CN114533774A - Natural compound for dispelling effects of alcohol and protecting liver and preparation process thereof - Google Patents
Natural compound for dispelling effects of alcohol and protecting liver and preparation process thereof Download PDFInfo
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- CN114533774A CN114533774A CN202210260830.2A CN202210260830A CN114533774A CN 114533774 A CN114533774 A CN 114533774A CN 202210260830 A CN202210260830 A CN 202210260830A CN 114533774 A CN114533774 A CN 114533774A
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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Abstract
The embodiment of the application provides a natural compound for relieving alcoholism and protecting liver and a preparation process thereof, and the natural compound for relieving alcoholism and protecting liver comprises the following components: the weight ratio of the polysaccharide of the dictyota dichotoma to the total flavone of the gynura divaricata is 5-10: 1. The provided anti-alcoholic compound can promote rapid anti-alcoholic and improve alcoholic liver injury conditions by improving oxidative stress caused by alcohol, and is a natural compound.
Description
Technical Field
The application belongs to the technical field of natural medicinal chemistry, and particularly relates to a natural compound for dispelling effects of alcohol and protecting liver and a preparation process thereof.
Background
With the continuous improvement of the life quality, the consumption of alcoholic beverages in life of people is remarkably improved, and excessive drinking can easily cause liver injury, such as alcoholic fatty liver, alcoholic hepatitis, even liver cancer and the like. At present, the number of deaths caused by alcohol-induced complications is increasing, and has caused a systemic hazard to human health.
The pathogenesis of complications caused by alcohol is complex, but oxidative stress generated in the process of alcohol metabolism is an important cause. Therefore, at present, the main therapeutic ideas of the drugs in clinical practice mainly include improving the anti-oxidation ability of the organism, inhibiting inflammatory reaction, regulating the liver function, and the like. A large amount of different medicines need to be taken according to the treatment thought, and the clinical requirements cannot be well met.
Dictyopteris divaricata (Dictyopteris divaricata) is a seaweed of Dictyotaceae of Phaeophyta, is widely distributed in China, and is a high-quality source of green raw materials for functional foods and cosmetics. The extract is rich in Seaweed Polysaccharides (SP), such as water-soluble acidic and neutral polysaccharides consisting of D-glucose, D-mannose, D-galactose, D-glucuronic acid and other monosaccharides, and has multiple biological activities of antioxidation, anti-inflammation, immunoregulation and the like, but the research on the aphanizomenon by scholars at home and abroad is less, and the effect of the extract are only reported in documents.
Therefore, the development of a natural compound product with the functions of relieving alcoholism and protecting liver based on dictyota dichotoma is urgently needed.
Disclosure of Invention
The embodiment of the application provides a natural compound for relieving alcoholism and protecting liver and a preparation process thereof, which can overcome the technical problem of insufficient development of the natural compound for relieving alcoholism in the prior art.
In an embodiment, a first aspect of the present application provides a natural compound for alleviating hangover and protecting liver, including: the weight ratio of the dictyostelium divaricatum polysaccharide (DDSP) to the Gynura divaricata (L.) DC) total flavone is 5-10: 1.
By adopting the scheme, the provided anti-alcoholic compound can promote rapid anti-alcoholic and improve the alcoholic liver injury condition by improving oxidative stress caused by alcohol, and is a natural compound.
In a second aspect, the present application provides a process for preparing a natural compound for alleviating hangover and protecting liver, comprising:
mixing, and mixing with polysaccharide of Ascophyllum divaricatum and total flavone of radix Gynurae Divaricatae to obtain the compound for relieving hangover.
The preparation method of the polysaccharide of the aphanizomenon comprises the following steps:
performing suction filtration, namely taking the dictyota dichotoma powder, adding deionized water, maintaining the pressure at 200-70 ℃ and the pressure at 400MPa for 0.1-0.2h, and performing suction filtration to obtain a crude extract;
removing protein, taking the crude extract, removing protein, centrifuging to remove precipitate;
precipitating with ethanol, collecting supernatant, precipitating with 2-3 times of ethanol, collecting precipitate, and washing with acetone;
purifying, concentrating the washing, crystallizing, filtering, and drying to obtain Dictyopteris divaricata polysaccharide.
The preparation method of the gynura divaricata total flavone comprises the following steps:
extracting with ethanol, pulverizing radix Gynurae Divaricatae, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain ethanol extract;
eluting, concentrating the ethanol extract, loading onto macroporous resin column, gradient eluting with ethanol water solution, collecting 35-55% ethanol eluate, and drying to obtain Gynura divaricata total flavone.
By adopting the scheme, the yield of the obtained anti-alcoholism compound is higher, and the byproducts are less.
Alternatively, in the preparation of polysaccharide from dictyospham, the deproteinization step can be performed by Sevage method.
Optionally, in the preparation of the dictyococcus divaricatus polysaccharide, the ethanol precipitation step is performed with ethanol with a volume fraction of 60-90%.
Optionally, in the preparation of the polysaccharide from the aphanidermia furiosa, the purification step comprises maintaining the temperature of the concentrated solution at 20-30 ℃, adding 2-3 times of ethanol with volume fraction of more than 95%, adding 1-2% by mass of seed crystal, stirring and crystallizing.
Further, in the preparation of the dictyota dichotoma polysaccharide, the purification step adopts freeze drying to obtain the dictyota dichotoma polysaccharide, and the freeze drying needs to cool a filtered object to 1-3 ℃ and then place the filtered object in a freeze drying box at-40 ℃ for drying.
Further, in the preparation of the polysaccharide from the dictyota dichotoma, the purification step adopts vacuum rotary evaporation concentration, the concentration temperature is 70-100 ℃, the vacuum degree is 18-25KPa, and the concentration is carried out until the mass concentration is 80-85%.
By adopting the scheme, the obtained extract can be further purified, the effective components are fully enriched, and the anti-alcohol effect is better.
Optionally, in the preparation of the gynura divaricata total flavonoids, ethanol is adopted in the ethanol extraction step, the volume fraction is 60-90%, and ethanol with the volume fraction of 40-95% is adopted in the elution step.
Optionally, the mixing step further comprises adding Balanophora Harlandii (Balanophora harlandii) extract, and the Pleuropterus Fulvioides polysaccharide, Gynura divaricata total flavone and Balanophora japonica extract at a weight ratio of 5-10:1: 0.5-1.
Optionally, the preparation process of the natural compound for alleviating hangover and protecting liver further comprises the preparation of a Balanophora japonica extract, wherein the preparation of the Balanophora japonica extract comprises the following steps:
extracting with ethanol, pulverizing dried Balanophora japonica Makino, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain Balanophora japonica Makino ethanol extract;
extracting, namely adding water into the ethanol extract of the Balanophora japonica Makino to suspend, and extracting for 1-3 times by using an organic solvent to obtain an Balanophora japonica Makino extract;
and (3) performing chromatography, namely loading the Balanophora japonica Makino extract on an ODS (ODS) chromatographic column, eluting with methanol with the volume fraction of 15-20% and 4-6 times of the column volume, eluting with methanol with the volume fraction of 22-25% and 7-10 times of the column volume, collecting the eluate, and concentrating and drying to obtain the Balanophora japonica Makino extract.
Further, in the preparation of the Balanophora japonica Makino extract, ethanol with the volume fraction of 60-90% is adopted in the ethanol extraction step, and the weight ratio of the Balanophora japonica Makino to the ethanol is 1: 10-20.
Further, in the preparation of the Balanophora japonica Makino extract, the organic solvent in the extraction step is one or more of ethyl acetate, n-butanol and tributyl phosphate.
Further, in the preparation of the Balanophora japonica Makino extract, ethanol with the volume fraction of 90% is adopted in the ethanol extraction step, and the weight ratio of Balanophora japonica Makino to ethanol is 1: 15.
By adopting the scheme, the alcohol extraction can be promoted, the active ingredients in the Balanophora japonica Makino can be contained as much as possible, and the higher alcohol extraction rate is ensured.
Compared with the prior art, the natural compound for relieving alcoholism and protecting liver provided by the embodiment of the application can inhibit triglyceride and cholesterol rising caused by alcohol, antagonize alcoholic oxidation and has a remarkable protection effect on acute alcoholic liver injury. The natural compound for relieving alcoholism and protecting liver prepared by the process provided by the application has more remarkable effects of relieving alcoholism and protecting liver.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a flow chart of a process for preparing polysaccharide from Dictyopteris divaricata according to an embodiment of the present application;
fig. 2 is a flow chart of the preparation of gynura divaricata total flavonoids in one embodiment of the present application.
Detailed Description
Embodiments of the present invention will be described in detail below. The following embodiments are merely used to more clearly illustrate the technical solutions of the present application, and therefore, the following embodiments are only used as examples, and the protection scope of the present application is not limited thereby.
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 application belongs; the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application; the terms "including" and "having," and any variations thereof, in the description and claims of this application and the description of the above figures are intended to cover non-exclusive inclusions.
In the description of the embodiments of the present application, the technical terms "first", "second", and the like are used only for distinguishing different objects, and are not to be construed as indicating or implying relative importance or implicitly indicating the number, specific order, or primary-secondary relationship of the technical features indicated. In the description of the embodiments of the present application, "a plurality" means two or more unless specifically defined otherwise.
Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. It is explicitly and implicitly understood by one skilled in the art that the embodiments described herein can be combined with other embodiments.
In the description of the embodiments of the present application, the term "and/or" is only one kind of association relation describing an associated object, and means that three kinds of relations may exist, for example, a and/or B, may mean: a exists alone, A and B exist simultaneously, and B exists alone. In addition, the character "/" herein generally indicates that the former and latter related objects are in an "or" relationship.
Example 1
As shown in fig. 1 and fig. 2, the preparation of natural anti-hangover and hepatoprotective compounds:
mixing, and mixing the polysaccharide of the dictyota dichotoma and the panax notoginseng flavones according to the weight ratio of 5:1 to obtain the compound for relieving alcoholism.
The preparation method of the polysaccharide of the aphanizomenon comprises the following steps:
performing suction filtration, namely taking the dictyota dichotoma powder, adding deionized water, pressurizing at 300MPa and 55 ℃, maintaining for 0.2h, and performing suction filtration to obtain a crude extract;
removing protein, taking the crude extract, removing protein, centrifuging to remove precipitate;
precipitating with ethanol, collecting supernatant, precipitating with 2 times of ethanol, collecting precipitate, and washing with acetone;
purifying, concentrating the washing, crystallizing, filtering, and drying to obtain Dictyopteris divaricata polysaccharide.
The preparation method of the gynura divaricata total flavone comprises the following steps:
extracting with ethanol, pulverizing radix Gynurae Divaricatae, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain ethanol extract;
eluting, concentrating the ethanol extract, loading onto macroporous resin column, gradient eluting with ethanol water solution, collecting 45% ethanol eluate, and drying to obtain Gynura divaricata total flavone.
In the preparation of the aphanidermia dichotoma polysaccharide, a protein removing step can adopt a Sevage method to remove protein, an ethanol precipitation step has the volume fraction of 90% of ethanol, a purification step comprises maintaining the temperature of a concentrated solution at 30 ℃, adding 3 times of ethanol with the volume ratio of 95%, adding seed crystals with the mass percentage of 2%, stirring and crystallizing, a freeze drying step is adopted to obtain the aphanidermia dichotoma polysaccharide, the freeze drying step is to cool an object obtained by filtering to 2 ℃, then placing the object in a freeze drying box with the temperature of-40 ℃ for drying, and in the preparation of the aphanidermia dichotoma polysaccharide, a purification step adopts vacuum rotary evaporation and concentration, the concentration temperature is 100 ℃, the vacuum degree is 22KPa, and the concentration is concentrated to the mass concentration of 80%; in the preparation of the gynura divaricata total flavone, ethanol is adopted in the ethanol extraction step, the volume fraction is 90%, and ethanol with the volume fraction of 95% is adopted in the elution step.
Example 2
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: wherein, in the preparation of the dictyospham polysaccharide, the volume fraction of ethanol in the ethanol precipitation step is 60%, the purification step comprises maintaining the temperature of the concentrated solution at 20 ℃, adding ethanol with the volume ratio of 2 times and the volume fraction of 95%, adding seed crystals with the mass percentage of 1%, stirring and crystallizing, the purification step adopts freeze drying to obtain the dictyospham polysaccharide, the freeze drying needs to cool an object obtained by filtering to 3 ℃, then placing the object in a freeze drying box at-40 ℃ for drying, in the preparation of the dictyospham polysaccharide, the purification step adopts vacuum rotary evaporation and concentration, the concentration temperature is 70 ℃, the vacuum degree is 18KPa, and the concentration is 82% by mass; in the preparation of the gynura divaricata total flavone, ethanol is adopted in the ethanol extraction step, the volume fraction is 60 percent, and ethanol with the volume fraction of 40 percent is adopted in the elution step.
Example 3
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: in the preparation of the aphanidermia dichotoma polysaccharide, the volume fraction of ethanol in the ethanol precipitation step is 70%, the purification step comprises maintaining the temperature of the concentrated solution at 25 ℃, adding ethanol with the volume ratio of 3 times and the volume fraction of 95%, adding seed crystals with the mass percentage of 2%, stirring and crystallizing, the purification step adopts freeze drying to obtain the aphanidermia dichotoma polysaccharide, the freeze drying needs to cool an object obtained by filtration to 1 ℃, then placing the object in a freeze drying box at the temperature of minus 40 ℃ for drying, the purification step adopts vacuum rotary evaporation and concentration, the concentration temperature is 80 ℃, the vacuum degree is 20KPa, and the concentration is 85% by mass; in the preparation of the gynura divaricata total flavone, ethanol is adopted in the ethanol extraction step, the volume fraction is 75%, and ethanol with the volume fraction of 60% is adopted in the elution step.
Example 4
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: in the preparation of the aphanidermia dichotoma polysaccharide, the volume fraction of ethanol in the ethanol precipitation step is 80%, the purification step comprises maintaining the temperature of the concentrated solution at 30 ℃, adding ethanol with the volume ratio of 3 times and the volume fraction of 95%, adding seed crystals with the mass percentage of 2%, stirring and crystallizing, the purification step adopts freeze drying to obtain the aphanidermia dichotoma polysaccharide, the freeze drying needs to cool an object obtained by filtration to 2 ℃, then placing the object in a freeze drying box at the temperature of-40 ℃ for drying, the purification step adopts vacuum rotary evaporation and concentration, the concentration temperature is 90 ℃, the vacuum degree is 25KPa, and the concentration is 85% by mass; in the preparation of the gynura divaricata total flavone, ethanol is adopted in the ethanol extraction step, the volume fraction is 90%, and ethanol with the volume fraction of 95% is adopted in the elution step.
Example 5
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: wherein, in the mixing step, the dictyococcus divaricatus polysaccharide and the gynura divaricata total flavone are mixed according to the weight ratio of 7: 1.
Example 6
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: wherein, in the mixing step, the dictyococcus divaricatus polysaccharide and the gynura divaricata total flavone are mixed according to the weight ratio of 10: 1.
Example 7
Preparing a natural compound for relieving alcoholism and protecting liver:
this embodiment is substantially the same as embodiment 1 except that: wherein, the mixing step also comprises adding aeginetia indica extract, and the aeginidopsis tetragonolobus polysaccharide, the gynura divaricata total flavone and the aeginetia indica extract, wherein the weight ratio of the dosage is 5:1: 1;
the preparation method of the Balanophora japonica Makino extract comprises the following steps:
extracting with ethanol, pulverizing dried Balanophora japonica Makino, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain Balanophora japonica Makino ethanol extract;
extracting, namely adding water into the ethanol extract of the Balanophora japonica Makino to suspend, and extracting for 1-3 times by using an organic solvent to obtain an Balanophora japonica Makino extract;
performing chromatography, namely loading the Balanophora japonica Makino extract on an ODS (ODS) chromatographic column, eluting with 15% methanol in volume fraction of 5 times of the column volume, eluting with 24% methanol in volume fraction of 8 times of the column volume, collecting eluate, concentrating and drying to obtain the Balanophora japonica Makino extract;
wherein, the ethanol extraction step adopts 90 percent ethanol by volume fraction, the weight ratio of the Balanophora japonica Makino to the ethanol is 1:15, and the organic solvent in the extraction step adopts a mixture of ethyl acetate and tributyl phosphate according to the weight ratio of 1: 2.
Example 8
Preparing a natural compound for relieving alcoholism and protecting liver:
this example is substantially the same as example 7 except that: in the preparation of the Balanophora japonica Makino extract, ethanol with the volume fraction of 60% is adopted in the ethanol extraction step, and the weight ratio of Balanophora japonica Makino to the ethanol is 1: 10.
Example 9
Preparing a natural compound for relieving alcoholism and protecting liver:
this example is substantially the same as example 7 except that: in the preparation of the Balanophora japonica Makino extract, ethanol with the volume fraction of 75% is adopted in the ethanol extraction step, and the weight ratio of Balanophora japonica Makino to the ethanol is 1: 15.
Example 10
Preparing a natural compound for relieving alcoholism and protecting liver:
this example is substantially the same as example 7 except that: in the preparation of the Balanophora japonica Makino extract, ethanol with the volume fraction of 90% is adopted in the ethanol extraction step, and the weight ratio of Balanophora japonica Makino to ethanol is 1: 20.
Example 11
Preparing a natural compound for relieving alcoholism and protecting liver:
this example is substantially the same as example 7 except that: wherein the mixing step also comprises adding aeginetia indica extract, and the weight ratio of the aeginidopsis indica polysaccharide, the gynura divaricata total flavone and the aeginetia indica extract is 7:1: 0.8.
Example 12
The preparation of the natural compound for relieving alcoholism and protecting liver comprises the following steps:
this example is substantially the same as example 7 except that: wherein the mixing step also comprises adding aeginetia indica extract, and the weight ratio of the aeginidopsis indica polysaccharide, the gynura divaricata total flavone and the aeginetia indica extract is 10:1: 0.5.
Example 13
This example is intended to demonstrate the inhibitory effect of the extracts obtained in examples 1-12 on the increase in the body mass index of liver in mice induced by alcohol.
Experimental animals: a mouse of Kunming species in a standard experiment is taken, and the body mass (20 +/-1) g is obtained.
The method comprises the following steps: the mice are taken and firstly fed with common feed adaptively under the experimental environment, observed for 5 days, 10 mice are randomly taken as normal groups, 10 mice are randomly taken as control groups, and the rest mice are taken as experimental groups.
Starting experiments in groups, feeding common feed, performing intragastric administration on test groups, wherein the administration dose is 100mg/kg, and after 2 hours, 4.8g/kg of alcohol is administered once, and water is not forbidden after fasting; normal group was gavaged with normal saline; the control group and the test group were administered with 4.8g/kg of alcohol at one time. After 16h, the liver was separated, rinsed twice with a pre-cooled PBS (0.1M, pH 7) solution, and the organ surface water was blotted with filter paper, weighed, and the organ index was calculated. The calculation formula is as follows: organ index (%) ═ organ weight (g)/body weight (g) × 100%.
The test components were divided into 12 groups of 10 mice each, and the respective administration was as follows: test groups 1 to 12 were administered with the compounds having anti-hangover and hepatoprotective effects obtained in examples 1 to 12, respectively.
The results are shown in table 1, comparing the test group with the normal group: p is less than 0.01; the p of the test group is less than 0.05 compared with the control group.
TABLE 1 inhibitory Effect of the Compounds obtained in examples 1 to 12 on the increase of liver index due to alcohol
As is clear from the data in Table 1, the compounds prepared in examples 1 to 12 have a good inhibitory effect on the liver body mass index, and the administration of the compounds before the administration of alcohol can significantly reduce the liver body mass index of mice.
In addition, it can be found from table 1 that, after the aeginetia indica extract is introduced in examples 7 to 12, the inhibition effect on the mouse liver body mass index is similar to that of examples 1 to 6 only by using the dictyospham polysaccharide and the gynura divaricata total flavone, and it is considered that the addition of the aeginetia indica extract does not affect the inhibition effect on the liver body mass index of the obtained compound. Next, it is understood from examples 1 and 7 that the best inhibitory effect is obtained when only the polysaccharide of gaeumannomyces divaricata and the total flavone of gynura divaricata are used in a ratio of 5:1 by weight of the former to the latter, and the best inhibitory effect is obtained when the polysaccharide of gaeumannomyces divaricata, the total flavone of gynura divaricata and the extract of Balanophora japonica are used in a ratio of 5:1:1 by weight of the three components.
Example 14
This example is to verify the inhibitory effect of the extracts obtained in examples 1-12 on the increase of TG, TC, ALT and AST levels in the serum of mice induced by alcohol.
Experimental animals: a mouse of Kunming species in a standard experiment is taken, and the body mass (20 +/-1) g is obtained.
The method comprises the following steps: the mice are taken and firstly fed with common feed adaptively under the experimental environment, observed for 5 days, 10 mice are randomly taken as normal groups, 10 mice are randomly taken as control groups, and the rest mice are taken as experimental groups.
The experiment is started in groups, adaptive feeding is carried out for 5 days, the mice freely eat common feed and drinking water during the experiment, the experimental group is administrated once at 100mg/kg, the experimental group and the control group are administrated once at 4.8g/kg of alcohol after 2h, the mice are fasted without water prohibition, the eyeball blood is taken after 16h, serum is prepared at 3000rpm multiplied by 10min, and TG, TC, ALT and AST indexes are measured.
The test components were divided into 12 groups of 10 mice each, and the respective administration was as follows: test groups 1 to 12 were administered with the compounds having anti-hangover and hepatoprotective effects obtained in examples 1 to 12, respectively.
The results are shown in table 2, comparing the test group with the normal group: p is less than 0.01; the p of the test group is less than 0.05 compared with the control group.
TABLE 2 inhibition of elevated TC, TG, ALT, AST levels by the compounds obtained in examples 1-12
The data in table 2 show that the compounds prepared in examples 1-12 have good liver body mass index inhibition effect, and the administration of the compounds before the administration of alcohol can obviously reduce the levels of cholesterol (TC), Triglyceride (TG), glutamic-pyruvic transaminase (AST) and glutamic-oxalacetic transaminase (ALT) in mice, which indicates that the compounds provided by the application can protect liver cells and reduce inflammatory response.
In addition, it can also be found from table 2 that, compared with the application of the dictyophora divaricata polysaccharide and the gynura divaricata total flavone in examples 1 to 6, the reduction effect of AST and ALT levels in mice is more obvious and the reduction effect of TC and TG levels is slightly improved after the introduction of the zizania rubra extract in examples 7 to 12. Next, it is understood from examples 1 and 7 that the best inhibitory effect is obtained when only the polysaccharide of gaeumannomyces divaricata and the total flavone of gynura divaricata are used in a ratio of 5:1 by weight of the former to the latter, and the best inhibitory effect is obtained when the polysaccharide of gaeumannomyces divaricata, the total flavone of gynura divaricata and the extract of Balanophora japonica are used in a ratio of 5:1:1 by weight of the three components.
Example 15
This example is to demonstrate the antagonistic effect of the extracts obtained in examples 1, 5, 6, 7, 11, 12 on the reduction of the levels of T-AOC and GSH in serum of mice induced by alcohol.
Experimental animals: a mouse of Kunming species in a standard experiment is taken, and the body mass (20 +/-1) g is obtained.
The method comprises the following steps: the mice are taken and firstly fed with common feed adaptively under the experimental environment, observed for 5 days, 10 mice are randomly taken as normal groups, 10 mice are randomly taken as control groups, and the rest mice are taken as experimental groups.
The experiment is started in groups, adaptive feeding is carried out for 5 days, the mice freely eat common feed and drinking water during the experiment, the experimental group is administrated once at 100mg/kg, the experimental group and the control group are administrated once at 4.8g/kg of alcohol after 2h, water is not forbidden when fasting, eyeball blood is taken after 6h, serum is prepared at 3000rpm multiplied by 10min, and T-AOC and GSH indexes in the serum are measured.
The test components were divided into 6 groups of 10 mice each, and the respective administration was as follows: test groups 1 to 6 were administered with the compounds having anti-hangover and hepatoprotective effects obtained in examples 1, 5, 6, 7, 11, and 12, respectively.
The results are shown in table 3, comparing the test group with the normal group: p is less than 0.01; the p of the test group is less than 0.05 compared with the control group.
TABLE 3 antagonism of alcohol-induced reduction of T-AOC and GSH levels by compounds of the present application
| Group of | T-AOC(mM) | GSH(μM) |
| Normal group | 1.92±0.12 | 36.93±5.01 |
| Control group | 1.67±0.29 | 25.04±3.97 |
| Test group 1 | 1.89±0.17 | 37.15±5.58 |
| Test group 2 | 1.81±0.14 | 36.53±2.88 |
| Test group 3 | 1.78±0.21 | 38.65±3.35 |
| Test group 4 | 1.90±0.38 | 36.02±5.46 |
| Test group 5 | 1.87±0.26 | 37.45±4.59 |
| Test group 6 | 1.85±0.23 | 37.32±3.75 |
As is clear from the data in Table 3, the mice treated with the compounds obtained in examples 1, 5, 6, 7, 11 and 12 were able to significantly increase the total serum antioxidant capacity (T-AOC), antagonize the alcohol-induced decrease in the serum Glutathione (GSH) level of mice, and reduce the damage to the liver.
Example 16
This example is to demonstrate the antagonistic effect of the extracts obtained in examples 1, 5, 6, 7, 11 and 12 on drunkenness in mice.
Experimental animals: a mouse of Kunming species in a standard experiment is taken, and the body mass (20 +/-1) g is obtained.
The method comprises the following steps: the mice are taken and firstly fed with common feed adaptively under the experimental environment, observed for 5 days, 10 mice are randomly taken as normal groups, 10 mice are randomly taken as control groups, and the rest mice are taken as experimental groups.
Starting experiments in groups, feeding adaptively for 5 days, wherein the mice freely eat common feed and drinking water during the experiment period, the experiment group is dosed with 100mg/kg once, the control group and the experiment group are dosed with 5.4g/kg of alcohol once after 2 hours, the disappearance of the overturning reflection of the mice is observed, namely drunkenness is obtained, the number of drunkenness mice is counted, and the drunkenness rate is calculated: the drunkenness rate is the number of drunkenness mice/experimental mice multiplied by 100%. The time from the administration of alcohol to the mice until the rolling reflection of the mice disappears is the drunk time, and the time for the mice to recover the rolling reflection after being drunk is the sober-up time. And (3) judging drunkenness of the mouse: the mouse was placed in a cage in a back-to-back orientation, and if the mouse was held in a back-down posture for more than 30 seconds, the flip-flop was considered to have disappeared.
The test components were divided into 6 groups of 10 mice each, and the respective administration was as follows: test groups 1 to 6 were administered with the compounds having anti-hangover and hepatoprotective effects obtained in examples 1, 5, 6, 7, 11, and 12, respectively.
The results are shown in table 4, comparing the test group with the normal group: p is less than 0.01; the p of the test group is less than 0.05 compared with the control group.
Table 4 antagonism of intoxication in mice by compounds of the present application
As can be seen from the data in table 4, the compounds obtained in examples 1, 5, 6, 7, 11 and 12 can significantly reduce the drunkenness rate of mice, prolong the drunkenness time and shorten the sobering time, thereby achieving the purpose of sobering up. By combining with analysis of other measurement index results, the compound provided by the application can reduce body mass index of mouse liver, inhibit the increase of TG and TC levels in serum caused by alcohol, antagonize the increase of ALT and AST levels in serum caused by alcohol, and has the effects of dispelling the effects of alcohol and protecting liver. The compound provided by the application can improve the T-AOC content and the GSH level in mouse serum, namely, can activate an antioxidant system of an organism and inhibit oxidative stress caused by ethanol to a certain extent, thereby realizing the liver protection effect.
Finally, it should be noted that: the above embodiments are only used for illustrating the technical solutions of the present application, and are not limited thereto; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may be modified, or some or all of the technical features may be equivalently replaced; these modifications and substitutions do not cause the corresponding technical solutions to depart from the scope of the technical solutions of the embodiments of the present application, and are intended to be covered by the claims and the specification of the present application. In particular, the features mentioned in the embodiments can be combined in any manner as long as there is no structural conflict. This application is not intended to be limited to the particular embodiments disclosed herein but is to cover all embodiments that may fall within the scope of the appended claims.
Claims (10)
1. A natural compound for relieving alcoholism and protecting liver, which is characterized by comprising: the weight ratio of the polysaccharide of the dictyota dichotoma to the total flavone of the gynura divaricata is 5-10: 1.
2. A process for preparing the natural sobering-up liver-protecting compound according to claim 1, comprising:
mixing, mixing the polysaccharide and total flavone of Gynura divaricata to obtain the compound for relieving hangover,
the preparation method of the polysaccharide of the aphanizomenon comprises the following steps:
performing suction filtration, namely taking the dictyota dichotoma powder, adding deionized water, maintaining the pressure at 200-70 ℃ and the pressure at 400MPa for 0.1-0.2h, and performing suction filtration to obtain a crude extract;
removing protein, taking the crude extract, removing protein, centrifuging to remove precipitate;
precipitating with ethanol, collecting supernatant, precipitating with 2-3 times of ethanol, collecting precipitate, and washing with acetone;
purifying, concentrating the washing, crystallizing, filtering, drying to obtain Dictyopteris divaricata polysaccharide,
the preparation method of the gynura divaricata total flavone comprises the following steps:
extracting with ethanol, pulverizing radix Gynurae Divaricatae, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain ethanol extract;
eluting, concentrating the ethanol extract, loading onto macroporous resin column, gradient eluting with ethanol water solution, collecting 35-55% ethanol eluate, and drying to obtain Gynura divaricata total flavone.
3. The process for preparing natural compounds for alleviating hangover and protecting liver as claimed in claim 2, wherein the deproteinization step is carried out by Sevage method in the preparation of Neurospora divaricata polysaccharide.
4. The process for preparing a natural compound for alleviating hangover and protecting liver as claimed in claim 3, wherein the volume fraction of ethanol in the alcohol precipitation step is 60-90% in the preparation of the aphanidermia dichotoma polysaccharide.
5. The process for preparing natural compounds for alleviating hangover and protecting liver as claimed in claim 4, wherein in the preparation of the dictyococcus divaricatus polysaccharide, the purification step includes maintaining the temperature of the concentrated solution at 20-30 ℃, adding ethanol with a volume ratio of 2-3 times and a volume fraction of more than 95%, adding seed crystals with a mass percentage of 1-2%, stirring and crystallizing.
6. The process for preparing a natural compound for alleviating hangover and protecting liver as claimed in claim 5, wherein the purification step employs vacuum rotary evaporation concentration at 70-100 ℃ under a vacuum degree of 18-25KPa in the preparation of the dictyococcus divaricatus polysaccharide.
7. The process for preparing natural compounds for alleviating hangover and protecting liver as claimed in claim 6, wherein in the preparation of the aphanidermia dichotoma polysaccharide, the purification step is to obtain the aphanidermia dichotoma polysaccharide by freeze-drying, wherein the freeze-drying is to cool the filtered product to 1-3 ℃, and then dry the product in a freeze-drying oven at-40 ℃.
8. The process for preparing natural compounds for alleviating hangover and protecting liver as claimed in claim 2, wherein in the preparation of gynura divaricata total flavonoids, ethanol is used in the ethanol extraction step, the volume fraction is 60-90%, and ethanol is used in the elution step, the volume fraction is 40-95%.
9. The preparation process of the natural compound for alleviating hangover and protecting liver as claimed in claim 2, wherein the mixing step further includes adding aeginetia indica extract, and the weight ratio of the dictyosphaera leucotaxifolia polysaccharide, the panax notoginseng flavones and the aeginetia indica extract is 5-10:1: 0.5-1.
10. The process for preparing a natural compound for alleviating hangover and protecting liver according to claim 8, further comprising preparing an aeginetia indica extract, wherein the preparing of the aeginetia indica extract comprises the following steps:
extracting with ethanol, pulverizing dried Balanophora japonica Makino, extracting with ethanol, concentrating the extractive solution, and removing solvent to obtain Balanophora japonica Makino ethanol extract;
extracting, namely adding water into the ethanol extract of the Balanophora japonica Makino to suspend, and extracting for 1-3 times by using an organic solvent to obtain an Balanophora japonica Makino extract;
and (3) performing chromatography, namely loading the Balanophora japonica Makino extract on an ODS (ODS) chromatographic column, eluting with methanol with the volume fraction of 15-20% and 4-6 times of the column volume, eluting with methanol with the volume fraction of 22-25% and 7-10 times of the column volume, collecting the eluate, and concentrating and drying to obtain the Balanophora japonica Makino extract.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210260830.2A CN114533774A (en) | 2022-03-16 | 2022-03-16 | Natural compound for dispelling effects of alcohol and protecting liver and preparation process thereof |
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Citations (4)
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| CN103446165A (en) * | 2013-09-12 | 2013-12-18 | 中国海洋大学 | Application of guluronic acid oligomers in preparing medicines, health products or dietary supplements for preventing and treating alcoholic liver injury |
| CN108771690A (en) * | 2018-08-23 | 2018-11-09 | 嘉应学院医学院 | It is a kind of that there is hypoglycemic or effect for reducing blood fat red winter Balaenoptera borealis Lesson extract and its preparation method and application |
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| CN103446165A (en) * | 2013-09-12 | 2013-12-18 | 中国海洋大学 | Application of guluronic acid oligomers in preparing medicines, health products or dietary supplements for preventing and treating alcoholic liver injury |
| CN108771690A (en) * | 2018-08-23 | 2018-11-09 | 嘉应学院医学院 | It is a kind of that there is hypoglycemic or effect for reducing blood fat red winter Balaenoptera borealis Lesson extract and its preparation method and application |
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