CN102934769B - Bee pollen mixture buccal tablet for alleviating hangovers - Google Patents

Abstract

The invention provides a bee pollen mixture buccal tablet for alleviating hangovers, relating to the technical field of processes for extracting active substances from animals and plants, especially bee pollens and bee glue and combining the active substances to prepare royal jelly lyophilized powder. The buccal tablet is mainly formed by lyophilized powder of the bee pollen water extract which is obtained by a bee pollen water extraction method and has molecular weight less than 5000D, ethanol extracted bee glue, royal jelly lyophilized powder, sorbitol, magnesium stearate and menthol. The buccal tablet has obvious effect of preventing alcoholic fatty liver, does not have side effect, is low in cost and is simple and practicable.

Description

For the Bee Pollen intermixture buccal tablet relieved the effect of alcohol

Technical field

The present invention relates to from animals and plants, particularly from Bee Pollen water extract, propolis and royal jelly, be mixed with the technology field of buccal tablet.

Background technology

China is the country of making wine in the world the earliest, and spirits culture is permeated in the many culture of society, 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 of the alcoholic hepatitis occurred therefrom also constantly increases, and in China, has become the second largest hepatopathy after virus hepatitis; Due 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.

Alcoholic strength heat and gas are strong, insobriety, and the wine poison is accumulated, and the many internal organs of liver taste are impaired and cause impairment of both QI and YIN.Chinese traditional treatment is when take clear relieving alcoholism, clearing heat and eliminating phlegm, supplementing qi and nourishing yin as main; U.S. FDA has been ratified 3 medicines for the alcoholism treatment, i.e. ester is adjoined in naltrexone, Acamprosate and holder.Find in animal experiment and clinical research, as ubenimex, Nimodipine, emetine etc. also have, to a certain degree improve crapulent effect.Bee Pollen water extract, propolis and royal jelly have the metabolic active component of a lot of adjusting organism physiologies, have the effects such as clearing heat and detoxicating, prevention fatty liver, the deterioration of inhibition liver cell, reduction onset of liver cancer rate.By the research of molecular mechanisms of action that Bee Pollen, propolis and royal jelly are relieved the effect of alcohol, for researching and developing new solution drinks medicine, there is profound influence.

Current plant alcohol intoxication-alleviating preparation roughly comprises take the ethanol absorption inhibitor that Araliaceae is representative, take the liver metabolism promoter that the root of kudzu vine, FI puerariae be 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 to alcohol metabolism, also can play certain prevention and result for the treatment of to various chemical damages, is the good medicine of a class facilitating alcohol metabolism and protecting liver.

The AML incidence of disease is cumulative year after year trend, and the liver diseases caused by drinking is day by day serious, therefore finds efficient, safe solution very urgent, by the research relieving alcoholism and protecting liver, mainly by following several approach, can reach:

1, reach the relieving alcoholism and protecting liver effect by raising alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) activity

Research about antialcoholic drugs at present 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.Gastric mucosa is the first barrier of alcohol metabolism, and gastric mucosa is impaired, and can cause entering the septicemia element increases, and increases the weight of the bioavilability of ethanol in human body, so improve ADH activity in stomach, strengthens ethanol first-pass metabolism under one's belt, can alleviate the load to liver.In the situation that the single low alcohol consumption is first taken medicine and is drunk, can improve ADH activity in liver, the ethanol of degrading rapidly, effectively stop the absorption of ethanol, reduces blood alcohol concentration, to reach the purpose of relieving the effect of alcohol.The root of kudzu vine, FI puerariae, Semen Hoveniae (Fructus Hoveniae) are the most representative solution drinks of traditional Chinese medicine medicines.Find after deliberation, the active ingredient of their performance antialcoholism actions is Flavonoid substances, trifoliate orange yellow party (Semen Hoveniae (Fructus Hoveniae) and rheum officinale) on the impact of alcoholic liver injury in rats stomach ADHmRNA experiment in, thereby the trifoliate orange yellow party is proved and can improves alcohol and alleviate at the first-pass metabolism of stomach the load that alcohol causes liver.Trifoliate orange yellow party and Qinggan Huoxue Recipe (radix bupleuri, the root of large-flowered skullcap, the red sage root, turtle shell, the root of kudzu vine) 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 liver, ethanol is 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 great drinker, improve its ADH activity and can slow down the oxidative stress 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), show that Semen Hoveniae (Fructus Hoveniae) water extraction liquid can effectively strengthen acute alcoholism liver tissues of rats ADH, ALDH activity, thereby the oxidation Decomposition metabolism of accelerating alcohol in liver, reduce its toxic metabolite (being mainly acetaldehyde) to the toxic action of liver, reach certain liver effect of protecting.Yet, only improve ADH, the ALDH activity can not slow down the ethanol toxic action of metabolism in vivo, also tackle the key enzyme that the alcohol metabolism overall process is relevant, as CYP2E1, CAT carry out system research, guarantee ethanol, the thorough metabolism of acetaldehyde and the balance of vivo oxidation reducing condition.

2, reach the relieving alcoholism and protecting 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 reaction is brought out the damage that principal character occurs to become with fat liver organization.The factor relevant to oxidative stress has: MDA (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX).The content of MDA embodies level of lipid, and the activity of SOD and GSH-PX embodies the ability of Scavenger of ROS, and they are the common counters in relieving alcoholism and protecting liver research.Exist the balance of antioxidant system and enzymatic oxidation system in liver, after absorption alcohol, this balance will be broken, the activity decreased of SOD and GSH-PX.So in protective, activity, the removing interior free yl of antioxidant system is significant by the hepatic injury due to oxidative stress to control.At present relevant meals show with the research of medicine on alcohol metabolism impact and anti-ALD: the antioxidant content in black tea can suppress the interior enzyme system of body that causes in the alcohol metabolism process and the antioxidation activity reduction of non-enzyme system, contributes to body to maintain normal redox state.Aldehydes matter in the fermentation Fructus Hordei Vulgaris extract can be removed free radical and the active oxygen produced by alcohol dehydrogenase enzyme system and CYP2E1 system, slow down the hepatic injury that response to oxidative stress causes, the water extract of thyme and ginger can effectively reduce the content of enzymatic oxidation material in liver and improve the level of polyphenoils.

3, by suppressing the NF-kB activation and lower 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 that regulates and controls inflammation and immune response, Apoptosis and infection and stress reaction, and COX-2 is through stimulating the induced 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, carbohydrate 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, fibrillatable.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 lowering the COX-2 expression, realize the target of protection hepatic tissue.

4, by reducing level of endotoxin and CD-14, express

In blood plasma, the level of endotoxin rising is also the key factor that causes ALD, but in modern relieving alcoholism and protecting liver goods, how research affects the very few of level of endotoxin about goods, and bicyclic alcohols, as the novel Antihepatitis medicament of Chinese independent development, has been used for the treatment of slow virus hepatitis.Current research shows, bicyclic alcohols can reduce Endotoxin Levels and lower CD-14 and express, 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 protection liver organization 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, as ADH, ALDH, CYP2E1, CAT; The oxidative stress factor of influence, as MDA, SOD, GSH-PX; Signal conducts crucial nuclear factor, as NF-κ B, COX-2; Level of endotoxin and acceptor CD-14 expression etc. thereof.

Therefore, the research about the relieving alcoholism and protecting liver mechanism of action also should launch around these key factors.Simultaneously, China's hepatinica product that relieve the effect of alcohol, in health products, mainly with Chinese herbal medicine, be main, the exploitation of relevant dietary ingredient seldom, therefore according to the theory of traditional Chinese medicine " integration of drinking and medicinal herbs ", the relieving alcoholism and protecting 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, find the effectively damaging action of alleviation of alcohol to liver of Bee Pollen and study through us, successful relieves the effect of alcohol, its molecular mechanism adjusting stronger with it of relieving the effect of alcohol is multiple, and to have a gene of antialcoholism action relevant, be expected to be developed as a kind of health food that alleviates the alcohol chemical damage.

Bee Pollen is to be gathered the stamen of the sources of nectar and pollen by honeybee, the dough of through processing, carrying back nest, its nutritional labeling is very abundant, not only contains rich in protein, carbohydrate, lipid, vitamin, organic acid, mineral matter, nucleic acid, also contains enzyme and other biological active material.Bee Pollen is usually used in health food and nutraceutical, its chemical composition studied, so that the Bee Pollen product of the more kinds of exploitation, take full advantage of Bee Pollen Resource, be a current developing direction.Bee Pollen is the elite of plant, is described as " nutriment the most completely ", " concentrated nutrition library ", is considered to the most promising food resource of 21 century.

The Bee Pollen Resource of China is very abundant, and output is large, and kind is many, but the Bee Pollen product is less, and the health products of declaring only have kind more than 20, and how the development of new functional food, be very worth research and discussion.Yet, study and grasp the Changing Pattern of the crucial physical and chemical indexs such as protein, ash content, moisture and Determination of Vitamin C of Bee Pollen, be still the focus of present research; Along with the increase of people to pollens nutrition product demand, the basic research of China's pollen and application study will deepen continuously, and will create more value to the mankind.In recent years, pollen has some progress at medicine, food, health food, cosmetic field; Such as, pollen treatment prostatitis, tumour, hepatitis, angiocardiopathy, and the pollen wet goods research of pollen polysaccharide Sulfation, pollen metabolism, pollen pharmaceutical carrier, pollen recombinant protein, pollen fermentation food, pollen anticorrisive agent, SCF-CO 2, the exploitation of China's Bee Pollen has wide market.

Bee Pollen, propolis and royal jelly are the raw materials of the traditional integration of drinking and medicinal herbs in the Chinese medicine treasure-house, as traditional medicine, it is thousands of year that the history of its dietotherapeutic has exceeded, and the Shennong's Herbal from 2400, Compendium of Material Medica are to all on the books in the Pharmacopoeia of the People's Republic of China of today.

Modern study proves, pollen polysaccharide can improve quantity and the function of body T lymphocyte and macrophage, improves sero-immunity Protein G level; the enhanced machine body immunity function; thereby play tumor suppression and antitumaous effect, and can effectively stop radiotherapy, chemotherapy damage, protection body.The multiple Antioxidative Factors such as Flavonoid substances, polyphenol compound, carotenoid in Bee Pollen, can work in coordination with mutually the performance antioxidation; Bee Pollen, with its efficient antioxidation activity, suffers from the disease relevant to radical damage and plays an important role aspect anti-ageing the control mankind; Therefore, Bee Pollen is the functional food that a kind of suitable ideal has antioxidation.Bee pollen nutrition is comprehensive, composition is extremely complicated, contain life required complete nutrients matter and various bioactivators, have and regulate immunologic function, growth promotion, antifatigue, anti-aging, softening blood vessel, reducing blood lipid, the various biological functions such as short hematopoiesis, beauty treatment and weight reducing, cellular immunity, humoral immunity are all had to obvious humidification, but utilize the Bee Pollen aspect research of relieving the effect of alcohol to remain blank.

Propolis has very strong physiological regulatory action, propolis is the material of the resinae from different plants of being collected by honeybee, just by the mankind, be used as drug therapy from the ancient times, now it remains the most frequently used treatment article of Balkan Peninsula country, is applied to wound, burns, has a sore throat, the treatment such as gastric ulcer.Over nearly 10 years, the researcher has carried out comprehensive research to pharmacologically active and the chemical composition of propolis, finds that propolis has pharmacologically active widely, as antibiotic property, antimycotic, antiviral, anti-inflammatory, protect the liver, anti-oxidant, anticancer etc.Propolis flavone is one of main active ingredient contained in propolis, and its pharmacological action is very extensive, and the effects such as anti-inflammatory, antiallergy, carcinogenesis, anti-hyperplasia, antiviral and anti-cell differentiation are arranged; Can alleviate active oxygen radical to biomembranous decay and preservation SOD; Contained MDA in hepatic tissue be can reduce in a large number, and mouse liver SOD, CAT activity obviously improved; The prompting propolis flavone has stronger removing oxygen radical, oxidation resistant effect.Mahran etc. study discovery, and bee glue water solution is the hepatocellular damage of dose dependent ground protection, and its mechanism may be that water extracts of propolis has repair to hepatocellular injury.Lin etc. study discovery, and the hepatic injury that propolis causes alcohol also has protective effect.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, anti-inflammatory, antitumor, hypotensive, regulate blood-fat and blood sugar, strengthen immunity; protect cardiovascular, sedation anesthesia, the protection liver; anti-ageing, improve the effects such as memory function, its preparation has been widely used in clinical.But the research that propolis extracted with alcohol is applied to relieve the effect of alcohol, remain blank at present, and by this research in several years, we have made the molecular mechanisms of action that propolis relieves the effect of alcohol substantially clear, for researching and developing new solution drinks medicine based theoretical.

Royal jelly is 5～15 age in days worker bee lingual glands and upper palatine gland secreted milky or faint yellow emulsion liquid, is whole puberty of queen bee nit and drone larvae unique food in earlier stage, is similar to mammiferous milk, claims again bee milk, royal jelly, royal jelly.Attention and attention to royal jelly, at first be to have found that feeding the young honeybee of raising honey can only grow up to worker bee, general 3 0～4 0 days of life-span, the longest 7～8 months, can grow up to queen bee and feed the young honeybee of raising royal jelly, life-span reaches 3～5 years, and these phenomenon prompting people utilize royal jelly as the food of promoting longevity, but its nature and role mechanism is not understood.By 1888 years, Germanization scholar A Von Planta just first component to royal jelly carries out generality research, so far people have started to inquire into the profoundness of royal jelly, but royal jelly is comprised to comprehensive research of biology, chemical pharmacology and clinical trial, or nearest decades are inchoate, and people also do not study clear to some the micro-bioactivators in royal jelly fully so far.At present, the about 9 large classes of fixed function factor: they are peptides and proteins classes; Functional sweetener; Functional grease; Free radical scavenger; Vitamin; Active trace element; Active polysaccharide; Flavone compound and micro-ecological factor are as lactic acid bacteria etc.Find according to people's research, contain these abundant function factors in royal jelly, the mankind are had to extremely strong alimentary health-care function and medical function.

Modern study shows: royal jelly can alleviate hepatocellular damaging action in the ALD course of disease, liver organization after damage is had to 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 mitochondria and lipid peroxidation injury, thus the level of reduction serum AST and ALT; Royal jelly has reduced the expression of TNF-α in liver cell, and TNF-α has mediated hepatocellular inflammatory reaction as inflammatory factor, multiple pathology and the physiological activities such as immune response, hepatocellular apoptosis and amplification liver fibrosis.The reason that analysing royal jelly TNF-α reduces may be that royal jelly 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 subject to its regulation and control.Royal jelly can press down the expression of COX-2 and TNF-α in the forming process of AML, and alcoholic liver injury is had to certain protective effect.Royal jelly also all has significant improvement effect to hepatic fibrosis-renal tubular ectasia syndrome serological index and the hepatic pathology morphology of CC14 hepatic fibrosis rats, but to forming liver tissue fibrosis after liver fibrosis, improve not obviously, its possible mechanism is the royal jelly inflammation that can improve hepatic tissue, reduce the synthetic of ECM and suppress the secretion of TNF-α.And Bee Pollen, royal jelly and propolis have natural orthofunction and uniformity, the triplicity result of use can be better.

The medical care effect of Bee Pollen, propolis and royal jelly in recent years more and more by people cognition, but the material that exists a class to relieve the effect of alcohol in Bee Pollen, propolis and royal jelly is just not known.Through us, up-to-date research is found in Bee Pollen also to exist a class to have the material that promotes alcohol metabolism, to promoting body to relieve the effect of alcohol, prevent and treat the fatty liver aspect, has remarkable effect.The Chinese patent that is 201010607398.7 as number of patent application discloses by water extracting method, and Bee Pollen, through broken wall, immersion, water extraction, ultrafiltration, is obtained to the Bee Pollen water-soluble substances, and proof Bee Pollen water-soluble substances has the effect for the treatment of fatty liver.

Buccal tablet, be a class without peptic digest, directly by oral mucosa, absorb and enter blood, adopt soluble material in oral cavity to carry out for main coating material the tablet that dressing is made.Can prevent acidity and enzyme to the destruction of some drugs or prevent the intense stimulus of medicine to stomach.

Current various dinner party is more and more, and the incidence of disease of the various diseases brought out owing to drinking improves year by year, and the healthy problem that excessive drinking is brought out has become the importance that affects modern's quality of life.

Summary of the invention

The object of the invention is to invent a kind of Bee Pollen intermixture buccal tablet that can efficiently relieve the effect of alcohol.

Freeze-dried powder, bee glue powder, royal jelly freeze-dried powder, D-sorbite, dolomol and menthol that the present invention mainly is less than the Bee Pollen water extract of 5000D by the molecular weight of obtaining with the Bee Pollen water extracting method form.

Described freeze-dried powder, bee glue powder, royal jelly freeze-dried powder, D-sorbite, dolomol and menthol alcoholic solution account for respectively 50～70%, 10～25%, 10～25%, 5～10%, 0.2～0.6%, 0.1～0.4% of buccal tablet gross mass.

Through research, find that 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 antialcoholic drugs simultaneously, compare with stomach due to dose and differ 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 used after meal preferably adopts the mode without stomach.

Through research; finding has many bioactivators under one's belt easily by stomach acids destroy in Bee Pollen, propolis and royal jelly; and can directly absorb by hypoglossis mucous membrane; to promoting the body aspect of relieving the effect of alcohol to there is remarkable effect; so make buccal tablet, can protect better bioactivator to exempt from destruction, improve the result of use of product.

The present invention utilizes Bee Pollen water extract, bee glue powder and the royal jelly freeze-dried powder mixture effect of relieving the effect of alcohol remarkable, and has no side effect, and expense is cheap, simple.Bee Pollen water extract, propolis and royal jelly etc. can be brought into play the pharmacological action of relieving the effect of alcohol on gene level, and inventor's research has solved the molecule mechanism that mixture relieves the effect of alcohol substantially, for this reason, files an application the present invention.

The accompanying drawing explanation

Fig. 1 is natural health control group liver tissue slices figure.

Fig. 2 is natural health control 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 liver tissue slices figure.

Fig. 6 is gavage buccal tablet group renal tissue slice map.

The specific embodiment

The so-called Bee Pollen of the present invention refers to the pollen of gathering through honeybee as camellia powder, rape pollen, lotus powder etc.

One, prepare the water extract freeze-dried powder that Bee Pollen relieves the effect of alcohol:

1, Bee Pollen removal of impurities, drying, make water content drop to 4～8%.

2, adopt the mode of air-flow crushing broken wall by broken wall of melissa pollen.

3, soak the Bee Pollen of broken wall with the normal pressure and temperature pure water, process 10～40 minutes with low-frequency ultrasonic waves simultaneously, control the temperature of ultrasonic extract at 10～40 ℃.

4, turn refrigerated centrifugation with 8000～15000 and remove solid impurity, get supernatant.

5, by supernatant under the environment temperature of 1～5 ℃ with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight and be the following Bee Pollen water-soluble substances of 5000D.

6, the freeze-dried powder lower than 6% Bee Pollen water extract by above-mentioned Bee Pollen water extract freeze drying 24～48 hours preparation water content.

Two, prepare the propolis ethanol extract dry powder that relieves the effect of alcohol:

1, after propolis-18 ℃ freezing 20-40 hour, pulverize.

2, under condition of normal pressure, by concentration, be that 50%～90% edible alcohol mixes by the weight ratio of 1～10:1 with propolis, process 10～40 minutes with low-frequency ultrasonic waves, then soak 1～4 day, and keep 10～40 ℃ of room temperatures.

3, turn refrigerated centrifuge with 8000～16000 after the immersion and remove solid impurity in 10～15 minutes, get the supernatant that ethanol dissolves propolis.

4, by supernatant at room temperature with 5000D molecular sieve ultra-filtration and separation, obtain the propolis extracted with alcohol that molecular weight is less than 5000D,

5, the ethanol distillation recover reclaims ethanol and obtains the dry thing that the propolis solid content is greater than 90%.

Three, prepare royal jelly freeze-dried powder:

By in fresh royal jelly and freeze drier dry 36～48 hours, the preparation water content was less than 6% freeze-dried powder.

Four, prepare the buccal tablet that Bee Pollen, propolis and royal jelly mixture relieve the effect of alcohol:

1, granulating process: after the boiling pot is preheating to 40～60 ℃, first freeze-dried bee pollen, bee glue powder, royal jelly freeze-dried powder, D-sorbite are added in the boiling pot, arrange according to the following table parameter, open peristaltic pump spray purified water and carry out one-step palletizing.Granulation adds dolomol and menthol alcoholic solution to mix after finishing, after sub-cooled, and the buccal tablet of Bee Pollen water extract, propolis and royal jelly mixture that preparation is relieved the effect of alcohol.

Typical several quality proportioning table: (unit: kg)

?	Freeze-dried powder	Bee glue powder	Royal jelly freeze-dried powder	D-sorbite	Dolomol	Menthol
							Proportioning 1	70	11	10	8.1	0.6	0.3
Proportioning 2	63	16	14	6.2	0.5	0.3
							Proportioning 3	65	12	16	6.4	0.4	0.2

Processing parameter:

Parameter	EAT	Leaving air temp	The air inducing frequency	Peristaltic pump	Atomizing pressure
						During whitewashing	60-80℃	40-55℃	40Hz	150rpm	0.2MPa
After whitewashing	65-80℃	50-60℃	35Hz	150rpm	0.2MPa

2, tablet forming technique: adopt ZP1100 tablet press machine (29 punching) to carry out compressing tablet experiment, technological parameter: 30～40 rev/mins of rotating speeds; Operating pressure 7～15kN; Sheet weighs 0.4～0.6g.

Three, application:

(1) subjects:

The clean level of the Wistar in 22 week age of a gavage male rat, 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 after body weight 1% gavage strong, colourless liquor distilled from sorghum.

(2) using dosage:

Using dosage: by the amount gavage of every rat 50-150mg every day buccal tablet of the present invention, connect and take 30～60 days.

(3) comparing result:

1, rats in test groups liver and renal tissue section result are relatively:

(1) by nature control group liver and renal tissue section, see Fig. 1,2.

Be polygon from Fig. 1,2 visible male white rat liver liver cell clear in structure, hepatic sinusoid is obvious, and the hepatic cell cords marshalling does not have obvious fat vacuole in liver cell; Kidney group section mesonephric glomerulus structure is clear, and the Bowman's capsule gap is obviously moderate, not obviously damage.

(2) by gavage white wine model group liver and renal tissue section, see Fig. 3,4.

From Fig. 3,4 visible liver cell obscure boundaries, become in circle cavity is arranged, the hepatic sinusoid pressurized narrows down, liver rope arrangement disorder; The glomerulus structure is unclear, and damage is serious, the Bowman's capsule distortion.

(3) gavage buccal tablet liver and renal tissue section, be shown in Fig. 5,6.

From Fig. 5,6 visible liver lobuli hepatis clear in structure, hepatic sinusoid is evenly distributed, and liver cell is polygon, marshalling; The renal glomerulus structure is obviously improved, and the Bowman's capsule gap is obvious, has the vestige of obvious glomerulus reparation.

2, on an empty stomach gavage buccal tablet of the present invention intervene after to the comparative analysis of rats in test groups Main Blood Biochemical Index.

Adopt the blood sampling of anesthesia abdominal aorta, detect Main Blood Biochemical Index by Toshiba's fully automatic blood bio-chemical detector after getting serum.

Comparative analysis is as following table:

Group	The nature group	Model group	Wine mixes
				ALT	73.00±16.67	91.25±29.33	51.71±13.74
ALP	102.50±11.90	146.75±21.70	96.57±19.75
				LDH	962.75±96.05	909.75±326.58	1653.00±277.08
CHO	1.23±0.08	1.52±0.26	1.21±0.14
				TG	1.25±0.18	1.84±0.37	1.15±0.19
BUN	5.89±0.65	4.78±0.71	5.72±0.48
				TP	61.81±0.31	58.49±1.58	61.67±2.20
ALB	15.50±0.43	14.88±0.53	16.38±0.59

Through the SPSS statistics, model group glutamic-pyruvic transaminase (ALT) is extremely remarkable with gavage buccal tablet group of the present invention difference; Transaminase extensively exists in histocyte, and especially the strongest with activity in the tissue such as liver, cardiac muscle, brain, kidney, when normal, plasma content is very low, and ALT mainly is present in liver, and when the hepatic tissue disease damage, ALT is released in a large number blood and can causes that in blood, transaminase raises.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 bone, and the ALP in liver is with the transhipment of material and drain relevant.And the diabetes B patient causes metabolism obstacles of blood glucose due to insulin secretion obstacle or insulin resistance, also cause the obstacle of lipid metabolism simultaneously and, at intrahepatic deposition, even form fatty liver, and make liver damage cause the rising of serum levels of ALP level.Through the SPSS statistics, model group alkaline phosphatase and other two experimental group difference are extremely remarkable; Illustrate that buccal tablet of the present invention can significantly improve hepatocellular injury, reduce the alkaline phosphatase enzyme level, rising and the hepatocellular damage of alkaline phosphatase (ALP) are closely related, illustrate that the excessive drinking of this experimental model group rat is very serious to the liver cell injury, and can improve the bad reaction that long-term excessive drinking causes after the gavage buccal tablet, alleviate hepar damnification.

Through the SPSS statistics, natural control group and model group lactic dehydrogenase (LDH) are extremely remarkable with gavage buccal tablet group of the present invention difference; Lactic dehydrogenase is glucolytic key enzyme, after gavage buccal tablet of the present invention, the lactic dehydrogenase enzyme level is very high, illustrate and promote glycolysis closely related, that is to say that buccal tablet of the present invention has stronger short glycolysis under this experiment condition, being beneficial to that liver relieves the effect of alcohol and glycometabolism.

Diabetes B is to take the class disease that insulin resistance and islet beta cell function obstacle be principal character, and wherein the loss of beta Cell of islet and dysfunction play a decisive role in the generation of disease and development.Research in recent years shows, cholesterol metabolic and islet beta 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, make the content of cell inner cholesterol maintain a dynamic equilibrium.After the metabolism of beta Cell of islet inner cholesterol gets muddled, can affect the glycometabolism of β cell by number of ways, finally induce the β Apoptosis.Therefore, the cholesterol metabolic disorder may be a new mechanism that causes diabetes B patient β cell dysfunction.Through SPSS, add up, model group cholesterol (CHO) and other two experimental group significant differences, illustrate that excessive drinking is the arch-criminal who causes hypercholesterolemia, and gavage buccal tablet of the present invention has remarkable improvement effect, significantly lower the cholesterol levels caused due to excessive drinking and raise, the generation that prevention is brought out to diabetes B due to excessive drinking is significant.

Fat toxicity due to high triglyceride (HTG) has caused extensive concern in recent years, and it can cause and increase the weight of insulin resistance (IR), and the biological effect of insulin is reduced.Glycogen is synthetic by participating in for liver, decomposition of glycogen and gluconeogenesis play a part direct and important in maintaining glucostasis, particularly G-6-Pase plays a crucial role in glycogen generates, hepatic insulin resistance also shows as the fat metabolic disorder, as triglycerides discharges, increases and the liver fat sex change.Through the SPSS statistics, model group triglycerides (TG) and natural control group significant difference, extremely remarkable with gavage buccal tablet group of the present invention difference; From the experiment situation, serum high triglyceride level appears in model group, illustrates that excessive drinking is the one of the main reasons that produces hypertriglyceridemia, is one of paathogenic factor of bringing out diabetes B; And take buccal tablet of the present invention, the Triglyceride Metabolism brought out due to excessive drinking is had to good regulating and controlling effect.

Urea concentration, except being subject to Influence on kidney, also is subject to the impact of protein and organism metabolism state in diet, as bad as high-protein diet, gastrointestinal function etc.Because kidney has powerful reserve capabillity and compensatory capacity, impaired early stage or slight when impaired at glomerulus, blood urea nitrogen still can maintain normal level, only at the severe renal bead, damages, when general glomerular filtration rate(GFR reduction by 50% is following, blood urea nitrogen concentration just obviously raises.Through SPSS statistics, model group urea nitrogen and natural control group and buccal tablet group significant difference; Under this experiment condition, the model group rat is due to excessive excessive drinking, drunk serious, causing feed intake to descend and protein metabolism disturbance, make urea nitrogen not rise counter falling, is minimum in all experimental group, and gavage buccal tablet experimental group of the present invention, metabolism to protein is improved effect, and urea nitrogen levels gains momentum, and approaches with the urea nitrogen levels of natural control group.

Through the SPSS statistics, model group total protein and natural control group and buccal tablet group significant difference, model group albumin and buccal tablet group difference are extremely remarkable, natural control group albumin and buccal tablet group significant difference; Above these change the long-term excessive drinking of explanation can cause the body protein dysbolism, and can bring into play very strong improvement effect to the body protein dysbolism caused due to excessive drinking after gavage buccal tablet of the present invention.

By the analysis to blood parameters, we are not difficult to draw such conclusion: in the excessive drinking model, propolis, pollen and royal jelly mixture buccal tablet not only can help body to relieve the effect of alcohol, and can recover rapidly the metabolism of body to three large materials, improve multinomial blood parameters, make organism metabolism return to normal condition.

3, lozenge of the present invention is expressed and is changed comparative analysis oligogene in the rats in test groups liver metabolism after intervening, as following table:

By biochip technology, study the important metabolic pathway of many impacts in the model group rat liver expressing gene of finding 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 eubolism, and after the intervention of gavage buccal tablet, the variation of this gene expression has obtained good correction, from 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 due to long-term excessive drinking in example table, the reparation of tissue damage has very strong improvement effect, research by us finds that from gene level the gavage buccal tablet can relieve the effect of alcohol effectively, improve the metabolism of liver.

3.1 the impact that the gavage buccal tablet is expressed key gene relevant to glycolipid metabolism in liver after intervening

Group

Dci

Ehhadh

Fasn

Fbp2

G6pc

Scd

Scd1

Model group/natural group

↓3.72

↓2.74

↑4.86

↓4.57

↓5.62

↑5.04

↑5.65

Buccal tablet group/model group

↑3.16

↑3.29

↓3.8

↑2.73

↑2.81

↓3.16

↓2.74

Notes: in table, ↓ 3.72 expression Dci genes model group in liver cell mRNA is lowered 3.72 times with the expression ratio of natural control group, 3.16 times of the expression ratio rises of ↑ 3.16 expression Dci gene gavage buccal tablet groups and model group.

3.1.1 Dci: ten dichloro alkane coacetylase δ isomerases, anti-enoyl CoA isomerase, mitochondria, participate in the fat metabolism, the aliphatic acid beta-oxidation.

Under this experiment condition the excessive drinking model group with natural control group than 3.72 times of the expression of lowering ten dichloro alkane coacetylase δ isomerase genes, illustrating that the model group rat is indulged in excessive drinking for a long time can be by lowering this gene expression, mitochondria aliphatic 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.

Under this experiment condition, gavage buccal tablet group has raised 3.16 times of this genes with the model group ratio, can promote the aliphatic acid beta-oxidation of liver organization, and 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 Ehhadh: difunctional peroxidase (peroxisomalbifunctionalenzyme), enoyl-CoA hydratase, peroxisome, participate in the fat metabolism, fatty acid metabolism.Peroxisome is unique place that saturated/undersaturated very-long-chain fatty acid carries out beta-oxidation, D-3 hydroxyl acyl CoA dehydratase/D-3 hydroxyl acyl CoA dehydrogenase is that D-bifunctional protein (D-BP) is a kind of enzyme of the participation peroxisome aliphatic acid beta-oxidation of reported first, in the beta-oxidation of polyunsaturated fatty acid, 2-methyl branch aliphatic acid and D-isomers hydroxy fatty acid, plays vital effect.Under diabetic disease states the rising of blood plasma very-long-chain fatty acid and levels of polyunsaturated fatty acids whether with the beta-oxidation of peroxisome aliphatic acid, particularly the change of D-BP activity is relevant.Eukaryotic mitochondria and peroxisome all carry out the beta-oxidation of aliphatic acid, carbon chain lengths is less than the mainly beta-oxidation energy supply in mitochondria of aliphatic acid of 20 carbon, and carbon chain lengths is greater than saturated/unsaturated very-long-chain fatty acid, dimethyl branched chain fatty acid and the mainly oxidation in peroxisome of bile acid biosynthesis metabolic intermediate of 2O carbon, then the Short-Chain Fatty Acids of generation proceeds to the mitochondria exhaustive oxidation.Second step and the three-step reaction of peroxisome aliphatic acid beta-oxidation approach, be that alkene acyl CoA generates the reaction of 3-ketoacyl CoA through hydration, dehydrogenation, by a kind of bifunctional protein catalysis with alkene acyl CoA hydrase/3-hydroxyl acyl CoA dehydrogenase activity, two kinds of bifunctional proteins have been confirmed in peroxisome to exist: L-bifunctional protein (L-BP) and D-bifunctional protein.The trans alkene acyl of L-BP catalysis CoA is converted into L-type 3-hydroxyl acyl CoA, and then dehydrogenation is 3-ketoacyl CoA; And the intermediate product that the trans alkene acyl of D-BP catalysis CoA hydration generates is D type 3-hydroxyl acyl CoA, and take it as its dehydrogenation of substrate catalysis.Substratspezifitaet research shows, L-BP mainly works in the oxidation of saturated fatty acid, non-branched chain fatty acid; And D-BP is not only unique enzyme relevant with the alkene acyl CoA oxidation of polyunsaturated fatty acid, 2-methyl branch aliphatic acid, and D-BP defect patient's research is also found, D-BP participates in utmost point long-chain straight chain fatty acid, for example Major Enzymes of lignoceric acid beta-oxidation.In liver, the content of D-BP and L-BP is almost equal, in every gram protein, approximately containing 2.5mg, illustrates that two kinds of bifunctional proteins play a part in the beta-oxidation of hepatocellular peroxisome aliphatic acid of equal importance.In the higher organism body, there are two kinds of multifunctional enzymes to there is L-3-hydroxyl acyl CoA dehydrogenase activity, be respectively the L-BP be present in peroxisome, and there are three functional proteins of enoyl-CoA hydratase/L-3-hydroxyl acyl CoA dehydrogenase/3-ketoacyl-CoA thiolase activity in mitochondria.Known, during diabetes, the aliphatic acid beta-oxidation activity of mitochondria and peroxisome increases simultaneously, research is found, the beta-oxidation of diabetes rat liver peroxisome aliphatic acid is active also to raise with liver L-3-hydroxyl acyl CoA dehydrogenase activity simultaneously, and when diabetes are described, the beta-oxidation of mitochondria and peroxisome linear saturated fatty acids, L-isomers hydroxy fatty acid strengthens; But, the protein content and the enzymatic activity that have research to observe D-BP significantly reduce, thereby the beta-oxidation of polyunsaturated fatty acid, branched chain fatty acid and D-isomers long-chain hydroxy fatty acid probably weakens, cause the intermediate product D-isomers of unsaturated very-long-chain fatty acid, the accumulation of branched chain fatty acid.This discovery conforms to the phenomenon that arachidonic level obviously raises with the patients with NIDDM blood plasma very-long-chain fatty acid of observing, and prompting D-BP activity decreased may be that while causing diabetes, blood plasma fatty acid is composed one of abnormal reason.

Under this experiment condition the excessive drinking model group with natural control group than 2.74 times of the expression of lowering difunctional peroxidase gene, illustrating that the model group rat is indulged in excessive drinking for a long time can be by lowering this gene, aliphatic acid beta-oxidation ability in peroxisome and mitochondria is weakened, promoting liver fat to become, is one of the main molecules mechanism of bringing out the liver fat change of indulging in excessive drinking for a long time.

Under this experiment condition, gavage buccal tablet group has raised 3.29 times of difunctional peroxidase genes with the model group ratio, can promote the aliphatic acid beta-oxidation in liver organization peroxisome and mitochondria, to preventing and treating the fatty liver caused of indulging in excessive drinking, positive effect being arranged, is that one of main molecules mechanism of liver fat change is brought out in the long-term excessive drinking of gavage buccal tablet control.

3.1.3 Fasn: fatty acid synthetase participates in fatty acid biological synthetic.In mammal, fatty acid synthetase plays vital effect in aliphatic acid is synthetic, be responsible for from the beginning all catalytic steps of synthetic a kind of long-chain palmitic acid (palmitate) of acetyl coenzyme A and malonyl coenzyme A, the expression of gene directly affects the fatty acid synthetase number and has great importance to controlling the body fat deposition.

This experiment condition drag group leader's phase indulge in excessive drinking make in liver cell the fatty acid synthetase gene with natural control group than 4.86 times of up-regulateds, illustrate: it is the arch-criminal who causes liver cell fat excess deposition that model group can make synthetic the increasing of fatty acid synthetase, is to form one of fatty liver molecule mechanism.

Under this experiment condition, gavage buccal tablet group and model group are lowered 3.8 times of the gene expressions of fatty acid synthetase, and illustrate: the buccal tablet group can reduce hepatic tissue aliphatic acid synthetic by reducing the synthetic of fatty acid synthetase, significant to alleviating the liver fat change.

3.1.4 Fbp2: fructose-1,6-diphosphatase 2, the regulatory enzyme of gluconeogenesis, catalyzing hydrolysis fructose-1,6-diphosphonic acid is fructose-6-phosphoric acid and Phos.Involved in sugar metabolism, gluconeogenesis.

This experiment condition drag group leader's phase indulges in excessive drinking and makes Fbp2 gene in liver cell lower 4.57 times with natural control group than expressing, and illustrate: model group, by lowering the expression of this gene, descends model group gluconeogenesis ability, consistent with the weight fat liver.

Under this experiment condition, gavage buccal tablet group and model group raise 2.73 times of Fbp2 gene expressions, illustrate: the buccal tablet group can be improved hepatocellular gluconeogenesis level by raising Fbp2 gene expression, alleviates liver cell fat and becomes.

3.1.5 G6pc: G-6-Pase participates in glycogen metabolism and synthetic.G-6-Pase is an important enzyme controlling body glucose stable state, and it is the rate-limiting enzyme of gluconeogenesis and decomposition of glycogen final step reaction. so the variation of its activity directly has influence on the output of endogenous sugar.The hyperglycaemia of diabetes B is excessively exported and is closely related with the glycogen due to hepatic insulin resistance, so G6pc is a potential target spot for the treatment of diabetes.The hydrolysis of G-6-Pase catalysis G6P produces phosphoric acid and glucose, the i.e. rate-limiting reaction of gluconeogenesis approach and decomposition of glycogen approach.It is generally acknowledged, by the glucose phosphorylation of glucokinase enzymatic be the key of controlling blood glucose balance by the G6P hydrolysis of G6Pase catalysis.G6Pase is a kind of multienzyme complex that is combined in endoplasmic reticulum, and the hydrolysis of its catalysis occurs in endoplasmic reticulum.In mammal, the expression of G6PC is subject to the adjusting of hormone and trophic level, the hungry expression that can stimulate G6PC, and hyperglycemic factor also can stimulate the expression of G6PC by improving cAMP, insulin and ingest and reduce the expression of G6PC.

This experiment condition drag group leader's phase indulges in excessive drinking and makes G-6-Pase gene in liver cell lower 5.62 times with natural control group than expressing, illustrate: long-term excessive drinking can make model group rat G-6-Pase down regulation of gene expression, cause model group rat hepatocytes gluconeogenesis and decomposition of glycogen to weaken, the utilization of reduction to glucose, reduce the supply level of liver cell energy, be unfavorable for that liver cell recovers eubolism.

Under this experiment condition, gavage buccal tablet group and model group raise 2.81 times of the gene expressions of G-6-Pase, illustrate: the buccal tablet group can be by raising the expression of G-6-Pase, improve the utilization of liver cell to glucose, recover the liver cell eubolism, alleviate fat and become.

3.1.6 Scd: sterol coacetylase desaturase participates in fatty acid biological synthetic.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.

3.1.7 Scd1: sterol coacetylase desaturase 1 participates in fatty acid biological synthetic.Stearyl-coenzyme A desaturase 1 (Scd-1) is the biosynthetic rate-limiting enzyme of monounsaturated fatty acids, plays the center adjustment effect in fatty acid metabolism, is also one of genes of interest of leptin (Leptin) effect.Leptin and diabetes, obesity, fatty liver close relation are one of hot fields of metabolic disease in recent years.The diabetes B patient, usually with insulin resistance, hyperleptinaemia, approximately there is fatty liver in 50% diabetes B patient simultaneously.Scd-1st, the key enzyme that the catalysis saturated fatty acid changes to monounsaturated fatty acids, with the generation of a series of metabolic syndromes such as fat, fatty hepatic disease and insulin resistance, develop closely related, 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 result for the treatment of, in clinical medicine, have broad application prospects.Because Scd-1 mainly is present in endoplasmic reticulum, at present the detection of Scd in body-1 mainly adopted mensuration blood fat index of unsaturation (unrighted acid/saturated fatty acid ratio) or by molecular biology method, Scd in cell-1mRNA expression detected.In sum, Scd-1 has been play a part important in energetic supersession.By the research to Scd-1 and energetic supersession relation, can the mechanism of clearer understanding Scd-1 in energetic supersession, thereby be clinical target adjusting energy metabolism and avoid fat toxicity, a series of metabolic syndromes such as prevention or treatment are fat, fatty hepatic disease and diabetes provide basic.

Under this experiment condition, the excessive drinking model group raises 5.04 times of sterol coacetylase desaturase (Scd) gene expressions with natural control group ratio, sterol coacetylase desaturase 1(SCD-1) raise 5.65 times, illustrate: the two gene up-regulated is one of main molecule mechanism that causes alcohol fatty to become, lipotropic is the pathological index of most critical in the process of relieving the effect of alcohol, processing mode validity 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 reach the state of liver fibrosis.

Under this experiment condition, gavage buccal tablet group is lowered 3.16 times of sterol coacetylase desaturase (Scd) gene expressions with the model group ratio, downward sterol coacetylase desaturase 1(Scd-1) gene expression is 2.74 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 that buccal tablet of the present invention is prevented and treated one of main molecule mechanism of fatty liver.

3.2 after the gavage buccal tablet in liver to hepatic injury, repair the impact that relevant key gene is expressed

Group	Fgf1	Il7r	Pla2g2a	Pla2g2c	Tgfb2
						Model group/natural group	↑4.76	↓2.75	↑2.76	↑2.34	↑3.63
Buccal tablet group/model group	↓2.15	↑2.52	↓4.04	↓2.39	↓2.58

Notes: in table, ↑ 4.76 mean that Fgf1 genes model group in liver cell mRNA raises 4.76 times with the expression ratio of natural control group, and the expression ratio of ↓ 2.15 expression Fgf1 gene gavage buccal tablet groups and model group is lowered 2.15 times.

3.2.1 Fgf1: desmocyte growth factor-21 (acidity) participates in the regulation and control process, cell cycle, signal transduction, cell proliferation, Cell Differentiation.Fgf1 improves Angiogenesis, may contribute to newly-generated cardiac muscle cell's existence, and research shows Fgf1 and p38 map kinase, is the protein that promotes cardiomyocyte proliferation and associated angiogenesis, and promotes the generation of fibrillatable and scar.

This experiment condition drag group raises 4.76 times of Fgf1 gene expressions with natural control group than model group, illustrates: model group, by raising Fgf1 gene expression, promotes the liver fibrosis process.

Under this experiment condition, the buccal tablet group, illustrates than lowering 2.15 times of Fgf1 genes with model group: buccal tablet can, by lowering Fgf1 gene expression, slow down the liver cell progression of fibrosis caused due to long-term excessive drinking after intervening.

3.2.2 Il7r: interleukin-17, acceptor, participate in the regulating DNA restructuring, immune response, the cell surface receptor signal connects conduction, antimicrobial humoral response.

This experiment condition drag group has been lowered 2.75 times of interleukin-17 genes with natural control group than model group, illustrate: the long-term excessive drinking of model group rat causes liver cell immune response ability to descend, the expression of lowering the interleukin-17 gene is exactly to mean to increase the weight of hepatocellular inflammatory reaction, promotes liver cell fat to become and Fibrotic process.

Under this experiment condition, the buccal tablet group raises 2.52 times of interleukin-17 genes with the model group ratio, illustrate: after buccal tablet is intervened, strengthen hepatocellular immune response ability by the expression of raising the interleukin-17 gene, alleviate the inflammation reaction, to preventing and treating the fatty liver and the fibrillatable that cause due to long-term excessive drinking, positive effect is arranged.

3.2.3 Pla2g2a: phospholipase A2, base 2A, calcium relies on, and participates in fat catabolism, and cell membrane has destruction, raises the destruction that this gene can increase the weight of the liver cell film, increases the weight of hepatic injury.

3.2.4 Pla2g2c: phospholipase A2, base 2C, act on similar to Pla2g2a.

This experiment condition drag group leader's phase indulges in excessive drinking and makes Pla2g2a gene and Pla2g2c gene in liver cell raise respectively 2.76 times and 2.34 times with natural control group than expressing, illustrate: model group is because long-term excessive drinking can be by raising Pla2g2a and Pla2g2c gene expression, increase the weight of the destruction to liver plasma membrane, cause the rising of serum transaminase.

Under this experiment condition, gavage buccal tablet group and model group are than lowering 4.04 times and 2.39 times of Pla2g2a and Pla2g2c gene expressions, illustrate: the buccal tablet group can be by lowering Pla2g2a and Pla2g2c gene expression, alleviate the destruction to liver plasma membrane, promote the reparation of hepatocellular injury.

3.2.5 Tgfb2: transforming grouth factor beta 2, participate in cell death, just regulating and controlling process, regulating cell propagation, regulation and control immune response, dopamine biosynthesis.Studies confirm that to only have Tgf-β energy HSC stimulated rubber polymer fibrillation in cell factor, other cell factors are a proliferation of HSC stimulated.In process of hepatic fibrosis, KC produces a large amount of Tgf-β by the mode of autocrine or paracrine, and it not only can promote that HSC is converted into fibroblast, promotes the activation of HSC, and can strengthen the synthetic of ECM, suppresses its degraded.Tgf-β can increase the expression of the upper pdgf receptor of HSC simultaneously, promotes HSC autocrine Tgf-β, PDGF, obviously suppresses the apoptosis of activation HSC.PDGF and Tgf-β have formed the Autocrine of activation HSC, are the important mechanisms of the sustained activation of HSC.

This experiment condition drag group and natural control group ratio, the up-regulated of Tgfb2 gene 3.63 times, illustrate: long-term excessive drinking causes hepatocellular injury, raise the expression of Tgfb2 gene, HSC stimulated rubber polymer fibrillation, promote the formation of liver fibrosis, consistent with the section result.

Model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition, the expression of Tgfb2 gene has been lowered 2.58 times, illustrate: lozenge of the present invention can be prevented and treated the process of liver fibrosis by lowering this gene expression, great to safeguarding liver cell eubolism Functional Significance.

3.3 the impact that buccal tablet is expressed key gene relevant to controlling normal activity in liver after intervening

Group	Igfbp2	Lepr	Srd5a1	Sult2a2
					Model group/natural group	↓3.12	↓2.19	↑4.0	↑5.16
Buccal tablet group/model group	↑7.45	↑4.95	↓2.6	↓3.42

Notes: in table, ↓ 3.12 expression Igfbp2 genes model group in liver cell mRNA is lowered 3.12 times with the expression ratio of natural control group, 7.45 times of the expression ratio rises of ↑ 7.45 expression Igfbp2 gene gavage buccal tablet groups and model group.

3.3.1 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, favourable to the prevention fatty liver.

Study and confirm, IGFs is relevant with the formation development of liver fibrosis, 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 with insulin-like growth factor binding protein (IGFBPs) compound that relative molecular mass is 150kb or 50kb in body fluid.The single chain polypeptide that IGF-l is comprised of 70 amino acid residues, mainly synthetic at liver, be the extensive cell mitogen existed 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 acceptor (IGF-lR) mediation, and the validity that IGF-l is combined with IGF-lR is subject to the adjusting of IGFBPs.Liu Lixins etc. studies show that, IGFBP2 (IGFBP2) is expressed obviously and strengthened in IGF-β l induces the HSC-T6 of activation, and prompting IGF and acceptor thereof participate in the formation of liver fibrosis.Due to the biologically active that IGFBPs and IGFs have high-affinity and can regulate IGFs, these are protein-bonded induces generation also can affect the genesis of liver fibrosis.Yet separately there are some researches show, IGF-1 has the effect of anti-hepatic fibrosis in body, can suppress the activation of HSC, its may with improve superoxide dismutase and catalatic activity, improve the ability that liver cell is removed free radical, the stimulus of minimizing HS activation etc. are relevant.

This experiment condition drag group has been lowered 3.12 times of IGFBP2 genes with natural control group than model group, illustrate: model group is by lowering the expression of IGFBP2 gene, cell cultured supernatant transforms to the fibrillatable direction, promotes liver cell fat to become and fibrillatable;

Under this experiment condition, gavage buccal tablet group of the present invention raises 7.45 times of IGFBP2 genes with the model group ratio, illustrate: buccal tablet can be by raising the expression of IGFBP2 gene after intervening, slow down or prevent and treat the Fibrotic process of liver cell, the control alcoholic fatty liver is had to positive effect.

3.3.2 Lepr: leptin receptor is the important intermediary that leptin (1eptin) plays a role, Lepr gene mutation and people's obesity, and the fat deposition of the animals such as common ox, pig is relevant, participates in the energy reserve metabolism, by the JAK2/STAT3 signal transduction.Leptin has energy balance and the fat effect of regulating, and regulation and control breeding, glucose dynamic equilibrium is arranged, promoting bone growing, wound healing and immune effect.Leptin, by leptin receptor, activates JAK2/STAT3, PI3K and MAPK signal cascade.Leptin resistance can be used as the pathophysiological mechanism of an obesity, and it is expressed and lowers is the unresponsive performance of Leptin signaling.

This experiment condition drag group is lowered 2.19 times of leptin receptor genes with natural control group than model group, illustrates: model group, by lowering leptin receptor gene, has been strengthened leptin resistance, to the Leptin signaling delay of response, increases the weight of liver fat and becomes.

Under this experiment condition, gavage buccal tablet group of the present invention raises 4.95 times of leptin receptor genes with the model group ratio, illustrate: buccal tablet can be by improving OB-RmRNA after intervening, improve leptin receptor content, strengthen the respond to leptin, to alleviating the liver fat change, positive effect is arranged.

3.3.3 Srd5a1: steroids 5 alpha-reductases, α peptide 1, distribution endoplasmic reticulum, microsome, 3-C-5 α steroids 4 dehydrogenases, participate in cell signalling, Sex Determination, Cell Differentiation.Can be converted into another kind of androgen-dihydrotestosterone by the catalysis testosterone, therefore play a significant role in property differentiation and androgen physiology, this enzyme has 1,2 two isomers of Srd5a.Mainly be expressed in skin and adipose tissue and liver, but the catalysis overwhelming majority the 4th, and on 5 carbon atoms, the reduction reaction of unsaturation steroids, make 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 with tissue local, have the GC concentration of physiologically active closely related.The Srd5a1 activity strengthened in liver causes serum cortisol (COR) clearance rate to increase, and then compensatory hypothalamo-pituitary-adrenal axis (HPA) increased activity, COR secretes increase, further cause abdomen type obesity and MS(MS to refer to a series of metabolic disorders and the gathering of cardiovascular and cerebrovascular hazards on same individuality, be the syndrome of the multiple metabolic disorder clinical signs such as obesity, hypertension, dyslipidemia and IGT).

This experiment condition drag group and natural control 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, makes model group MS occur, finally makes liver cell that fat occurs and becomes.

After intervening with buccal tablet under this experiment condition with intervene before the model group ratio, the expression of Srd5a1 gene has been lowered 2.6 times, illustrate: buccal tablet of the present invention can be by lowering this gene expression, the metabolic syndrome that improvement brings due to excessive drinking, this has explained the reason why buccal tablet of the present invention can prevent and treat metabolic syndrome from another point of view.

3.3.4 Sult2a2: sulfotransferase family, endochylema, 2A, dehydrobenzene (DHEA) is preferential, and the member 2, participate in the fat metabolism, steroid metabolism, cholic acid catabolism.Sulfate transferase (sulfotransferase, Sult) be body catalysis multiple in and the key enzyme of exogenous compound sulphation metabolism, sulfating reaction is at various allogenes and endogenous thing, in biotransformation as medicine, food additives, carcinogenic substance, steroid hormone, neurotransmitter, be a kind of important association reaction, sulfate transferase superfamily mainly is responsible for this reaction of catalysis.The sulfonyl (SO3-) of Sult catalysis 3 AMPs-5 phosphoric acid sulfate (PAPS) is transferred to allogene and endogenous thing, reduces its biologically active and increases its excretion rate.Mankind Sult expresses in a lot of histoorgans, is main detoxication enzyme in human body.The research in recent years discovery, the generation of some diseases may be extremely relevant with Sult.Within 1987, at first in rat, clone Sult 2A2, originally thought that it was a kind of Senescence Marker Protein, after this cloning and identification goes out Sult gene family member in succession.Up to the present, at least find that there is 9 Sult families and 14 subfamilies.The Sult superfamily divides two classes, and a class is to be positioned at film on golgiosome in conjunction with Sult, mainly peptide class, protein, lipid and polysaccharide is carried out to sulphation; Another kind of is cytoplasm Sult, mainly to steroids, bile acid and neurotransmitter sulphation.Sulfating reaction is the important channel of sex hormone metabolism inactivation, and women's body inner estrogen mainly contains E2 and E1, and E2 and E1 form the steroidal sulfuric ester of non-activity by sulphation, OES (E1S) and sulfuric acid estradiol (E2S).People recognize that the sulfuration steroids is not only final excretory product gradually in recent years, have also participated in other many important biological respinses, as sulphation steroid hormone dehydrobenzene (DHEA) is a kind of fundamental mechanism of body modulability hormonal activity.The sulfuration cholesterol is relevant with many bioprocess, as cholesterol biosynthesis regulation and control, capacitation, fibrin ferment and plasmin activity, mechanism of Protein Kinase C.And the sulfate transferase of catalysis vulcanization reaction is being controlled the ratio of substrate sulphation form and non-sulfuric acid form to a certain extent, thereby affect many bioprocess.There is important Practical significance so inquire into the adjusting of sulfate transferase.

This experiment condition drag group and natural control group ratio, the up-regulated of Sult2a2 gene 5.16 times, illustrate: model group, by improving Sult2a2 gene expression, promotes liver cell to sex hormone and cholesterol metabolic, promotes liver cell that fat occurs and becomes and hypercholesterolemia.

After intervening with buccal tablet under this experiment condition with intervene before the model group ratio, the expression of Sult2a2 gene has been lowered 3.42 times, illustrate: buccal tablet of the present invention can be by lowering Sult2a2 gene expression, improvement, because the liver cell that excessive drinking brings is unbalance to sex hormone and cholesterol metabolic, prevents and treats hypercholesterolemia and fat change.

3.4 the impact that the gavage buccal tablet is expressed key gene relevant to controlling normal activity in liver after intervening

Group	Dusp1	Hspb1	Irs2	Socs2
					Relieve the effect of alcohol model group/natural group	↓6.18	↓2.64	↓21.42	↓33.58
Buccal tablet group/the model group of relieving the effect of alcohol	↑2.76	↑3.22	↑3.0	↑10.73

Notes: in table, ↓ 6.18 expression Dusp1 genes model group in liver cell mRNA is lowered 6.18 times with the expression ratio of natural control group, 2.76 times of the expression ratio rises of ↑ 2.76 expression Dusp1 gene gavage buccal tablet groups and model group.

3.4.1 Dusp1: dual specificity phosphatase enzyme 1(MKP-1), participate in the Argine Monohydrochloride dephosphorylation, reply oxidative pressure, the cell cycle.Studies confirm that, silk splits in the systemic inflammatory reaction that plain activated protein kinase phosphatase Dusp1 causes at gramnegative bacterium lipopolysaccharides (LPS) and plays important regulating and controlling effect, thereby the activation that can suppress p38MAPK prevents that inflammatory mediator from excessively discharging.Studies have shown that, 17 beta estradiols (E2) can be by increasing cardiac muscle cell MKP-1 protein expression, and the cardiac muscle cell ERK1/2 activity that inhibition endothelin-1 (ET-1) is induced increases, thereby suppress the myocardial hypertrophy reaction that ET-1 induces.This enzyme is liver specificity bidirectional modulation phosphatase, and the MAPKs dephosphorylation is had than high specific, and the activity of MAPKs is regulated important meaning is arranged; Inflammation reaction is the main inducing that causes liver fibrosis, lowers this gene expression and is conducive to liver and controls Fibrotic process, significant to improving liver function.

This experiment condition drag group and natural control group ratio, the expression of Dusp1 gene has been lowered 6.18 times, illustrates: model group, by lowering the expression of Dusp1 gene, activates the p38MAPK approach, promotes that inflammatory mediator excessively discharges, the inflammatory reaction of enhance hepatocyte.

Model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition, the up-regulated of Dusp1 gene 2.76 times, illustrate: buccal tablet of the present invention can be by raising the expression of this gene, suppress the p38MAPK approach, alleviate hepatocellular inflammatory reaction, inflammatory reaction is the main inducing that causes liver fibrosis, and reducing inflammation is exactly the fibrillatable of preventing liver organization.

3.4.2 Hspb1: much research shows, no matter body in stress situation or normal physiological state, all can participate in complicated and important biological process.As sHSP and other albumen are interacted, stop unnecessary interaction between albumen, maintain various cell proteins stable; Carry out the maintenance of film, and relevant with the mobility of film fat; There is driving functions in core, or the Molecular regulator process, or protected protein; The stabilized cell skeleton, participate in regulating growth and the differentiation of cell, suppresses the apoptosis of cell; Also there is the functions such as immunology; In the pathophysiological process of the physiological status that maintains body and some disease, all play a significant role.

This experiment condition drag group and natural control group ratio, the expression of Hspb1 gene has been lowered 2.64 times, illustrates: model group, by lowering the expression of Hspb1 gene, descends to damaging the hepatocellular ability of maintenance, is unfavorable for hepatocellular reparation.

Alcoholic fatty liver model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition, 3.22 times of the up-regulateds of Hspb1 gene, illustrate: after buccal tablet is intervened, by the expression of raising the Hspb1 gene, improve the hepatocellular ability of maintenance, promote the reparation of hepatocellular injury.

3.4.3 Irs2: IRS 2, Irs2 expresses at liver and pancreatic beta cell, the participation glucose metabolism, signal transduction, positive regulating cell propagation, the Irs2 protein expression is lowered and is made the active attenuating of PI3-K, can increase the weight of the liver insulin resistance.Use gene Knockout to confirm that the defect of Irs2 gene can cause the discoveries such as insulin resistance, Kubota, after knocking out Irs2, liver shows obvious insulin resistance, and to the insulin sensitivity of skeletal muscle without effect.

This experiment condition drag group has been lowered 21.42 times of IRS 2 genes with natural control group than model group, illustrate: the model group rat has been lowered 21.42 times because excessive drinking causes IRS 2 genes, must make IRS synthesize is greatly affected, obviously can have influence on the sensitiveness of liver cell to insulin response, impel model group to produce serious insulin resistance, bring out a series of metabolic syndromes, finally form fatty liver.

After intervening with buccal tablet under this experiment condition with the model group ratio, 3.0 times of the expression of Irs2 gene have been raised, illustrate: gavage buccal tablet of the present invention can improve the sensitiveness of liver cell to insulin response by the expression of raising the Irs2 gene, and buccal tablet of the present invention can play certain alleviation Insulin Resistance.

3.4.4 Socs2: Suppressor of Cytokine Signaling is 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.There are 8 members in this family containing the SH2 domain, i.e. CIS and Socs-1～7, this structure directly with the phosphotyrosine interaction, be that inhibiting key plays in Socs family.Extensively, to fat deposition, skeletal development, central nervous system effect, immune response, cancer occurs all to play an important role in Socs 2 effects.Socs 2 can regulate the hormone secretions such as insulin, prolactin in addition, and the cytokine signaling paths such as GH/1GF, insulin, interleukin are risen and promote or inhibitory action.Cytokine profiles is by lipometabolic balance in complicated signal network regulating cell and body, and most research at present concentrates 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 control group ratio, the expression of Socs2 gene has been lowered 33.58 times, illustrate: model group excessive drinking for a long time can cause metabolism " brake " regulator control system in liver cell seriously malfunctioning, will inevitably exert an influence to normal fat metabolism and insulin action in liver cell, form dysbolism, finally impel liver cell fat to become.

After intervening with buccal tablet of the present invention under this experiment condition with intervene before the model group ratio, the up-regulated of Socs2 gene 10.73 times, illustrate: lozenge of the present invention can be by improving the expression of Socs2 gene, in the liver cell that reparation causes due to excessive drinking, " brake " regulator control system is malfunctioning, significant to improving liver cell fat metabolism and insulin action.

By the mutation analysis in model group gene expression, be not difficult to find that the consequence of these variations causes Fatty synthesis to rise exactly, the fatty acid oxidation ability descends, and gluconeogenesis weakens, and produces and is similar to insulin and leptin resistance effect, and liver occurs that serious fat becomes; And the change of the gene expression caused due to long-term excessive drinking after gavage buccal tablet of the present invention is adjusted, the liver organization function is restored, and fatty liver is effectively controlled, so take buccal tablet of the present invention, can either bring into play the effect of relieving the effect of alcohol, can prevent and treat well fatty liver again, be worthy to be popularized.

In a word, by above test, show that product of the present invention can effectively participate in the body process of relieving the effect of alcohol, effectively control fat is in generation and the accumulation of liver.At present China Bee Pollen, propolis and royal jelly mix relieve the effect of alcohol aspect research at the early-stage, by experiment, find Bee Pollen water extraction liquid, propolis and royal jelly mix after to the antialcoholism action better effects if, the medicine relieved the effect of alcohol for exploitation proposes new approaches.Bee Pollen, propolis and royal jelly are all China's Traditional health care products, can improve hepatocyte activity, improve multiple blood parameters, and the fatty liver that prevention causes due to long-term excessive drinking is significant to the health care of safeguarding and improve the excessive drinking of Chinese.Bee Pollen, propolis and royal jelly can be used as for the first-selected health products that relieve the effect of alcohol, promotion and application in addition.

Claims (1)

1. the Bee Pollen intermixture buccal tablet for relieving the effect of alcohol, comprise bee glue powder, royal jelly freeze-dried powder, D-sorbite, dolomol and menthol, characterized by further comprising the freeze-dried powder that the molecular weight of obtaining with the Bee Pollen water extracting method is less than the Bee Pollen water extract of 5000D; The freeze-dried powder of described Bee Pollen water extract, bee glue powder, royal jelly freeze-dried powder, D-sorbite, dolomol and menthol account for respectively 50～70%, 10～25%, 10～25%, 5～10%, 0.2～0.6%, 0.1～0.4% of buccal tablet gross mass; Preparation method's step of Bee Pollen water extract is: 1) Bee Pollen removal of impurities, drying make water content drop to 4～8%; 2) adopt the mode of air-flow crushing broken wall by broken wall of melissa pollen; 3) soak the Bee Pollen of broken wall with the normal pressure and temperature pure water, process 10～40 minutes with low-frequency ultrasonic waves simultaneously, control the temperature of ultrasonic extract at 10～40 ℃; 4) turn refrigerated centrifugation with 8000～15000 and remove solid impurity, get supernatant; 5) by supernatant under the environment temperature of 1～5 ℃ with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight and be the following Bee Pollen water-soluble substances of 5000D;

The preparation method of bee glue powder is: 1) after propolis-18 ℃ freezing 20-40 hour, pulverize; 2) under condition of normal pressure, by concentration, be that 50%～90% edible alcohol mixes by the weight ratio of 1～10:1 with propolis, process 10～40 minutes with low-frequency ultrasonic waves, then soak 1～4 day, and keep 10～40 ℃ of room temperatures; 3) turn refrigerated centrifuge with 8000～16000 after the immersion and remove solid impurity in 10～15 minutes, get the supernatant that ethanol dissolves propolis; 4) by supernatant at room temperature with 5000D molecular sieve ultra-filtration and separation, obtain the propolis extracted with alcohol that molecular weight is less than 5000D; 5) the ethanol distillation recover reclaims ethanol and obtains the dry thing that the propolis solid content is greater than 90%.

Images