阅读说明：本技术 岩青兰的应用 (Application of Dracocephalum rupestre ) 是由 李会芳 程生辉 王智深 郎霞 于 2021-02-01 设计创作，主要内容包括：本发明公开了岩青兰的应用。本发明岩青兰应用于制备解酒的产品中或制备用于防治酒精性肝损伤的产品中。本发明通过实验表明岩青兰高、中、低剂量组醒酒时间均显著缩短,证明本发明岩青兰茶有一定的解酒作用,且效果呈剂量依赖性。本发明岩青兰对酒精性肝损伤具有保护作用,其对酒精性肝损伤大鼠血清中AST、TC、TG具有不同程度的降低作用。本实验为岩青兰的临床运用提供了实验依据,为其进一步开发和利用奠定了基础,其作用机制有待于进一步深入研究。(The invention discloses application of Dracocephalum rupestre Hance. The dracocephalum rupestre linn is applied to preparing a product for relieving alcoholism or a product for preventing and treating alcoholic liver injury. Experiments show that the sobering-up time of the high, medium and low dosage groups of the glaucophyllum cochinchinensis is obviously shortened, and the glaucophyllum cochinchinensis tea has a certain sobering-up effect and the effect is dose-dependent. The dracocephalum rupestre Hance has a protection effect on alcoholic liver injury, and has reduction effects on AST, TC and TG in serum of rats with alcoholic liver injury to different degrees. The experiment provides experimental basis for the clinical application of the dracocephalum rupestre hance, lays a foundation for the further development and utilization of the dracocephalum rupestre hance, and the action mechanism of the dracocephalum rupestre hance is to be further deeply researched.)

1. Use of Dracocephalum rupestre in any one of:

(A1) as a medicine for relieving alcoholism;

(A2) preparing the product for relieving alcoholism.

2. Use according to claim 1, characterized in that: the product for relieving alcoholism comprises medicines and/or health products.

3. A product for relieving alcoholism is characterized in that: the product for alleviating hangover comprises the Dracocephalum rupestre as claimed in claim 1.

4. An anti-hangover product according to claim 3, characterized in that: when the product for relieving alcoholism is a medicine, the product is a pharmaceutically acceptable dosage form.

5. Use of Dracocephalum rupestre in any one of:

a) as a medicament for the prevention and treatment of alcoholic liver injury;

b) preparing a product for preventing and treating alcoholic liver injury.

6. Use according to claim 5, characterized in that: the product for preventing and treating alcoholic liver injury comprises medicines and/or health products.

7. A product for preventing and treating alcoholic liver injury, which is characterized in that: the product for preventing and treating alcoholic liver injury comprises the Dracocephalum rupestre Hance as claimed in claim 5.

8. The product for preventing and treating alcoholic liver injury according to claim 7, wherein: when the product for preventing and treating alcoholic liver injury is a medicament, the product is a pharmaceutically acceptable dosage form.

Technical Field

The invention relates to application of Dracocephalum rupestre Hance, and belongs to the field of traditional Chinese medicine application.

Background

Dracocephalum rupestre Hance, also called Maojian grass, is a plant of the genus Dracocephalum of the family Labiatae (Lamiaceae). The plant is perennial herb, the plant height is 15-42 cm, the leaves are triangular and oval, and the edge is provided with circular saw teeth; in the flowering period of 7-9 months, the inflorescence of the cymbidium floribunda is purple blue, and the whole grass has fragrance. In China, the Dracocephalum rupestris is mainly distributed in Hebei, Shanxi, Qinghai, inner Mongolia, Liaoning and other places and grows in grasslands, grass slopes or sparse forests with the elevation of 650-2400 m. According to the record of Chinese medicine dictionary, the herb of corydalis saxicola bunting can be used for replacing tea and mainly treating wind-heat common cold, headache, cold and heat, cough and icterohepatitis. In addition to being used as a traditional medicinal material, the corydalis saxicola bunting has been developed into tea drinks in Shanxi et al. The research finds that the extract of the dracocephalum rupestris L belonging to the same genus has the pharmacological effects of resisting oxidation, resisting tumors, resisting high altitude anoxia, protecting the cardiovascular system and the like.

Alcoholism refers to a condition in which a large amount of alcohol or alcoholic beverages are taken in a short time, leading to the central nervous system being excited and then converted into an inhibited state. Alcoholism can be divided into periods of excitability, ataxia and lethargy. Some manifestations of the excitation period include excitation, euphoria, language increase and the like, and even the manifestations are rough and have aggressive behaviors; some are silent and few. Ataxia is manifested by clumsy movements, unsteady gait, unclear mouth and teeth, blurred vision, nausea, vomiting, etc. Patients in the narcolepsy stage can have the symptoms of narcolepsy, body temperature drop, blood pressure reduction and the like, even respiratory and circulatory paralysis occur, and serious patients can die.

Alcoholic liver injury (ALD) refers to toxic pathological damage of the liver caused by short-term heavy drinking or long-term heavy drinking, and is manifested as steatosis of liver cells at the initial stage, and further alcoholic hepatitis, which finally leads to hepatic fibrosis and alcoholic cirrhosis. Clinically, the medicine is mainly characterized by nausea, vomiting, hypodynamia, jaundice, liver swelling and tenderness. The primary feature was accumulation of Triglycerides (TG) in hepatocytes; the metaphase is a group of clinical pathological syndromes caused by massive necrosis of liver cells in a short term and can occur on the basis of the existence or non-existence of liver cirrhosis; the latter stage is primarily characterized by the accumulation of extracellular matrix proteins (e.g., collagen) produced by activated Hepatic Stellate Cells (HSCs).

Disclosure of Invention

The invention aims to provide application of dracocephalum rupestre which is proved by experiments to be capable of alleviating hangover and has a protection effect on alcoholic liver injury.

The invention provides application of the glaucomatous blue in the following 1) or 2):

(A1) as a medicine for relieving alcoholism;

(A1) preparing the product for relieving alcoholism.

In the application, the product for relieving alcoholism comprises a medicament and/or a health-care product.

The invention also provides a product for relieving alcoholism, which contains the dracocephalum rupestre.

In the product for relieving alcoholism, when the product for relieving alcoholism is a medicine, the product is a pharmaceutically acceptable dosage form.

In the invention, the dracocephalum rupestris can be prepared from a dracocephalum rupestris aqueous extract when being used as a medicine for relieving alcoholism.

The invention also provides the application of the glaucomatous blue in a) or b) as follows:

a) as a medicament for the prevention and treatment of alcoholic liver injury;

b) preparing a product for preventing and treating alcoholic liver injury.

In the application, the product for preventing and treating alcoholic liver injury comprises medicines and/or health-care products.

The invention further provides a product for preventing and treating the alcoholic liver injury, and the active ingredient of the product for preventing and treating the alcoholic liver injury is dracocephalum rupestre.

In the above products for preventing and treating alcoholic liver injury, when the products for preventing and treating alcoholic liver injury are medicines, the products are pharmaceutically acceptable dosage forms.

In the invention, the dracocephalum rupestris can be used as a medicament for preventing and treating alcoholic liver injury and can be a dracocephalum rupestris water decoction;

the product for preventing and treating alcoholic liver injury can be a product containing Dracocephalum rupestre Hance water decoction.

The invention has the following advantages:

experiments show that the sobering-up time of the high, medium and low dosage groups of the glaucophyllum cochinchinensis is obviously shortened, and the glaucophyllum cochinchinensis tea has a certain sobering-up effect and the effect is dose-dependent. The dracocephalum rupestre Hance has a protection effect on alcoholic liver injury, and has reduction effects on AST, TC and TG in serum of rats with alcoholic liver injury to different degrees. The experiment provides experimental basis for the clinical application of the dracocephalum rupestre hance, lays a foundation for the further development and utilization of the dracocephalum rupestre hance, and the action mechanism of the dracocephalum rupestre hance is to be further deeply researched.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Examples 1,

1 materials of the experiment

1.1 animals

60 Kunming mice, half male and female, body weight: 18-20g, purchased from the animal testing center of Shanxi university of medicine, animal qualification number-SCXK (jin) 2015-0001. The mice are normally adapted to be raised for 7 days, and the food intake and the water drinking are normal.

1.2 drugs and reagents

Tuqinglan (purchased from Shenchi county, Shanxi province); naloxone hydrochloride injection (Hainan Lingkang pharmaceutical Co., Ltd., lot number: 190101112); 50% absolute ethanol solution; physiological saline (0.9% sodium chloride, Shijiazhuang four drugs Co., Ltd., batch No.: 1701253204)

1.3 instruments

BCD-229KCK refrigerator (Hefei Mei Ling Ltd.); HH-2 digital display constant temperature water bath (Jie Rui Er electric appliances Co., Ltd., King Tan); PL-S40 model Consumer digital ultrasonic instrument; YP601N type electronic balance (division: 0.1g, for mouse weighing); PA224 type electronic precision balance (Shunhua constant scientific instruments, Inc. in Shanghai).

2 method of experiment

2.1 animal groups

50 Kunming mice are divided into 5 groups of 10 mice each, namely a model group, a positive group (naloxone hydrochloride injection 0.002g/kg, intraperitoneal injection) and a high (3g/kg), medium (1.5g/kg) and low (0.75g/kg) dosage group of dracocephalum rupestris. Wherein naloxone hydrochloride is suitable for acute alcoholism, wherein g/kg means the administration dosage g per kg body weight of animals, the same is as follows.

The high, medium and low dosage groups of the Dracocephalum rupestre adopt Dracocephalum rupestre water decoction, and the preparation method comprises: 252g of corydalis saxicola bunting medicinal material decoction pieces are soaked in 10 times of water for 1 hour, the first decoction is started, the decoction is changed into small fire decoction for 1 hour after the decoction is boiled by big fire, and the liquid medicine is taken after the decoction is finished. Decocting the residue with 8 times of water for the second time, decocting with slow fire for 1 hr after boiling, collecting medicinal liquid, mixing the medicinal liquids for 2 times, and concentrating to 840 ml. And then preparing the water decoctions of the dracocephalum rupestre with different concentrations, namely 3g/kg of the high dose group, 1.5g/kg of the medium dose group and 0.75g/kg of the low dose group, marking, and storing in a refrigerator at 4 ℃ for later use.

2.2 animal administration

Each group is fasted for 16h in advance, 2h after the last administration, each group is given corresponding 50% alcohol (20mL/kg), after 30min, the positive group is given naloxone hydrochloride (intraperitoneal injection, 0.002g/kg), and the high-dose group, the medium-dose group and the low-dose group of the dracocephalum rupestris are respectively administered by gastric lavage. The conditions of the mice that the righting reflection disappears and recovers are closely observed in the experimental process, and meanwhile, the records are made. The latent period of drunkenness: the time taken for the mice to begin alcohol administration until the mice appear to have a righting reflex disappear; sleep time: time spent until the mouse regains again after the mouse righting reflex disappears; the sobering-up time is as follows: time difference from alcohol administration to mice to re-recovery of mice.

2.3 statistical analysis

The experimental data are statistically tested by SPSS 22.0 statistical software, the measurement data are analyzed by single-factor variance, the counting data are tested by fisher, and the difference between groups is P less than 0.05, which has statistical significance.

3 results of the experiment

3.1 the disappearance of orthotropic reflex at different time points after the administration of wine in mice of different dose groups of Dracocephalum rupestris is shown in Table 1.

Table 1 number of mice in different dose groups of dracocephalum rupestris with disappearance of righting reflex at different time points after administration of alcohol (n ═ 10)

Note: p <0.05 compared to model group.

Due to the influence of various conditions in the experimental process, the number of mice which have the righting reflex disappear at part of time point after the mice in each group are given alcohol is only listed in table 1, after the mice are given 50% alcohol for 0.5h, 8 mice in the model group, 7 mice in the positive group, 4 mice in the high dose group, 2 mice in the medium dose group and 2 mice in the low dose group have the righting reflex disappear; after the mice are given 50% alcohol for 1.5h, the mice of the model group, the positive group and the high-dose group all have the phenomenon of disappearance of righting reflex, the mice of the medium-dose group are 7, the mice of the low-dose group are 9, and the rest mice have the symptoms of intoxication: sleepiness and unsteady action, half of mice in the positive group begin to recover the righting reflex after drinking, the mice in the high, medium and low dose groups begin to recover the righting reflex after drinking, the mice in the positive group completely recover after drinking for 7 hours, the mice in the high dose group recover the righting reflex for the most, the mice in the medium dose group gradually recover after drinking for 9 hours, the mice in the low dose group do not recover, the mice in the high and medium dose groups can basically eat food, the mice in the low dose group basically do not eat food, and in conclusion, the high and medium dose groups of the glaucomatodes according to the invention can reduce the number of the mice which disappear from the righting reflex through fisher test.

3.2 influence of the groups of Dracocephalum rupestris on the intoxication latency, sleep time and sober-up time of the mice, as shown in Table 2.

Table 2 influence of glaucomatous blue on intoxication latency, sleep time and sobering-up time of mice (n ═ 10)

Note: p <0.05, P <0.01 compared to model group.

As can be seen from the results in Table 2, the inebriation latency time of mice in the high, medium and low dose groups of Dracocephalum rupestre is not significantly different compared with the model group; compared with a model group, the sleeping time of the positive group and the high, medium and low dose groups of the Dracocephalum rupestris is obviously shortened (P is less than 0.01), and certain dose dependence exists; compared with the model group, the sobering-up time of the high, medium and low dosage groups of the Dracocephalum rupestris is obviously shortened (P is less than 0.01) in the positive group and the Dracocephalum rupestris, and certain dosage dependence also exists, so that the Dracocephalum rupestris has a certain sobering-up effect, and the effect is dosage dependence.

Examples 2,

1. Experimental Material

1.1 drugs and reagents

The corydalis saxicola bunting medicinal materials: purchased from Shenchi county of Shanxi province; dongbaogantai tablets: tongbao pharmaceutical industry, batch number: 180113, respectively; glutamic-pyruvic transaminase (ALT) detection kit: nanjing was established as a bioengineering institute, with a cargo number: c009-2, batch number: 20190425, respectively; glutamic-oxaloacetic transaminase (AST) detection kit: nanjing was established as a bioengineering institute, with a cargo number: c010-2, batch number: 20190319, respectively; total Cholesterol (T-CHO) detection kit: nanjing was established as a bioengineering institute, with a cargo number: a111-1, batch number: 20190223, respectively; triglyceride (TG) detection kit: nanjing was established as a bioengineering institute, with a cargo number: a110-1, batch number: 20190227.

1.2 instruments

Enzyme-labeled analyzer: beijing Prolang New technology Co., Ltd, type: DNM-9602;

electronic analytical balance, available from shunhui scientific instruments ltd, model: FFA 224;

micropipettes, available from semer feishel (shanghai) instruments ltd, model: 40270530, respectively;

electronic balance, available from Shanghai Acoustic weighing apparatus, Inc., model number; IC-1;

rotary evaporator, shanghai yanglong biochemical instrument factory, model: RE-2000B.

1.3 Experimental animals

SPF grade healthy SD rats, half female and half male, body weight: (180-220) g, purchased from Shanxi university of medicine animal center, and the certificate number: 20180006003154, rats were acclimatized for 1 week on regular choking with free water intake.

2, experimental method:

2.1 preparation of the drug

The preparation method of the water decoction of the corydalis saxicola: as in inventive example 1.

Preparation of positive control group medicament: taking Dongbaogantai tablets, preparing a solution with the concentration of 0.036g/ml by using distilled water, marking the dosage of the rat with the dosage of 0.36g/kg and the dosage of about 6 times of clinical adults, and storing the rat in a refrigerator at 4 ℃ for later use.

Preparation of 10% chloral hydrate solution: adding 50ml distilled water into 5g chloral hydrate to prepare 10% chloral hydrate solution, marking, and storing in a refrigerator at 4 ℃ for later use.

Preparation of 10% neutral formalin solution: weighing formaldehyde 220ml, sodium dihydrogen phosphate 8.8g, disodium hydrogen phosphate 14.3g, adding distilled water to obtain 2200ml 10% formalin solution, measuring pH to 7.2-7.4 to obtain neutral formalin solution, labeling, and storing in refrigerator at 4 deg.C.

2.2 Molding and dosing regimens

72 rats were randomly divided into 6 groups: blank group, model group, positive drug group (Dongbaogantai tablet), Tuqinglan high (3g/kg), medium (1.5g/kg) and low (0.75g/kg) dose group, and each group contains 12 animals. Except for the blank group, the other groups were separately gazed with 1ml/100g of 50% absolute ethyl alcohol (diluted with distilled water) in the morning. Respectively performing intragastric administration at a dose of 1ml/100g in the afternoon every day for 6 weeks, continuously performing intragastric administration for 2 times per week, and adjusting the liquor and dosage of each group according to body weight. After the experiment is finished at the 6 th week, the rats are fasted for 12 hours without water prohibition, 10% chloral hydrate is used for anesthesia (the administration dosage is 3ml/kg), blood is taken from abdominal aorta, whole blood is injected into a blank clean test tube, after centrifugation (3000r/min, 15min and 4 ℃), serum is separated, and ALT, AST, TC and TG in the serum to be detected are frozen and stored at the temperature of-80 ℃.

2.3ALT, AST, TC, TG determination method

2.3.1 microplate assay for ALT

Alanine Aminotransferase (ALT) acts on a substrate consisting of alanine and alpha-ketoglutaric acid at 37 deg.C and pH7.4 to produce pyruvic acid and glutamic acid. After reacting for 30min (fixed time), adding 2, 4-Dinitrophenylhydrazine (DNPH) hydrochloric acid solution, stopping the reaction, simultaneously adding DNPH and carbonyl in ketoacid to generate pyruvic acid phenylhydrazone, wherein the phenylhydrazone is reddish brown under the alkaline condition, reading the absorbance at 505nm and calculating the enzyme activity.

2.3.2 microplate assay for AST

AST can make alpha-ketoglutaric acid and aspartic acid exchange amino and keto groups to generate glutamic acid and oxaloacetic acid. Oxaloacetate can be spontaneously decarboxylated to pyruvate during the reaction. Pyruvic acid reacts with 2, 4-dinitrophenylhydrazine to generate 2, 4-dinitrophenylhydrazone which is reddish brown in alkaline solution. After color comparison, the activity unit of the enzyme can be obtained by looking up the standard curve.

2.3.3 colorimetric determination of TC

The cholesterol ester generates cholesterol and fatty acid under the action of cholesterol esterase. Cholesterol and oxygen react with carbon monoxide to produce 4-AAP and hydrogen peroxide. The hydrogen peroxide, the 4-AAP and the phenol generate red quinone compounds and water under the action of peroxidase. The shade of the generated quinone compound is in direct proportion to the content of the cholesterol, the absorbance values of the calibration standard tube and the sample tube are respectively measured, and the content of the cholesterol is calculated.

2.3.4 colorimetric TG determination

Triglyceride and water produce glycerol and fatty acid under the action of lipase. Glycerol and ATP are reacted by glycerol kinase to produce glycerol-3-phosphate and ADP. Glycerol-3-phosphate and oxygen gas generate hydroxyacetone phosphate and hydrogen peroxide under the action of glycerol-3-phosphate oxidase. The hydrogen peroxide, the 4-AAP and the parachlorophenol generate red quinone compounds under the action of peroxidase. The shade of the color of the generated quinone compound is in direct proportion to the content of the triglyceride, and the absorbance values of the standard tube and the sample tube are respectively measured, so that the content of the triglyceride in the sample can be calculated.

2.4 Observation index

2.4.1 general morphological observations: the rat body mass change, eating, drinking, mental state and behavior conditions are observed in the experimental process.

2.4.2 Biochemical indexes: and (3) detecting the levels of ALT, AST, TC and TG in serum by using a microplate reader.

2.4.3 statistical analysis: statistical analysis using a sps 19.0, data was calculated toData between groups were examined by t, and P was used<0.05 has statistical significance.

3. Results of the experiment

3.1 Effect of Dracocephalum rupestre on signs of alcoholic liver injury caused by alcohol

In the process of gavage, rats in the model group, the positive control group, the high, medium and low dose groups have hair explosion phenomena which are represented by sleepiness, dull state and drunkenness, the rats begin to return to the normal state after 1-2h, and the rats also have nose bleeding, vomit medicine and excessive bite. After administration, the mental state of the rats in the administration group is better than that in the model group, and the activity is obviously increased compared with that in the model group. After the experiment, all rats had significantly increased body weight.

3.2 Effect of Dracocephalum rupestre on serum ALT and AST levels in rats with alcoholic liver injury

TABLE 3 influence of Dracocephalum rupestre decoction on serum ALT and AST of rats with alcoholic liver injury ((n＝10)

Note: p <0.05 and P <0.01 in the blank group, and P <0.05 and P <0.01 in the model group.

The experimental results in table 3 show that: compared with a blank group, the ALT and AST levels in the serum of the rat of the model group are obviously increased, and the statistical significance is achieved (P is less than 0.05); compared with the model group, ALT level in the serum of the rats in the positive group and the high, medium and low dosage groups of the dracocephalum rupestris has a reduction trend, but has no statistical significance; the AST level in the serum of rats in the positive group and the high, medium and low dose groups of the dracocephalum rupestris is obviously reduced, the statistical significance is achieved (P is less than 0.05, and P is less than 0.01), and the comparison of the positive group data proves that the dracocephalum rupestris has a treatment effect on alcoholic liver injury.

3.2 Effect of Dracocephalum rupestre on serum TC and TG levels in rats with alcoholic liver injury

TABLE 4 influence of Dracocephalum rupestre decoction on serum TC, TG of alcoholic liver injury rat ((n＝10)

Note: p <0.05 and P <0.01 in the blank group, and P <0.05 and P <0.01 in the model group.

The experimental results in table 4 show that: compared with a blank group, the TC and TG levels in the serum of the rat of the model group are obviously increased, and the statistical significance is achieved (P is less than 0.05, and P is less than 0.01); compared with the model group, the TC level in the serum of the rat of the positive drug and the rat of the high-dose group of the dracocephalum rupestris of the invention is obviously reduced (P is less than 0.05, P is less than 0.01), and the TG level in the serum of the rat of the positive drug and the rat of the low-dose group of the dracocephalum rupestris of the invention is obviously reduced (P is less than 0.05, P is less than 0.01).

Discussion 4

The liver is the main part of ethanol metabolism and is also the main target organ of liver injury caused by ethanol. The liver has limited capacity of processing alcohol, and the liver cannot bear the alcohol in an amount exceeding the capacity of processing the alcohol by the liver, so that the alcoholic liver injury can be caused. Alcohol intragastric perfusion is a common molding method. According to the method for quantitatively and regularly administering the alcohol with the volume percentage concentration of 50% by the stomach perfusion method, the levels of ALT, AST, TC and TG in serum are obviously higher than those of a blank rat, and the success of model establishment is shown.

Serum ALT and AST concentrations are sensitive markers for diagnosing liver cell damage, and when liver cells are damaged, ALT in the liver cells is released into blood, so that the serum ALT concentration is increased, the serum ALT concentration and the ALT are lower in normal conditions, and only enzyme in intracellular fluid is released into the blood when the liver cells are damaged, and then the level is increased. TC and TG are normally transported to the blood in the form of low density lipoproteins, and when TC and TG levels rise, the liver encounters an obstacle in the metabolic process. The experimental research results show that the dracocephalum rupestris has different degrees of reducing effects on AST, TC and TG in the serum of a rat with alcoholic liver injury, and the dracocephalum rupestris has a certain protective effect on the alcoholic liver injury.