chewing gum, The form of any one of confection and beverage.

A kind of 14. yellow paint wood extracts for improving liver function, it contains polyphenol.

Yellow paint wood extract described in 15. claim 14, wherein the improvement of the liver function is appointing in the following What one or more：Be still drank after a night mitigation, hepatocyte protection, be protected from alcoholic liver injury and be protected from Hepatoxic substance and make Into hepatic injury.

Yellow paint wood extract described in 16. claim 14, wherein the improvement of the liver function is in the following Hepatic function improvement in any one or more of hepatopathy：Fatty liver, hepatitis, jaundice, liver cirrhosis and hepatocarcinoma.

Yellow paint wood extract described in 17. claim 14, wherein yellow paint wood extract is obtained in the following manner： Yellow paint wooden branch, leaf or both are added to selected from water, C₁To C₄In any solvent of lower alcohol and their mixture, with Extract 3-48 hours at 60 to 120 DEG C afterwards.

Yellow paint wood extract described in 18. claim 14, wherein the gross weight based on yellow paint wood extract is with 1.2- The amount of 10.8 weight % contains total polyphenols.

A kind of 19. compositionss for improving liver function, the compositionss for improving liver function are included according to claim Yellow paint wood extract any one of 14 to 18 is used as active component.

Compositionss described in 20. claim 19, the compositionss are also comprising one or more in the following：Dimension life It is plain B, vitamin C, Vitamin E, beta-carotene, Ca, Mg, Zn, lecithin, alanine, taurine, maltol, Fructose, oligomeric Sugar, Ganoderma, glutamate, Glu, shitosan, aspartic acid, Cordyceps, Semen Hoveniae (Fructus Hoveniae) extract, Japanese alder extract, Milk Thistle and Their mixture.

Compositionss described in 21. claim 20, wherein the compositionss are any form in the following：Liquid Body, suspension, powder, granule, tablet, capsule, pill and extract.

A kind of 22. functional foods for improving liver function, the functional food is comprising according to claim 19 Compositionss.

A kind of 23. food for improving liver function, the food includes compositionss according to claim 19.

The food for improving liver function described in 24. claim 23, wherein the food be selected from tea, fruit jelly, chewing gum, The form of any one of confection and beverage.

A kind of 25. methods for improving liver function, methods described includes that the yellow paint wood containing chlorogenic acid for applying effective dose is carried Take thing.

Purposes of the 26. wood extracts of the yellow paint containing chlorogenic acid in the medicine or food for improving liver function is prepared.

A kind of 27. methods for improving liver function, methods described includes that the wood of the yellow paint containing polyphenol for applying effective dose is extracted Thing.

Purposes of the 28. wood extracts of the yellow paint containing polyphenol in the medicine or food for improving liver function is prepared.