CN102934808B - Pollen pini water extract buccal tablet for alleviating hangovers - Google Patents

Abstract

The invention provides a pollen pini water extract buccal tablet for alleviating hangovers, relating to the technical field of processes for preparing buccal tablets from pollen pini water extracts. The buccal tablet is mainly formed by lyophilized powder of the pollen pini water extract which is obtained by a pollen pini water extraction method and has molecular weight less than 5000D, kudzu root powder, orange peel powder, palatinitol, sorbitol and magnesium stearate. The buccal tablet has obvious effect of alleviating hangovers, does not have side effect, is low in cost and is simple and practicable.

Description

For the pollen pini 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 pollen pini 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 the daily life necessary part already.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 of Fa Shenging also constantly increases therefrom, has become second largest hepatopathy after virus 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, wine poison 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 based on clear relieving alcoholism, clearing heat and eliminating phlegm, supplementing qi and nourishing yin; 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 as ubenimex, Nimodipine, emetine etc. and to a certain degree improve crapulent effect in animal experiment and the clinical research.The pollen pini water extract has the active component of a lot of adjusting physiological metabolism effects, has clearing heat and detoxicatingly, prevents and treats ethylism, and prevention alcoholic liver, fatty liver suppress liver cell and worsen, and reduce effects such as onset of liver cancer rate.By the research of molecular mechanisms of action that pollen pini is relieved the effect of alcohol, for researching and developing new solution drinks medicine, adopt Pollen Pini extract to reduce the Health cost of bringing of drinking from the medicines and health protection angle and have actual application value.

Present plant alcohol intoxication-alleviating preparation roughly comprises with the Araliaceae being the ethanol absorption inhibitor of representative, is the liver metabolism promoter of representative with the root of kudzu vine, FI puerariae, and the latter is more paid attention to because it has hepatoprotective effect concurrently.

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

1, reaches the relieving alcoholism and protecting liver effect by the activity that improves ADH, ALDH gene expression

Research about the medicine that relieves the effect of alcohol 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 road barrier of alcohol metabolism, and gastric mucosa is impaired, can cause entering the plain increase of septicemia, increases the weight of the bioavilability of ethanol in human body, so improve ADH activity in the stomach, strengthens ethanol first-pass metabolism under one's belt, can alleviate load of liver.Under the situation of single low alcohol consumption, take medicine earlier and drink again, can improve ADH activity in the liver, the ethanol of degrading rapidly effectively stops the absorption of ethanol, reduces the blood determining alcohol, 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 Huang side (Semen Hoveniae (Fructus Hoveniae) and rheum officinale) to the influence of alcoholic liver injury rat stomach ADHmRNA experiment in, thereby Zhi Huangfang is proved and can improves alcohol and alleviate the load that alcohol causes liver at the first-pass metabolism of stomach.Trifoliate orange Huang side and clearing liver blood circulation promoting 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.Under 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 great drinker, 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 Huang side as described above and clearing liver blood circulation promoting recipe 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 big.Discover that acute alcoholism liver tissues of rats ADH, ALDH specific activity model group after the intervention of Semen Hoveniae (Fructus Hoveniae) water extract all significantly increase (P＜0.05), show that Semen Hoveniae (Fructus Hoveniae) water extract 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 (mainly being acetaldehyde) to the toxic action of liver, reach certain liver effect of protecting.Yet, only improve ADH, the ALDH activity can not be slowed down the ethanol toxic action of metabolism in vivo, also tackle the relevant key enzyme of alcohol metabolism overall process, carry out system research 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 liver effect by slowing down the oxidative stress effect

As mentioned above, the three approach of ethanol 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 takes place to become with fat liver organization.The factor relevant with 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 removing active oxygen, and they are the common counters in the relieving alcoholism and protecting liver research.Exist the balance of antioxidant system and enzymatic oxidation system in the liver, this balance will be broken behind the absorption alcohol, and the activity of SOD and GSH-PX reduces.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 influence 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 antioxidation 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 thyme and ginger can effectively reduce the content of enzymatic oxidation material in the liver and improve the level of polyphenoils.

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 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 oxidative stress of liver and the release of the promotion various kinds of cell factor, thus pathological changes such as the steatosis of participation liver, 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 Huang side 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 liver goods, how research influences the very few of level of endotoxin about goods, and bicyclic alcohols has been used for the treatment of slow virus hepatitis as the novel anti hepatitis medicament of Chinese independent development.Current research shows that bicyclic alcohols can reduce the plasma endotoxin level 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, from suppressing the effect that Kupffer cell activation angle realizes the protection liver organization.

In sum, the pathogenesis of ALD is relevant with multiple factor, mainly comprises: the key enzyme in the alcohol metabolism process, as ADH, ALDH, CYP2E1, CAT; The oxidative stress factor of influence is 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, also should launch around these key factors about the relieving alcoholism and protecting liver Its Mechanisms.Simultaneously, China's relieving alcoholism and protecting liver medicine, many based on Chinese herbal medicine in the health products, 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, and through we discover pollen pini effectively alleviation of alcohol to the damaging action of liver, the effect of relieving the effect of alcohol is obvious, 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 alcohol chemical damage.

Pollen pini is the pollen of pine trees such as gymnosperm Pinaceae Pinus masson pine and Chinese pine, and China utilizes and edible pollen pini country the earliest in the world.The resource major part of global pollen pini is distributed in China, but as the exclusive living resources of China, the pollen of masson pine and Chinese pine is not also developed fully.The medical care effect of pollen pini in recent years more and more by people cognition, but the material that exists a class to relieve the effect of alcohol in the pollen pini is just not known, discover for many years that through us this class material is water soluble state in pollen pini, thereby separation and Extraction difficulty especially, we successfully separate recently, and have done the relieve the effect of alcohol research work of correlation molecule mechanism of pollen pini, the key gene that finds it to play a role, its effect of relieving the effect of alcohol is very desirable, for this reason, proposes this application for a patent for invention.

The root of kudzu vine contains kudzu root flavone class material, improves hepatocellular power of regeneration, recovers the normal liver function; promote choleresis, prevent fat in the liver accumulation, the reinforcement liver detoxification function; prevent alcohol to the damage of liver, zoopery proves that it has good protective action to liver.Dried orange peel is except the well-known appetizing of regulating the flow of vital energy, and outside the eliminating dampness and eliminating phlegm effect, also has the disappear effect of wine of stopping nausea and vomiting by lowering the adverse flow of QI, fast spleen.Compendium of Material Medica is drawn forefathers' experience and is said " treating the wine disease ".Make a general survey of successive dynasties side's book, have many Fang Zhongjun that relieves the effect of alcohol to use dried orange peel.Control insobriety as " General Records of Holy Universal Relief " dried orange peel soup, the wine poison amasss at stomach, or vomits and do not eat, and thirsty many drinks respectively 30 restrain with dried orange peel, the root of kudzu vine, honey-fried licorice root, gypsum, are the end, every clothes 9 grams, decocting temperature clothes.Above three share, and can improve the product antialcoholism function.

Buccal tablet, be a class without peptic digest, directly absorb by oral mucosa and enter blood, adopt that soluble material carries out the tablet that dressing is made for main coating material in the oral cavity.Can prevent acidity and enzyme to the destruction of some drugs or prevent that medicine is to the intense stimulus of stomach; We find to have in the pollen pini many bioactivators under one's belt easily by stomach acids destroy through research, and can directly absorb by hypoglossis mucous membrane, can protect bioactivator to exempt from destruction better so make buccal tablet, improve the result of use of product.Also absorbed a large amount of food in the time of through our finder's heavy drinking of research, particularly high innage fat food makes people's stomach be high full abdomen state, the emptying of its stomach generally needs about 2 hours, if take the medicine that relieves the effect of alcohol simultaneously, because comparing with stomach, dose differs too great disparity, be difficult to play a role and lost efficacy, if but adopt heavy dose of medicine that relieves the effect of alcohol, can be subjected to the restriction of gastric capacity again and can't implement, so the medicine that relieves the effect of alcohol of Shi Yonging preferably adopts the mode without stomach after meal, Here it is, and why we will develop the main cause of buccal tablet.

The research of China's pollen pini at present has been in the leading level in the world; external research also is in the primary research stages such as analysis of nutritional labeling; and a series of difficult problems such as China has solved well that pollen pini is fresh-keeping, broken wall and production; be that 201010607364.8 Chinese patent discloses and passes through water extracting method as number of patent application; realized large-scale production; and improved nutrient absorption rate behind the edible pollen pini greatly, for pollen pini industry leading position in the world provides powerful guarantee.

However external pollen product is but more extensive on using than China, and Sweden is applied to beauty treatment with pollen food, and Japan is used as nutriment with pollen food, and France is with the function food of pollen as the growth encourage child development.By comparison, though the developmental research of China's pollen pini has been carried out the more than ten years, a large amount of pollen pini products has appearred, but the comprehensive processing and utilization level is also not high, the product category of existing pollen pini is single on the market, and quality is uneven, and the degree of intensive processing is not high yet, exported product is rare especially, and the resource of valuable pollen pini is not well utilized.

Containing profuse nutritional labeling in the pollen pini, Wang Kaifa etc. study summary, and pollen pini contains 104 kinds of enzymes such as oxidoreducing enzyme, phosphatase, superoxide dismutase; Except macroelement, also contain various trace elements such as iodine, cobalt, selenium; Flavonoids, carrotene, nucleic acid, auxin etc.; Abundant phosphatide, have fat, protein and the metabolic effect of carbohydrate adjusted, effective disease such as atherosis, the obesity of prevention of arterial, the protection liver also promotes the liver cell generation, lecithin wherein can also promote brain development, strengthens memory; Also containing multivitamins such as A, B, C, D, E, can satisfy different levels crowd's nutritional need, is first-class natural health-care products, but utilizes the pollen pini aspect research of relieving the effect of alcohol to remain blank.

Pollen pini is the dietotherapeutic pollen kind in the Chinese medicine treasure-house, as the traditional medicinal material of China, 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.In pollen, also there is a class to have to promote the material of alcohol metabolism through our up-to-date discovering, has remarkable effect to promoting the body aspect of relieving the effect of alcohol.

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

Summary of the invention

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

The present invention includes the molecular weight obtained with the pollen pini water extracting method less than freeze-dried powder, kudzu-vine root powder, dried orange peel powder, hydroxyl isomaltulose, D-sorbite and the dolomol of the pollen pini water extract of 5000D.

Described freeze-dried powder, kudzu-vine root powder, dried orange peel powder, hydroxyl isomaltulose, D-sorbite and dolomol account for 10～30%, 10～20%, 1～5%, 45～60%, 5～10%, 0.2～0.4% of buccal tablet gross mass respectively.

Pollen pini is the pollen of pine trees such as gymnosperm Pinaceae Pinus masson pine and Chinese pine, and pollen includes various rich nutrient substances, has multiple BA.The pharmacological action that pollen pini water extract can relieve the effect of alcohol in gene level performance has solved its molecule mechanism of relieving the effect of alcohol substantially through inventor's research.Through test, the present invention adopts pollen pini water extract freeze-dried powder as the solution liquor raw material, total protein content 0.25-2% in the freeze-dried powder, and water content is lower than 8%, have no side effect, and expense is cheap, and is simple.Buccal tablet advantage of the present invention is that the water extract that is used for relieving the effect of alcohol in the pollen pini can directly absorb by hypoglossis mucous membrane, also enter blood rapidly, avoids hydrochloric acid in gastric juice to its destruction, and result for the treatment of is better.

Description of drawings

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

Fig. 2 is natural health group renal tissue slice map.

Fig. 3 is for irritating stomach liquor model group liver tissue slices figure.

Fig. 4 is for irritating stomach liquor model group renal tissue slice map.

Fig. 5 is for irritating appetite lozenge group liver tissue slices figure.

Fig. 6 is for irritating appetite lozenge group renal tissue slice map.

The specific embodiment

One, the preparation pollen pini water extract freeze-dried powder that relieves the effect of alcohol:

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

2, adopt the mode of air-flow pulverize and break cellular wall with the pollen pini broken wall.

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

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

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

6, above-mentioned pollen pini water extract freeze drying 24～48 hours preparation water content is lower than the freeze-dried powder of 8% pollen pini water extract.

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

1, granulating process:

It is standby D-sorbite to be made into 10～30% the aqueous solution.

After the pot that will seethe with excitement is preheating to 60～80 ℃, earlier freeze-dried powder, kudzu-vine root powder, dried orange peel powder and isomalt are added in the boiling pot, arrange according to the following table parameter, open peristaltic pump spray aqueous sorbitol solution and granulate.The end back adding dolomol of granulating mixes.

Typical several quality proportioning table: (unit: kg)

?	Freeze-dried powder	Kudzu-vine root powder	The dried orange peel powder	Hydroxyl isomaltulose	D-sorbite	Dolomol
							Proportioning 1	20	10	3	58.6	8	0.4
Proportioning 2	25	12	4	52.7	6	0.3
							Proportioning 3	30	12	5	46.6	6	0.4

The processing parameter table:

Parameter	EAT	Leaving air temp	The air inducing frequency	Peristaltic pump	Atomizing pressure
						During whitewashing	70-85℃	40-60℃	40Hz	200rpm	0.2MPa
After the whitewashing	85-90℃	50-65℃	35Hz	200rpm	0.2MPa

(2) tablet forming technique: adopt ZP1100 tablet press machine (29 dash) 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, use:

(1) subjects:

Irritate the stomach Wistar cleaning level male rat in 22 ages in week, body weight 344.43 ± 38.49; The nature control rats is irritated the stomach pure water by body weight 1% every day; The model group rat is irritated stomach strong, colourless liquor distilled from sorghum simulation by body weight 1% every day; Irritate to irritate after the stomach strong, colourless liquor distilled from sorghum by body weight 1% appetite lozenge experimental group rat every day and irritated the appetite lozenge in 8 hours.

(2) experimental group adopts the mode of irritating stomach lozenge of the present invention on an empty stomach to experimentize, and irritates stomach dosage:

50～100mg/ days, connect and took 30～60 days.

(3) comparing result:

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

(1) with nature control group liver and renal tissue section, sees 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 tangible fat vacuole in the liver cell; Kidney group section mesonephric glomerulus structure is clear, and the Bowman's capsule gap is obviously moderate, not obviously damage.

(2) will irritate the section of stomach liquor model group liver and renal tissue, see Fig. 3,4.

From Fig. 3,4 visible liver cell obscure boundaries, becoming in the circle has cavity, and 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) irritate the section of stomach pollen pini water extract buccal tablet liver and renal tissue, see 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, irritate on an empty stomach after the stomach lozenge intervention of the present invention the main blood parameters comparative analysis of rats in test groups.

The method of testing of blood parameters: adopt the blood sampling of anesthesia abdominal aorta, detect main blood parameters by Toshiba's fully automatic blood bio-chemical detector after getting serum.

Comparative analysis such as following table:

Group	Naturally organize	Model group	Irritate stomach pine extract
				ALT	73.00±16.67	91.25±29.33	54.00±8.32
ALP	102.50±11.90	146.75±21.70	89.29±8.92
				TG	1.25±0.18	1.84±0.37	1.31±0.48
ALB	15.50±0.43	14.88±0.53	16.07±0.51

Through the SPSS statistics, model group glutamic-pyruvic transaminase (ALT) is extremely remarkable with filling stomach pollen pini buccal tablet group difference; Transaminase extensively exists in the histocyte, and the strongest with activity in the tissue such as liver, cardiac muscle, brain, kidney especially, just often plasma content is very low, and ALT mainly is present in liver, and ALT is released into blood in a large number can cause that transaminase raises in the blood when hepatic tissue is sick when decreasing.This experiment filling appetite lozenge 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 the liver is with the transhipment of material and drain relevant.And the diabetes B patient also causes the obstacle of lipid metabolism to reach at intrahepatic deposition simultaneously because insulin secretion obstacle or insulin resistance 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 alkaline phosphatase and other two experimental group difference are extremely remarkable; Illustrate that the pollen pini buccal tablet can 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 irritating the appetite lozenge, alleviate hepar damnification.

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 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 keeping glucostasis, particularly G-6-Pase plays a crucial role in glycogen generates, the liver insulin resistance also shows as the fat metabolic disorder, and discharging as triglycerides increases and the liver fat sex change.Through the SPSS statistics, model group triglycerides (TG) and filling stomach buccal tablet group of the present invention significant difference; From the experiment situation, serum high triglyceride level appears in model group, illustrates that long-term excessive drinking is the main cause that produces hypertriglyceridemia, is one of paathogenic factor of bringing out diabetes B; And take buccal tablet of the present invention the triglycerides metabolic disorder of bringing out owing to excessive drinking had good regulating and controlling effect.

The model group albumin is extremely remarkable with filling appetite lozenge group difference, can find also that from the variation of albumin level pollen pini water extract buccal tablet can regulate and control the protein metabolism of body, that is to say by the analysis to blood parameters, be not difficult to draw such conclusion: long-term excessive drinking back body three big metabolisms all occur significantly unusual, and can bring out diabetes, and pollen pini water extract buccal tablet not only can help body to relieve the effect of alcohol, and can recover body rapidly to the metabolism of three big materials, improve multinomial blood parameters, make organism metabolism return to normal condition.

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

Discover by biochip technology the important metabolic pathway of many influences 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 eubolism, and the variation of this gene expression has obtained good correction after the intervention of filling appetite lozenge, 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 filling appetite lozenge to forming owing to long-term excessive drinking from following example table, the reparation of tissue damage has very strong improvement effect, finding to irritate the appetite lozenge by our research from gene level can relieve the effect of alcohol effectively, improves the metabolism of liver.

3.1 the influence of after the intervention of filling appetite lozenge key gene relevant with glycolipid metabolism in the liver being expressed

Group

Dci

Fasn

Fbp2

G6pc

Pla2g2a

Scd

Scd1

Model group/organize naturally

↓3.72

↑4.86

↓4.57

↓5.62

↑2.76

↑5.04

↑5.65

Buccal tablet group/model group

↑3.8

↓2.44

↑2.46

↑2.29

↓8.02

↓2.69

↓2.81

Annotate: 3.8 times of the expression ratio rises of appetite lozenge groups and model group are irritated in 3.72 times of the expression ratio downward modulations of ↓ 3.72 expression Dci genes model group and natural control group in liver cell mRNA, ↑ 3.8 expression Dci genes.

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

Excessive drinking model group and natural control group are than 3.72 times of the expression of downward modulation ten dichloro alkane coacetylase 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, mitochondria aliphatic acid beta-oxidation ability is weakened, liver fat is become unfavorable.

Irritate appetite lozenge group under this experiment condition and raised 3.8 times of this genes with the model group ratio, can promote the aliphatic acid beta-oxidation of liver organization, to preventing and treating fatty liver positive effect is arranged.

3.1.2 Fasn: fatty acid synthetase, it is synthetic to participate in fatty acid biological.In the mammal, fatty acid synthetase plays crucial effects 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, expression of gene directly affects the fatty acid synthetase number, and lipidosis has great importance to the control animal body.

This experiment condition drag group leader's phase indulges in excessive drinking and makes in the liver cell fatty acid synthetase gene and 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, causes fatty liver generation primary factor.

Irritate 2.44 times of the gene expressions of appetite lozenge group and model group downward modulation fatty acid synthetase under this experiment condition, illustrate: the buccal tablet group can be by reducing the synthetic of fatty acid synthetase, and it is synthetic to reduce hepatic tissue aliphatic acid, and is significant to the change of alleviation liver fat.

3.1.3 Fbp2: fructose-1,6-diphosphatase 2, the regulatory enzyme of gluconeogenesis, catalyzing hydrolysis fructose-1,6-diphosphonic acid are 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 that Fbp2 gene and natural control group illustrate than 4.57 times of down-regulated expressions in the liver cell: model group can descend model group gluconeogenesis ability by this expression of gene of downward modulation, and is consistent with the weight fat liver.

Irritate 2.46 times of appetite lozenge group and model group rise Fbp2 gene expressions under this experiment condition, illustrate: the buccal tablet group can promote hepatocellular gluconeogenesis level by raising Fbp2 gene expression, alleviates the change of liver cell fat.

3.1.4 G6pc: G-6-Pase participates in glycogen metabolism and synthetic.G-6-Pase is an important enzyme of control 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 with the glycogen due to the liver insulin resistance and is closely related, so G6pc is a potential target spot for the treatment of diabetes.The hydrolysis of G-6-Pase catalysis 6-glucose 1-phosphate1-produces phosphoric acid and glucose, the i.e. rate-limiting reaction of gluconeogenesis approach and decomposition of glycogen approach.It is generally acknowledged, be the key of control blood glucose balance by the glucose phosphorylation of glucokinase enzymatic with by the 6-glucose 1-phosphate1-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 subjected to the adjusting of hormone and trophic level, the hungry expression that can stimulate G6PC, and hyperglycemic factor is by improving the expression that cAMP also can stimulate G6PC, insulin and ingest and then reduce the expression of G6PC.

This experiment condition drag group leader's phase indulges in excessive drinking and makes in the liver cell G-6-Pase gene and natural control group than 5.62 times of down-regulated expressions, 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, reduction is to the utilization of glucose, namely reduce the supply level of liver cell energy, be unfavorable for that liver cell recovers eubolism.

Irritate 2.29 times of the gene expressions of appetite lozenge group and model group rise G-6-Pase under this experiment condition, illustrate: the buccal tablet group can be by raising the expression of G-6-Pase, improve liver cell to the utilization of glucose, recover the liver cell eubolism, alleviate fat and become.

3.1.5 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 liver cell Pla2g2a gene and natural control 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.

Irritate appetite lozenge group and 8.02 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 hepatocellular injury.

3.1.6 Scd: sterol coacetylase desaturase, it is synthetic to participate in fatty acid biological.Stearyl-coenzyme A desaturase (SCD) can be introduced two keys in fatty acid carbon chain cis-9 position, 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, 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 acids, 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 have substantial connection, are one of hot fields of metabolic disease in recent years.The diabetes B patient, usually with insulin resistance, high leptin mass formed by blood stasis, there is fatty liver in about 50% diabetes B patient simultaneously.Scd-the 1st, the key enzyme that the catalysis saturated fatty acid changes to monounsaturated fatty acids, closely related with generation, the development of a series of metabolic syndromes such as obesity, fatty hepatic disease and insulin resistance, 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, 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 (unrighted acid/saturated 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 energetic supersession.By the research to Scd-1 and energetic supersession relation, can the mechanism of clearer understanding Scd-1 in energetic supersession, thereby regulate energetic supersession and avoid fat toxicity for clinical target, a series of metabolic syndromes such as prevention or treatment obesity, fatty hepatic disease and diabetes provide the basis.

The excessive drinking model group raises 5.04 times of sterol coacetylase desaturase (Scd) gene expressions with natural control group ratio under this experiment condition, sterol coacetylase 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, 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 reaches the state of liver fibrosis.

Irritate appetite lozenge group and 2.69 times of model group ratio downward modulation sterol coacetylase desaturase (Scd) gene expressions under this experiment condition, downward modulation sterol coacetylase desaturase 1(Scd-1) gene expression is 2.81 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 the pollen pini buccal tablet is prevented and treated one of main molecule mechanism of fatty liver.

3.2 the influence of behind the filling appetite lozenge key gene relevant with hepatic injury, reparation and metabolism in the liver being expressed

Group

Il7r

Irs3

Srd5a1

Socs2

Tgfb2

Tnfsf10

Model group/organize naturally

↓2.75

↑2.47

↑4.0

↓33.58

↑3.63

↑2.92

Buccal tablet group/model group

↑3.73

↓2.01

↓4.37

↑8.02

↓2.34

↓2.15

Annotate: 3.73 times of the expression ratio rises of appetite lozenge groups and model group are irritated in 2.75 times of the expression ratio downward modulations of ↓ 2.75 expression Il7r genes model group and natural control group in liver cell mRNA, ↑ 3.73 expression Il7r genes.

3.2.1 Il7r: interleukin-17, acceptor participates in the regulating DNA reorganization, immune 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 control group than model group rat, illustrate: the model group rat is because long-term excessive drinking can cause liver cell immune response ability to descend, downward modulation interleukin-17 expression of gene means exactly can increase the weight of hepatocellular inflammatory reaction, can promote liver cell fat to become and Fibrotic process.

The buccal tablet group raises 3.73 times of interleukin-17 genes with the model group ratio under this experiment condition, illustrate: after buccal tablet is intervened, strengthen hepatocellular immune response ability by raising the interleukin-17 expression of gene, alleviate the liver inflammatory reaction, to preventing and treating the fatty liver and the fibrillatable that cause owing to long-term excessive drinking positive effect is arranged.

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

This experiment condition drag group has raised 2.47 times of IRS 3 genes with natural control group than model group rat, illustrate: the model group rat is by raising Irs3 gene expression, impel the IRS 3 of the synthetic more negative regulation IRSs 2 of liver cell, further increase the weight of hepatocellular insulin resistance, increase the weight of metabolic syndrome, promote fat to become.

Under this experiment condition with after the buccal tablet intervention with the model group ratio, reduced 2.01 times of Irs3 expression of gene, illustrated: irritating stomach buccal tablet of the present invention can alleviate hepatocellular insulin resistance by downward modulation Irs3 expression of gene, improve liver cell to the sensitiveness of insulin, alleviate liver cell fat and become.

3.2.3 Srd5a1: steroids 5 alpha-reductases, α peptide 1, distribution endoplasmic reticulum, microsome, 3-C-5 α steroids 4 dehydrogenases participate in cell signalling, and sex determines, the 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 adipose tissue and liver, but the reduction reaction of unsaturation steroids on the 4th, 5 carbon atom of the catalysis overwhelming majority makes activated glucocorticoid (GC) be converted into the metabolite of non-activity.Discover that the glucocorticoid effect is not only relevant with GC level in the blood circulation, 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 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 multiple metabolic disorder clinical signs such as obesity, hypertension, dyslipidemia and IGT).

This experiment condition drag group and natural control group ratio, the Srd5a1 expression of gene has raised 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 takes place.

Under this experiment condition with after the buccal tablet intervention with intervene before the model group ratio, the Srd5a1 expression of gene has been reduced 4.37 times, illustrate: buccal tablet of the present invention can be by this gene expression of downward modulation, improve the metabolic syndrome that brings owing to excessive drinking, this has explained the reason why the pollen pini buccal tablet can prevent and treat metabolic syndrome from another point of view.

3.2.4 Socs2: Suppressor of Cytokine Signaling was named according to 4O amino acid whose Socs frame by discoveries such as Starr R in 2,1997 years.Found that at present the negative molecule of regulating of this class 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 fat deposition, skeletal muscle growth, central nervous system effect, immune response, cancer takes place all to play an important role.Socs 2 can regulate hormone secretions such as insulin, prolactin in addition, and cytokine signaling paths such as GH/1GF, insulin, interleukin are played promotion or inhibitory action.The various kinds of cell factor is by lipometabolic balance in complicated signal network regulating cell and the body, and most researchs at present concentrate growth hormone (GH) to lipometabolic influence, 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 Socs2 expression of gene has been reduced 33.58 times, illustrate: long-term excessive drinking can cause metabolism " brake " regulator control system in the liver cell seriously malfunctioning, will inevitably exert an influence to normal fat metabolism and insulin action in the liver cell, form dysbolism, finally impelling liver cell fat to become, is that one of main molecules mechanism of fatty liver is brought out in long-term excessive drinking.

Under this experiment condition with after the buccal tablet intervention of the present invention with intervene before the model group ratio, the Socs2 expression of gene has raised 8.02 times, illustrate: buccal tablet of the present invention can be by improving the Socs2 expression of gene, repair because interior " brake " regulator control system of the liver cell that excessive drinking causes is malfunctioning, significant to improving liver cell fat metabolism and insulin action.

3.2.5 Tgfb2: transforming grouth factor beta 2, participate in cell death, just regulating and control process, regulating cell propagation, regulation and control immune response, dopamine biosynthesis.Studies confirm that to have only Tgf-β energy HSC stimulated rubber polymer fibrillation in the cell factor, other cell factors are a proliferation of HSC stimulated then.In the liver fibrosis forming process, 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 that HSC goes up pdgf receptor simultaneously, promotes HSC autocrine Tgf-β, PDGF, obviously suppresses the apoptosis of activation HSC.PDGF and Tgf-β have constituted the autocrine circulation of activation HSC, are the important mechanisms of the sustained activation of HSC.

This experiment condition drag group and natural control group ratio, the Tgfb2 expression of gene has raised 3.63 times, illustrates: long-term excessive drinking causes hepatocellular injury, raise the Tgfb2 expression of gene, HSC stimulated rubber polymer fibrillation promotes the formation of liver 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 Tgfb2 expression of gene has been reduced 2.34 times, illustrate: buccal tablet of the present invention can pass through the process of this gene expression control liver fibrosis of downward modulation, and is significant to safeguarding liver cell eubolism function.

3.2.6 Tnfsf10: TNF surpasses family, and the member 10, participates in the cell guiding apoptosis, is just regulating and control NF-κ B cascade.

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

Model group ratio with experimental group after the buccal tablet intervention and before intervening under this experiment condition; the Tnfsf10 expression of gene has been reduced 2.15 times; illustrate: buccal tablet of the present invention can be by downward modulation Tnfsf10 expression of gene; suppress the activation of NF-κ B in the rat body; alleviate the peroxidating of liver cell inner lipid and inflammatory reaction; alleviate liver cell fat change and fibrillatable take place, play the target of protection hepatic tissue.

In a word, show that by above test 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 pollen pini relieve the effect of alcohol aspect research at the early-stage, by experiment, find that the pollen pini buccal tablet is effective to antialcoholism action, the medicine that relieves the effect of alcohol for exploitation proposes new approaches.Pollen pini is China's Traditional health care product, can improve hepatocyte activity, improves blood parameters, and prevention is because the fatty liver that long-term excessive drinking causes, the health care of safeguarding and improve China common people's excessive drinking is significant.Pollen pini can be used as at the first-selected health products that relieve the effect of alcohol, promotion and application in addition.

Claims (1)

1. for the pollen pini water extract buccal tablet that relieves the effect of alcohol, it is characterized in that mainly being formed less than freeze-dried powder, kudzu-vine root powder, dried orange peel powder, hydroxyl isomaltulose, D-sorbite and the dolomol of the pollen pini water extract of 5000D by the molecular weight of obtaining with the pollen pini water extracting method; Described freeze-dried powder, kudzu-vine root powder, dried orange peel powder, hydroxyl isomaltulose, D-sorbite and dolomol account for 10～30%, 10～20%, 1～5%, 45～60%, 5～10%, 0.2～0.4% of buccal tablet gross mass respectively.

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