CN111808877A

CN111808877A - Production method, composition and preparation of acetaldehyde dehydrogenase

Info

Publication number: CN111808877A
Application number: CN202010925634.3A
Authority: CN
Inventors: 任哲; 王一飞; 王巧利; 刘俊伟
Original assignee: Guangzhou Jinan Biomedicine Research and Development Base Co Ltd
Current assignee: Guangzhou Jinan Biomedicine Research and Development Base Co Ltd
Priority date: 2020-09-07
Filing date: 2020-09-07
Publication date: 2020-10-23

Abstract

The invention provides a production method, a composition and a preparation of acetaldehyde dehydrogenase for alleviating hangover and protecting liver. A production method of acetaldehyde dehydrogenase for alleviating hangover and protecting liver comprises the following steps: inoculating the engineering bacteria of Escherichia coli into a fermentation culture medium, and performing fermentation culture at 25-37 deg.C for 6-8 h; collecting the thallus, carrying out ultrasonication, and separating and purifying ALDH2 protein. The invention can not only quickly relieve discomfort after drinking, but also effectively protect and nourish intestines, stomach and liver, and has good development potential.

Description

Production method, composition and preparation of acetaldehyde dehydrogenase

Technical Field

The invention relates to the technical field of biology, and in particular relates to a production method, a composition and a preparation of acetaldehyde dehydrogenase.

Background

At present, the antialcoholic products on the market are mainly chemical drugs. Most of the chemicals are components such as stimulants, vitamins and amino acids, and can quickly achieve the effects of dispelling the effects of alcohol and sobering up, but most of the chemical anti-alcohol mechanisms are only capable of promoting alcohol decomposition to achieve a temporary relieving effect, the liver protection effect is difficult to achieve, and the toxic and side effects are large.

Disclosure of Invention

The present invention overcomes at least one of the deficiencies of the prior art described above,

an object of the present invention is to provide a method for producing acetaldehyde dehydrogenase, comprising the steps of:

s1, inoculating an escherichia coli engineering bacterium expressing an acetaldehyde dehydrogenase gene into a fermentation culture medium for culture;

s2, collecting thalli, and separating and purifying after carrying out ultrasonic crushing.

Further, fermenting and culturing for 6-8h at 25-37 ℃.

Further, fermenting and culturing for 6-8h at 25 ℃.

The invention also aims to provide an acetaldehyde dehydrogenase composition for relieving alcoholism, which comprises the following components in parts by weight: 1-10 parts of acetaldehyde dehydrogenase obtained by the production method of the acetaldehyde dehydrogenase, 40-80 parts of kudzu root extract, 10-50 parts of dried orange peel extract, 10-50 parts of fructus amomi extract, 20-40 parts of flos puerariae extract, 15-30 parts of chinaroot greenbrier rhizome extract, 10-30 parts of small bean flower extract, 10-30 parts of elecampane extract, 10-30 parts of radix asparagi extract, 10-30 parts of moutan bark extract, 10-30 parts of rhizoma alismatis extract and 10-30 parts of liquorice extract.

Further, the composition comprises the following components in parts by weight: 5 parts of acetaldehyde dehydrogenase obtained by the production method of the acetaldehyde dehydrogenase, 60 parts of kudzu root extract, 30 parts of dried orange peel extract, 30 parts of fructus amomi extract, 40 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 20 parts of small bean flower extract, 15 parts of elecampane extract, 15 parts of radix asparagi extract, 10 parts of moutan bark extract, 10 parts of rhizoma alismatis extract and 10 parts of liquorice extract.

The invention also aims to provide an anti-alcohol liver-protecting acetaldehyde dehydrogenase composition preparation, which comprises the anti-alcohol liver-protecting acetaldehyde dehydrogenase composition.

Furthermore, the composition also comprises pharmaceutically acceptable auxiliary materials.

Further, the coating also comprises at least one of excipient, binder, bulking agent, lubricant, antioxidant, preservative or thickening agent.

Further, the preparation is an oral preparation.

Furthermore, the preparation can be prepared into tablets, powder, pills, capsules, granules or oral liquid.

Compared with the prior art, the technical scheme of the invention has the beneficial effects that:

the invention provides an acetaldehyde dehydrogenase composition with anti-intoxication and anti-alcoholism effects and a preparation method thereof, wherein the acetaldehyde dehydrogenase composition comprises the following components: acetaldehyde dehydrogenase, a kudzu root extract, a dried orange peel extract, a fructus amomi extract, a pueraria flower extract, a white poria extract, a small bean flower extract, a costus root extract, an asparagus root extract, a tree peony bark extract, an alisma orientale extract and a liquorice root extract are combined according to different proportions to fully play the synergistic effect. Animal experiments show that the composition prepared by the invention has obvious anti-intoxication and anti-alcoholism effects, can fundamentally remove acetaldehyde, has the functions of harmonizing the stomach and protecting the liver, and achieves the purposes of anti-alcoholism and protecting the liver. The composition has simple preparation process, can be subjected to large-scale industrial production, and has wide application prospect in preparation of health care products or foods with the effects of resisting and neutralizing the effect of alcoholic drinks.

The acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcoholic drinks can fundamentally remove acetaldehyde, has the effects of harmonizing the stomach and protecting the liver, and achieves the purposes of neutralizing the effect of alcoholic drinks and protecting the liver. No chemical reagent and toxic substance are used in the extraction and preparation processes, so that the preparation is safe and effective, has definite treatment purpose and good development potential.

Drawings

FIG. 1 is an electrophoretogram of PCR products of ALDH2 of the present invention.

FIG. 2 shows the induction of expression of ALDH2 in the present invention.

FIG. 3 shows the induction of ALDH2 expression at different temperatures in the present invention.

FIG. 4 shows the solubility analysis and identification of ALDH2 protein in the present invention.

FIG. 5 shows the differentiation and purification of ALDH2 protein in the present invention.

FIG. 6 is a standard curve of the activity of the enzyme of the present invention.

FIG. 7 shows the effect of acetaldehyde dehydrogenase (ALDH2) composition of the present invention on the intoxication and sobering of mice.

Detailed Description

The drawings are for illustrative purposes only and are not to be construed as limiting the patent; instruments and medicines used in the application are all commercially available if not specifically stated; it will be understood by those skilled in the art that certain well-known structures and descriptions thereof may be omitted.

The invention provides a production method of acetaldehyde dehydrogenase, which comprises the following steps:

As an improvement of the specific embodiment, the fermentation culture is carried out for 6-8h at 25-37 ℃.

As an improvement of the specific embodiment, the fermentation culture is carried out for 6-8h at 25 ℃.

The invention also provides an acetaldehyde dehydrogenase composition for relieving alcoholism, which comprises the following components in parts by weight: 1-10 parts of acetaldehyde dehydrogenase obtained by the production method of the acetaldehyde dehydrogenase, 40-80 parts of kudzu root extract, 10-50 parts of dried orange peel extract, 10-50 parts of fructus amomi extract, 20-40 parts of flos puerariae extract, 15-30 parts of chinaroot greenbrier rhizome extract, 10-30 parts of small bean flower extract, 10-30 parts of elecampane extract, 10-30 parts of radix asparagi extract, 10-30 parts of moutan bark extract, 10-30 parts of rhizoma alismatis extract and 10-30 parts of liquorice extract.

As an improvement of the specific embodiment, the composition comprises the following components in parts by weight: 5 parts of acetaldehyde dehydrogenase obtained by the production method of the acetaldehyde dehydrogenase, 60 parts of kudzu root extract, 30 parts of dried orange peel extract, 30 parts of fructus amomi extract, 40 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 20 parts of small bean flower extract, 15 parts of elecampane extract, 15 parts of radix asparagi extract, 10 parts of moutan bark extract, 10 parts of rhizoma alismatis extract and 10 parts of liquorice extract.

The invention also provides an alcohol-relieving liver-protecting acetaldehyde dehydrogenase composition preparation, which comprises the alcohol-relieving liver-protecting acetaldehyde dehydrogenase composition.

As an improvement of the specific embodiment, the composition also comprises pharmaceutically acceptable auxiliary materials.

As a refinement of a particular embodiment, at least one of an excipient, a binder, a filler, a lubricant, an antioxidant, a preservative, or a thickener is further included.

As a modification of the specific embodiment, the formulation is an oral formulation.

As an improvement of the specific embodiment, the preparation is in the form of tablets, powder, pills, capsules, granules or oral liquid.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

Preparing acetaldehyde dehydrogenase materials:

1. preparation of ALDH2 Gene

The ALDH2 gene fragment which is optimized and stored before the laboratory is used as a template, NcoI enzyme cutting sites and Xho I enzyme cutting sites are respectively introduced into the upstream and the downstream, and the ALDH2 gene fragment is amplified (figure 1). The reaction products were identified by electrophoresis on a 1% agarose gel. The PCR product ALDH2 was recovered and purified by gel. Plasmid extraction and gel recovery steps were performed according to the instructions of the OMEGA gel recovery kit.

FIG. 1 is an electrophoretogram of the PCR product of ALDH 2. The results showed a product length of 1500 bp, consistent with the expected results.

1.2, constructing pet-28a-aldh2 recombinant expression plasmid:

2. induced expression and purification of acetaldehyde dehydrogenase protein

2.1, transformation of the recombinant plasmid pET-28a-aldh 2:

the correctly identified recombinant plasmid pET-28a-aldh2 was transformed into competent cells of E.coli BL21(DE 3). Positive monoclonal strains were screened on resistant (kanamycin at 100. mu.g/ml) plates.

2.2 screening of ALDH2 expression bacteria and optimization of optimal expression conditions

A plurality of randomly selected monoclonal colonies were inoculated into 5ml of LB medium (LB medium formulation: tryptone: 10g/L, yeast extract: 5g/L, sodium chloride: 10g/L, NaOH-adjusted medium pH at 7.4, containing 100. mu.g/ml kanamycin) and cultured at 37 ℃ and 180 rpm until OD600 = 0.6-0.8. Taking 1ml as an uninduced control, adding IPTG (isopropyl-beta-D-thiogalactoside) to a final concentration of 1mmol/L, inducing at 37 ℃ for 6h, centrifuging to collect thalli, and performing 10% SDS-PAGE analysis after thalli precipitation treatment. The results show that the selected 4 single clones can express the target protein (figure 2), and the protein expression has no obvious difference at different temperatures (figure 3), so that the protein expression is carried out under the condition that the protein expression is favorable for soluble protein expression at 25 ℃, and the protein is mainly expressed in a soluble mode after the identification of solubility analysis (figure 4). Therefore, the supernatant after ultrasonication is subjected to protein separation and purification by using a nickel column protein purification system, and finally the desired target protein is obtained in 300mM imidazole eluent (figure 5), and the enzyme activity per unit volume is 1886.9U/L after enzyme activity detection (Table 1).

FIG. 2 shows the induction of expression of ALDH 2. 1: an untransformed control; 2.3, 4, 5: 4 single clones were picked and induced to express. The results show that the protein expression is obvious at about 55 kd.

2.3 determination of Induction temperature

FIG. 3 shows the induction of ALDH2 expression at different temperatures. 1. Untransformed control 2, 25 ℃ induced expression; 3. inducing expression at 30 ℃; 4. expression was induced at 37 ℃. The result shows that the protein can generate higher target protein amount at 25-37 ℃, and the low temperature is favorable for a prokaryotic expression system to induce and generate soluble protein, so that the subsequent experiment is carried out at 25 ℃.

2.4 solubility analysis and identification

FIG. 4 shows the ALDH2 protein solubility assay. 1. Untransformed control 2, supernatant after induced expression ultrasonication; 3. inducing expression, ultrasonic breaking and depositing. The results showed that a portion of the ALDH2 protein was expressed in soluble form (a distinct protein band at 55kd in the supernatant after ultrasonication).

2.5 purification of ALDH2 protein

FIG. 5 shows the ALDH2 protein differentiation purification. 1. Identifying supernatant after induced expression and ultrasonic disruption; 2. elution with 50mM imidazole; 3. 75mM imidazole; 4. 300mM imidazole. The results showed a clear band of interest in the 300mM eluate, which was assayed at a protein concentration of 183 mg/L.

2.6 detection of ALDH2 Activity

The activity detection is carried out by using Nanjing-derived ALDH2 enzyme activity detection kit, and a standard curve (U/200 ul) is established by using Shanghai-derived leaf ALDH2 enzyme activity standard substance as shown in figure 6. The activity of the extracted ALDH2 protease is shown in Table 1, and the unit volume activity of the protease is 1886.9U/L.

TABLE 1 ALDH2 enzyme activity assay

Absorbance Change Δ A (340 nm)	Unit volume enzyme activity (U/L)
		0.0392	1886.9

Example 1

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 10 parts of acetaldehyde dehydrogenase, 80 parts of kudzu root extract, 50 parts of dried orange peel extract, 50 parts of fructus amomi extract, 40 parts of pueraria flower extract, 30 parts of chinaroot greenbrier rhizome extract, 30 parts of small bean flower extract, 30 parts of costustoot extract, 30 parts of radix asparagi extract, 30 parts of moutan bark extract, 30 parts of rhizoma alismatis extract and 30 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 2

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 5 parts of acetaldehyde dehydrogenase, 50 parts of kudzu root extract, 30 parts of dried orange peel extract, 20 parts of fructus amomi extract, 20 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 15 parts of small bean flower extract, 20 parts of costus root extract, 15 parts of cochinchnese asparagus root extract, 15 parts of tree peony bark extract, 20 parts of rhizoma alismatis extract and 10 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 3

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 3 parts of acetaldehyde dehydrogenase, 40 parts of kudzu root extract, 20 parts of dried orange peel extract, 30 parts of fructus amomi extract, 20 parts of pueraria flower extract, 15 parts of chinaroot greenbrier rhizome extract, 10 parts of small bean flower extract, 10 parts of costus root extract, 10 parts of cochinchnese asparagus root extract, 10 parts of tree peony bark extract, 10 parts of rhizoma alismatis extract and 10 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 4

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 6 parts of acetaldehyde dehydrogenase, 60 parts of kudzu root extract, 20 parts of dried orange peel extract, 30 parts of fructus amomi extract, 35 parts of pueraria flower extract, 25 parts of chinaroot greenbrier rhizome extract, 25 parts of small bean flower extract, 20 parts of costus root extract, 20 parts of cochinchnese asparagus root extract, 15 parts of tree peony bark extract, 10 parts of rhizoma alismatis extract and 15 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 5

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 4 parts of acetaldehyde dehydrogenase, 50 parts of kudzu root extract, 20 parts of dried orange peel extract, 20 parts of fructus amomi extract, 30 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 25 parts of small bean flower extract, 15 parts of costustoot extract, 15 parts of radix asparagi extract, 10 parts of moutan bark extract, 10 parts of rhizoma alismatis extract and 15 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 6

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 4 parts of acetaldehyde dehydrogenase, 45 parts of kudzu root extract, 10 parts of dried orange peel extract, 15 parts of fructus amomi extract, 30 parts of pueraria flower extract, 15 parts of chinaroot greenbrier rhizome extract, 15 parts of small bean flower extract, 10 parts of costus root extract, 10 parts of cochinchnese asparagus root extract, 15 parts of tree peony bark extract, 10 parts of rhizoma alismatis extract and 15 parts of liquorice extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract.

Example 7

An acetaldehyde dehydrogenase composition for resisting and neutralizing the effect of alcohol, which is prepared from the following raw materials in parts by weight: 5 parts of acetaldehyde dehydrogenase, 50 parts of kudzu root extract, 30 parts of dried orange peel extract, 20 parts of fructus amomi extract, 20 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 15 parts of small bean flower extract, 20 parts of costus root extract, 15 parts of cochinchnese asparagus root extract, 20 parts of oriental waterplantain rhizome extract and 10 parts of liquorice root extract. The preparation method of the extracts of the components comprises the following steps: soaking the medicinal materials in water for 1-2 hr, and decocting with water twice, wherein the amount of water added is 3-5 times of the weight of the medicinal materials each time; during decoction, the mixture is boiled by strong fire and then decocted by slow fire until the ratio of liquid to dregs in the medicament is 0.5-1: 1; filtering the obtained medicinal liquid with gauze, mixing filtrates, concentrating, and drying to obtain extract. During the second decoction, the following raw materials in parts by weight can be added: 10 parts of wild jujube, 5 parts of ginseng, 10 parts of mung bean, 10 parts of adenophora stricta, 10 parts of persimmon calyx, 10 parts of asparagus, 10 parts of duckweed, 10 parts of silkworm excrement, 10 parts of pogostemon cablin, 10 parts of semen hominis, 10 parts of ginger, 10 parts of linearstripe rabdosia herb, 10 parts of fistular onion stalk and 10 parts of chrysanthemum.

According to the traditional Chinese medicine, the treatment is based on the basic principle of relieving alcoholism, clearing heat and eliminating dampness, wherein the alcoholism is eliminated throughout the treatment, and methods such as reducing phlegm and eliminating dampness, promoting the circulation of qi and removing stagnancy, tonifying qi and strengthening spleen, warming middle-jiao and activating blood, lowering adverse qi and harmonizing stomach are mostly adopted. The traditional Chinese medicine composition takes the kudzuvine root, the kudzuvine flower and the rice bean flower as monarch drugs and is good at relieving alcoholism and activating the spleen, the kudzuvine flower and the kudzuvine root are traditional alcoholism relieving drugs, and the kudzuvine flower has the most prominent effect of relieving alcoholism and activating the spleen. Pueraria root, radix Puerariae has the action of expelling pathogenic factors from the muscles and skin and relieving damp-heat from the exterior. The ministerial drugs use the amomum fruit to eliminate dampness and promote qi circulation, warm the middle-jiao to stop diarrhea, the ginger helps to invigorate spleen qi, and eliminate dampness and activate the middle-jiao to invigorate the spleen; poria, Alismatis rhizoma, herba Spirodelae, herba Agastaches, herba Rabdosiae Lophanthoidis, and semen Phaseoli Radiati are used for promoting diuresis and eliminating dampness, and guiding wine dampness to relieve constipation; radix aucklandiae, pericarpium Citri Reticulatae, calyx kaki and semen Hoveniae are used to regulate qi, relieve stagnation, regulate middle energizer, eliminate dampness and phlegm, and promote the dispelling of alcohol and qi in the body; silkworm excrement and ginseng are used for tonifying middle-jiao, strengthening spleen, replenishing primordial qi, tonifying qi, strengthening body resistance and nourishing stomach, and radix adenophorae and radix asparagi are used for replenishing yin liquid; the spina date seed is used for nourishing the heart and soothing the nerves, the fistular onion stalk is used for activating yang and refreshing the mind, and the mint and the chrysanthemum enter the liver channel to clear away the heat and improve the eyesight; the liquorice is used for harmonizing the medicines and has the effects of detoxifying and promoting diuresis, soothing liver and purging heat, dispelling alcohol dampness and removing stagnation by combining the medicines; the health-care tea has the effects of reducing blood pressure, blood fat and blood sugar, resisting aging and fatigue, relaxing bowel, protecting liver, nourishing stomach, beautifying and moistening skin, improving human immunity, protecting liver, harmonizing stomach, promoting urination, clearing heat, detoxifying, relieving summer heat, relieving restlessness, beautifying and moistening skin, clearing lung, relieving cough, dispelling alcohol effect, dispelling wind, cooling blood, quickly and safely dispelling alcohol effect, has no toxic or side effect on human bodies, and has obvious effect.

Reference of drug efficacy:

kudzu root, wind-heat-dispersing medicine: relieving muscles and fever, promoting eruption, promoting fluid production to quench thirst, invigorating yang and relieving diarrhea (1. fever exterior syndrome, strong pain on neck and back 2. measles without adequate eruption 3. fever thirst, yin deficiency and thirst 4. fever diarrhea, spleen deficiency and diarrhea)

Dried orange peel, qi-regulating herbs: regulate qi to invigorate spleen, dry dampness and resolve phlegm. (1, syndrome of qi stagnation in spleen and stomach 2. emesis, hiccup 3. phlegm-dampness, cold-phlegm cough 4. thoracic obstruction)

Amomum fruit, dampness-resolving herbs: resolve dampness, promote qi circulation, warm middle energizer to arrest diarrhea, prevent abortion. (1. syndrome of dampness obstruction in middle energizer and qi stagnation in spleen and stomach 2. vomiting and diarrhea due to deficiency-cold in spleen and stomach 3. aversion to pregnancy due to qi stagnation and threatened abortion)

Pueraria flower, wind-heat-dispersing medicine: relieving alcoholism, activating spleen and regulating stomach. Can be used for treating excessive drinking, headache, dizziness, polydipsia, emesis, and chest and diaphragm fullness

Tuckahoe, diuretic detumescence drugs: induce diuresis and drain dampness, invigorate spleen and calm heart. (1. edema 2. phlegm and fluid 3. spleen deficiency diarrhea 4. palpitation insomnia)

Alisma orientale, diuresis-inducing and edema-alleviating herbs: induce diuresis to drain dampness and purge heat. (1. edema, dysuria, diarrhea 2. stranguria, spermatorrhea)

Aucklandia root, qi-regulating herbs: promoting qi circulation, relieving pain, invigorating spleen, and promoting digestion. (1. spleen and stomach qi stagnation syndrome 2. diarrhea and dysentery with tenesmus 3. abdominal pain and hypochondriac pain, jaundice, hernia pain 4. thoracic obstruction)

Chizuki bean flower has the effects of clearing heat, quenching thirst, sobering up and detoxifying. (malaria, dysentery, diabetes, headache due to injury or drinking, haemorrhoids and fistula, erysipelas, furuncle)

Licorice, a qi-tonifying drug: invigorating spleen and replenishing qi, eliminating phlegm and relieving cough, relieving spasm and pain, clearing away heat and toxic materials, and harmonizing the recipe. (1. heart-qi deficiency, intermittent pulse, palpitation, 2. spleen-qi deficiency syndrome, 3. cough and asthma, 4. epigastric and abdominal spasm, acute pain, 5. toxic heat, sore and abscess, sore throat, medicine and food poisoning, 6. harmonize medicine property)

Spina date seed, heart-nourishing and nerve-soothing medicine: nourishing heart, benefiting liver, tranquilizing, arresting sweating, and promoting fluid production. (1. palpitation insomnia 2. spontaneous perspiration, night sweat)

Mung bean, heat-clearing and toxicity-removing herbs: clearing away heat and toxic material, relieving summer-heat, and inducing diuresis (1. carbuncle, swelling, sore, toxin 2. summer-heat polydipsia 3. medicine and food poisoning 4. edema, dysuria)

Bitter apricot seeds, antitussive and antiasthmatic herbs: relieving cough and asthma, loosening bowel to relieve constipation (1. cough and asthma 2. constipation due to intestinal dryness)

Adenophora tetraphylla, yin-tonifying herbs: nourishing yin to clear away the lung-heat, benefiting the stomach and promoting the production of body fluid, invigorating qi and resolving phlegm (1. lung yin deficiency syndrome 2. stomach yin deficiency syndrome)

Calyx kaki, qi-regulating herbs: directing qi downward to stop hiccup (hiccup)

Asparagus, yin-tonifying herbs: to nourish yin, moisten dryness, clear lung and promote fluid production. (1. Lung yin deficiency syndrome 2. Kidney yin deficiency syndrome 3. fever impairment of body fluids caused by inappetence, thirst and constipation due to intestinal dryness)

Duckweed, wind-heat-dispersing drug: inducing sweat to relieve exterior syndrome, promoting eruption, relieving itching, inducing diuresis, and relieving swelling (1. wind-heat type common cold 2. measles without adequate eruption 3. rubella pruritus 4. edema oliguria)

Silkworm excrement, wind-cold-dampness-dispelling medicine: dispelling wind-damp, harmonizing stomach and resolving dampness (1. rheumatism 2. vomiting and diarrhea and spasm 3. rubella, eczema)

Agastache rugosus, dampness-resolving herbs: resolving dampness, arresting vomiting, relieving summer-heat (1. damp obstruction of middle energizer 2. vomiting 3. summer-heat dampness or early stage of damp-warm disease)

The ginseng, semen homalomenae, invigorates the stomach; promoting the secretion of saliva; sobering up; and (5) detoxifying. Mainly treats inappetence; thirst due to febrile disease; drunkenness; swollen and sore throat; sores and itching due to wind-toxin.

Ginseng, qi tonics: invigorating primordial qi, invigorating spleen, benefiting lung, promoting fluid production, tranquilizing mind, and improving intelligence (1. primordial qi deficiency syndrome 2. lung spleen heart kidney qi deficiency syndrome 3. fever with qi deficiency and body fluid consumption

Ginger, wind-cold-dispersing herbs: relieving exterior syndrome, dispelling cold, warming middle energizer, relieving vomiting, warming lung, and relieving cough (1. wind-cold type common cold 2. spleen and stomach cold syndrome 3. cough due to lung cold)

Herba Rabdosiae Lophanthoidis (herba Gei Repentis), herba Rabdosiae Lophanthoidis has effects of clearing heat, promoting diuresis, eliminating jaundice, eliminating dampness, cooling blood and removing blood stasis

Fistular onion stalk, wind-cold-dispersing medicine: inducing sweat to relieve exterior syndrome, dispelling cold and activating yang (1. wind-cold type common cold 2. exuberance of yin and yang)

Chrysanthemum, wind-heat-dispersing herbs: dispelling wind-heat, suppressing liver-yang, clearing liver-heat, improving eyesight, clearing heat and detoxicating (1. wind-heat type common cold, warm disease onset 2. liver-yang vertigo, liver-wind excess syndrome 3. conjunctival congestion and coma 4. sore and carbuncle pyogenic infections)

Mint, wind-heat-dispersing medicine: dispelling wind-heat, clearing head and eyes, relieving sore throat, promoting eruption, dispersing stagnated liver qi, and activating qi-flowing (1. wind-heat type common cold, warm disease onset 2. wind-heat type headache, conjunctival congestion with lacrimation, sore throat 3. measles without adequate eruption, rubella pruritus 4. stagnation of liver-qi, chest distress and hypochondriac pain)

Animal test conditions were as follows:

1. experimental Material

ALDH2 composition (ALDH 2+ Chinese traditional medicine): kudzu root, dried orange peel, fructus amomi, flos puerariae lobatae, white poria, small bean flower, costustoot, radix asparagi, moutan bark, rhizoma alismatis, liquorice and the like are purchased from Dashenlin pharmacy, extracts are prepared according to the original formula proportion, and are uniformly mixed with acetaldehyde dehydrogenase (ALDH2) for later use. According to the conversion method of the drug for animal pharmacology experiments and the conventional dosage of adults, the drug administration dosage is designed according to the crude drug of 10 g/kg of mouse body weight in the experiments. The drug concentration was adjusted with distilled water in an administration volume of 0.1mL (drug solution)/10 g (mouse body weight) before the experiment.

The animal experiments were as follows:

60 SPF-level Kunming mice with the weight of 18-22g are taken and provided by the experimental animal center of Guangdong province. After 1 week of adaptive feeding, mice were randomly divided into 6 groups (blank control group, model group, ALDH2 single-dose group, ALDH2 multiple-dose group, ALDH2 composition (ALDH 2+ chinese medicine) single-dose group, ALDH2 composition (ALDH 2+ chinese medicine) multiple-dose group), 10 mice per group. Mice were fasted without water deprivation for 12 h prior to treatment. The experimental group mice were gavaged with 56% vol niu bangshan Erguotou at 15 ml/kg body weight, and then administered with the following drugs: the test results sequentially comprise a model group, an ALDH2 single-dose group, an ALDH2 multi-dose group, an ALDH2 single-dose group and an ALDH2 multi-dose group.

Two modes of administration were used in the experiment. The first administration mode is three times of administration before, during and after drinking wine, 100ul of aqueous extract each time; wherein the administration time before drinking is recorded as-30 min, the administration time in drinking is recorded as 0min, and the administration time after drinking is recorded as 30 min. The second way is to administer the drug just once before the wine. The behavioral indexes of each group are measured at 30min after drinking, the observation lasts for 4h, the activity of each group of mice is observed, and the number of drunk mice, the number of dead mice, the drinking time, the disappearance time of righting reflex and the recovery time of righting reflex are recorded. The drunk time is the righting reflex recovery time-righting reflex disappearance time;

as can be seen from table 2 and fig. 7, 10 mice in the model group were all drunk, the drunk rate was 100%, and only 2 mice were sobered within 4 hours, and the sobered rate was 20%. The alcoholism rate of the ALDH2 multi-dose group is only 40%, and the alcoholism mice are fully sobered within 4h, and the sobering rate is 100%. The alcoholism rate of the ALDH2 single-dose group is 100%, and the alcoholism rate of the drunk mice is 50% within 4 h. The alcoholism rate of the ALDH2 composition multi-dose group is 40%, and the alcoholism rate within 4h is 100% which is the same as that of the ALDH2 multi-dose group, but compared with the ALDH2 multi-dose group, the alcoholism time of the alcoholism mice in the ALDH2 composition multi-dose group is obviously reduced to 87.75 +/-8.77 min. In addition, the single-dose group of the ALDH2 composition showed a 30% increase in the anti-hangover rate compared to the single-dose group of ALDH 2. The drunk time is also obviously reduced from 210.38 +/-26.69 min to 114.625 +/-36.15 min. The ALDH2 composition can obviously reduce the drunk time of drunk mice and accelerate sobering up.

TABLE 2 Effect of acetaldehyde dehydrogenase (ALDH2) composition on intoxication status and time in mice

Group of	Quantity (only)	Drunk number (only)	Drunkenness rate	Death number (only)	Sobering-up quantity of drunk mouse	Percentage of sobering up	Drunk time (min) of drunk mouse
								Blank group	10	0	0	0	-	-	-
Model set	10	10	100%	0	2	20%	236.50±7.01
								ALDH2 multi-dose group	10	4	40%	0	4	100%	137.25±27.11**
ALDH2 single administration group	10	10	100 %	0	5	50%	210.38±26.69
								Multiple administration of ALDH2 compositionMedicine group	10	4	40%	0	4	100%	87.75±8.77***
ALDH2 composition single administration group	10	10	100%	0	8	80%	114.625±36.15*

It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. An acetaldehyde dehydrogenase composition for relieving alcoholism, which is characterized by comprising the following components in parts by weight: 1-10 parts of acetaldehyde dehydrogenase, 40-80 parts of kudzu root extract, 10-50 parts of dried orange peel extract, 10-50 parts of fructus amomi extract, 20-40 parts of pueraria flower extract, 15-30 parts of white poria extract, 10-30 parts of small bean flower extract, 10-30 parts of elecampane extract, 10-30 parts of radix asparagi extract, 10-30 parts of moutan bark extract, 10-30 parts of rhizoma alismatis extract and 10-30 parts of liquorice extract.

2. The acetaldehyde dehydrogenase composition for alleviating hangover according to claim 1, wherein the composition comprises, in parts by weight: 5 parts of acetaldehyde dehydrogenase, 60 parts of kudzu root extract, 30 parts of dried orange peel extract, 30 parts of fructus amomi extract, 40 parts of pueraria flower extract, 20 parts of chinaroot greenbrier rhizome extract, 20 parts of small bean flower extract, 15 parts of costustoot extract, 15 parts of cochinchnese asparagus root extract, 10 parts of tree peony bark extract, 10 parts of rhizoma alismatis extract and 10 parts of liquorice extract.

3. An anti-hangover and hepatoprotective acetaldehyde dehydrogenase composition preparation, comprising the anti-hangover and hepatoprotective acetaldehyde dehydrogenase composition according to any one of claims 1 to 2.

4. The composition preparation for alleviating hangover and protecting liver acetaldehyde dehydrogenase as claimed in claim 3, further comprising pharmaceutically acceptable excipients.

5. The composition preparation of claim 3, further comprising at least one of an excipient, a binder, a filler, a lubricant, an antioxidant, a preservative or a thickener.

6. The preparation of the composition for relieving alcoholism and protecting liver acetaldehyde dehydrogenase of claim 3, wherein the preparation is an oral preparation.

7. The composition preparation for relieving alcoholism and protecting liver acetaldehyde dehydrogenase as claimed in claim 3, wherein the dosage form of the preparation is tablet, powder, pill, capsule, granule or oral liquid.