CN111297917A - Probiotic preparation for relieving acute alcoholism and preparation method and application thereof - Google Patents
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Abstract
The invention relates to a probiotic preparation for relieving acute alcoholism and a preparation method and application thereof, wherein the probiotic preparation is prepared by taking lactobacillus rhamnosus and bulgaricus as raw materials and carrying out proper proportioning, the lactobacillus rhamnosus has the effect of producing acetaldehyde dehydrogenase, the bulgaricus has the effects of producing ethanol dehydrogenase and acetaldehyde dehydrogenase, and vitamin coenzyme with redox coenzyme is added, so that the probiotic preparation for relieving acute alcoholism is finally prepared. Can be directly taken after drinking, and can be used for relieving hepatosis caused by ammonia after drinking or acetaldehyde after drinking, and can also be used for preparing medicine for treating alcoholic hepatic injury.
Description
Technical Field
The invention belongs to the technical field of probiotics, and particularly relates to a probiotic preparation for relieving acute alcoholism, and a preparation method and application thereof.
Background
The world health organization reports that the number of alcohol related cause deaths is about 300 million per year. Prolonged alcohol abuse can lead to a range of physical ailments. Acute or chronic alcohol intake can cause liver diseases (alcoholic hepatitis, steatosis, steatohepatitis and the like), the lipogenesis can be increased by drinking, and the oxidation of fatty acid is inhibited, wherein the alcoholic liver steatosis is more than 90 percent, and most commonly, the excessive accumulation of lipid drops in liver cells is mainly caused.
After the alcohol is taken, part of the alcohol is firstly metabolized under the action of gastric mucosa alcohol dehydrogenase, most of the alcohol is mainly metabolized in the liver, and the liver is the main place of alcohol metabolism. There are three major metabolic pathways for alcohol in hepatocytes: oxidative metabolism of dehydrogenase; cytochrome P450 enzyme systems, most commonly CYP2E 1; the peroxidase system metabolizes. The dehydrogenases in the liver are primarily responsible for alcohol metabolism. Alcohol is first metabolized by Alcohol Dehydrogenase (ADH) to produce toxic acetaldehyde, which is metabolized by acetaldehyde dehydrogenase (ALDH) to non-toxic acetic acid.
The alcohol-induced liver disease is associated with hepatotoxicity and toxic acetaldehyde produced thereby, the former enzyme ADH is generally present in substantially equal amounts in humans, but the latter enzyme ALDH is more abundant, the absence of ALDH prevents complete decomposition of ethanol into carbon dioxide and water, but remains in the body in the form of acetaldehyde, which is a major cause of intoxication, the major virulence factor responsible for alcoholic liver injury caused by excessive drinking, dizziness, headache, nausea, vomiting caused by drinking, and is primarily a result of acetaldehyde poisoning.
Foreign research on this aspect is early. Adawi et al fed liver-injured mice with Lactobacillus reuteri R2LC, Lactobacillus rhamnosus DSM6594, Lactobacillus fermentum 8704:3, Lactobacillus reuteri 108, Lactobacillus plantarum DSM9843+ arginine in 1997 found that the group Lactobacillus plantarum DSM9843+ arginine reduced the release of liver active enzymes (total bilirubin, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase). In 2009, short term supplementation of bifidobacterium bifidum and lactobacillus plantarum 8PA3 was reported to improve gut health in alcoholics by research performed at the university of medical science, north russia and the university of lewis vere, usa. Probiotics have potential health effects on alcoholics and may contribute to improving the activity of liver enzymes.
Further deepening the research at home and abroad. Animal experiments are utilized in the artificial liver treatment training center of Beijing Youyan hospital in 2000 to prove the protective effect of lactobacillus on the damage of gastric mucosa and liver caused by alcohol. Animal experiments such as Xiaolixian of Yangzhou university in 2007 prove the liver protection effect of lactobacillus after drinking. The experiments of experiments on 3 strains of lactic acid bacteria (s. thermophilus GP1, e.faecium ST1, l.rhamnosus green straw) obtained by screening in 2008 from the royal comedy crystal of the university of Yangzhou prove that the lactic acid bacteria have the characteristic of protecting liver injury in acute alcoholic liver injury mice.
At present, related technologies exist in China, and both patents CN201110432768.2 and CN201210005581.9 relate to a liquid beverage which has the defects that the survival time of probiotics in liquid is short, prebiotics and vitamins are lacked, the effect of relieving acute alcoholism is not obvious, and the beverage can only play a role after 12 hours after use. The invention overcomes the defects of the technology, has long survival time and can relieve acute alcoholism.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a probiotic preparation for relieving acute alcoholism and a preparation method thereof. The probiotic preparation can obviously prolong the drunkenness tolerance time, shorten the sleep time, reduce the mass concentration of ethanol and acetaldehyde, and has a relatively obvious hangover alleviating effect on acute alcoholism. The probiotic preparation can be directly taken after drinking, can relieve hepatotoxicity caused by ammonia after drinking or relieve hepatotoxicity caused by acetaldehyde after drinking, and can also be used for preparing a medicament for treating alcoholic liver injury.
The technical scheme adopted by the invention is as follows:
a probiotic preparation for relieving acute alcoholism comprises Lactobacillus rhamnosus powder and Saccharomyces bulgaricus powder.
Further preferably, the raw material components of the probiotic preparation for relieving acute alcoholism comprise:
5-15 parts of lactobacillus rhamnosus;
5-15 parts of Bujiadi yeast.
Further preferably, the raw material components of the probiotic preparation for relieving acute alcoholism comprise:
10 parts of lactobacillus rhamnosus;
10 parts of Bucadiazus yeast.
The raw material components of the probiotic preparation for relieving acute alcoholism further preferably comprise auxiliary materials, wherein the auxiliary materials comprise one or more of pyrroloquinoline quinone, xylooligosaccharide, inulin and resistant dextrin;
the added mass of the auxiliary materials accounts for 80% of the mass of the probiotic preparation.
The Lactobacillus rhamnosus is Lactobacillus rhamnosus XM106, which has been obtained in 20173Moon cake10Daily preservation in the microbiological laboratory of the national institute of Life sciences of the university of science and technology of China, the address of the preservation unit: anhui province Hefei city Jinzhai Lu 96, with accession number XM 106;
the Bujiadi yeast is the XM211 yeast Bujiadi yeast which has been obtained in 20173Moon cake10Daily preservation in the microbiological laboratory of the national institute of Life sciences of the university of science and technology of China, the address of the preservation unit: anhui province Hefei city Jinzhai Lu No. 96 with the preservation number ofXM211；
The viable count of the lactobacillus rhamnosus powder is 1.5 multiplied by 1010CFU/g, viable count of Bujiadi yeast powder is 1.5 × 1010CFU/g。
The lactobacillus rhamnosus powder is prepared by adopting the following method:
taking lactobacillus rhamnosus XM106, carrying out anaerobic culture in a fermentation medium, centrifuging, and drying to obtain lactobacillus rhamnosus XM106, wherein the fermentation temperature is 36 ℃, and the fermentation time is 9 h;
the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water.
The Bujiadi saccharomycete powder is prepared by the following method:
taking the saccharomyces bulgaricus XM211, performing aerobic culture in a fermentation medium, centrifuging, and drying to obtain the saccharomyces bulgaricus XM211, wherein the fermentation temperature is 30 ℃, and the optimal fermentation time is 22 h;
the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance water.
The preparation method of the probiotic preparation for relieving acute alcoholism comprises the following steps:
(1) respectively taking fermentation liquor obtained after anaerobic culture of lactobacillus rhamnosus XM106 and fermentation liquor obtained after aerobic culture of saccharomyces bulgaricus XM211, respectively carrying out centrifugal treatment, removing supernate, and collecting bacterial sludge;
(2) uniformly mixing lactobacillus rhamnosus bacterial mud and saccharomyces bulgaricus bacterial mud according to the mass ratio of 1:1, adding a freeze-drying protective agent, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
(3) and (3) adding other raw material components into the bacterium powder obtained in the step (2), and fully and uniformly mixing to obtain the probiotic preparation for relieving acute alcoholism.
In the step (2), the formula of the freeze-drying protective agent comprises: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose;
the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3: 1;
the freeze-drying mode is that pre-freezing is carried out at minus 25 ℃, then sublimation drying is carried out at minus 15 ℃, then desorption drying is carried out at 4 ℃, and the freeze-drying time is 18-22 h.
The probiotic preparation for relieving acute alcoholism is applied to the preparation of the drug for treating alcoholic liver injury.
The probiotic preparation of the present invention can be prepared into various dosage forms, such as powder, capsule, etc., according to the conventional method of the prior art.
The invention has the beneficial effects that:
(1) according to the probiotic preparation for relieving acute alcoholism, the lactobacillus rhamnosus and the bujiadi yeast are adopted as raw materials and are properly proportioned, the lactobacillus rhamnosus has the effect of producing acetaldehyde dehydrogenase, the bujiadi yeast has the effects of producing ethanol dehydrogenase and acetaldehyde dehydrogenase, vitamin coenzyme with the effect of redox coenzyme is also added, and the probiotic preparation for relieving acute alcoholism is finally prepared. Can be directly taken after drinking, and can be used for relieving hepatosis caused by ammonia after drinking or acetaldehyde after drinking, and can also be used for preparing medicines for treating alcoholic hepatic injury.
(2) The preparation method of the probiotic preparation has the advantages of strong practicability, simple operation, short period, simple and convenient steps, low cost, high yield, suitability for large-scale production and the like.
Detailed Description
The present invention will be described in detail with reference to specific examples. In the following examples, 1 part by weight represents 1 g.
Example 1
The embodiment provides a probiotic preparation for relieving acute alcoholism, which comprises the following raw material components:
lactobacillus rhamnosus XM106 powder (viable count is 1.5 × 10)10CFU/g), 10 parts by weight;
bacterial powder of Saccharomyces bulgaricus XM211 (viable count is 1.5 × 10)10CFU/g), 10 parts by weight;
2 parts of pyrroloquinoline quinone;
18 parts of xylo-oligosaccharide;
5 parts of inulin;
75 parts of resistant dextrin.
The probiotic preparation for relieving acute alcoholism is further prepared by the following method:
(1) taking lactobacillus rhamnosus XM106, carrying out anaerobic fermentation culture at 36 ℃ for 9h in a fermentation medium to obtain a fermentation liquid, centrifuging the fermentation liquid by a centrifugal machine, removing supernatant, and collecting lactobacillus rhamnosus XM106 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water;
taking the saccharomyces bulgaricus XM211, ventilating in a fermentation medium, carrying out aerobic fermentation culture at 30 ℃ for 22h to obtain fermentation liquor, centrifuging by a centrifuge, removing supernatant, and collecting saccharomyces bulgaricus XM211 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance of water;
(2) uniformly mixing lactobacillus rhamnosus XM106 bacterial sludge and Bujiadi yeast XM211 bacterial sludge according to the mass ratio of 1:1, adding a freeze-drying protective agent, wherein the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3:1, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
the formula of the freeze-drying protective agent comprises the following components: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose; the freeze-drying mode is that pre-freezing is carried out for 1h at minus 25 ℃, then sublimation drying is carried out for 18h at minus 15 ℃, and then resolution drying is carried out for 3h at 4 ℃;
(3) and (3) adding pyrroloquinoline quinone, xylooligosaccharide, inulin and resistant dextrin into the bacterial powder obtained in the step (2), and fully and uniformly mixing to obtain the powder of the probiotic preparation for relieving acute alcoholism.
The prepared powder is filled into aluminum plastic bags, each bag is 5g, and the number of viable bacteria is about 800 hundred million.
Example 2
The embodiment provides a probiotic preparation for relieving acute alcoholism, which comprises the following raw material components:
lactobacillus rhamnosus XM106 powder (viable count is 1.5 × 10)10CFU/g), 5 parts by weight;
bacterial powder of Saccharomyces bulgaricus XM211 (viable count is 1.5 × 10)10CFU/g), 15 parts by weight;
2 parts of pyrroloquinoline quinone;
18 parts of xylo-oligosaccharide;
5 parts of inulin;
75 parts of resistant dextrin.
The probiotic preparation for relieving acute alcoholism is further prepared by the following method:
(1) taking lactobacillus rhamnosus XM106, carrying out anaerobic fermentation culture at 36 ℃ for 9h in a fermentation medium to obtain a fermentation liquid, centrifuging the fermentation liquid by a centrifugal machine, removing supernatant, and collecting lactobacillus rhamnosus XM106 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water;
taking the saccharomyces bulgaricus XM211, ventilating in a fermentation medium, carrying out aerobic fermentation culture at 30 ℃ for 22h to obtain fermentation liquor, centrifuging by a centrifuge, removing supernatant, and collecting saccharomyces bulgaricus XM211 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance of water;
(2) uniformly mixing lactobacillus rhamnosus XM106 bacterial sludge and Bujiadi yeast XM211 bacterial sludge according to the mass ratio of 1:3, adding a freeze-drying protective agent, wherein the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3:1, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
the formula of the freeze-drying protective agent comprises the following components: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose; the freeze-drying mode is that pre-freezing is carried out for 2h at minus 25 ℃, then sublimation drying is carried out for 14h at minus 15 ℃, and then resolution drying is carried out for 2h at 4 ℃;
(3) and (3) adding pyrroloquinoline quinone, xylooligosaccharide, inulin and resistant dextrin into the bacterial powder obtained in the step (2), and fully and uniformly mixing to obtain the powder of the probiotic preparation for relieving acute alcoholism.
The prepared powder is filled into aluminum plastic bags, each bag is 5g, and the number of viable bacteria is about 800 hundred million.
Example 3
The embodiment provides a probiotic preparation for relieving acute alcoholism, which comprises the following raw material components:
lactobacillus rhamnosus XM106 powder (viable count is 1.5 × 10)10CFU/g), 15 parts by weight;
bacterial powder of Saccharomyces bulgaricus XM211 (viable count is 1.5 × 10)10CFU/g), 5 parts by weight;
2 parts of pyrroloquinoline quinone;
18 parts of xylo-oligosaccharide;
5 parts of inulin;
75 parts of resistant dextrin.
The probiotic preparation for relieving acute alcoholism is further prepared by the following method:
(1) taking lactobacillus rhamnosus XM106, carrying out anaerobic fermentation culture at 36 ℃ for 9h in a fermentation medium to obtain a fermentation liquid, centrifuging the fermentation liquid by a centrifugal machine, removing supernatant, and collecting lactobacillus rhamnosus XM106 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water;
taking the saccharomyces bulgaricus XM211, ventilating in a fermentation medium, carrying out aerobic fermentation culture at 30 ℃ for 22h to obtain fermentation liquor, centrifuging by a centrifuge, removing supernatant, and collecting saccharomyces bulgaricus XM211 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance of water;
(2) uniformly mixing lactobacillus rhamnosus XM106 bacterial sludge and Bujiadi yeast XM211 bacterial sludge according to the mass ratio of 3:1, adding a freeze-drying protective agent, wherein the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3:1, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
the formula of the freeze-drying protective agent comprises the following components: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose; the freeze-drying mode is that pre-freezing is carried out for 1h at minus 25 ℃, then sublimation drying is carried out for 18h at minus 15 ℃, and then resolution drying is carried out for 1h at 4 ℃;
(3) and (3) adding pyrroloquinoline quinone, xylooligosaccharide, inulin and resistant dextrin into the bacterial powder obtained in the step (2), and fully and uniformly mixing to obtain the powder of the probiotic preparation for relieving acute alcoholism.
The powder is filled into a cylindrical capsule shell which is prepared by taking gelatin as a main raw material, the volume of the capsule is 0.6ml, and the viable count of each capsule is about 150 hundred million.
Example 4
The embodiment provides a probiotic preparation for relieving acute alcoholism, which comprises the following raw material components:
lactobacillus rhamnosus XM106 powder (viable count is 1.5 × 10)10CFU/g), 10 parts by weight;
bacterial powder of Saccharomyces bulgaricus XM211 (viable count is 1.5 × 10)10CFU/g), 10 parts by weight;
1.4 parts by weight of pyrroloquinoline quinone;
1.44 parts by weight of xylo-oligosaccharide;
inulin, 4 parts by weight;
60 parts of resistant dextrin.
The probiotic preparation for relieving acute alcoholism is further prepared by the following method:
(1) taking lactobacillus rhamnosus XM106, carrying out anaerobic fermentation culture at 36 ℃ for 9h in a fermentation medium to obtain a fermentation liquid, centrifuging the fermentation liquid by a centrifugal machine, removing supernatant, and collecting lactobacillus rhamnosus XM106 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water;
taking the saccharomyces bulgaricus XM211, ventilating in a fermentation medium, carrying out aerobic fermentation culture at 30 ℃ for 22h to obtain fermentation liquor, centrifuging by a centrifuge, removing supernatant, and collecting saccharomyces bulgaricus XM211 bacterial sludge; the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance of water;
(2) uniformly mixing lactobacillus rhamnosus XM106 bacterial sludge and Bujiadi yeast XM211 bacterial sludge according to the mass ratio of 1:1, adding a freeze-drying protective agent, wherein the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3:1, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
the formula of the freeze-drying protective agent comprises the following components: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose; the freeze-drying mode is that pre-freezing is carried out for 1h at minus 25 ℃, then sublimation drying is carried out for 18h at minus 15 ℃, and then resolution drying is carried out for 1h at 4 ℃;
(3) and (3) adding pyrroloquinoline quinone, xylooligosaccharide, inulin and resistant dextrin into the bacterial powder obtained in the step (2), and fully and uniformly mixing to obtain the powder of the probiotic preparation for relieving acute alcoholism.
The prepared powder is filled into aluminum plastic bags, each bag is 5g, and the number of viable bacteria is about 800 hundred million.
Examples of the experiments
1. The liver protection characteristics of the probiotic preparation for relieving acute alcoholism of the probiotic preparation of example 1 of the invention are tested
(1) Experimental animals: healthy Kunming mice, provided by the animal experiment center of university of medical science in Anhui.
(2) Experimental methods
Mice were randomly divided into 5 groups, high, medium and low dose groups (0.8, 0.4, 0.2g/kg body weight) and blank, model groups, 10 per group, fasted for 12h, weighed and recorded. Respectively feeding low, medium and high doses of probiotic preparation and physiological saline into stomach, feeding liquor into stomach after 30min according to the alcohol dose of 0.4mL/10g body weight, and obtaining the drunk mouse (whether the mouse is drunk is based on whether the righting reflex disappears, the mouse is lightly put into a squirrel cage with the back facing downwards after stomach feeding, and if the posture of the mouse with the back facing downwards is kept for more than 30s, the righting reflex disappears, namely the drunk mouse is obtained).
The influence of the probiotic preparation on the biochemical indexes of the liver part of the drunken mice is detected, and the result is shown in table 1.
Table 1-effect of the probiotic formulation on AST, ALT, ADH and SOD activity of liver of intoxicated mice at different additive doses
AST and ALT are important indexes for clinically judging whether liver function abnormality exists. In this experiment, ALT and AST activities were significantly increased (P <0.01) in the model group mice compared to the blank group, indicating that alcohol caused some damage to liver function. Compared with the model group, the ALT and AST activities in the serum of the low, medium and high 3 probiotic preparation administration groups are obviously reduced (P is less than 0.01), which shows that the probiotic preparation has a certain protective effect on liver cell injury caused by acute alcoholism. SOD activity and ADH activity are also important biochemical indexes for evaluating liver functions, and in our experiments, 3 doses of probiotic preparations can also obviously increase SOD activity and ADH activity in liver tissues (P is less than 0.01); further suggesting that the probiotic preparation can prevent the mouse liver injury oxidative stress condition caused by acute alcoholism and protect the liver of the mouse. The data show that the probiotic preparation low-dose group can better reduce the ALT and AST activity and increase the SOD and ADH activity (P <0.05), and compared with the probiotic preparation low-dose group, the probiotic preparation middle-high dose group has more remarkable ALT and AST reduction and SOD and ADH activity increase (P < 0.01).
And (4) conclusion: the probiotic preparation can generate liver cell protection effect through various ways of influencing ethanol metabolism, lipid peroxidation and the like. In experiments, the probiotic preparation is also found to be capable of obviously prolonging the drunkenness tolerance time, shortening the sleep time, reducing the mass concentration of ethanol and acetaldehyde and further generating a certain detoxification effect on acute alcoholism of drunkenness mice. The experimental result fully shows that the probiotic preparation has a relatively obvious liver protection effect on the acute alcoholism mice.
2. The anti-alcoholism effect of the probiotic preparation for relieving acute alcoholism disclosed in the embodiment 1 of the invention is tested
(1) Experimental animals: healthy Kunming mice, provided by the animal experiment center of university of medical science in Anhui.
(2) Experimental methods
Preparing a positive control medicament: weighing 100mg of naloxone hydrochloride, and preparing 50mg/mL of normal saline for later use during experiments.
60 mice were randomly divided into a model group, a probiotic preparation 2g/kg/d group (low dose group), a probiotic preparation 4g/kg/d group (medium dose group), a probiotic preparation 6g/kg/d group (high dose group), and naloxone hydrochloride 5mg/kg/d group (positive drug group), and 12 mice in each group. Each group is subjected to intragastric molding by using 20mL/kg/d of 56% alcohol, after intragastric administration by using alcohol, the low, medium and high dose components of the probiotic preparation are respectively subjected to intragastric administration, the positive drug group is subjected to intraperitoneal injection of naloxone hydrochloride, the activity condition of each group of mice is observed, the righting reflex disappearance time is taken as the intoxication latency period, and the righting reflex recovery time is taken as the sobering time. The results are shown in Table 2.
Table 2-anti-hangover effect of probiotic formulation acute alcoholism mouse model at different additive doses
As can be seen from table 2, the probiotic preparation can effectively prolong the hangover latency and shorten the hangover time in the aspect of relieving alcoholism, which is related to the improvement of hepatic anti-alcoholism enzyme (ADH, ALDH) activity.
The probiotic preparation can obviously prolong the drunkenness tolerance time, shorten the sleep time and reduce the mass concentration of ethanol and acetaldehyde. The experimental result fully shows that the probiotic preparation has a relatively obvious hangover alleviating effect on mice suffering from acute alcoholism.
The present invention is not limited to the above-mentioned preferred embodiments, and any other products in various forms can be obtained by anyone in the light of the present invention, but any changes in the shape or structure thereof, which have the same or similar technical solutions as those of the present application, fall within the protection scope of the present invention.
Claims (10)
1. A probiotic preparation for relieving acute alcoholism is characterized in that the raw material components comprise lactobacillus rhamnosus powder and saccharomyces bulgaricus powder.
2. The probiotic formulation according to claim 1, characterized in that the raw material components comprise:
5-15 parts of lactobacillus rhamnosus powder;
5-15 parts of Bujiadi saccharomycete powder.
3. The probiotic formulation according to claim 1, characterized in that the raw material components comprise:
10 parts of lactobacillus rhamnosus powder;
10 parts of Bujiadi saccharomycete powder.
4. The probiotic preparation according to claim 1, further comprising an adjuvant comprising one or more of pyrroloquinoline quinone, xylo-oligosaccharide, inulin, resistant dextrin;
the added mass of the auxiliary materials accounts for 80% of the mass of the probiotic preparation.
5. The probiotic preparation according to claim 1, wherein the lactobacillus rhamnosus is lactobacillus rhamnosus XM106, which has been deposited at the microbiology laboratory of the life sciences college of science and technology university of china at 3 and 10 months in 2017 at the address of the deposition unit: anhui province Hefei city Jinzhai Lu 96, with accession number XM 106;
the saccharomyces bulgaricus is saccharomyces bulgaricus XM211, which is preserved in microbiological laboratories of national institute of Life sciences of university of science and technology in 2017, 3 months and 10 days, and the preservation unit address is as follows: anhui province Hefei city Jinzhai Lu 96, with accession number XM 211;
the viable count of the lactobacillus rhamnosus powder is 1.5 multiplied by 1010CFU/g, viable count of Bujiadi yeast powder is 1.5 × 1010CFU/g。
6. The probiotic preparation according to claim 1, characterized in that the lactobacillus rhamnosus bacterial powder is prepared by the following method:
taking lactobacillus rhamnosus XM106, carrying out anaerobic culture in a fermentation medium, centrifuging, and drying to obtain lactobacillus rhamnosus XM106, wherein the fermentation temperature is 36 ℃, and the fermentation time is 9 h;
the fermentation medium comprises the following components in percentage by weight: 3.0 percent of glucose, 0.5 percent of yeast extract, 0.5 percent of casein peptone, 0.2 percent of citric acid diamine, 0.4 percent of sodium acetate, 0.06 percent of magnesium sulfate, 0.05 percent of span-80 and the balance of water.
7. The probiotic preparation for alleviating acute alcoholism as claimed in claim 6, wherein said Bugardisia fungus powder is prepared by the following method:
taking the saccharomyces bulgaricus XM211, performing aerobic culture in a fermentation medium, centrifuging, and drying to obtain the saccharomyces bulgaricus XM211, wherein the fermentation temperature is 30 ℃, and the optimal fermentation time is 22 h;
the fermentation medium comprises the following components in percentage by weight: 3.0% glucose, 2.5% peptone, 2.0% corn steep liquor, 2.0% potassium nitrate, 0.12% monopotassium phosphate, 0.08% magnesium sulfate and the balance water.
8. The method for preparing the probiotic preparation for relieving acute alcoholism according to any one of claims 1-7, characterized by comprising the following steps:
(1) respectively taking fermentation liquor obtained after anaerobic culture of lactobacillus rhamnosus XM106 and fermentation liquor obtained after aerobic culture of saccharomyces bulgaricus XM211, respectively carrying out centrifugal treatment, removing supernate, and collecting bacterial sludge;
(2) uniformly mixing lactobacillus rhamnosus bacterial mud and saccharomyces bulgaricus bacterial mud according to the mass ratio of 1:1, adding a freeze-drying protective agent, and freeze-drying in a freeze dryer to obtain white powdery bacterial powder;
(3) and (3) adding other raw material components into the bacterium powder obtained in the step (2), and fully and uniformly mixing to obtain the probiotic preparation for relieving acute alcoholism.
9. The preparation method according to claim 8, wherein in the step (2), the formulation of the lyoprotectant comprises: 5% of skimmed milk powder, 5% of trehalose, 3% of konjac polysaccharide and 5% of sucrose;
the ratio of the adding mass of the freeze-drying protective agent to the total mass of the bacterial sludge is 3: 1;
the freeze-drying mode is that pre-freezing is carried out at minus 25 ℃, sublimation drying is carried out at minus 15 ℃, and then resolution drying is carried out at 4 ℃; the freeze-drying time is 18-22 h.
10. Use of a probiotic formulation for alleviating acute alcoholism as defined in any one of claims 1 to 7 in the manufacture of a medicament for the treatment of alcoholic liver injury.
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