CN114129613A - Application of rabdosia lophanthide in preventing or treating alcohol dependence - Google Patents
Application of rabdosia lophanthide in preventing or treating alcohol dependence Download PDFInfo
- Publication number
- CN114129613A CN114129613A CN202010916751.3A CN202010916751A CN114129613A CN 114129613 A CN114129613 A CN 114129613A CN 202010916751 A CN202010916751 A CN 202010916751A CN 114129613 A CN114129613 A CN 114129613A
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- China
- Prior art keywords
- alcohol
- rabdosia lophanthide
- rabdosia
- product
- lophanthide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Abstract
The invention relates to application of rabdosia lophanthide in preventing or treating alcohol dependence. The invention researches for the first time to find that the rabdosia lophanthide can obviously reduce the position preference effect of mice on alcohol, interfere the formation of alcohol-induced behavior sensitization, reduce the drinking capacity of the mice, reduce the alcohol preference degree, effectively improve the anxiety-like behavior of the mice after acute alcohol withdrawal, does not influence the absorption and metabolism of the alcohol and has excellent prevention and treatment effect on alcohol dependence.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of rabdosia lophanthide in preventing or treating alcohol dependence.
Background
The alcohol has strong mental activity, and can generate subjective feelings of being happy, forgetting to worry, relaxing the whole body, relieving fatigue, revivifying the spirit and the like after drinking. Alcohol is a recognized social lubricant, interpersonal interaction and compensation are more and more in the modern society, and the consumption amount of alcohol is increased sharply. However, prolonged unregulated drinking necessarily leads to alcohol-related health problems such as acute alcoholism (intoxication), alcoholic liver disease and alcohol dependence (addiction). Acute alcoholism refers to abnormal body function after a drinker takes a large amount of alcohol at one time, and is manifested by self-control force reduction, speech ambiguity, limb weakness and the like, and serious patients can die; the alcoholic liver disease is a liver disease caused by long-term excessive drinking, is manifested as fatty liver at the initial stage, and further can develop into alcoholic hepatitis, hepatic fibrosis and liver cirrhosis, and can induce extensive hepatocyte necrosis even cause liver failure when the alcohol is seriously drunk; unlike acute alcoholism and alcoholic liver disease, alcohol dependence is a physical or psychological disorder caused by repeated drinking, manifested by intense and compulsive drinking craving (mental dependence), and when drinking is stopped, withdrawal symptoms (physical dependence) such as anxiety, dysphoria, depression, palpitation, and even tremor can be seen. Although alcoholism is viewed as a bad habit worldwide, the number of alcohol-dependent patients is increasing day by day. Since alcohol can cause serious damage to various tissues and organs of the human body, especially the liver, the treatment of alcohol dependence is the first step in the treatment of alcohol-related diseases. However, for the treatment of alcohol dependence, there are very limited drugs available, and the opioid receptor antagonists naltrexone and the GABA receptor agonist calcium acamprosate are FDA approved for the prevention of over-drinking and relapse from alcohol withdrawal. Yet another drug approved for the prevention of alcohol consumption is disulfiram, which reduces alcohol consumption by increasing the alcohol sensitivity of the drinker.
The existing medicines for treating alcohol dependence have serious adverse reactions, poor patient compliance and a plurality of limitations on clinical use, and cannot meet the requirements of a plurality of current alcohol dependence patients.
Rabdosia lophanthide is a plant of Rabdosia of Labiatae, is a mainstream variety in Guangdong, wherein Isodon lophanthide var. gerardianus (Benth) H.Hara) is listed as a new food raw material catalog by national Weijie in 2013, has the effects of clearing away dampness and heat, removing jaundice and benefiting gallbladder, cooling blood and removing stasis, and is frequently used for treating diseases such as acute icteric hepatitis, acute cholecystitis, pharyngolaryngitis, traumatic injury stasis and the like. Research shows that the rabdosia lophanthide mainly contains flavonoid, terpenoid, phenolic acid, volatile oil, coumarin and other components.
The recent research shows that the rabdosia lophanthide aqueous extract can inhibit the mouse ear inflammatory reaction caused by xylene and is used for treating carbon tetrachloride (CCl)4) Acute liver injury of mice caused by alcohol, acute liver injury of mice caused by D-galactosamine (D-CalN), acute liver injury of mice caused by acetaminophen, acute liver injury of mice caused by Canavalia gladiata protein A (ConA) and immunological liver injury of mice caused by lipopolysaccharide plus BCG (LPS + BCG) have protection effects. However, the alcohol dependence of rabdosia lophanthide has not been studied so far.
Disclosure of Invention
Based on the situation, the invention aims to provide the application of the rabdosia lophanthide in preventing or treating the alcohol dependence.
The specific technical scheme is as follows:
application of herba Rabdosiae Lophanthoidis in preparing product for preventing or treating alcohol dependence is provided.
The invention also aims to provide application of the rabdosia lophanthide in preparing a product for improving anxiety-like behaviors caused by acute alcohol withdrawal.
The invention also aims to provide the application of the rabdosia lophanthide in preparing the product for giving up alcohol.
In some of these embodiments, the rabdosia lophanthide is a crude rabdosia lophanthide or a rabdosia lophanthide extract.
In some of these embodiments, the rabdosia lophanthide extract is a water extract or an alcohol extract of rabdosia lophanthide. The rabdosia lophanthide alcohol extract is a rabdosia lophanthide alcohol extract.
In some of these embodiments, the product is a nutraceutical, a functional food, or a pharmaceutical.
Another object of the present invention is to provide a product for preventing or treating alcohol dependence, which comprises linearstripe rabdosia herb or linearstripe rabdosia herb extract, and auxiliary materials.
In some of these embodiments, the rabdosia lophanthide is rabdosia lophanthide tea.
In some of these embodiments, the Rabdosia lophanthide tea is prepared by the steps comprising: sequentially withering, rolling, fermenting, air cooling, carding and drying the fresh leaves of the rabdosia lophanthide.
In some of these embodiments, the Rabdosia lophanthide tea is prepared by the steps comprising:
(a) withering: withering fresh leaves of the rabdosia lophanthide;
(b) rolling: rolling the withered leaves obtained in the step (a) for 20 +/-10 min;
(c) fermentation: fermenting the twisted leaves at 40 + -5 deg.C for 6 + -1 h;
(e) air cooling: spreading the fermented leaves to a thickness of (1 + -0.2) cm;
(f) strip tidying: treating the air-cooled leaves by carding at 240 +/-20 ℃ for 30 +/-5 min, and spreading for cooling;
(g) and (3) drying: and drying the tea leaves to obtain the rabdosia lophanthide tea.
In some of these embodiments, the rabdosia lophanthide extract is a water extract or an alcohol extract of rabdosia lophanthide. In some of the embodiments, the alcohol extract of rabdosia lophanthide is an ethanol extract of rabdosia lophanthide, and further, the preparation of the ethanol extract of rabdosia lophanthide comprises the following steps: taking rabdosia lophanthide, adding 50-80 wt% of ethanol water solution, extracting, filtering, and concentrating under reduced pressure to obtain the rabdosia lophanthide. Further, the ethanol aqueous solution is 60-80 wt% of ethanol aqueous solution, and the extraction time is 10-40 min, preferably 20-30 min. Furthermore, the mass of the ethanol water solution is 10-30 times of that of the rabdosia lophanthide.
In some embodiments, the preparation process of the rabdosia lophanthide water extract comprises: taking rabdosia lophanthide, decocting for 1-3 times, and concentrating the obtained decoction into thick paste.
In some of the examples, the relative density of the thick paste at 50-60 ℃ is 1.1-1.25.
In some embodiments, the time for each decoction with water is 0.5-2 h, preferably 0.8-1.2 h.
The invention also aims to provide a product for preventing or treating alcohol dependence, wherein the active ingredients of the product are prepared from rabdosia lophanthide and at least one of sweet osmanthus, Chinese date, momordica grosvenori and medlar.
In some of these embodiments, the active ingredients of the product are prepared from linearstripe rabdosia herb and sweet osmanthus flower.
In some of these embodiments, the active ingredients of the product are prepared from Rabdosia lophanthide, Zizyphi fructus, and Lycii fructus.
In some of these embodiments, the active ingredients of the product are prepared from linearstripe rabdosia herb, jujube, sweet osmanthus flower and wolfberry fruit.
Compared with the prior art, the invention has the following beneficial effects:
the invention adopts animal models with mental dependence of various addictive drugs to carry out experimental research on the rabdosia lophanthide, discovers for the first time that the rabdosia lophanthide has good function of preventing and treating alcohol dependence and is beneficial to promoting alcohol withdrawal. The above-mentioned function of herba Rabdosiae Lophanthoidis for preventing and treating alcohol dependence does not affect blood alcohol concentration, and is not produced by affecting blood alcohol absorption and metabolism, and its mechanism is completely different from conventional mechanism for relieving alcoholism and protecting liver. The effect of the linearstripe rabdosia herb for preventing and treating alcohol dependence is embodied in that: the rabdosia lophanthide can obviously reduce the position preference effect of mice on alcohol, interfere the formation of behavior sensitization induced by alcohol dependence, effectively improve anxiety-like behaviors of the mice after acute alcohol withdrawal and reduce alcohol preference degree, thereby realizing good effect of preventing and treating alcohol dependence.
Drawings
FIG. 1 is a graph of the effect of Rabdosia Lophanthoidis on alcohol-induced conditioned place preference
A: a base value forward measurement stage; b:a testing stage;##P<0.01vs control group;**P<0.01vs model set;
FIG. 2 is a graph showing the effect of Rabdosia serra on sensitization of alcohol-induced behavior
A: a base value forward measurement stage; b: acute alcohol stimulation phase; c: a testing stage;##P<0.01vs control group;*P<0.05、**P<0.01vs model set;
FIG. 3 is the effect of Rabdosia lophanthide on the blood alcohol concentration in mice
FIG. 4 is a graph showing the effect of Rabdosia lophanthide on the drinking capacity and alcohol preference of mice
A: a base value of drinking capacity; b: the drinking capacity; c: alcohol preference base value; d: alcohol preference degree;**P<0.01vs model set;
FIG. 5 is a graph showing the effect of Rabdosia lophanthide on drinking in mice in the dark
**P<0.01vs model set;
FIG. 6 is the effect of Rabdosia Lophanthoidis on dark and light box experiment after acute alcohol withdrawal of mice
A: entering a latent period of a dark box; b: the residence time in the open box; c: the number of crossings between light and dark boxes;##P<0.01vs control group;*P<0.01、**P<0.01vs model set.
Detailed Description
Experimental procedures according to the invention, in which no particular conditions are specified in the following examples, are generally carried out under conventional conditions, or under conditions recommended by the manufacturer. The various chemicals used in the examples are commercially available.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, article, or device that comprises a list of steps is not limited to only those steps or modules listed, but may alternatively include other steps not listed or inherent to such process, method, article, or device.
The "plurality" referred to in the present invention means two or more. "and/or" describes the association relationship of the associated objects, meaning that there may be three relationships, e.g., a and/or B, which may mean: a exists alone, A and B exist simultaneously, and B exists alone. The character "/" generally indicates that the former and latter associated objects are in an "or" relationship.
One embodiment of the invention provides application of rabdosia lophanthide in preparing a product for preventing or treating alcohol dependence.
In another embodiment of the invention, the application of rabdosia lophanthide in preparing a product for improving anxiety-like behavior caused by acute alcohol withdrawal is provided.
The invention also provides application of the rabdosia lophanthide in preparing a product for giving up alcohol.
In some embodiments, the rabdosia lophanthide is a rabdosia lophanthide crude drug, namely a whole herb dried product.
In some embodiments, the rabdosia lophanthide is a rabdosia lophanthide extract, which may be, for example, a water extract of rabdosia lophanthide or an alcohol extract of rabdosia lophanthide, obtained by conventional methods in the industry.
The product for preventing or treating alcohol dependence can be a health-care product and also can be a functional food, and the product can also be the combination of rabdosia lophanthide and other common health-care foods (such as sweet osmanthus, Chinese date, fructus momordicae, medlar and the like) as long as the activity of the rabdosia lophanthide for preventing and treating alcohol dependence is not influenced.
The embodiment also provides a product for improving mouthfeel and preventing or treating alcohol dependence, and the active ingredients of the product are prepared from rabdosia lophanthide and at least one of sweet osmanthus, Chinese date, momordica grosvenori and medlar. Wherein the rabdosia lophanthide can be combined with at least one of sweet osmanthus, Chinese date, fructus momordicae and medlar in any proportion.
In some embodiments, the active ingredients of the product are prepared from ingredients including linearstripe rabdosia herb and sweet osmanthus flower. The rabdosia lophanthide and the sweet osmanthus flower can be combined in any mass proportion, for example, the rabdosia lophanthide and the sweet osmanthus flower can be selected from 1-100: 0.5 to 100, 50 to 100:0.5 to 1 or 1 to 20:1 to 20, further selected from 1 to 5: 1-5, specifically selected from: 80:0.5, 100:1, 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1, 5: 1. The osmanthus fragrans of the invention can be selected from dried osmanthus fragrans or osmanthus fragrans extracts (such as water extracts, alcohol extracts or osmanthus fragrans oil), and can also be selected from other conventional processed products which do not influence the activity of osmanthus fragrans.
In some embodiments, the active ingredients of the product are prepared from linearstripe rabdosia herb, jujube, and wolfberry. The rabdosia lophanthide, the Chinese date and the Chinese wolfberry can be combined in any mass proportion, such as 1-5: 1-5: 1-5, specifically selected from: 1:1:1, 1:2:1, 1:3:1, 1:4:1, 1:5:1, 2:1:1, 3:1:1, 4:1:1, 5:1:1, 1:1:2, 1:1:3, 1:1:4, 1:1: 5.
In some embodiments, the active ingredient of the product is prepared from linearstripe rabdosia herb, sweet osmanthus flower, Chinese date, and Chinese wolfberry. The rabdosia lophanthide, the sweet osmanthus flower, the Chinese date and the Chinese wolfberry can be combined in any mass ratio, for example, the rabdosia lophanthide, the sweet osmanthus flower, the Chinese date and the Chinese wolfberry can be selected from 0.5-10: 1-5, and specifically selected from: 1:1:1:1, 1:2:1:1, 1:3:1:1, 1:4:1:1, 1:5:1:1, 2:1:1:1, 3:1:1:1, 4:1:1:1, 5:1:1:1, 6:1:1:1, 7:1:1:1, 1:1:2:1, 1:1:3:1, 1:1:4:1, 1:1:5:1, 1:1:1:2, 1:1:1:3, 1:1:1:4, 1:1:1: 5.
In some embodiments, the active ingredients of the product are prepared from linearstripe rabdosia herb, sweet osmanthus flower, and luo han guo. The rabdosia lophanthide, the sweet osmanthus flower and the momordica grosvenori can be combined in any mass ratio, for example, the rabdosia lophanthide, the sweet osmanthus flower and the momordica grosvenori can be selected from 1-100: 0.5-5: 1-100, further from 80-100: 0.5-2: 20-40 or from 1-5: 1-5, and specifically can be selected from: 100:0.5:20, 1:1:1, 1:2:1, 1:3:1, 1:4:1, 1:5:1, 2:1:1, 3:1:1, 4:1:1, 5:1:1, 1:1:2, 1:1:3, 1:1:4, 1:1: 5.
The products according to the invention may also be in the form of medicaments.
The embodiment also provides a product for preventing or treating alcohol dependence, which comprises rabdosia lophanthide or rabdosia lophanthide extract and auxiliary materials. Furthermore, the rabdosia lophanthide is rabdosia lophanthide tea, and the rabdosia lophande tea is tea prepared by using rabdosia lophande fresh leaves as raw materials and according to a conventional black tea preparation method, and the tea comprises withering, rolling and fermenting treatment, and further can comprise air cooling, strip tidying and drying treatment.
In some embodiments, the rabdosia lophanthide extract is a water extract or an alcohol extract of rabdosia lophanthide. Further, the rabdosia lophanthide extract is a rabdosia lophanthide water extract; the preparation process of the yellow grass water extract comprises the following steps: taking rabdosia lophanthide, decocting for 1-3 times, and concentrating the obtained decoction into thick paste. Further, the relative density of the thick paste at 50-60 ℃ is 1.1-1.25.
The product of the embodiment is not limited in dosage form, and the food can be any conventional dosage form such as beverage, tea bag, tea, capsule, tablet, granule, pill, syrup, paste and the like; the health care product can be any conventional dosage form such as oral liquid, tea, capsules, tablets, granules, pills, syrup, paste, emulsion or suspension; the medicine can be any conventional dosage forms such as tea, oral liquid, capsules, tablets, granules, injection, pills, syrup, powder, paste, emulsion or suspension.
In this embodiment, the adjuvant is any adjuvant acceptable in the preparation of medicines, health products and foods, and is selected from any one or more of excipients, fillers, bulking agents, binders, humectants, disintegrating agents, slow-dissolving agents, absorption accelerators, adsorbents, diluents, solubilizers, emulsifiers, lubricants, wetting agents, suspending agents, flavoring agents and perfumes.
The present invention will be described in further detail with reference to specific examples.
Example 1
Preparing an aqueous extract of rabdosia lophanthide: taking 100g of rabdosia lophanthide whole herb dried product, adding 600ml of water, decocting for 2 times, wherein the decocting time is 1 hour each time, combining the decoctions, and concentrating to obtain thick paste with the relative density of 1.1-1.25 (50-60 ℃) for later use (1g of rabdosia lophanthide crude drug is equivalent to 0.1g of rabdosia lophanthide extract).
1. Experimental reagent
Alcohol: (intraperitoneal injection dose is 2g/kg, concentration is 15%);
0.9% physiological saline;
the aqueous extract of linearstripe rabdosia herb prepared in example 1 (the modeling is intraperitoneal injection, and the treatment is intragastric administration).
2. Experimental animals and groups
C57BL/6J mice, male, 150, at least 60 mice which can be used for experiments are screened by condition position preference experiment (CPP) culling, and the mice are divided into 6 groups of 10 mice; the normal control group (normal saline), the single linearstripe rabdosia herb group (linearstripe rabdosia herb crude drug 1.0g/kg + normal saline), the alcohol dependence model group (normal saline + alcohol), the linearstripe rabdosia herb low dose group (linearstripe rabdosia herb crude drug 0.25g/kg + alcohol), the linearstripe rabdosia herb medium dose group (linearstripe rabdosia herb crude drug 0.5g/kg + alcohol), and the linearstripe rabdosia herb high dose group (linearstripe rabdosia herb crude drug 1.0g/kg + alcohol) are arranged in sequence. According to the experimental result measured in the stage of basic value forward measurement, eliminating the over-moved or under-moved mice, and randomly and uniformly grouping the remaining mice: the 60 mice were divided into 6 groups of 10 mice each, the groups being identical to those described above.
3. Test apparatus
CPP video experimental system (guangzhou feidi biotechnology limited): CPP box (400mm X150 mm X200 mm).
4. Test procedure
4.1 conditional site preference test
A conditional situational preference (CPP) animal model is a common means for evaluating alcohol reward effect and dependence effect, and is an animal model for researching drug reward effect established according to the Barplov conditioned reflex theory, and after the reward effect of an addictive drug is associated with a specific environment, an experimental animal generates the reward effect in the environment related to the reappearance of the addictive drug. A general experiment comprises three stages: a base value pretest phase, a conditional location preference training phase and a test phase. On day 1 of the experiment, the isolation door in the CPP box was opened to allow the mice to explore freely between the black and white boxes. The residence time in the black and white boxes within 15min was recorded, and the preference of each mouse for CPP in the black and white boxes was determined. Mice with a natural preference for black or white boxes (dwell >500s) were culled according to basal values. The experiment comprises 4 training rounds on 2-9 days. According to the basic value, a non-preferential case of the mouse is used as a medicine accompanying case, a white case is used as the medicine accompanying case in the experiment, and a black case is used as the non-medicine accompanying case. Injecting normal saline into abdominal cavity on 2, 4, 6 and 8 days of training for 5min, placing into non-accompanied medicine box, staying for 30min, and taking out. Alcohol is given for 5min on 3 rd, 5 th, 7 th and 9 th days of training, and then the medicine is put into the medicine-accompanying box, stays for 30min and then is taken out. The normal control group and the model group of mice are given gavage normal saline 30min before training on the 2 nd to 9 th days, and the rabdosia lophanthide group is given gavage with different doses 30min before training on the 2 nd to 9 th days.
After 4 rounds of training, the test was performed on day 10 with no drug administration on the day of the test. And (4) removing the middle isolation door, putting the mouse into the middle isolation door, freely selecting the mouse between the two boxes, and recording the residence time of the mouse in the medicine accompanying box and the non-medicine accompanying box within 15min respectively.
4.2 behavioral sensitization test
The behavior sensitization experiment is divided into four stages: a basal value pretest phase, an acute alcohol induction phase, a sensitization phase and a test phase. (1) Preliminary stage of basal values (1 st to 2d) in order to let mice adapt to intraperitoneal injection and establish basal activity values. Before the experiment, the mice are adapted to a test room for at least 1h, weighed and injected with normal saline in an abdominal cavity according to the body weight, immediately placed in the center of an open box, freely explored for 15min in the open box, and recorded with the activity times as the basic activity value of the mice. (2) Acute alcohol stimulation (3 d): after the mice of the alcohol dependence model group, the rabdosia lophanthide low dose group, the rabdosia lophanthide medium dose group and the rabdosia lophande high dose group are adapted to the test environment, alcohol (2.0g/kg) is injected into the abdominal cavity according to the body weight, normal control group and the rabdosia lophande single group are injected with normal saline in equal volume, the normal control group and the rabdosia lophande single group are immediately placed in the center of an open box, the mice freely explore, and the activity times of the mice are recorded. (3) Sensitization stage (4 th to 13 th): after the mice in each group were administered normal saline or Rabdosia lophanthide for 30min by gavage every day, alcohol (2.5g/kg) was intraperitoneally injected 1 time per day, and the mice were returned to their cages immediately after the injection. (4) Test phase (14 d), also called challenge experiment: following 1h acclimation to the test environment, mice were challenged i.p. with a small dose of alcohol (2.0g/kg), placed immediately in the center of the open box, mice were freely explored for 15min, and total activity was recorded as per sample 3 d. The toilet water was used to wipe the device after each animal was tested (alcohol was used for wiping, but diluted toilet water was used instead of alcohol to avoid the effect of alcohol on the test).
4.3 automatic headspace-gas chromatography-mass spectrometry for determining blood alcohol concentration
After preparing the standard solution and the blood sample, precisely measuring the standard solution or the sample solution in a headspace bottle, adding a tertiary butanol internal standard solution, and immediately sealing the headspace bottle by a silica gel sealing gasket by using a jaw device. And (5) placing the sample into an automatic headspace constant temperature furnace for constant temperature, and then measuring.
4.4 Two bottle experiment (Two bottle choice)
After the mice adapt to the environment for 1 week, the mice are raised in a single cage and are fed with two bottles, wherein one bottle is filled with pure water, the other bottle is filled with 10% alcohol, and the mice freely forage for 3 weeks; water and alcohol were weighed at 4:30-6:00 pm each day and fresh water and alcohol were replaced every other day. To prevent mice from developing a site preference, the two bottle sites were alternated daily. According to the 24-hour alcohol consumption and alcohol preference of the mice in the last 3 days of alcohol drinking training, the mice with over-high or over-low alcohol consumption are removed, and the rest mice are randomly divided into 3 groups. The administration by intraperitoneal injection is carried out 1 time a day for 1 week after the weighing of the water bottle is finished. Taking the experimental data of the last 3 days into statistics, and calculating the drinking capacity according to a formula: (consumption of alcohol solution × mass ratio of 10% alcohol)/body weight in g/kg/24 h; alcohol preference calculation formula: alcohol consumption/(alcohol consumption + water consumption). To reduce the effect of stress on the behavior of alcohol consumption, mice were stroked before the start of the experiment, each for 1min each day for 3 consecutive days.
4.5 Drinking experiment in darkness (Drink in dark)
Alcoholism behaviour was defined by the national institute of alcoholism and alcoholism as a short time of heavy drinking leading to blood alcohol concentrations in excess of 0.08%. Alcoholism is more common among young people and is a risk factor for the future development of alcohol abuse and dependence. The experiment of drinking in the dark is a common animal model for simulating human behaviour of alcoholism, i.e. behaviour of drinking at high concentration in large quantities in a short time.
According to the circadian rhythm of the drinking behavior of animals, the experiment is usually carried out after the lamp is turned off, water is changed into 20% alcohol, the animals are allowed to drink for 2-4 h continuously, and the drinking volume/body weight ratio of the animals in a short time is measured. The natural high-drinking strain mouse (C57BL/6J mouse) can drink a large amount of alcohol to make the blood alcohol concentration more than or equal to 0.1%, and produce drunk behavior. This model is commonly used to study neurochemical pathways and neural circuits of alcohol dependence or rehydration behavior. According to the circadian rhythm of the mice drinking, more drinking was made at night and less drinking was made in the daytime, and the experiment was started after turning off the light for 1 h. The water bottle is removed, only one bottle filled with 20% alcohol is provided, the mouse drinks the wine for 4 hours by itself, the weight of the bottle before and after drinking is weighed, the experiment is continuously tested for 1 week, and the result of the last 3d experiment is taken for statistics. Alcohol consumption calculation formula: (consumption amount of alcohol solution × mass ratio of 20% alcohol)/body weight. To determine whether the aqueous extract of Rabdosia lophanthide affects the drinking behavior of mice, each group of mice was given the corresponding drug or saline by intraperitoneal injection 30min before the start of the experiment, once a day for 1 week.
4.6 light and dark box experiment (light/dark box test)
The light and dark box experiment is a common animal model for evaluating anxiety-like behavior. The light and dark box is divided into a light box and a dark box by a baffle plate with a circular hole with the diameter of 4cm at the center, and a 40w bulb is provided above the light box by 60 cm. The experimenter puts the mouse into the light box with the back facing the dark box, and records the latency period of the mouse passing from the light box into the dark box, the residence time in the light box and the number of times of passing between the light box and the dark box within 5 min. The device was wiped with floral water after each animal experiment was completed. And in 7 days before the experiment, mice in the single rabdosia lophanthide group and the rabdosia lophanthide low-dose group, the medium-dose group and the high-dose group are gavaged with rabdosia lophanthide, and normal control groups and the alcohol dependence model group are gavaged with physiological saline with the same volume, and are subjected to light and dark box experiments in 7 days after administration.
5. Statistical analysis process
Statistical software is adopted for analysis, and experimental data are expressed by mean +/-standard error. Except that the results of the alcohol-induced behavior sensitization experiment adopt single-factor repeated measurement analysis of variance, the rest data adopt single-factor analysis of variance to carry out statistics. Carrying out homogeneity of variance test, and if the variances are uniform, carrying out pairwise comparison between groups by using an LSD method; if the variance is not uniform, comparisons between groups are made using the Dunnett's T3 method. P <0.05 was determined to be statistically significant.
6. Test results
6.1 Rabdosia Lophanthoides Lophant attenuates alcohol-induced conditioned place preference
The conditional position preference experiment is to establish an animal model for researching the reward effect of the drug according to Pavlov's conditioned reflex theory, and after the reward effect of the addictive drug is associated with a specific environment, the experimental animal generates the reward effect in the environment related to the reappearance of the addictive drug. The results of the baseline pre-test phase experiments showed no significant difference in baseline values among the groups of mice (p-0.7656, fig. 1A). After 4 rounds of normal saline alcohol training for 8 days, compared with the training before, the preference degree of the normal control group for the companion drug case has no obvious difference, which indicates that the normal saline does not produce reward effect; compared with a normal control group, the detention time of the pure rabdosia lophanthide treatment group in the concomitant drug box has no obvious difference, the detention time of the alcohol treatment group (namely the model group) in the concomitant drug box is obviously prolonged, the preference degree of the alcohol treatment group (namely the model group) to the concomitant drug box is obviously improved (P <0.01, figure 1B), the fact that the pure rabdosia lophanthide treatment does not cause the preference degree of mice to the concomitant drug box is shown, the alcohol generates obvious reward stimulation effect, after training, the mice establish the relevance between the alcohol reward effect and the concomitant drug box, and the alcohol induction conditional position preference model is successfully prepared. Compared with an alcohol dependence model group, the preference degree of mice treated by the rabdosia lophanthide medium and high dose groups on the concomitant drug box is obviously reduced (P <0.01, figure 1B), and the rabdosia lophanthide interferes with the establishment of the association of the alcohol reward effect and a specific environment.
6.2 Effect of Rabdosia Lophanthoides on alcohol-induced behavioral sensitization
Repeated, intermittent treatments with psychotropic drugs can significantly increase the individual's responsiveness to the drug, which manifests itself in animals with significantly increased spontaneous activity. Induction of behavioral sensitization by addictive drugs is associated with compulsive medication. Baseline pretest phase experimental results showed no significant difference in the number of mouse activities between groups (p-0.4057, fig. 2A). In the acute alcohol stimulation stage, the number of activities of the mice stimulated by alcohol is obviously increased in the model group, the rabdosia serra low dose group, the medium dose group and the high dose group, which shows that the alcohol causes the behavior sensitization of the mice, and the model is successfully established (P is less than 0.01, and figure 2B). In the sensitization training stage, the phenomenon of the activity increase of sensitized mice caused by alcohol stimulation is obviously reduced by applying the rabdosia lophanthide before the alcohol injection (P <0.05, P <0.01, figure 2C), which indicates that the rabdosia lophanthide interferes with the formation of alcohol-induced behavioral sensitization.
6.3 Effect of Rabdosia Lophanthoides on blood alcohol concentration in mice
To evaluate whether Rabdosia lophanthide acts by affecting the absorption and metabolism of alcohol, blood was collected after the behavioral test (after each group of mice was gavaged with physiological saline or Rabdosia lophanthide drug, alcohol (2.5g/kg) was intraperitoneally injected, and mouse serum was collected after 1 hour for assay analysis), and the whole blood alcohol concentration was determined by headspace gas chromatography-mass spectrometry. The results showed no significant difference in blood alcohol concentration between groups (p-0.4057, fig. 3). The rabdosia lophanthide does not influence the blood alcohol concentration, and the rabdosia lophanthide does not inhibit the acquisition of alcohol-induced behavior sensitization by influencing alcohol absorption and metabolism.
6.4 Effect of Rabdosia Lophanthoides on mouse alcohol consumption and alcohol preference
The experimental results of the alcohol consumption and alcohol preference of the mice are shown in figure 4, the alcohol consumption and the alcohol preference basic value of the mice among the groups have no obvious difference, and after the mice are trained to drink, compared with a blank control group, the alcohol consumption and the alcohol preference of the mice can be reduced to different degrees in the rabdosia lophanthide medium-high dose group and the rabdosia lophanthide high-dose group (P is less than 0.05, and P is less than 0.01).
6.5 Effect of Rabdosia Lophanthoides on alcohol consumption in mice in the dark
The mouse dark drinking experiment is an animal model which is commonly used for simulating the behavior of drinking a large amount of alcohol in a short time (namely the behavior of alcoholism) in human beings. Experimental results show that after 1 hour of lamp turning off, the drinking capacity of the mice reaches 4.575 +/-0.7259 g/kg within only 4 hours, and the drinking capacity of the mice can be obviously reduced by high dose of rabdosia lophanthide (P is less than 0.01, and figure 5).
6.6 Effect of Rabdosia Lophanthoides on light and dark box experiment after acute alcohol withdrawal of mice
The light and dark box is also a common animal model for evaluating anxiety states in animals. Experimental results fig. 6 shows that the model mice had significantly reduced Latency into the dark box (Latency) (P <0.01, fig. 6A), significantly reduced residence time in the light box (Duration) (P <0.01, fig. 6B), and significantly reduced number of crossings between the light and dark boxes (Transitions) (P <0.01, fig. 6C), compared to the control mice, while the high dose of rabdosia serrata was given to reverse these changes to varying degrees (P <0.05, P <0.01), significantly improving anxiety-like behavior of the mice.
In addition, the rabdosia lophanthide does not inhibit the action of acetaldehyde deaminase in the liver and the activity of monoamine oxidase in the body, thereby avoiding the damage of acetaldehyde to the liver and adverse reactions such as blood pressure rise, heart rate increase and the like caused by the accumulation of monoamine substances such as norepinephrine and the like, and improving the safety of treatment and the compliance of patients.
Example 2
The preparation method of the rabdosia lophanthide ethanol extract comprises the following steps: adding a certain amount of herba Rabdosiae Lophanthoidis into 20 times of 70% ethanol, extracting for 25min, filtering, and concentrating under reduced pressure to obtain the final product.
Example 3
The embodiment provides a product for preventing and treating alcohol dependence, and the preparation method comprises the following steps: collecting dried herba Rabdosiae Lophanthoidis 100g and flos Osmanthi Fragrantis 100g, decocting with 1000ml water for 1 hr for 2 times, mixing decoctions, and concentrating.
Example 4
This example provides an herb of Rabdosia Lophanthoidis tea for preventing and treating alcohol dependence
Weighing according to the following formula, and preparing according to the tea process
Rabdosia lophanthide 1000g
The preparation method comprises the following steps:
(a) picking: selecting fresh leaves of rabdosia lophanthide;
(b) withering: placing the fresh leaves in a withering trough for withering treatment;
(c) rolling: putting the withered tea leaves into a rolling machine for rolling for 20 min;
(d) fermentation: fermenting the rolled tea leaves at 40 ℃ for 6 h;
(e) air cooling: spreading the fermented tea leaves for cooling, wherein the spreading thickness is 1 cm;
(f) strip tidying: the air-cooled tea leaves are subjected to strip tidying treatment, the strip tidying temperature is 240 ℃, the strip tidying time is 30min, and the tea leaves are spread for cooling and natural moisture regaining after strip tidying is finished, so that the moisture on the leaf surfaces is redistributed;
(g) and (3) drying: drying the tea leaves after carding to obtain the linearstripe rabdosia herb tea.
Example 5
The present example provides a tablet for preventing and treating alcohol dependence, which is prepared according to the following formulation and tablet process;
the preparation method comprises the following steps: decocting the three medicines with 10000ml of water for 2 times, wherein the decocting time is 1 hour each time, combining the decoctions, concentrating until the relative density is 1.12-1.25 (50-60 ℃), preparing the concentrated solution and a proper amount of auxiliary materials into granules, drying, uniformly mixing, pressing into 1000 tablets, and coating sugar coat or film coat to obtain the traditional Chinese medicine.
Example 6
This example provides a granule for preventing and treating alcohol dependence with good taste, which is prepared by weighing the following formulation and preparing the same according to the preparation method
The preparation method comprises the following steps: decocting the linearstripe rabdosia herb with 4000ml of water for 2 times, each time for 1 hour, combining decoctions, filtering, concentrating the filtrate until the relative density is 1.12-1.25 (55-60 ℃), granulating the concentrated solution with a proper amount of auxiliary materials, drying, adding bay oil, uniformly mixing, and preparing 1000g of granules.
Example 7
This example provides a tea preparation for preventing and treating alcohol dependence with good taste, which is prepared by weighing the following ingredients according to the following formulation and processing
The preparation method comprises the following steps: decocting herba Rabdosiae Lophanthoidis and fructus Siraitiae Grosvenorii with 5000ml water for 2 times, each for 1 hr, mixing decoctions, filtering, concentrating the filtrate to relative density of 1.20(50 deg.C), adding ethanol to make ethanol content reach 60%, standing for precipitation, collecting supernatant, recovering ethanol and concentrating to appropriate amount, adding flos Osmanthi Fragrantis oil and appropriate amount of adjuvants, mixing, pressing into blocks, and drying.
Example 8
The embodiment provides a convenient-to-take and good-taste tea bag for preventing and treating alcohol dependence, which is weighed according to the following formula and prepared according to the tea bag process
The preparation method comprises the following steps: mixing the above four materials, and packaging into small bags to obtain teabag.
Example 9
Preparation of capsules
Weighing according to the following formula, and preparing according to capsule process
The preparation method comprises the following steps: taking the raw rabdosia lophanthide medicinal materials to prepare the rabdosia lophanthide water extract according to the method of the embodiment 1, concentrating, drying, powdering, adding a proper amount of auxiliary materials, mixing uniformly, and encapsulating to obtain the rabdosia lophanthide medicinal capsule.
Example 10
Preparation of oral liquid
Weighing according to the following formula, and preparing according to the process of oral liquid
The preparation method comprises the following steps: adding 8000ml of water into the rabdosia lophanthide crude drug, decocting twice, merging decoction, filtering, concentrating filtrate under reduced pressure to obtain clear paste with the relative density of 1.10-1.30 (50-60 ℃), cooling, adding ethanol to ensure that the alcohol content reaches 50% -80%, standing, filtering, recovering ethanol from the filtrate, concentrating the filtrate to obtain clear paste with the relative density of 1.10-1.30 (50-60 ℃), adding sweet osmanthus oil and proper amount of water, mixing uniformly, adjusting the pH value, refrigerating, filtering, adding water to 1000ml, stirring uniformly, subpackaging and sterilizing to obtain the traditional Chinese medicine.
Example 11
Preparation of beverages
Weighing according to the following formula, and preparing according to the beverage process
The preparation method comprises the following steps: soaking the above two crude drugs in 60L water for 30min, heating and decocting for 1 hr, filtering, adding appropriate amount of adjuvants, adding water to 100L, packaging, and sterilizing.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. Application of herba Rabdosiae Lophanthoidis in preparing product for preventing or treating alcohol dependence is provided.
2. Application of rabdosia lophanthide in preparing a product for improving anxiety-like behaviors caused by acute alcohol withdrawal.
3. Application of herba Rabdosiae Lophanthoidis in preparing product for stopping alcohol.
4. The use according to any one of claims 1 to 3, wherein said Rabdosia lophanthide is Rabdosia lophanthide crude drug or Rabdosia lophanthide extract.
5. The use according to any one of claims 1 to 3, wherein the product is a health product, a functional food or a medicament.
6. A product for preventing or treating alcohol dependence is characterized by comprising rabdosia lophanthide or rabdosia lophanthide extract and auxiliary materials.
7. The product of claim 6, wherein said Rabdosia serra extract is an aqueous or alcoholic Rabdosia serra extract; and/or the rabdosia lophanthide is rabdosia lophanthide tea.
8. The product of claim 7, wherein said rabdosia lophanthide extract is an aqueous rabdosia lophanthide extract; the preparation process of the rabdosia lophanthide water extract comprises the following steps: taking rabdosia lophanthide, decocting for 1-3 times, and concentrating the obtained decoction into thick paste.
9. The product according to claim 8, wherein the time for each decoction with water is 0.5-2 hours.
10. A product for improving mouthfeel and preventing or treating alcohol dependence is characterized in that the active component of the product is prepared from rabdosia lophanthide and at least one of sweet osmanthus, Chinese date, fructus momordicae and medlar.
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| CN202010916751.3A CN114129613B (en) | 2020-09-03 | 2020-09-03 | Application of rabdosia lophanthide in preventing or treating alcohol dependence |
| AU2020103237A AU2020103237A4 (en) | 2020-09-03 | 2020-11-04 | Application of Rabdosia Serra in Preventing or Treating Alcohol Dependence |
| PCT/CN2021/092060 WO2022048176A1 (en) | 2020-09-03 | 2021-05-07 | Use of rabdosia serra, and rabdosia serra composition |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101612193A (en) * | 2009-07-17 | 2009-12-30 | 吉林紫鑫药业股份有限公司 | A kind of pharmaceutical composition that is used for the treatment of cholecystitis, cholangitis |
| CN104940284A (en) * | 2015-06-25 | 2015-09-30 | 广西梧州制药（集团）股份有限公司 | Novel application of rabdosia lophanthide in preparing hyperlipidaemia treating and/or preventing medicine |
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2020
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- 2020-11-04 AU AU2020103237A patent/AU2020103237A4/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101612193A (en) * | 2009-07-17 | 2009-12-30 | 吉林紫鑫药业股份有限公司 | A kind of pharmaceutical composition that is used for the treatment of cholecystitis, cholangitis |
| CN104940284A (en) * | 2015-06-25 | 2015-09-30 | 广西梧州制药（集团）股份有限公司 | Novel application of rabdosia lophanthide in preparing hyperlipidaemia treating and/or preventing medicine |
Non-Patent Citations (1)
| Title |
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| 邓乔华等: "溪黄草资源现状及产业化发展的策略", 《2013年中国药学大会暨第十三届中国药师周论文集》 * |
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