CN102920737A - Bee pollen water extract buccal tablet used for preventing alcoholism - Google Patents

Abstract

The invention discloses a bee pollen water extract buccal tablet used for preventing alcoholism, relating to the technical field of preparation of the buccal tablet from a bee pollen water extract. The buccal tablet mainly consists of freeze-dried bee pollen water extract powder, sorbitol, starch slurry, magnesium stearate and menthol crystal, wherein the freeze-dried bee pollen water extract powder is obtained by using a bee pollen water extraction method and has the molecular weight being smaller than 5000D. The buccal tablet disclosed by the invention has obvious effect for preventing alcoholism, has no side effects, and is low in cost and simple and easy to implement.

Description

For the bee pollen water extract buccal tablet that relieves the effect of alcohol

Technical field

The present invention relates to from plant, particularly prepare the Technology field of buccal tablet from the bee pollen water extract.

Background technology

China is the earliest the country of making wine in the world, 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 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.

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.The bee pollen water extract has the active component of a lot of adjusting physiological metabolism effects, has heat-clearing and toxic substances removing, and the prevention fatty liver suppresses hepatocyte and worsens, and reduces the effects such as onset of liver cancer rate.By the research of molecular mechanisms of action that bee pollen is relieved the effect of alcohol, has far-reaching value for researching and developing new solution drinks medicine.

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, mainly can reach by following several approach by the research relieving alcoholism and protecting the liver:

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.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 bioavailability of ethanol in human body, so improve ADH activity in the stomach, strengthens ethanol first pass metabolism under one's belt, can alleviate the load to liver.In the situation of single low alcohol consumption, take medicine first and drink again, can improve ADH activity in the liver, the ethanol of degrading rapidly effectively stops the absorption of ethanol, reduces blood alcohol concentration, to reach the purpose of relieving the effect of alcohol.Radix Puerariae, Flos puerariae lobatae, Semen Hoveniae (Fructus Hoveniae) are the most representative solution drinks of Chinese medicine medicines.Find after deliberation, the effective ingredient of their performance antialcoholism actions is Flavonoid substances, the trifoliate orange yellow party 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 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, 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 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, and the activity of SOD and GSH-PX embodies the ability of Scavenger of ROS, 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, helps body to keep normal redox state.Aldehydes matter in the fermentation Fructus Hordei Vulgaris extract can be removed free radical and the active oxygen that is produced by alcohol dehydrogenase enzyme system and CYP2E1 system, slow down the hepatic injury that response to oxidative stress causes, the water extract of Herba thymi vulgaris and Rhizoma Zingiberis Recens can effectively reduce the content of enzymatic oxidation material in the liver and improve the level of antioxidant.

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 bee pollen 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.

Bee pollen is the stamen that is gathered the sources of nectar and pollen by Apis, the dough of carrying back nest through processing, its nutritional labeling is very abundant, not only contains rich in protein, saccharide, lipid, vitamin, organic acid, mineral, nucleic acid, also contains enzyme and other biological active substance.Bee pollen is usually used in health food and nutraceutical, its chemical constituent 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 " the most completely nutriment ", " 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 product of declaring only have kind more than 20, and how the development of new functional food is very worth research and discussion.Yet research is also grasped the Changing Pattern of the crucial physical and chemical indexs such as protein, ash, moisture and Determination of Vitamin C of bee pollen, is 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 applied research will deepen continuously, and will give human more value of creating.In recent years, pollen has some progress at medicine, food, health food, cosmetic field; Such as, pollen treatment prostatitis, tumor, hepatitis, cardiovascular disease, and the pollen wet goods research of pollen polysaccharide Sulfation, pollen metabolism, pollen pharmaceutical carrier, pollen recombiant protein, pollen fermentation food, pollen antiseptic, SCF-CO 2, the exploitation of China's bee pollen has wide market.

Bee pollen is the dietotherapeutic pollen kind in the Chinese medicine treasure-house, as Chinese traditional medicine, the history of its dietotherapeutic has exceeded thousands of years, Shennong's Herbal before 2400, Compendium of Material Medica are all on the books to the Pharmacopoeia of the People's Republic of China of today, are regarded as " elixir " with its magical effect especially among the people.Find in pollen, also to exist a class to have the material that promotes alcohol metabolism through our up-to-date research, have remarkable effect to promoting the body aspect of relieving the effect of alcohol.China is along with the rapid development of economy at present, and various dinner parties are more and more, and the sickness rate of the various diseases that brings out owing to drinking improves year by year, and the healthy problem that excessive drinking is brought out has become affects modern's importance of quality of life.

Modern study proves that pollen polysaccharide can improve quantity and the function of body T lymphocyte and macrophage, improves seroimmunity Protein G level; the enhancing human 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 the bee pollen can be worked in coordination with mutually the performance antioxidation; Bee pollen is with its efficient antioxidant activity, suffers from the disease relevant with radical damage and plays an important role aspect the defying age in that control is human; 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, resisting fatigue, anti-aging, vessel softening, blood fat reducing, the various biological functions such as short hemopoietic, beauty treatment and weight reducing, cellular immunization, humoral immunization all there are obvious potentiation, but utilize the bee pollen aspect research of relieving the effect of alcohol to remain blank.

Find that through research 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 antialcoholic drugs, differ too great disparity because dose is compared with stomach, 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 preferably adopts the mode without stomach.

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.

Find through the inventor's for many years research that this class material exists and be water soluble state in the bee pollen, thereby separation and Extraction is difficult especially, we successfully separate recently, and done the bee pollen correlation molecule research on mechanism work of relieving the effect of alcohol, the key gene that finds it to play a role, its effect of relieving the effect of alcohol is ideal, for this reason, proposes this application for a patent for invention.

Summary of the invention

The object of the invention is to invent a kind of bee pollen water extract buccal tablet that can efficiently relieve the effect of alcohol.

The present invention includes the molecular weight obtained with the bee pollen water extracting method forms less than lyophilized powder, Sorbitol, starch slurry, magnesium stearate and the Mentholum of the bee pollen water extract of 5000D.

Described lyophilized powder, Sorbitol, starch slurry, magnesium stearate and Mentholum alcoholic solution account for respectively 50～85%, 5～20%, 5～25%, 0.2～0.4%, 0.1～0.4% of buccal tablet gross mass.

Bee pollen is the stamen that is gathered the sources of nectar and pollen by Apis, the dough of carrying back nest through processing, its nutritional labeling is very abundant, not only contains rich in protein, saccharide, lipid, vitamin, organic acid, mineral, nucleic acid, also contains enzyme and other biological active substance.Bee pollen water extraction liquid can be brought into play the pharmacological action of relieving the effect of alcohol at gene level, and the inventor's research has solved its molecule mechanism of relieving the effect of alcohol substantially, for this reason, files an application patent of the present invention.

Through test, the present invention adopts bee pollen water extract lyophilized powder as the solution liquor raw material, total protein content 0.25-2% in the lyophilized powder, and water content is lower than 8%, have no side effect, and expense is cheap, and is simple.Product advantage of the present invention is that the water extract that is used for relieving the effect of alcohol in the bee pollen can directly absorb by hypoglossis mucous membrane, also enter rapidly blood, avoids gastric acid to its destruction, and therapeutic effect is better.

Find many bioactive substances are arranged under one's belt easily by stomach acids destroy in the bee pollen 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.

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 gavage Chinese liquor model group liver tissue slices figure.

Fig. 4 is gavage Chinese liquor 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 pass through the pollen that Apis is gathered such as Flos Camelliae Japonicae powder, Pollen Brassicae campestris, Flos Nelumbinis pollen etc.

One, the preparation bee pollen water extract lyophilized powder that relieves the effect of alcohol:

1, bee pollen remove impurity, drying make water content drop to 4～8%.

2, adopt the mode of comminution by gas stream breaking cellular wall with broken wall of melissa pollen.

3, soak the bee pollen of breaking cellular wall with the normal pure water of normal pressure, processed 10～40 minutes with low-frequency ultrasonic waves simultaneously, the temperature of control supersound extraction liquid is at 10～40 ℃.

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

5, with described supernatant under 1～5 ℃ ambient temperature with 5000D molecular sieve ultra-filtration and separation, obtain molecular weight and be the following bee pollen water-soluble substances of 5000D.

6, above-mentioned bee pollen water extract lyophilization 24～48 hours preparation water content is lower than the lyophilized powder of 8% bee pollen water extract.

Two, the preparation bee pollen buccal tablet that relieves the effect of alcohol:

1, granulating process:

Sorbitol is made into 10～30% solution for standby.

After the pot that will seethe with excitement is preheating to 40～60 ℃, first lyophilized powder, Sorbitol, starch slurry are added in the boiling pot, arrange according to the following table parameter, open peristaltic pump spray sorbitol solution and granulate.Granulate and finish rear magnesium stearate and the Mentholum alcoholic solution mix homogeneously of adding, cold drying is for subsequent use.

Typical several quality proportioning table: (unit: kg)

?	Lyophilized powder	Sorbitol	Starch slurry	Magnesium stearate	The Mentholum alcoholic solution
						Proportioning 1	85	8	6.4	0.3	0.3
Proportioning 2	80	6	13.5	0.3	0.2
						Proportioning 3	70	9	20.6	0.2	0.2

Processing parameter:

Parameter	Inlet temperature	Leaving air temp	The air inducing frequency	Peristaltic pump	Atomizing pressure
						During whitewashing	60-80℃	40-60℃	40Hz	200rpm	0.2MPa
After the whitewashing	65-80℃	50-60℃	35Hz	200rpm	0.2MPa

(2) 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.

Three, 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 after the body weight 1% gavage strong, colourless liquor distilled from sorghum.

(2) experimental group adopts the mode of empty stomach gavage buccal tablet of the present invention to test using dosage:

Gavage preparation: pressed the Oral Administration in Rats buccal tablet 40～80mg/ days, and connected and took 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.

Be polygon from Fig. 1,2 visible male white rat liver hepatocyte clear in structure, hepatic sinusoid is obvious, and the hepatic cords marshalling does not have obvious fat vacuole in the hepatocyte; Kidney group 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 gavage bee pollen water extraction liquid buccal tablet liver 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, on an empty stomach after the gavage buccal tablet intervention of the present invention to the comparative analysis of rats in test groups Main Blood Biochemical Index.

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	The buccal tablet group
				ALT	73.00±16.67	91.25±29.33	64.29±15.39
ALP	102.50±11.90	146.75±21.70	90.43±6.83
				LDH	962.75±96.05	909.75±326.58	1523.86±430.29
CHO	1.23±0.08	1.52±0.26	1.27±0.16
				TG	1.25±0.18	1.84±0.37	1.11±0.25
BUN	5.89±0.65	4.78±0.71	5.11±0.54
				ALB	15.50±0.43	14.88±0.53	16.09±0.54

Through the SPSS statistics, model group glutamate pyruvate transaminase (ALT) is extremely remarkable with gavage bee pollen buccal tablet group difference; The transaminase extensively exists in the histiocyte, and especially the strongest with activity in the tissue such as liver, cardiac muscle, brain, kidney, plasma content is very low when normal, and ALT mainly is present in liver, and ALT is released in a large number blood and can causes that the transaminase raises in the blood 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 also causes the obstacle of lipid metabolism to reach at intrahepatic deposition simultaneously because insulin secretion obstacle or insulin resistant cause metabolism obstacles of blood glucose, even forms fatty liver, and makes liver damage cause the rising of serum levels of ALP level.Through the SPSS statistics, model group alkali phosphatase and other two experimental grouies difference are extremely remarkable; Illustrate that the bee pollen buccal tablet can improve hepatocyte injury, reduce the alkaline phosphatase enzyme level, rising and the hepatocellular damage of alkali phosphatase (ALP) are closely related, illustrate that the excessive drinking of this experimental model group rat is very serious to the hepatocyte injury, and can improve the untoward reaction that long-term excessive drinking causes behind the gavage buccal tablet, alleviate hepar damnification.

Through the SPSS statistics, natural matched group and model group lactic acid dehydrogenase (LDH) are extremely remarkable with gavage buccal tablet group of the present invention difference; Lactic acid dehydrogenase is glucolytic key enzyme, the lactic dehydrogenase enzyme level is very high behind the gavage bee pollen water extraction liquid buccal tablet, 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, be beneficial to that liver relieves the effect of alcohol and carbohydrate metabolism.

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 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 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 (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 reduce the cholesterol levels that causes owing to indulging in excessive drinking and raise, significant to the generation that prevention is brought out type 2 diabetes mellitus owing to indulging in excessive drinking for a long time.

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 (TG) and natural matched 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 type 2 diabetes mellitus; And take buccal tablet of the present invention the Triglyceride Metabolism that brings out owing to excessive drinking had good regulating and controlling effect.

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.Through SPSS statistics, model group blood urea nitrogen and natural matched group significant difference; Under this experiment condition, the model group rat is because excessive excessive drinking, drunk serious, cause feed intake to descend and protein metabolism disturbance, so that blood urea nitrogen does not rise counter falling, be minimum in all experimental grouies, and gavage buccal tablet experimental group of the present invention, metabolism to protein is improved effect, and urea nitrogen levels gains momentum.

Add up through SPSS, model group albumin level and gavage buccal tablet group of the present invention difference is extremely remarkable, can find that from the albumin level variation gavage buccal tablet can to because long-term excessive drinking causes albumin level lowly to have regulating and controlling effect, namely can improve the protein in body metaboilic level.

Above result shows: buccal tablet of the present invention is to having sizable therapeutical effect because the rat liver fat that excessive drinking causes becomes, and from blood parameters and liver tissue slices, the improvement effect of buccal tablet of the present invention is very clear and definite.

3, after the buccal tablet intervention of the present invention major gene in the rats in test groups liver metabolism express is changed comparative analysis, such as following table:

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

Ehhadh

Fasn

Fbp2

LpL

Scd

Scd1

Model group/naturally organize

↓3.72

↓2.74

↑4.86

↓4.57

↓2.82

↑5.04

↑5.65

Buccal tablet group/model group

↑3.52

↑2.96

↓2.67

↑2.41

↑3.26

↓7.79

↓8.42

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, 3.52 times of the expression ratio rises of ↑ 3.52 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, mitochondrion fatty acid beta-oxidation ability is weakened, strengthen liver fat and become.

Gavage buccal tablet group has raised 3.52 times of this genes with the model group ratio under this experiment condition, can promote the fatty acid beta-oxidation of liver organization, and the fatty liver that the control excessive drinking is caused has positive effect.

3.1.2 Ehhadh: difunctional peroxidase (peroxisomalbifunctionalenzyme), enoyl-CoA hydratase, peroxisome participates in lipid 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 fatty acid beta-oxidation of reported first, plays vital effect in the beta-oxidation of polyunsaturated fatty acid, 2-methyl branch fatty acid and D-isomer hydroxy fatty acid.Under the diabetic disease states rising of blood plasma very-long-chain fatty acid and levels of polyunsaturated fatty acids whether with the beta-oxidation of peroxisome fatty acid, particularly the change of D-BP activity is relevant.Eukaryotic mitochondrion and peroxisome all carry out the beta-oxidation of fatty acid, carbon chain lengths is less than the mainly beta-oxidation energy supply in mitochondrion of fatty acid of 20 carbon, and carbon chain lengths greater than 2O carbon saturated/unsaturated very-long-chain fatty acid, dimethyl branched chain fatty acid and the mainly oxidation in peroxisome of bile acid biosynthesis metabolic intermediate, then the Short-Chain Fatty Acids of generation changes the mitochondrion exhaustive oxidation over to.Second step and the three-step reaction of peroxisome fatty acid beta-oxidation approach, it is alkene acyl CoA generates 3-ketoacyl CoA through hydration, dehydrogenation reaction, 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 to exist in the peroxisome: 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 studies show that L-BP mainly works in the oxidation of satisfied 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 fatty acid, and D-BP defective patient's research also found, D-BP participates in utmost point long-chain straight chain fatty acid, for example Major Enzymes of lignoceric acid beta-oxidation.The content of D-BP and L-BP is almost equal in liver, approximately contains 2.5mg in every gram protein, and it is of equal importance to illustrate that two kinds of bifunctional proteins play a part in the beta-oxidation of hepatocellular peroxisome fatty acid.There are two kinds of multifunctional enzymes to have L-3-hydroxyl acyl CoA dehydrogenase activity in the higher organism body, be respectively the L-BP that is present in the peroxisome, and have three functional proteins of enoyl-CoA hydratase/L-3-hydroxyl acyl CoA dehydrogenase/3-ketoacyl-CoA thiolase activity in the mitochondrion.Known, the fatty acid beta-oxidation activity of mitochondrion and peroxisome increases simultaneously during diabetes, research is found, the beta-oxidation of diabetes rat liver peroxisome fatty acid is active also to raise simultaneously with liver L-3-hydroxyl acyl CoA dehydrogenase activity, and the beta-oxidation of mitochondrion and peroxisome linear saturated fatty acids, L-isomer hydroxy fatty acid strengthened when diabetes were described; 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-isomer long-chain hydroxy fatty acid probably weakens, and causes the intermediate product D-isomer 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 one of reason that the blood plasma fatty acid spectrum is unusual when causing diabetes.

Excessive drinking model group and natural matched group are than 2.74 times of the expression of the difunctional peroxidase gene of downward modulation under this experiment condition, illustrate the model group rat indulge in excessive drinking for a long time can by the downward modulation this gene, make that fatty acid beta-oxidation ability weakens in peroxisome and the mitochondrion, 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.

Gavage buccal tablet group has raised 2.96 times of difunctional peroxidase genes with the model group ratio under this experiment condition, can promote the fatty acid beta-oxidation in liver organization peroxisome and the mitochondrion, to preventing and treating the fatty liver that causes 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 buccal tablet control.

3.1.3 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, causes fatty liver generation primary factor.

2.67 times of the gene expressions of gavage buccal tablet group and model group downward modulation fatty acid synthetase under this experiment condition illustrates: the buccal tablet group can be by reducing the synthetic of fatty acid synthetase, and it is synthetic to reduce the hepatic tissue fatty acid, and is significant to alleviating the liver fat change.

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

This experiment condition drag group leader's phase indulges in excessive drinking and makes that Fbp2 gene and natural matched group illustrate than 4.57 times of down-regulated expressions in the hepatocyte: model group can descend model group glyconeogenesis ability by the expression of this gene of downward modulation, and is consistent with the weight fat liver.

Gavage buccal tablet group and model group raise 2.41 times of Fbp2 gene expressions under this experiment condition, illustrate: the buccal tablet group can promote hepatocellular glyconeogenesis level by raising Fbp2 gene expression, alleviates hepatocyte fat and becomes.

3.1.5 LpL: lipoprotein lipase, participate in fatty acid metabolism, be one of lipometabolic key enzyme.LpL is synthetic at the rough endoplasmic reticulum of parenchyma, and new synthetic LpL non-activity after glycosylation, just changes into active LpL.The heparin sulfate glycoprotein that is present in the cell membrane outer surface makes enzyme keep a kind of unvital enrichment stage, stimulated the LpL that obtains activating in the rear blood plasma by heparin, be distributed in the lipoprotein that contains triacylglycerol, it mainly is the triacylglycerol that decomposes Chylomicron and very low density lipoprotein (VLDL), and combination and being attached in these remnant lipoproteins, may absorb as liver the signal of these granules.LpL acts on lipoprotein, mainly decomposes the triacylglycerol in the lipoprotein, is to remove the rate-limiting enzyme of triacylglycerol in the blood plasma, and impels lipoprotein to shift cholesterol, phospholipid and apolipoprotein.LpL also has to be increased Chylomicron and is attached to ability on the LP receptor, promotes the Chylomicron picked-up.

This experiment condition drag group and 2.82 times of natural matched group ratio downward modulation lipoprotein lipase gene expressions, illustrate: model group can make the lipoprotein lipase content decrease, causes triacylglycerol to decompose and reduces, the change of acceleration model group rat liver fat.

Gavage buccal tablet group and model group illustrate than 3.26 times of up-regulateds under this experiment condition: the buccal tablet group can make that lipoprotein lipase is synthetic in the experimental group rat hepatocytes increases, and is conducive to hepatocyte and reduces fat, and alleviates liver fat and becomes.

3.1.6 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.

3.1.7 Scd1: sterin coenzyme A desaturase 1, it is synthetic to participate in fatty acid biological.Stearyl-coenzyme A desaturase 1 (Scd-1) is the biosynthetic rate-limiting enzyme of monounsaturated fatty acid, plays 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 sterin coenzyme A desaturase (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 7.79 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 8.42 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 bee pollen buccal tablet control fatty liver.

3.2 behind the gavage buccal tablet in the liver with hepatic injury, repair the impact of relevant key gene expression

Group	Il7r	Pla2g2a	Tgfb2	Tnfsf10
					The model group/naturally of relieving the effect of alcohol group	↓2.75	↑2.76	↑3.63	↑2.92
Buccal tablet group/wine model group	↑4.58	↓2.45	↓5.68	↓2.82

Annotate: in the table, 2.75 times of the expression ratio downward modulations of ↓ 2.75 expression Il7r genes model group and natural matched group in hepatocyte mRNA, 4.58 times of the expression ratio rises of ↑ 4.58 expression Il7r gene gavage buccal tablet groups and model group.

3.2.1 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 2.75 times of interleukin-17 genes with natural matched group than model group, illustrate: the model group rat is because long-term excessive drinking can cause 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, can promote hepatocyte fat to become and Fibrotic process.

The buccal tablet group raises 4.58 times of interleukin-17 genes with the model group ratio under this experiment condition, illustrate: after 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.

3.1.2 Pla2g2a: phospholipase A2, basic 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.

This experiment condition drag group leader's phase indulges in excessive drinking and makes in the hepatocyte Pla2g2a gene and natural matched group than 2.76 times of up-regulateds, illustrate: long-term excessive drinking can be by raising Pla2g2a gene expression, increase the weight of the destruction to liver plasma membrane, cause the rising of serum transaminase.

Gavage buccal tablet group and 2.45 times of model group ratio downward modulation Pla2g2a gene expressions under this experiment condition, illustrate: the buccal tablet group can alleviate the destruction to liver plasma membrane by downward modulation Pla2g2a gene expression, promotes the reparation of hepatocyte injury.

3.2.3 Tgfb2: transforming grouth factor beta 2, participate in cell death, just regulating and control process, regulating cell propagation, regulation and control immunne response, dopamine biosynthesis.Studies confirm that to only have Tgf-β energy HSC stimulated rubber polymer fibril in the cytokine, other cytokines are a proliferation of HSC stimulated then.In the 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 consisted of the Autocrine of activation HSC, are the important mechanisms of the sustained activation of HSC.

This experiment condition drag group and natural matched group ratio, the up-regulated of Tgfb2 gene 3.63 times, illustrate: long-term excessive drinking causes hepatocyte injury, raise the expression of Tgfb2 gene, HSC stimulated rubber polymer fibril promotes the formation of hepatic fibrosis, and the result is consistent with section.

Model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition, the down-regulated expression of Tgfb2 gene 5.68 times, illustrate: buccal tablet of the present invention can be prevented and treated by reducing this gene expression the process of hepatic fibrosis, and is great to safeguarding hepatocyte homergy Functional Significance.

3.2.4 Tnfsf10: tumor necrosis factor surpasses family, and the member 10, participates in the guiding apoptosis, is just regulating and control NF-κ B cascade.

This experiment condition drag group and natural matched group ratio, the up-regulated of Tnfsf10 gene 2.92 times, illustrate: long-term excessive drinking is just being regulated and control NF-κ B cascade by raising the expression of Tnfsf10 gene, promote hepatocyte inner lipid peroxidating and inflammatory reaction, increase the weight of the process of hepatic fibrosis.

Model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition; the down-regulated expression of Tnfsf10 gene 2.82 times; illustrate: buccal tablet of the present invention can be by the expression of downward modulation Tnfsf10 gene; suppress the activation of NF-κ B in the rat body; alleviate the peroxidating of hepatocyte inner lipid and inflammatory reaction; alleviate hepatocyte fat change and fibrosis occur, play the target of protection hepatic tissue.

3.3 the impact of after the buccal tablet intervention key gene relevant with the control normal activity in the liver being expressed

Group

Igfbp6

Irs2

Irs3

Prlr

Socs2

Srd5a1

The model group/naturally of relieving the effect of alcohol group

↓3.16

↓21.42

↑2.47

↑12.62

↓33.58

↑4.0

Buccal tablet group/wine model group

↑2.15

↑2.36

↓3.52

↓9.78

↑8.84

↓4.15

Annotate: in the table, 3.16 times of the expression ratio downward modulations of ↓ 3.16 expression Igfbp6 genes model group and natural matched group in hepatocyte mRNA, 2.15 times of the expression ratio rises of ↑ 2.15 expression Igfbp6 gene gavage buccal tablet groups and model group.

3.3.1 Igfbp6: IGFBP (insulin-like growth factor binding protein) 6, regulating cell growth, signal transduction, negative regulation cell proliferation.Rise is conducive to Igf1 and plays a role, and control fatty liver and hepatic fibrosis are had fine effect.

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, IGFBP (insulin-like growth factor binding protein) 6 (IGFBP6) 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 3.16 times of IGFBP (insulin-like growth factor binding protein) 6 genes with natural matched group than model group rat, illustrate: model group has promoted hepatocyte fat to become and fibrosis by the expression of downward modulation IGFBP (insulin-like growth factor binding protein) 6 gene.

The buccal tablet group raises 2.15 times of IGFBP (insulin-like growth factor binding protein) 6 genes with the model group ratio under this experiment condition, illustrate: can be by raising the expression of this 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.

3.3.2 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 insulin resistant, bring out a series of metabolism syndromes, the final fatty liver that forms is that one of key molecule mechanism of insulin resistant is brought out in long-term excessive drinking.

Under this experiment condition with after the buccal tablet intervention with the model group ratio, 2.36 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.

3.3.3 Irs3: IRS 3, Irs3 expresses at liver and pancreatic beta cell, participates in glucose metabolism, and at present research generally believes that Irs3 can be combined on the Insulin receptor INSR, plays the negativity regulating action to Irs1, Irs2.

This experiment condition drag group has raised 2.47 times of IRS 3 genes with natural matched group than model group, illustrate: model group is by raising Irs3 gene expression, impel the IRS 3 of the synthetic more negative regulation IRSs 2 of hepatocyte, further increase the weight of hepatocellular insulin resistant, increase the weight of metabolism syndrome, promoting fat to become, is that one of key molecule mechanism of insulin resistant is brought out in long-term excessive drinking.

Under this experiment condition with after the buccal tablet intervention with the model group ratio, reduced 3.52 times of the expression of Irs3 gene, illustrated: gavage buccal tablet of the present invention can by the expression of downward modulation Irs3 gene, be alleviated hepatocellular insulin resistant, improve hepatocyte to the sensitivity of insulin, alleviate hepatocyte fat and become.

3.3.4 Prlr: hprl receptor, participate in the steroid biosynthesis, anti-apoptotic, the cell surface signal transduction activates tyrosine kinase, JAK2 albumen.PRL combines by the hprl receptor with the target cell membrane surface, starts the JAK2/STAT5 signal transduction path, finally activates trans acting factor STAT5.Modern study proof liver is one of main position of the synthetic immune effector molecule of body, and many members of JAK/STAT signal path express abnormal JAK/STAT signal transduction in hepatic tissue can cause multiple hepatic injury.

This experiment condition drag group has raised 12.62 times of prolactin receptor genes with natural matched group than model group, illustrate: model group is by raising prolactin receptor gene, the final trans acting factor STAT5 that activates is obstructed the transduction of JAK/STAT signal path, causes the generation of hepatic injury.

Under this experiment condition with after the buccal tablet intervention with the model group ratio, 9.78 times of the expression of prolactin receptor gene have been reduced, illustrate: gavage buccal tablet of the present invention can improve the JAK/STAT signal transduction pathway by the downward modulation prolactin receptor gene, and is favourable to the liver organization of repairing damage.

3.3.5 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: long-term excessive drinking 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 key molecule mechanism of insulin resistant 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 8.84 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.

3.3.6 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 4.15 times, illustrate: buccal tablet of the present invention can be by this gene expression of downward modulation, improve the metabolism syndrome that brings owing to excessive drinking, this has explained the reason why the bee pollen buccal tablet can prevent and treat metabolism syndrome from another point of view.

In a word, show the product of the present invention process of can the effective participation body relieving the effect of alcohol by above test, effectively control fat is in generation and the accumulation of liver.At present China bee pollen relieve the effect of alcohol aspect research at the early-stage, by experiment, find that bee pollen water extraction liquid is effective to antialcoholism action, the medicine that relieves the effect of alcohol for exploitation proposes new approaches.Bee pollen is China's Traditional health care product, can improve hepatocyte activity, improves multiple blood parameters, and prevention is because the fatty liver that long-term excessive drinking causes, the health care of safeguarding and improve the excessive drinking of Chinese is significant.Bee pollen can be used as for the first-selected health product that relieve the effect of alcohol, in addition promotion and application.

Claims (2)

1. for the bee pollen water extract buccal tablet that relieves the effect of alcohol, it is characterized in that mainly being formed by lyophilized powder, Sorbitol, starch slurry, magnesium stearate and the Mentholum of the molecular weight of obtaining with the bee pollen water extracting method less than the bee pollen water extract of 5000D.

2. described bee pollen water extract buccal tablet be used to relieving the effect of alcohol according to claim 1, it is characterized in that described lyophilized powder, Sorbitol, starch slurry, magnesium stearate, and the Mentholum alcoholic solution accounts for respectively 50～85%, 5～20%, 5～25%, 0.2～0.4%, 0.1～0.4% of buccal tablet gross mass.

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