CN103054906A - Propolis ethanol extract for alleviating hangover and preparation method thereof, and application of propolis ethanol extract in producing buccal tablets - Google Patents
Propolis ethanol extract for alleviating hangover and preparation method thereof, and application of propolis ethanol extract in producing buccal tablets Download PDFInfo
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Abstract
The invention relates to a propolis ethanol extract for alleviating a hangover and a preparation method thereof, and application of the propolis ethanol extract in producing buccal tablets, belonging to the technical field of techniques for extracting effective substances from propolis. The method comprises the following steps: freezing propolis, pulverizing, removing impurities, soaking at normal temperature under normal pressure, carrying out ultrasonic extraction, carrying out refrigerated centrifugation to remove solid impurities, taking the ethanol solution, and finally, separating out the propolis ethanol extract with the molecular weight of lower than 5000D at the ambient temperature of 1-5 DEG C; and distilling the ethanol to recover a bioactive dried substance with the solid content of higher than 90% and the molecular weight of lower than 5000D, and mixing the bioactive dried substance with royal jelly freeze-dried powder, isomalt, magnesium stearate, menthol and the like to obtain the buccal tablets for alleviating a hangover. The invention has the advantages of obvious hangover alleviating effect, no side effect and low cost, and is simple and easy to implement.
Description
Technical field
The present invention relates to from animals and plants and secretions thereof, particularly from propolis, extract the Technology field of active substance.
Background technology
China is the earliest the country of making wine in the world, and spirits culture is permeated in many Social Culture, becomes already the requisite part of daily life.Along with people's material, culture and growth in the living standard, the excessive consumption of alcohol social public health problem that become international, the incidence rate of the alcoholic hepatitis that occurs therefrom also constantly increases, and has become second largest hepatopathy after viral hepatitis in China; Owing to reasons such as culture, historical or individual's hobbies, eliminate bad alcohol drinking patterns fully and still need and want the long-term endeavour of entire society.
Present various dinner party is more and more, because the sickness rate of the caused various hepatic disease of excessive drinking improves year by year, alcoholic fatty liver has become affects modern's major issue of quality of life, and society is in the urgent need to the appearance of the natural disintoxicating product of novel cheapness.
Alcoholic strength heat and gas are strong, and insobriety, alcoholism are accumulated, and the many internal organs of liver taste are impaired and cause impairment of both QI and YIN.Chinese traditional treatment is worked as take removing summer-heat alcoholism, removing heat-phlegm, supplementing QI and nourishing YIN as main; Drugs approved by FDA 3 medicines that are used for the alcoholism treatment, i.e. ester is adjoined in naltrexone, acamprosate and holder.Find also have such as ubenimex, nimodipine, emetine etc. and to a certain degree improve crapulent effect in animal experiment and the clinical research.
Propolis has very strong physiological regulatory action, propolis is the resinae material from different plants of being collected by Apis, just be used as Drug therapy by the mankind from the ancient times, it remains the most frequently used treatment article of Balkan Peninsula country now, is applied to the treatments such as wound, burn, sore throat, gastric ulcer.Researcher has carried out comprehensive research to pharmacologically active and the chemical constituent of propolis over nearly 10 years, finds that propolis has widely pharmacologically active, such as antibiotic property, antifungal, antiviral, antiinflammatory, protect the liver, antioxidation, anticancer etc.Propolis flavone is one of main effective ingredient contained in the propolis, and its pharmacological action is very extensive, and the effects such as antiinflammatory, antiallergic, carcinogenesis, anti-hypertrophy, antiviral and anti-cell differentiation are arranged; Can alleviate reactive oxygen free radical to biomembranous decay and preservation SOD; Can reduce in a large number contained MDA in the hepatic tissue, and it is active obviously to improve mouse liver SOD, CAT; The prompting propolis flavone has stronger removing oxygen-derived free radicals, antioxidative effect.Mahran etc. study discovery, and propolis solution is the hepatocellular damage of dose dependent ground protection, and its mechanism may be that water extracts of propolis has repair to hepatocyte injury.Lin etc. study discovery, and propolis also has protective effect to the hepatic injury that ethanol causes.Ethanol extracts of propolis can make serum transaminase and TG level significantly reduce, and the hepatic steatosis degree obviously descends.In a word, that propolis has is antibiotic, antiviral, antiulcer, antiinflammatory, antitumor, blood pressure lowering, regulates blood-fat and blood sugar, enhancing immunity; the protection cardiovascular, sedation anesthesia, the liver protecting; defying age improves the effects such as memory function, and its preparation has been widely used in clinical.But the research that propolis extracted with alcohol is applied to relieve the effect of alcohol remains blank at present, and we have made the molecular mechanisms of action that propolis relieves the effect of alcohol substantially clear by this research in several years, for researching and developing new solution drinks medicine based theoretical.
Present plant alcohol intoxication-alleviating preparation roughly comprises the ethanol absorption inhibitor take Araliaceae as representative, the liver metabolism promoter take Radix Puerariae, Flos puerariae lobatae as representative, and the latter is more paid attention to because it has hepatoprotective effect concurrently; In addition, Semen Hoveniae (Fructus Hoveniae) can accelerate the decomposition of body ethanol and acetaldehyde by activating the enzyme relevant with alcohol metabolism, also can play certain prevention and therapeutic effect to various chemical liver injury, is the good medicine of a class facilitating alcohol metabolism and protecting liver.
The alcoholic liver disease sickness rate is cumulative year after year trend, and the hepatic disease that causes by drinking is day by day serious, therefore finds efficient, safe solution very urgent, generally believes at present that by studying relieving alcoholism and protecting the liver mainly can reach by following several approach:
1, reaches the relieving alcoholism and protecting the liver effect by raising ethanol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) activity
The at present research about antialcoholic drugs focuses mostly in improving the active aspect of ADH, comprises the first pass metabolism (FPM) of stomach ADH and the oxidative metabolism of liver ADH.Radix Puerariae, Flos puerariae lobatae, Semen Hoveniae (Fructus Hoveniae) are the most representative solution drinks of Chinese medicine medicines, the effective ingredient of finding after deliberation their performance antialcoholism actions is Flavonoid substances, trifoliate orange yellow party (Semen Hoveniae (Fructus Hoveniae) and Radix Et Rhizoma Rhei) on the impact of alcoholic liver injury in rats stomach ADHmRNA experiment in, thereby the trifoliate orange yellow party is proved and can improves ethanol and alleviate the load that ethanol causes liver at the first pass metabolism of stomach.Trifoliate orange yellow party and Qinggan Huoxue Recipe (Radix Bupleuri, Radix Scutellariae, Radix Salviae Miltiorrhizae, Carapax Trionycis, Radix Puerariae) all can improve the activity of hepatic tissue ADH and the expression of mRNA.In the situation of long-term heavy drinking, cause the active decline of ADH in the liver, ethanol is then mainly by the CYP2E1 metabolism, but this approach can produce a large amount of acetaldehyde and active oxygen, and active oxygen makes redox state unbalance, produces response to oxidative stress and brings out the liver organization damage.Therefore, for long-term alcoholic, improve its ADH activity and can slow down the oxidative stress that is produced by the CYP2E1 metabolism, also can play liver-protective effect.Trifoliate orange yellow party and Qinggan Huoxue Recipe as described above can play antialcoholism action, also can play liver protection effect simultaneously.In addition, acetaldehyde is one of principal element of alcoholic liver injury, and its metabolism is slow and toxicity is large.Research finds that acute alcoholism liver tissues of rats ADH, the ALDH specific activity model group after the intervention of Semen Hoveniae (Fructus Hoveniae) water extraction liquid all significantly increases (P＜0.05), it is active to show that Semen Hoveniae (Fructus Hoveniae) water extraction liquid can effectively strengthen acute alcoholism liver tissues of rats ADH, ALDH, thereby the oxidation Decomposition metabolism of accelerating alcohol in liver, reduce its toxic metabolite (mainly being acetaldehyde) to the toxic action of liver, reach certain hepatoprotective effect.Yet, only improve ADH, the ALDH activity can not be slowed down the in vivo toxic action of metabolism of ethanol, also tackle the relevant key enzyme of alcohol metabolism overall process, carry out systematic study such as CYP2E1, CAT, guarantee ethanol, the thorough metabolism of acetaldehyde and the balance of vivo oxidation reducing condition.
2, reach the relieving alcoholism and protecting the liver effect by slowing down the oxidative stress effect
As mentioned above, ethanol 3 approach of metabolism in liver all can produce free radical and active oxygen, and the initiated oxidation stress brings out the damage that principal character occurs to become with fat liver organization.The factor relevant with oxidative stress has: malonaldehyde (MDA), superoxide dismutase (SOD), glutathion peroxidase (GSH-PX).The content of MDA embodies level of lipid, the active strong and weak ability that embodies Scavenger of ROS of SOD and GSH-PX, and they are the common counters in the relieving alcoholism and protecting the liver research.Exist the balance of antioxidant system and enzymatic oxidation system in the liver, this balance will be broken the activity decreased of SOD and GSH-PX behind the absorption ethanol.So activity, the removing interior free yl of antioxidant system is significant by the hepatic injury due to the oxidative stress to control in the protective.At present relevant meals and medicine studies show that alcohol metabolism impact and anti-ALD: the antioxidant content in the black tea can suppress the interior enzyme system of body that causes in the alcohol metabolism process and the antioxidant activity reduction of non-enzyme system, help body to keep normal redox state, and research in this respect remain blank out for propolis.
3, by suppressing the NF-kB activation and reduce the COX-2 expression to alleviate lipid peroxidation and inflammatory reaction
Current research shows, oxidative stress makes NF-κ B, COX-2 activation, NF-κ B is a kind of a kind of transcription factor of regulating and control inflammation and immunoreation, apoptosis and infection and stress, and COX-2 is through stimulating the inducible enzyme of rapid generation, is the key link of inflammatory reaction.COX-2 and synthetic product thereof can pass through to disturb the metabolism of fat, saccharide and protein, increase the weight of the release of oxidative stress and the promotion cytokine profiles of liver, thus the pathological changes such as the steatosis of participation liver, inflammation, fibrosis.This two factor is explored, become the new starting point that the research alcohol metabolism causes hepatic injury.Tea polyphenols, trifoliate orange yellow party all can alleviate lipid peroxidation and inflammatory reaction by the activation that suppresses NF-κ B in the rat body and the mechanism of action of reducing the COX-2 expression, realize the target of protection hepatic tissue.
4, express by reducing level of endotoxin and CD-14
The level of endotoxin rising also is the key factor that causes ALD in the blood plasma, but in the modern relieving alcoholism and protecting the liver goods, how research affects the very few of level of endotoxin about goods, and bicyclol has been used for the treatment of slow virus hepatitis as the novel Antihepatitis medicament of Chinese independent development.Current research shows that bicyclol can reduce Endotoxin Levels and downward modulation CD-14 expresses, and has the effect of protection alcoholic liver injury, the formation of control ALD.Therefore, can express by reducing level of endotoxin and CD-14, realize the effect of the liver protecting tissue from suppressing Kupffer cell activation angle.
In sum, the pathogenesis of ALD is relevant with many factors, mainly comprises: the key enzyme in the alcohol metabolism process, such as ADH, ALDH, CYP2E1, CAT; The oxidative stress factor of influence is such as MDA, SOD, GSH-PX; Signal conducts crucial nuclear factor, such as NF-κ B, COX-2; Level of endotoxin and receptor CD-14 expression etc. thereof.
Therefore, the research about the relieving alcoholism and protecting the liver mechanism of action also should launch around these key factors.Simultaneously, China's hepatinica product that relieve the effect of alcohol, be main mainly with Chinese herbal medicine in the health product, the exploitation of relevant dietary ingredient seldom, therefore according to the theory of Chinese medicine " integration of edible and medicinal herbs ", the relieving alcoholism and protecting the liver composition that searches out the high effect nontoxic evil from meals has innovative significance, also nutritional intervention and the drug development for the collaborative anti-ALD of many target spots provides theoretical foundation, and through we study find propolis effectively alleviation of alcohol to the damaging action of liver, successful relieves the effect of alcohol, its relieve the effect of alcohol molecular mechanism and its stronger adjusting be multiple, and to have a gene of antialcoholism action relevant, is expected to be developed as a kind of health food that alleviates the ethanol chemical liver injury.
Liver is the detoxifcation organ that body weight for humans is wanted, and the alcohol metabolism process is mainly carried out in liver.Take the photograph in a large number for a long time people's ethanol, can surpass liver normal physiological metabolic detoxification ability, cause the many important physiology of body, biochemistry and metabolism disorder, the major injury hepatocyte causes alcoholic liver disease of ZANG-organs (ALD).Alcoholic fatty liver (AFLD) is one of 3 kinds of histopathology forms of ALD, and pathogenesis is very complicated, and main manifestations is steatosis.Liver fat degeneration and fatty infiltration and disorders of lipid metabolism, closely related such as the synthetic increase of decline, the triglyceride of fatty acid oxidation rate, fatty minimizing of exporting and the mobilization of liver external fat etc.Ethanol is the following aspects to the damage main manifestations of liver particularly to body: 1, enzymatic activity indicates the changing function in the metabolic processes of body basis: alcohol metabolism has 3 majors avenues of approach in the hepatocyte, and one of them is ethanol dehydrogenase (ADH) path.In the Ethanol Oxidation, oxygen consumption increases in the liver, be reduced with a large amount of nadide (NAD) in the cytoplasm, form reducibility coenzyme I(NADH), the ratio that causes NAD+/NADH descends and unbalance, changed the redox state of cell, severe jamming the metabolism of cell, thereby tricarboxylic acid cycle is suppressed; In addition, because the mitochondria enzyme activities such as succinate dehydrogenase (SDH) weaken the oxidative phosphorylation ability is reduced, affect the breathing of cell, increase the weight of hepatocellular damage; Above-mentioned variation is to cause the topmost reason of AFLD.
Someone likens into two locusts on rope of hyperlipemia to fatty liver and coronary heart disease because obesity, hyperlipemia, diabetes and excessive drinking be fatty liver commonly encountered diseases because of, and these factors are close with atherosclerosis and coronary heart disease too.Obvious myocardial damage and hemodynamics disturbance are arranged, the T cellular immune function decreased, the liver biopsy studies show that 91.4% has hepatic cell fattydegeneration in these cases.Generally speaking, the ethanol fatty liver belongs to the reversibility disease, and early diagnosis and in time treatment often can recover normal.Long-term edible high lipid food or heavy drinking, ethanol can directly be poisoned hepatocyte, affects its structure and function; Many people are exactly because insobriety is just suffered from alcoholic hepatitis, fatty liver causes liver damage.And hepatic injury can cause triglyceride (TG) to descend, and triglyceride is the fat molecule that long-chain fatty acid and glycerol form.Be the maximum lipid of people's in-vivo content, most tissues all can utilize the triglyceride catabolite to supply with energy, is the main source of energy i (in vivo).The significant quantities of fat that fatty liver can cause eating absorbs minimizing, and bile secretion is not enough, can not decompose the fat of eating into, increases the weight of the state of an illness thereby fat forms fat drop at liver.
Propolis is that Apis gathers the plumelet of glue source plant or the resinoid of callus secretion, and mixes a kind of colloid substance with stickiness that secretions, pollen and the Cera Flava etc. of self body of gland mix.Propolis is rich in various bioactivators, especially contains a large amount of Flavonoid substances, has multiple medicinal health care function, is described as " purple gold ".Propolis inevitably can be sneaked into the impurity such as wood flour, bee body, weeds, sandstone in recovery process, before being used for food, medicine etc., must be through purifying, otherwise can not take.
The effective ingredient of the different gained of method for extracting propolis is not identical yet, and conventional extraction has at present: ethanol extraction method, water extraction method, super critical extraction, ultrasound wave assisted Extraction are followed the example of, enzyme extraction method, boron polyelectrolyte method, microwave-assisted extraction method.Find that by research Different Extraction Method, the different solvent that extracts can show different pharmacological effects, show to extract with pharmacological effect to have obvious dependency.By comparing SCF-CO 2 method, not only apparatus expensive, the difficult grasp of operating condition, and extract yield is low and bacteriostasis is low than the alcohol steep method, but the method extract safety, noresidue, pollution-free have certain DEVELOPMENT PROSPECT, and be main still for laboratory research at present.The organic solvent that organic solvent extraction is commonly used has ethanol, ether, acetone, ethyl acetate, n-butyl alcohol etc., and wherein ethanol is the most commonly used.Alcohol steep method equipment needed thereby is simple; reagent is buied easily; operate also very convenient; with ethanol as extracting solvent; can be easy to make the propolis dewaxing, and can protect some polyphenols, thereby obtain to be rich in the refined propolis of polyphenols; so ethanol is the most frequently used extraction solvent, particularly 70% and 80% ethanol.Although sometimes also extract with 95% ethanol, compare with water-alcohol solution, water-alcohol solution extract can make Cera Flava quickly free out, also be rich in a large amount of polyphenols in the extracting solution simultaneously.Ethanol with variable concentrations extracts propolis; and measure the absorption spectrum of Flavonoid substances in the variable concentrations extracting solution; the result is presented at the 290nm place; in the propolis extract of 80% ethanol extraction the highest absworption peak is arranged; this also just means and wherein contains maximum Flavonoid substances, mainly is the luxuriant and rich with fragrance alcohol of camphane, acacetin and isosakuranetin.But maximum Flavonoid substances that the ethanol extraction with 60% goes out is isosakuranetin, Quercetin and kaempferol, and 70% ethanol can extract maximum pinocembrins and sakuranetin; Can extract some polyphenols soluble in water with pure water, methanol, normal hexane, chloroform sometimes also are used as extractant and use.The research of organic solvent lixiviate propolis total flavones is mainly processed the aspect expansion such as ancillary method (such as microwave treatment, ultrasonic Treatment) around its influence factor (such as concentration of alcohol, extraction time, solid-liquid ratio, extraction temperature) and other both at home and abroad.The ultrasound wave assisted extraction can shorten extraction time greatly, has become in recent years the focus of propolis total flavones Study on extraction.Mostly studied around aspects such as ultrasonic treatment time, ultrasonic powers, it is less to change the research report that extraction affects on propolis total flavones about ultrasonic frequency, and research is not deep enough yet in the past.
Lac regis apis is 5～15 age in days worker bee lingual glands and upper palatine gland secreted milky or faint yellow emulsion liquid, is whole period of development of queen bee nit and drone larva unique food in early stage, is similar to mammiferous milk, claims again Lac regis apis, royal jelly, Lac regis apis.Attention and attention to Lac regis apis, at first be that the young honeybee of having found feed Mel can only grow up to worker bee, general 30～40 days of life-span, the longest 7～8 months, and the young honeybee of feed Lac regis apis can grow up to queen bee, life-span reaches 3～5 years, and these phenomenon prompting people utilize Lac regis apis as the food of life lengthening, but its nature and role mechanism is not understood.By 1888, Germanization scholar A Von Planta just first component to Lac regis apis carries out generality research, so far people have begun to inquire into the profoundness of Lac regis apis, but Lac regis apis is comprised comprehensive research of biology, chemical pharmacology and clinical experiment, or nearest decades are inchoate, and people also do not study clear to some the micro-bioactive substances in the Lac regis apis fully so far.At present, fixed functional factor totally 9 large classes: they are peptides and proteins classes; Functional sweetener; Functional grease; Free radical scavenger; Vitamin; Active trace element; Active polysaccharide; Flavone compound and little ecological factor such as lactobacillus etc.Research according to people is found, contains these abundant functional factors in the Lac regis apis, and the mankind are had extremely strong alimentary health-care function and medical function.
Modern study shows: Lac regis apis can alleviate hepatocellular damaging action in the ALD course of disease, liver organization after the damage there is the effect that promotes regeneration, its mechanism of action may be by reducing or suppress the generation of COX-2, avoid excessive inflammatory reaction, reach and alleviate mitochondrion and lipid peroxidation injury, thereby reduce the level of serum AST and ALT; Lac regis apis has reduced the expression of TNF-α in hepatocyte, and TNF-α has mediated hepatocellular inflammatory reaction as inflammatory factor, multiple pathology and the physiological activities such as immunoreation, hepatocellular apoptosis and amplification hepatic fibrosis.The reason that analysing royal jelly TNF-α reduces may be that Lac regis apis is natural oxygen free radical scavenger, can reduce the activation of the nuclear factor that ROS causes, thereby reduces the expression of the TNF-α gene that is subjected to its regulation and control.Lac regis apis can press down the expression of COX-2 and TNF-α in the forming process of alcoholic liver disease, alcoholic liver injury is had certain protective effect.Lac regis apis also all has significant improvement effect to hepatic fibrosis serological index and the hepatic pathology morphology of CC14 hepatic fibrosis rats, but improve not obviously to forming liver tissue fibrosis after the hepatic fibrosis, its possible mechanisms is the Lac regis apis inflammation that can improve hepatic tissue, reduce the synthetic of ECM and suppress the secretion of TNF-α.And Lac regis apis and propolis have natural orthofunction and concordance, and the two is combined with effect can be better.
Buccal tablet, be a class without peptic digestion, directly absorb by oral mucosa and enter blood, adopt that soluble material carries out the tablet that coating is made for main coating material in the oral cavity.Can prevent acidity and enzyme to the destruction of some drugs or prevent that medicine is to the intense stimulus of stomach; We find to have in the Lac regis apis many bioactive substances under one's belt easily by stomach acids destroy through research, and can directly absorb by hypoglossis mucous membrane, can protect better bioactive substance to exempt from destruction so make buccal tablet, improve the result of use of product.People have also absorbed a large amount of food in heavy drinking, particularly high innage fat food makes people's stomach be high full abdomen state, the emptying general needs of its stomach are about 2 hours, if take simultaneously the medicine that relieves the effect of alcohol, because comparing with stomach, dose differs too great disparity, be difficult to play a role and lost efficacy, if but adopt heavy dose of antialcoholic drugs, can be subject to the restriction of gastric capacity again and can't implement, so the antialcoholic drugs that uses after meal must adopt the insoluble processing mode of stomach, Here it is, and why we will develop the main cause of buccal tablet.
Summary of the invention
The present invention's the first purpose is to invent a kind of propolis extracted with alcohol that can efficiently fundamentally relieve the effect of alcohol.
The present invention be used for the propolis extracted with alcohol relieve the effect of alcohol for the molecular weight that adopts ethanol and extract from propolis less than the solid content of 5000D greater than the dry thing of 90% biological activity.
The inventor is through evidence, and the molecular weight that extracts from propolis has the effect of relieving the effect of alcohol preferably less than the bioactive substance of the dissolve with ethanol of 5000D.
Second purpose of the present invention is the preparation method that proposes above-mentioned propolis extracted with alcohol.
The inventive method may further comprise the steps:
1) propolis is placed under-18 ℃ the ambient temperature conditions and pulverize after freezing 20～40 hours;
2) under the normal temperature and pressure be 50%～90% edible ethanol with volumetric concentration with pulverize after propolis mix by the weight ratio of 1～10 ︰ 1, prior to low-frequency ultrasonic waves, under 10～40 ℃ of temperature, processed 10～40 minutes, be airtight immersion 1～4 day under 10～40 ℃ the condition in ambient temperature then;
3) get the rear mixture of immersion and turned frozen centrifugation 10～15 minutes with 8000～16000, remove solid impurity, get the supernatant of dissolve with ethanol propolis;
4) with supernatant under 1～5 ℃ of ambient temperature conditions with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight less than the propolis extracted with alcohol of 5000D;
5) above-mentioned molecular weight is placed the ethanol distillation recover less than the propolis extracted with alcohol of 5000D, boils off edible ethanol, obtain molecular weight less than the solid content of 5000D greater than the dry thing of 90% biological activity.
At present research is found to affect the active factor of propolis extraction and is mainly contained temperature, pH value, extraction time, solvent species, solvent strength, soak time, extraction time etc.The propolis that different extraction processes and solvent extract, its effective ingredient can be different, and extracting method is different, and the composition of extraction, content, solvent all may affect its function performance, cause kind of a species diversity, and then different pharmacological actions occurs.
After the present invention is freezing with propolis propolis is become fragile, accomplish when guaranteeing to pulverize propolis that fineness is even, be convenient to the extraction of effective ingredient; Low-frequency ultrasonic waves before soaking is processed except can realizing maximized separation, ethanol temperature in the time of can also keeping extracting is no more than 40 ℃, guarantee to extract in the alcoholic solution isolated material and keep active, and guarantee that the effumability composition keeps to greatest extent in the propolis; The present invention adopts airtight immersed with being beneficial to the effectively separation of bioactive substance in the propolis, dissolving and maintenance activity, reduces the oxidation deactivation of antioxidant in the propolis; It is can access maximized dissolving in order to ensure ethanol soluble substance in the propolis that suitable propolis and the ratio of ethanol are provided; The rear frozen centrifugation of immersion is that the temperature when guaranteeing centrifugalize is lower, guarantees that Cera Flava is separated out better in the propolis ethanol extract, makes the propolis that extracts purer, obtains abundant active substance when being beneficial to next step ultrafiltration; Get supernatant 5000D molecular sieve ultra-filtration and separation, obtaining molecular weight is the following propolis ethanol soluble substance of 5000D, at first find that through research the following propolis ethanol soluble substance of molecular weight 5000D all has the effect of relieving the effect of alcohol, next guarantees that extracting solution is pure clear, transparent, make extract pure, it is one of most critical measure of guaranteeing ethanol soluble substance purity under 1～5 ℃ of condition that temperature is extracted in control; Obtain the propolis solid content greater than 90% dry thing with ethanol distillation recover Recycled ethanol, remove ethanol and be the application for the ease of next step.
To sum up, the present invention adopts the ethanol extraction method of above improvement, but extraction efficiency is higher, and speed is faster, particularly can significantly keep the effective composition that relieves the effect of alcohol, and the product expression of producing goes out the very strong effect of relieving the effect of alcohol.
The 3rd purpose of the present invention is to propose for the propolis extracted with alcohol that relieves the effect of alcohol in the application of producing buccal tablet.
Make the buccal tablet that relieves the effect of alcohol for the preparation of propolis with described propolis extracted with alcohol, Lac regis apis lyophilized powder, hydroxyl isomaltulose, Sorbitol, magnesium stearate and Mentholum be used to relieving the effect of alcohol; Described propolis extracted with alcohol, Lac regis apis lyophilized powder, hydroxyl isomaltulose, Sorbitol, magnesium stearate and Mentholum alcoholic solution account for respectively 15～30%, 5～10%, 55～70%, 5～15%, 0.2～0.8%, 0.1～0.4% of buccal tablet gross mass.
Through test, it is remarkable to utilize propolis extracted with alcohol to merge the Lac regis apis lyophilized powder effect of relieving the effect of alcohol, and has no side effect, and expense is cheap, simple.Propolis is that Apis gathers the plumelet of glue source plant or the resinoid of callus secretion, and mixes a kind of colloid substance with stickiness that secretions, pollen and the Cera Flava etc. of self body of gland mix.Propolis is rich in various bioactivators, especially contains a large amount of Flavonoid substances, has multiple medicinal health care function, is described as " purple gold ".Lac regis apis is 5-1 5 age in days worker bee lingual glands and secreted milky or the faint yellow emulsion liquid of upper palatine gland, is whole period of development of queen bee nit and the drone larva food in early stage, is similar to mammiferous milk, claims again Lac regis apis, royal jelly, Lac regis apis.Research is found, contains abundant functional factor in the Lac regis apis, and the mankind are had extremely strong alimentary health-care function and medical function.Propolis extracted with alcohol merges Lac regis apis can bring into play the pharmacological action of relieving the effect of alcohol at gene level, but to being that what material works present research seldom in propolis and the Lac regis apis, its molecule mechanism of relieving the effect of alcohol is imperfectly understood.
But the present inventor's research has solved propolis substantially merges the molecule mechanism that Lac regis apis relieves the effect of alcohol.The present invention utilizes the method for ethanol extraction is isolated therapeutic effect to relieving the effect of alcohol composition from propolis, merge the Lac regis apis production buccal tablet that relieves the effect of alcohol.
As the advantage of buccal tablet be easy to use, absorb soon, mucosa directly absorbs and enters liver and relieve the effect of alcohol effective, cheap; The rear gastric of drinking is full abdomen state, can have a negative impact to the effect of relieving the effect of alcohol, and can overcome well this unfavorable condition and be made into buccal tablet, and the general solution wine product that buccal tablet can be used as ordinary people uses, and is good and cheap.
Find many bioactive substances to be arranged under one's belt easily by stomach acids destroy in propolis extracted with alcohol and the Lac regis apis through research, just can make well bioactive substance in the buccal tablet exempt from the destruction of gastric acid, the result of use of raising product so make buccal tablet.
Description of drawings
Fig. 1 is natural health matched group liver tissue slices figure.
Fig. 2 is natural health matched group renal tissue slice map.
Fig. 3 is excessive drinking model group liver tissue slices figure.
Fig. 4 is excessive drinking model group renal tissue slice map.
Fig. 5 is gavage buccal tablet group of the present invention liver tissue slices figure.
Fig. 6 is gavage buccal tablet group of the present invention renal tissue slice map.
The specific embodiment
One, the preparation molecular weight less than the solid content of 5000D greater than the dry thing of 90% biological activity:
1, propolis is placed under-18 ℃ the ambient temperature conditions and pulverize after freezing 20～40 hours.
2, under the normal temperature and pressure be 50%～90% edible ethanol with volumetric concentration with pulverize after propolis mix by the weight ratio of 1～10 ︰ 1, prior to low-frequency ultrasonic waves, under 10～40 ℃ of temperature, processed 10～40 minutes, be airtight immersion 1～4 day under 10～40 ℃ the condition in ambient temperature then.
3, get the rear mixture of immersion and turned frozen centrifugation 10～15 minutes with 8000～16000, remove solid impurity, get the supernatant of dissolve with ethanol propolis.
4, with supernatant under 1～5 ℃ of ambient temperature conditions with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight less than the propolis extracted with alcohol of 5000D.
5, above-mentioned molecular weight is placed the ethanol distillation recover less than the propolis extracted with alcohol of 5000D, boils off edible ethanol, obtain molecular weight less than the solid content of 5000D greater than the dry thing of 90% biological activity.
Two, the preparation propolis merges the Lac regis apis buccal tablet that relieves the effect of alcohol:
1, typical several quality proportioning table: (unit: kg)
| ? | Propolis powder | Lac regis apis lyophilized powder | Hydroxyl isomaltulose | Sorbitol | Magnesium stearate | Mentholum solution |
| Proportioning 1 | 30 | 6 | 55.5 | 8 | 0.3 | 0.2 |
| Proportioning 2 | 25 | 8 | 58.6 | 8 | 0.3 | 0.1 |
| Proportioning 3 | 15 | 10 | 66.6 | 8 | 0.3 | 0.1 |
2, granulating process:
It is that 10～30% aqueous sorbitol solution is for subsequent use that Sorbitol is made into mass percent.
After the boiling pot is preheating to 40～60 ℃, first propolis extracted with alcohol, Lac regis apis lyophilized powder, hydroxyl isomaltulose are added in the boiling pot, arrange according to the following table parameter, open peristaltic pump spray aqueous sorbitol solution and carry out one-step palletizing.Granulate after the end, sub-cooled adds magnesium stearate and Mentholum solution mix homogeneously.
| Parameter | Inlet temperature | Leaving air temp | The air inducing frequency | Peristaltic pump | Atomizing pressure |
| During whitewashing | 50-80℃ | 40-60℃ | 40Hz | 200rpm | 0.2MPa |
| After the whitewashing | 60-85℃ | 45-60℃ | 35Hz | 200rpm | 0.2MPa |
3, tablet forming technique: adopt ZP1100 tablet machine (29 punching) to carry out tabletting experiment, technological parameter: 30～40 rev/mins of rotating speeds; Operating pressure 7～15kN; Sheet weighs 0.4～0.6g.
Four, use:
(1) subjects:
The Wistar cleaning level male rat in ages in gavage 22 week, body weight 344.43 ± 38.49.
Nature control rats every day is by body weight 1% gavage pure water.
Model group rat every day is by body weight 1% gavage strong, colourless liquor distilled from sorghum simulation.
Gavage buccal tablet experimental group rat every day is by 8 hours gavage buccal tablets of the present invention after the body weight 1% gavage strong, colourless liquor distilled from sorghum.
(2) using dosage of buccal tablet:
50～150mg/ days, connect gavage 30～60 days.
(3) comparing result:
1, rats in test groups liver and renal tissue section result is relatively:
(1) with nature matched group liver and renal tissue section, sees Fig. 1,2.
The hepatocyte clear in structure is polygon from Fig. 1,2 visible male white rat liver tissue slices, and hepatic sinusoid is obvious, and the hepatic cords marshalling does not have obvious fat vacuole in the hepatocyte; Renal tissue section mesonephric glomerulus structure is clear, and the glomerular capsule gap is obviously moderate, not obviously damage.
(2) with gavage Chinese liquor model group liver and renal tissue section, see Fig. 3,4.
From Fig. 3,4 visible hepatocyte obscure boundaries, becoming in the circle has cavity, and the hepatic sinusoid pressurized narrows down, liver rope arrangement disorder; The glomerule structure is unclear, and damage is serious, the glomerular capsule distortion.
(3) Fig. 5,6 is seen in the animal livers of gavage buccal tablet of the present invention and renal tissue section.
From Fig. 5,6 visible liver lobules of liver clear in structure, hepatic sinusoid is evenly distributed, and hepatocyte is polygon, marshalling; The renal glomerulus structure is obviously improved, and the glomerular capsule gap is obvious, has the vestige of obvious glomerule reparation.
2, after the buccal tablet intervention to each rats in test groups Main Blood Biochemical Index comparative analysis:
Adopt the blood sampling of anesthesia ventral aorta, detect Main Blood Biochemical Index by Toshiba's fully automatic blood bio-chemical detector after getting serum.
Comparative analysis such as following table:
| Group | Naturally organize | Model group | Wine glue group |
| ALT | 73.00±16.67 | 91.25±29.33 | 41.20±6.12 |
| AST | 109.00±16.51 | 124.00±23.47 | 107.00±19.30 |
| ALP | 102.50±11.90 | 146.75±21.70 | 68.00±19.46 |
| CHO | 1.23±0.08 | 1.52±0.26 | 1.21±0.16 |
| TG | 1.25±0.18 | 1.84±0.37 | 1.08±0.17 |
| BUN | 5.89±0.65 | 4.78±0.71 | 5.66±0.41 |
| ALB | 15.50±0.43 | 14.88±0.53 | 16.04±1.10 |
Through the SPSS statistics, gavage buccal tablet group of the present invention glutamate pyruvate transaminase (ALT) is extremely remarkable with model group difference, with natural matched group significant difference; Model group glutamic oxaloacetic transaminase, GOT (AST) and gavage buccal tablet group of the present invention significant difference; The transaminase extensively exists in the histiocyte, especially the strongest with activity in the tissues such as liver, cardiac muscle, brain, kidney, plasma content is very low when normal, and ALT mainly is present in liver, AST mainly is present in heart, and ALT, AST are released in a large number blood and can cause in the blood that the transaminase raises when the hepatic tissue disease damage.This experiment gavage buccal tablet group serum transaminase vigor is very low, illustrates that buccal tablet is very strong to the repair effect of liver.
Healthy Human Serum ALP is mainly from liver and skeleton, and the ALP in the liver is with the transhipment of material and drain relevant.And the type 2 diabetes mellitus patient since insulin secretion obstacle or insulin resistant cause, cause metabolism obstacles of blood glucose, also cause the obstacle of lipid metabolism simultaneously and at intrahepatic deposition, even form fatty liver, and make liver damage cause the rising of ALP.Through the SPSS statistics, model group alkali phosphatase and other two experimental grouies difference are extremely remarkable, natural matched group alkali phosphatase and gavage buccal tablet group significant difference; Illustrate that buccal tablet can improve hepatocyte injury, reduce the alkaline phosphatase enzyme level, and long-term excessive drinking can increase the weight of hepatocellular damage.
Type 2 diabetes mellitus is the class disease take insulin resistant and islet beta cell function obstacle as principal character, and wherein losing with dysfunction of beta Cell of islet plays a decisive role in the generation of disease and development.Studies show that in recent years, cholesterol metabolism and β cell function are closely related.Under normal circumstances, absorption and outflow that beta Cell of islet can be controlled the cell inner cholesterol by series of receptors and the transport molecule of its surface expression are so that the content of cell inner cholesterol maintains a dynamic equilibrium.After the metabolism of beta Cell of islet inner cholesterol gets muddled, can affect by number of ways the carbohydrate metabolism of β cell, finally induce the β apoptosis.Therefore, the cholesterol metabolism disorder may be a new mechanism that causes type 2 diabetes mellitus patient β cell dysfunction.Add up through SPSS, model group cholesterol and natural matched group and gavage buccal tablet group significant difference, explanation is aspect the regulation and control cholesterol metabolism, gavage buccal tablet of the present invention can significantly be reduced because the cholesterol levels that excessive drinking causes raises, and is significant to the generation that prevents to bring out type 2 diabetes mellitus owing to long-term excessive drinking.
Fat toxicity due to the high triglyceride (HTG) has caused extensive concern in recent years, and it can cause and increase the weight of insulin resistant (IR), and the biological effect of insulin is reduced.Glycogen is synthetic by participating in for liver, glycogenolysis and glyconeogenesis play a part direct and important in keeping glucostasis, particularly G-6-Pase plays a crucial role in glycogen generates, it is unusual that hepatic insulin resistance also shows as lipid metabolism, and discharging such as triglyceride increases and the liver steatosis.Through the SPSS statistics, model group triglyceride and natural matched group and gavage buccal tablet group difference are extremely remarkable; From the experiment situation, serum high triglyceride level appears in model group, and long-term excessive drinking meeting production hypertriglyceridemia is described, is one of paathogenic factor of bringing out type 2 diabetes mellitus; And gavage buccal tablet of the present invention has good regulating and controlling effect to the Triglyceride Metabolism that brings out owing to excessive drinking.
Urea concentration also is subject to the impact of protein and organism metabolism state in the diet except being subjected to Influence on kidney, bad etc. such as high protein diet, gastrointestinal function.Because kidney has powerful reserve capabillity and compensatory capacity, impaired early stage or slight when impaired at glomerule, blood urea nitrogen still can maintain normal level, only damages at the severe renal bead, general glomerular filtration rate reduces by 50% when following, and blood urea nitrogen concentration is obviously rising.Add up through SPSS, model group blood urea nitrogen and natural matched group and gavage buccal tablet group significant difference, under this experiment condition, the model group rat is because excessive excessive drinking, drunk serious, causing feed intake to descend and protein metabolism disturbance, so that blood urea nitrogen does not rise counter falling, is minimum in all experimental grouies, and gavage buccal tablet experimental group, metabolism to protein is improved effect, and urea nitrogen levels gains momentum, and approaching with the urea nitrogen levels of natural matched group.
Through the SPSS statistics, model group albumin and gavage buccal tablet group difference are extremely remarkable, natural matched group albumin and gavage buccal tablet group significant difference; More than after these change explanation long-term excessive drinking are caused protein metabolism disturbance, and gavage buccal tablet of the present invention is to because the protein metabolism disturbance that long-term excessive drinking causes has very strong regulating and controlling effect.
By the analysis to blood parameters, we are not difficult to draw such conclusion in the excessive drinking model, propolis and Lac regis apis mixture not only can help body to relieve the effect of alcohol, and can recover rapidly body to the metabolism of three large materials, improve multinomial blood parameters, make organism metabolism return to normal condition.
3, after the gavage buccal tablet intervention of the present invention major gene in the rats in test groups liver metabolism is expressed the variation comparative analysis:
Find by biochip technology research the important metabolic pathway of many impacts in the model group rat liver expressing gene of long-term excessive drinking (such as with the related gene that relieves the effect of alcohol, Genes Associated with Lipid Metabolism, hepar damnification reparation gene) expression of key gene all shows the situation that is unfavorable for the body homergy, and the variation of this gene expression has obtained good correction after the intervention of gavage buccal tablet, we are not difficult to find that the ability that this being similar to " error correction " expressed is just embodying the liver metabolism obstacle of gavage buccal tablet to forming owing to long-term excessive drinking from lower example table, the reparation of tissue injury has very strong improvement effect, research by us finds that from gene level the gavage buccal tablet can relieve the effect of alcohol effectively, improves the metabolism of liver.
3.1 the impact of after the intervention of gavage buccal tablet key gene relevant with glycolipid metabolism in the liver being expressed
| Group | Dci | Fasn | Scd | Scd1 |
| Model group/naturally organize | ↓3.72 | ↑4.86 | ↑5.04 | ↑5.65 |
| Buccal tablet group/model group | ↑3.48 | ↓2.86 | ↓18.21 | ↓24.32 |
Annotate: in the table, 3.72 times of the expression ratio downward modulations of ↓ 3.72 expression Dci genes model group and natural matched group in hepatocyte mRNA, 3.48 times of the expression ratio rises of ↑ 3.48 expression Dci gene gavage buccal tablet groups and model group.
3.1.1 Dci: ten dichloro alkane coenzyme A δ isomerases, anti-enoyl CoA isomerase, mitochondrion participates in lipid metabolism, the fatty acid beta-oxidation.
Excessive drinking model group and natural matched group are than 3.72 times of the expression of downward modulation ten dichloro alkane coenzyme A δ isomerase genes under this experiment condition, illustrate the model group rat indulge in excessive drinking for a long time can by the downward modulation this gene expression, mitochondrion fatty acid beta-oxidation ability is weakened, promoting liver fat to become, is that long-term excessive drinking forms one of main molecules mechanism of fatty liver.
Gavage buccal tablet group has raised 3.48 times of this genes with the model group ratio under this experiment condition, illustrate: gavage buccal tablet of the present invention can be by promoting the fatty acid beta-oxidation of liver organization, the fatty liver that the control excessive drinking is caused has positive effect, is one of lipotropic main molecules mechanism of gavage buccal tablet.
3.1.2 Fasn: fatty acid synthetase, it is synthetic to participate in fatty acid biological.In the mammal, fatty acid synthetase plays vital effect in fatty acid is synthetic, be responsible for all catalytic steps of S-acetyl-coenzyme-A and a kind of long-chain palmitic acid of malonyl coenzyme A de novo synthesis (cetylate), the expression of gene directly affects the fatty acid synthetase number control body fat deposition is had great importance.
This experiment condition drag group leader's phase indulges in excessive drinking and makes in the hepatocyte fatty acid synthetase gene and natural matched group than 4.86 times of up-regulateds, illustrate: it is the arch-criminal who causes hepatocyte fat excess deposition that model group can make synthetic the increasing of fatty acid synthetase, is to form one of fatty liver molecule mechanism.
2.86 times of the gene expressions of gavage buccal tablet group of the present invention and model group downward modulation fatty acid synthetase under this experiment condition, illustrate: gavage buccal tablet group of the present invention can be by reducing the synthetic of fatty acid synthetase, it is synthetic to reduce the hepatic tissue fatty acid, becomes significant to alleviating liver fat.
3.1.3 Scd: sterin coenzyme A desaturase, it is synthetic to participate in fatty acid biological.Stearyl-coenzyme A desaturase (SCD) can be introduced in fatty acid carbon chain cis-9 position two keys, is catalysis butterfat cis-9, trans-11CLA and trans-7, the key enzyme that cis-9 CLA is synthetic.
3.1.4 Scd1: sterin coenzyme A desaturase 1, it is synthetic to participate in fatty acid biological.Stearyl-coenzyme A desaturase 1(Scd-1) being the biosynthetic rate-limiting enzyme of monounsaturated fatty acid, playing the center adjustment effect in fatty acid metabolism, also is one of genes of interest of leptin (Leptin) effect.Leptin and diabetes, obesity, fatty liver close relation are one of hot fields of in recent years metabolic disease.The type 2 diabetes mellitus patient, usually with insulin resistant, hyperleptinaemia, there is fatty liver in about 50% type 2 diabetes mellitus patient simultaneously.Scd-the 1st, the key enzyme that the catalysis satisfied fatty acid changes to monounsaturated fatty acid, closely related with generation, the development of a series of metabolism syndromes such as obesity, fatty hepatic lesions and insulin resistant, thereby the expression of research Scd-1, regulation and control may provide a very important lab index for the diagnosis of clinical disorders of lipid metabolism relevant disease and the monitoring of therapeutic effect, has broad application prospects in clinical medicine.Because Scd-1 mainly is present in endoplasmic reticulum, at present the detection of Scd in the body-1 is mainly adopted and measured blood fat index of unsaturation (unsaturated fatty acid/satisfied fatty acid ratio) or by molecular biology method Scd in the cell-1mRNA expression is detected.In sum, Scd-1 has play a part important in energy metabolism.By the research to Scd-1 and energy metabolism relation, can the mechanism of clearer understanding Scd-1 in energy metabolism, thereby be clinical targeting adjusting energy metabolism and avoid fat toxicity, a series of metabolic syndromes such as prevention or treatment are fat, fatty hepatic lesions and diabetes provide basic.
The excessive drinking model group raises 5.04 times of the auxiliary A desaturase of sterin (Scd) gene expressions with natural matched group ratio under this experiment condition, sterin coenzyme A desaturase 1(SCD-1) raises 5.65 times, illustrate: the two gene up-regulated is one of main molecule mechanism that causes the alcohol fatty change, lipotropic is the pathological index of most critical in the process of relieving the effect of alcohol, the processing mode effectiveness just is whether can control fatty liver, model group has raised two gene, and serious fatty liver has appearred in model group liver section display model group rat, even reaches the state of hepatic fibrosis.
Gavage buccal tablet group and 18.21 times of model group ratio downward modulation sterin coenzyme A desaturase (Scd) gene expressions under this experiment condition, downward modulation sterin coenzyme A desaturase 1(Scd-1) gene expression is 24.32 times, illustrate: the buccal tablet group has significant improvement effect to the formed fatty liver of bad habit of excessive drinking, and consistent with the result of liver section, this change is one of main molecule mechanism of buccal tablet control fatty liver of the present invention.
3.2 behind the gavage buccal tablet in the liver with hepatic injury, repair the impact of relevant key gene expression
| Group | Igfbp2 | Il7r | Irs2 | Socs2 | Srd5a1 |
| The model group/naturally of relieving the effect of alcohol group | ↓3.12 | ↓3.21 | ↓21.42 | ↓33.58 | ↑4.0 |
| Buccal tablet group/wine model group | ↑3.62 | ↑4.55 | ↑5.21 | ↑6.92 | ↓5.86 |
Annotate: in the table, 3.12 times of the expression ratio downward modulations of ↓ 3.12 expression Igfbp2 genes model group and natural matched group in hepatocyte mRNA, 3.62 times of the expression ratio rises of ↑ 3.62 expression Igfbp2 gene gavage buccal tablet groups and model group.
3.2.1 Igfbp2: IGFBP2 participates in the regulating cell growth.Rise is conducive to Igf1 and plays a role, and enhances metabolism, and is conducive to glycolipid metabolism, and is favourable to the prevention fatty liver.
Research confirms that IGFs is relevant with the formation development of hepatic fibrosis, liver cirrhosis.Having isolated at present two kinds of IGFs is IGF-l and IGF-2.They act on target organ with autocrine, paracrine and endocrine mode.Nearly all endocrine IGF-l or IGF-2 all form relative molecular mass with insulin-like growth factor binding protein (IGFBPs) in body fluid be the complex of 150kb or 50kb.The single chain polypeptide that IGF-l is comprised of 70 amino acid residues, mainly synthetic at liver, be the extensive cell mitogen that exists of body and the promoter of differentiation and maturation, affect the adjusting of growth, differentiation and the metabolism of many organ inner cells.The above-mentioned functions of IGF-l is by IGF-l receptor (IGF-lR) mediation, and the effectiveness that IGF-l is combined with IGF-lR is subjected to the adjusting of IGFBPs.Liu Lixins etc. studies show that, IGFBP2 (IGFBP2) is expressed obviously in IGF-β l induces the HSC-T6 of activation and strengthened, and prompting IGF and receptor thereof participate in the formation of hepatic fibrosis.Because IGFBPs and IGFs have high-affinity and can regulate the biological activity of IGFs, these protein-bonded genesis of inducing generation also can affect hepatic fibrosis.Yet there are some researches show in addition, IGF-1 has the effect of anti-hepatic fibrosis in the body, can suppress the activation of HSC, its may with improve superoxide dismutase and catalatic activity, improve the ability that hepatocyte is removed free radical, the stimulating factor of minimizing HS activation etc. are relevant.
This experiment condition drag group has been reduced 3.12 times of IGFBP2 genes with natural matched group than model group rat, illustrate: model group is by the expression of downward modulation IGFBP2 gene, cell cultured supernatant transforms to the fibrosis direction, has promoted hepatocyte fat to become and fibrosis;
The buccal tablet group raises 3.62 times of IGFBP2 genes with the model group ratio under this experiment condition, illustrate: can be by the expression of regulation and control IGFBP2 gene after buccal tablet is intervened, slow down or prevent and treat the Fibrotic process of hepatocyte, the control alcoholic fatty liver is had positive effect.
3.2.2 Il7r: interleukin-17, receptor participates in the regulating DNA restructuring, immunne response, the cell surface receptor signal connects conduction, antimicrobial humoral response.
This experiment condition drag group has been reduced 3.21 times of interleukin-17 genes with natural matched group than model group, illustrate: the long-term excessive drinking of model group rat causes hepatocyte immunne response ability to descend, the expression of downward modulation interleukin-17 gene is exactly to mean to increase the weight of hepatocellular inflammatory reaction, promotes hepatocyte fat to become and Fibrotic process.
The buccal tablet group raises 4.55 times of interleukin-17 genes with the model group ratio under this experiment condition, illustrate: after the gavage buccal tablet is intervened, strengthen hepatocellular immunne response ability by the expression of raising the interleukin-17 gene, alleviate the inflammation reaction, to preventing and treating the fatty liver and the fibrosis that cause owing to long-term excessive drinking positive effect is arranged.
3.2.3 Irs2: IRS 2, Irs2 expresses at liver and pancreatic beta cell, the participation glucose metabolism, signal transduction, positive regulating cell propagation, the downward modulation of Irs2 protein expression makes the active attenuating of PI3-K, can increase the weight of the liver insulin resistant.Use gene Knockout to confirm that the defective of Irs2 gene can cause the discoveries such as insulin resistant, Kubota, knock out Irs2 after, liver shows obvious insulin resistant, and to the insulin sensitivity of skeletal muscle without effect.
This experiment condition drag group has been reduced 21.42 times of IRS 2 genes with natural matched group than model group, illustrate: the model group rat is because excessive drinking causes IRS 2 genes to reduce 21.42 times, IRS is synthesized to be greatly affected, obviously can have influence on hepatocyte to the sensitivity of insulin response, impel model group to produce serious insulin resistant, bring out a series of metabolism syndromes, the final fatty liver that forms is that one of main molecule mechanism of type-II diabetes and fatty liver is brought out in long-term excessive drinking.
Under this experiment condition with after the buccal tablet intervention with the model group ratio, 5.21 times of the expression of Irs2 gene have been raised, illustrate: gavage buccal tablet of the present invention can improve hepatocyte to the sensitivity of insulin response by the expression of raising the Irs2 gene, and buccal tablet of the present invention can play certain alleviation Insulin Resistance.
3.2.4 Socs2: Suppressor of Cytokine Signaling was named according to 4O amino acid whose Socs frame by discoveries such as Starr R in 2,1997 years.Found that at present this class negative regulator molecule has 6 kinds, most members participate in the negative feedback of JAK/STAT path and regulate.This family has 8 members to contain the SH2 domain, and namely CIS and Socs-1～7, and the direct and phosphotyrosine interaction of this structure is that inhibiting key plays in Socs family.Socs 2 effects are extensive, and to lipidosis, skeletal development, central nervous system's effect, immunne response, Carciuogenesis all plays an important role.Socs 2 can regulate the hormone secretions such as insulin, prolactin antagonist in addition, and the cytokine signaling paths such as GH/1GF, insulin, interleukin are played promotion or inhibitory action.Cytokine profiles is by lipometabolic balance in complicated signal network regulating cell and the body, and at present most researchs concentrate growth hormone (GH) on lipometabolic impact, and Socs 2 is crucial negative feedback inhibition factors of growth hormone signal path.
This experiment condition drag group and natural matched group ratio, the down-regulated expression of Socs2 gene 33.58 times, illustrate: for a long time excessive drinking of model group can cause metabolism " brake " regulator control system in the hepatocyte seriously malfunctioning, will inevitably exert an influence to normal lipid metabolism and insulin action in the hepatocyte, form dysbolismus, finally impelling hepatocyte fat to become, is that one of main molecule mechanism of type-II diabetes and fatty liver is brought out in long-term excessive drinking.
Under this experiment condition with after the buccal tablet intervention of the present invention with intervene before the model group ratio, the up-regulated of Socs2 gene 6.92 times, illustrate: buccal tablet of the present invention can be by improving the expression of Socs2 gene, repair because interior " brake " regulator control system of the hepatocyte that excessive drinking causes is malfunctioning, significant to improving hepatocyte lipid metabolism and insulin action.
3.2.5 Srd5a1: steroid 5 alpha-reductases, α peptide 1, distribution endoplasmic reticulum, microsome, 3-C-5 α steroid 4 dehydrogenases participate in cell signalling, Sex determination, cell differentiation.Can be converted into another kind of androgen-dihydrotestosterone by the catalysis testosterone, so play a significant role in property differentiation and androgen physiology, this enzyme has Srd5a 1,2 two isomers.Mainly be expressed in skin and fatty tissue and liver, but the reduction reaction of unsaturation steroid on the 4th, 5 carbon atom of the catalysis overwhelming majority makes activated glucocorticoid (GC) be converted into the metabolite of non-activity.Research finds, the glucocorticoid effect not only with blood circulation in the GC Horizontal correlation, also have the GC concentration of physiologically active closely related with tissue local.The Srd5a1 activity that strengthens in the liver causes serum cortisol (COR) clearance rate to increase, and then compensatory hypothalamo-pituitary-adrenal axis (HPA) increased activity, COR secretes increase, further cause abdominal obesity and MS(MS to refer to a series of metabolism disorders and the gathering of cardiovascular and cerebrovascular vessel risk factor on same individuality, be the syndrome of the multiple Developmental and Metabolic Disorder clinical signs such as obesity, hypertension, dyslipidemia and impaired glucose tolerance).
This experiment condition drag group and natural matched group ratio, the up-regulated of Srd5a1 gene 4.0 times, illustrate: model group causes the serum cortisol clearance rate to increase by improving Srd5a1 gene expression enhancing Srd5a1 activity, so that MS appears in model group, finally makes hepatocyte that fat occurs and becomes.
Under this experiment condition with after the buccal tablet intervention with intervene before the model group ratio, the down-regulated expression of Srd5a1 gene 5.86 times, illustrate: buccal tablet of the present invention can be by this gene expression of downward modulation, improve because the metabolism syndrome that excessive drinking brings, this has explained the reason why buccal tablet of the present invention can prevent and treat metabolism syndrome from another point of view, also is that buccal tablet control excessive drinking causes one of molecule mechanism of fatty liver.
In a word, show by above test: product of the present invention can effectively relieve the effect of alcohol, and the liver fat that the control excessive drinking causes becomes, and the good effect of buccal tablet.The research aspect that present China uses propolis and Lac regis apis to relieve the effect of alcohol is at the early-stage, by experiment, finding that propolis merges the Lac regis apis buccal tablet to relieving the effect of alcohol and prevent and treat the lipotropism that excessive drinking causes effective, is the antialcoholic drugs proposition new approaches of exploitation propolis and Lac regis apis dosage form.Propolis and Lac regis apis are China's Traditional health care products, can improve hepatocyte activity, improve multiple blood parameters, and the prevention fatty liver is significant to the liver health care of safeguarding and improve Chinese.Propolis and Lac regis apis product can be used as the first-selected health product that relieve the effect of alcohol, in addition promotion and application.
Claims (3)
1. be used for the propolis extracted with alcohol relieve the effect of alcohol, for adopt molecular weight that ethanol extracts from propolis less than the solid content of 5000D greater than the dry thing of 90% biological activity.
2. preparation method that is used for as claimed in claim 1 the propolis extracted with alcohol relieve the effect of alcohol is characterized in that may further comprise the steps:
1) propolis is placed under-18 ℃ the ambient temperature conditions and pulverize after freezing 20～40 hours;
2) under the normal temperature and pressure be 50%～90% edible ethanol with volumetric concentration with pulverize after propolis mix by the weight ratio of 1～10 ︰ 1, prior to low-frequency ultrasonic waves, under 10～40 ℃ of temperature, processed 10～40 minutes, be airtight immersion 1～4 day under 10～40 ℃ the condition in ambient temperature then;
3) get the rear mixture of immersion and turned frozen centrifugation 10～15 minutes with 8000～16000, remove solid impurity, get the supernatant of dissolve with ethanol propolis;
4) with supernatant under 1～5 ℃ of ambient temperature conditions with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight less than the propolis extracted with alcohol of 5000D;
5) above-mentioned molecular weight is placed the ethanol distillation recover less than the propolis extracted with alcohol of 5000D, boils off edible ethanol, obtain molecular weight less than the solid content of 5000D greater than the dry thing of 90% biological activity.
3. a propolis extracted with alcohol that is used for as claimed in claim 1 relieving the effect of alcohol is characterized in that making the buccal tablet that relieves the effect of alcohol for the preparation of propolis with described propolis extracted with alcohol, Lac regis apis lyophilized powder, hydroxyl isomaltulose, Sorbitol, magnesium stearate and Mentholum be used to relieving the effect of alcohol in the application of producing buccal tablet; Described propolis extracted with alcohol, Lac regis apis lyophilized powder, hydroxyl isomaltulose, Sorbitol, magnesium stearate and Mentholum alcoholic solution account for respectively 15～30%, 5～10%, 55～70%, 5～15%, 0.2～0.8%, 0.1～0.4% of buccal tablet gross mass.
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| CN106858491A (en) * | 2017-03-31 | 2017-06-20 | 甘肃宇翔蜂产业技术服务有限公司 | A kind of propolis royal jelly tablet |
| CN111388490A (en) * | 2020-04-17 | 2020-07-10 | 陕西巨子生物技术有限公司 | Ginsenoside composition with function of preventing and treating alcoholic fatty liver |
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