CN117547564A - Fennel extract composition and preparation method and application thereof - Google Patents
Fennel extract composition and preparation method and application thereof Download PDFInfo
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- CN117547564A CN117547564A CN202311846606.2A CN202311846606A CN117547564A CN 117547564 A CN117547564 A CN 117547564A CN 202311846606 A CN202311846606 A CN 202311846606A CN 117547564 A CN117547564 A CN 117547564A
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- fennel
- water
- extract
- volatile oil
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Classifications
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/235—Foeniculum (fennel)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- 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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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
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Abstract
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a fennel extract composition, a preparation method and application thereof. The invention provides a preparation method of a fennel extract composition, which comprises the following steps: mixing fructus Foeniculi powder with water, and steam distilling to obtain fructus Foeniculi volatile oil component, first extractive solution and first residue; mixing the first residues with water, and decocting to obtain a second extract; mixing and concentrating the first extract and the second extract, and purifying by a macroporous resin column to obtain a water-soluble component of fennel; mixing the fennel volatile oil component and the fennel water-soluble component to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4. The prepared compatible composition has remarkable prevention and treatment effects on gastric mucosal injury, and can be well applied to stomach nourishing healthy products.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a fennel extract composition, a preparation method and application thereof.
Background
Fennel (Foeniculum vuLgare) is a dried mature fruit of Foeniculum vulgare of Umbelliferae. The medicinal material has pungent and warm nature, has the effects of dispelling cold, relieving pain and regulating qi, and is collected in Chinese pharmacopoeia. Fructus Foeniculi contains various chemical components such as flavone, coumarin, phenolic acid, alkaloid, volatile oil, etc. Modern pharmacological researches have shown that the fennel has remarkable antibacterial, diuretic and other effects, and also has the biological activities of protecting liver, promoting bile flow, resisting bacteria and inflammation, resisting tumors, taking sex hormone as a sample and the like.
Physical illness from alcoholism or long-term overdrinking is one of the most common preventable diseases worldwide, resulting in about 330 tens of thousands of deaths each year, accounting for 6% of the total deaths worldwide. Alcohol can cause damage to a number of organs, mainly the liver, gastrointestinal tract and brain, where gastric mucosal damage is a highly chronic disease resulting from the long-term accumulation of alcohol. After entering the stomach, ethanol is metabolized and converted into acetaldehyde, and the acetaldehyde is combined with gastric protein to destroy the gastric mucosa structure, so that neutrophils infiltrate the gastric mucosa, and myeloperoxidase, oxygen free radicals, active oxidative metabolites and protease are released, and adhere to vascular endothelium to cause mucosal injury. Therefore, research and development of traditional Chinese medicine healthy products for preventing and treating gastric mucosal injury caused by alcohol and having small side effects are receiving more and more attention.
Disclosure of Invention
In view of the above, the present invention aims to provide a fennel extract composition, a preparation method and application thereof. The fennel extract composition prepared by the invention has obvious prevention and treatment effects on gastric mucosal injury caused by alcohol, and has no toxic or side effect.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of a fennel extract composition, which comprises the following steps:
mixing fructus Foeniculi powder with the first water, and steam distilling to obtain volatile oil component, first extractive solution and first residue;
mixing the first residues with the second water, and decocting and extracting to obtain a second extract;
mixing and concentrating the first extract and the second extract, and purifying by a macroporous resin column to obtain a water-soluble component of fennel;
mixing the fennel volatile oil component and the fennel water-soluble component to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4.
Preferably, the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 2:3.
Preferably, the weight ratio of the fennel powder to the first water is 1:8-12.
Preferably, the time of the steam distillation extraction is 4 to 6 hours.
Preferably, the weight ratio of the first medicine residues to the second water is 1:10-16.
Preferably, the times of the decoction and the extraction are 1-3 times, and the time of each decoction and the extraction is 1 hour.
Preferably, the specific gravity of the extract obtained by mixing and concentrating the first extracting solution and the second extracting solution is 1.05-1.20.
Preferably, the macroporous resin column is a D101 macroporous adsorption resin column.
The invention also provides a fennel extract composition obtained by the preparation method according to the technical scheme, and the fennel extract composition comprises a fennel volatile oil component and a fennel water-soluble component;
the weight percentage of trans-anethole in the common fennel volatile oil component is 75.0-90.0%;
the weight percentage of total flavonoids in the water-soluble components of the fennel is 13.0-20.0%, the weight percentage of syringin is 5.0-8.0%, and the weight percentage of quercetin 3-O-beta-D glucuronide is 4.0-6.0%.
The invention also provides application of the fennel extract composition in preparing a medicament for treating and/or preventing gastric mucosal injury.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a preparation method of a fennel extract composition, which comprises the following steps: mixing fructus Foeniculi powder with water, and steam distilling to obtain fructus Foeniculi volatile oil component, first extractive solution and first residue; mixing the first residues with water, and decocting to obtain a second extract; mixing and concentrating the first extract and the second extract, and purifying by a macroporous resin column to obtain a water-soluble component of fennel; mixing the fennel volatile oil component and the fennel water-soluble component to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4. The invention sequentially separates and purifies the fennel to obtain the volatile oil component and the water-soluble component by a method of jointly using steam distillation, water decoction extraction and macroporous resin purification, and further combines the fennel volatile oil component and the fennel water-soluble component, so that the fennel volatile oil component and the fennel water-soluble component have obvious synergistic effect on preventing and treating gastric mucosal injury.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the effect of Fennel extract composition (FWEO) on the morphology of alcohol-induced gastric mucosal lesion (AAGI) in mice;
FIG. 2 is a graph of the effect of FWEO on the histopathological changes of the gastric mucosa of AAGI mice (×200);
wherein A is a normal control group; b is a model group; c is omeprazole group (OEM, 26 mg/kg); d is water-soluble group of fennel (FWE, 100 mg/kg); e is fennel volatile oil component group (FEO, 260 mg/kg); f is FWEO group (50 mg/kg); g is FWEO group (100 mg/kg); h is FWEO group (200 mg/kg).
Detailed Description
The invention provides a preparation method of a fennel extract composition, which comprises the following steps:
mixing fructus Foeniculi powder with the first water, and steam distilling to obtain volatile oil component, first extractive solution and first residue;
mixing the first residues with the second water, and decocting and extracting to obtain a second extract;
mixing and concentrating the first extract and the second extract, and purifying by a macroporous resin column to obtain a water-soluble component of fennel;
mixing the fennel volatile oil component and the fennel water-soluble component to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4.
In the present invention, materials and equipment used are commercially available in the art unless otherwise specified.
The invention mixes the fennel powder and the first water for steam distillation extraction to obtain the volatile oil component, the first extract and the first dregs of a decoction of the fennel respectively.
In the invention, the weight ratio of the fennel powder to the first water is preferably 1:8-12.
In the present invention, the steam distillation extraction is preferably preceded by soaking, and the soaking time is preferably 3 to 5 hours, more preferably 4 hours.
In the present invention, the time for the steam distillation extraction is preferably 4 to 6 hours, more preferably 5 hours.
In the invention, the method preferably further comprises the step of drying the collected volatile oil to obtain a fennel volatile oil component after the steam distillation extraction, wherein the drying is preferably anhydrous sodium sulfate.
In the invention, the weight percentage of trans-anethole in the fennel volatile oil component is preferably 75.0-90.0%, more preferably 79.16%; the fennel volatile oil component preferably further comprises anisaldehyde, D-limonene, gamma-terpinene, fennel alkene and estragole.
After the first medicine residues are obtained, the first medicine residues and the second part of water are mixed for decoction and extraction to obtain a second extracting solution.
In the present invention, the weight ratio of the first residue to the second water is preferably 1:10-16, more preferably 1:12.
In the present invention, the number of times of the extraction by decoction is preferably 1 to 3, more preferably 2, and the time of each extraction by decoction is preferably 1 hour.
After the first extracting solution and the second extracting solution are obtained, the first extracting solution and the second extracting solution are mixed and concentrated and then purified by a macroporous resin column to obtain the water-soluble component of the fennel.
In the present invention, the concentration is preferably reduced pressure concentration, and the conditions of the reduced pressure concentration preferably include: the negative pressure is 0.09-0.10 MPa, and the temperature is 50-55 ℃.
In the present invention, the specific gravity of the extract obtained by mixing and concentrating the first extract and the second extract is preferably 1.05 to 1.20, more preferably 1.15.
In the invention, the concentration preferably further comprises mixing the obtained extract with a third part of water, wherein the weight ratio of the extract to the third part of water is preferably 1:3, and the extract is fully dissolved by the water to facilitate column separation and purification.
In the invention, the macroporous resin column is preferably a D101 macroporous adsorption resin column, and the diameter-to-height ratio of the D101 macroporous adsorption resin column is preferably 1:6-10.
In the present invention, the macroporous resin column purification preferably includes the steps of:
loading the mixed solution obtained by mixing the extract with water, and sequentially removing impurities and eluting to obtain an eluent;
concentrating and drying the eluent in sequence to obtain the fennel water-soluble component.
In the invention, the impurity removal preferably adopts 8-10 times of column water, the elution preferably adopts 5-8 times of column water, and the volume concentration of the ethanol solution is preferably 50%.
In the present invention, the concentration is preferably reduced pressure concentration, and the conditions of the reduced pressure concentration preferably include: the negative pressure is 0.09-0.10 MPa and the temperature is 50-55 ℃.
In the present invention, the drying is preferably vacuum drying, and the conditions of the vacuum drying preferably include: the negative pressure is 0.09-0.10 MPa, and the temperature is 55-60 ℃.
In the invention, the weight percentage of total flavonoids in the fennel water-soluble components is preferably 13.0-20.0%, more preferably 15.75%; the water-soluble component of the fennel preferably further comprises beta-daucosterol and coumarin.
In the invention, the weight percentage of syringin in the water-soluble component of the fennel is preferably 5.0-8.0%, more preferably 6.50%; the weight percentage of quercetin 3-O-beta-D glucuronide in the water-soluble component of the fennel is preferably 4.0-6.0%, more preferably 4.32%.
After the fennel volatile oil component and the fennel water-soluble component are obtained, the fennel volatile oil component and the fennel water-soluble component are mixed to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4.
In the invention, the weight ratio of the fennel volatile oil component to the fennel water-soluble component is preferably 2:3.
The method for mixing the fennel volatile oil component and the fennel water-soluble component has no special requirements, and the common method for the person skilled in the art is adopted.
The invention also provides a fennel extract composition obtained by the preparation method according to the technical scheme, and the fennel extract composition comprises a fennel volatile oil component and a fennel water-soluble component;
the weight percentage of trans-anethole in the common fennel volatile oil component is 75.0-90.0%;
the weight percentage of total flavonoids in the water-soluble components of the fennel is 13.0-20.0%, the weight percentage of syringin is 5.0-8.0%, and the weight percentage of quercetin 3-O-beta-D glucuronide is 4.0-6.0%.
In the present invention, the weight percentage of the trans-anethole is preferably 25.0 to 40.0%, more preferably 31.66%, based on the weight percentage of the fennel extract composition.
In the present invention, the total flavonoids are preferably 7.0 to 12.0% by weight, more preferably 9.45% by weight based on the weight of the fennel extract composition.
In the present invention, the weight percentage of syringin is preferably 3.0 to 4.8%, more preferably 3.90%, based on the weight percentage of the fennel extract composition.
In the present invention, the weight percentage of quercetin 3-O-beta-D glucuronide is preferably 2.4-3.6%, more preferably 2.59%, based on the weight percentage of the fennel extract composition.
The invention also provides application of the fennel extract composition in preparing a medicament for treating and/or preventing gastric mucosal injury.
In the present invention, the gastric mucosal lesion is preferably a gastric mucosal lesion caused by a chemical substance, more preferably a gastric mucosal lesion caused by alcohol.
In the present invention, the fennel extract composition may be further formulated for application to healthy products.
For further explanation of the present invention, the fennel extract composition, the preparation method and application thereof provided in the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1 preparation of Fennel extract composition
Pulverizing fructus Foeniculi material 2.0kg, sieving, soaking in 12 times of water for 4 hr, extracting by steam distillation for 5 hr, collecting volatile oil, drying with anhydrous sodium sulfate to obtain fructus Foeniculi volatile oil component (FEO) 38mL, and storing at-20deg.C for use.
Decocting the residues with 12 times of water for 2 times and 1 hr/time, mixing the extractive solutions (including the rest extractive solution after volatile oil extraction), concentrating under reduced pressure at 50deg.C under 0.09MPa to obtain fluid extract 810g with specific gravity of 1.15; dissolving the extract with 3 times of deionized water, loading onto D101 macroporous adsorbent resin column (diameter-height ratio 1:10), removing impurities with 10 column volumes of water, eluting with 8 column volumes of 50% ethanol, collecting 50% ethanol eluate, concentrating under reduced pressure of 0.09MPa, and vacuum drying under reduced pressure of 0.09MPa at 55deg.C to obtain brown yellow powder 66.87g, which is fructus Foeniculi water soluble component FWE.
Mixing the fennel volatile oil component FEO and the water-soluble component FWE according to the weight ratio of 2:3 to obtain the fennel extract composition FWEO.
1. The content of total flavonoids in FWE is determined by adopting a method of Chinese pharmacopoeia 2020 edition:
taking 10.0mg of rutin reference substance, precisely weighing, placing into 50mL volumetric flask, adding appropriate amount of methanol, heating in water bath to dissolve, cooling, adding methanol to scale, and shaking to obtain reference substance solution (containing rutin 0.2mg per 1 mL). Precisely measuring 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0mL of reference substance solution, respectively placing into 25mL volumetric flasks, respectively adding water to 6.0mL, adding 1.0mL of 5% sodium nitrite solution, uniformly mixing, standing for 6 minutes, adding 1.0mL of 10% aluminum nitrate solution, shaking uniformly, standing for 6 minutes, adding 10.0mL of 4% sodium hydroxide test solution, adding water to the scale, shaking uniformly, standing for 15 minutes, taking the corresponding reagent as a blank, measuring absorbance at 500nm wavelength according to ultraviolet-visible spectrophotometry (general rule 0401), taking absorbance as an ordinate, and taking reed as an ordinateThe butyl concentration is plotted on the abscissa, and a standard curve is drawn, and the obtained linear regression equation is y= 0.5259x-0.0007 (R 2 = 0.9986), there is a good linear relationship in the range of 0.2 to 1.2 mg. The results showed a total flavone content in FWE of 15.75%.
2. The content of syringin and quercetin-3-O-beta-D-glucuronide in FWE is determined by adopting a high performance liquid chromatography method:
chromatographic conditions: agilent Eclipse XDB-C18 chromatographic column (250 mm. Times.4.6 mm,5 μm); taking chromatographic grade acetonitrile (B) and 0.1% phosphoric acid solution (A) as mobile phases, and performing gradient elution (0-25 min,8% -20%; 25-30 min,20% -45%; 30-35 min,45% -45%; 35-40 min,45% -8%; 40-45 min,8% -8% B); the flow rate is 1.0mL/min, the column temperature is 30 ℃, the detection wavelength is 245nm, and the sample injection amount is 10 mu L. Drawing a standard curve: precisely weighing appropriate amount of syringin and quercetin-3-O-beta-D-glucuronide reference substances, diluting with 70% methanol water, fixing volume to 10mL volumetric flask, and shaking. Precisely sucking the syringin reference substance solutions 0.2, 0.5, 1.0, 2.0 and 5.0mL respectively in a 10mL volumetric flask, fixing the volume to the scale with 70% methanol, and shaking uniformly. Sampling according to chromatographic conditions, recording peak areas, and drawing a standard curve to obtain a regression equation: syringin: y=10464x+2.1828, r 2 =0.999, with good linearity in the range of 0.001-0.06 mg/mL; quercetin-3-O-beta-D-glucuronide: y= 19042x-2.7027, r 2 The linearity was good in the range of 0.0006-0.06mg/mL, =0.999. The measurement results showed that the content of syringin and quercetin-3-O-beta-D-glucuronide in FWE was 6.50% and 4.32%, respectively.
3. And (3) measuring the content of trans-anethole in the FEO by adopting a high performance liquid chromatography:
chromatographic conditions: a Shim-pack XC 18 column (4.6 mm. Times.250 mm,5 μm); chromatographic methanol (B) -water (A) is used as mobile phase. Gradient elution (0-19 min,70% -80%, 19-21 min,80% -95%, 21-30 min,95% -90%, 30-40 min,90% -70% B); the flow rate is 1.0mL/min, the column temperature is 30 ℃, the detection wavelength is 254nm, and the sample injection amount is 10 mu L. Drawing a standard curve: taking trans-anethole reference substance, precisely weighing, and dissolving in absolute ethanol to obtain concentration0.0096mg/mL of control solution. Precisely sucking the reference solutions 0.1, 0.2, 0.5, 1.0 and 2.0mL respectively in a 10mL volumetric flask, fixing the volume to scale with absolute ethanol, shaking, sampling according to chromatographic conditions, and recording peak area to obtain regression equation y=818233535.493x+20181.300, R 2 The linearity was good in the range of 0.000096 to 0.0096mg/mL, =0.999. The measurement result showed that the content of trans-anethole in FEO was 79.16%.
The compatible composition of FEO and FWE contains 31.66% of trans-anethole, 3.90% of syringin, 2.59% of quercetin 3-O-beta-D glucuronide and 9.45% of total flavonoids.
EXAMPLE 2 protective Effect of Fennel composition FWEO on mice with alcoholic gastric mucosal injury (AAGI)
The experimental method comprises the following steps: after 4 days of adaptive feeding, 96 male Kunming mice were randomly divided into a normal group (blank), a model group, a positive omeprazole group (OEM, 26 mg/kg), fennel Huang Tongzu (FWE, 100 mg/kg), fennel volatile oil group (FEO, 260 mg/kg), fennel extract composition group (FWEO, 50, 100, 200 mg/kg) 12 groups each according to body weight. Prior to administration, the corresponding concentrations were dissolved with 0.5wt% tween-80 solution: positive medicine is 2.6mg/mL, FWE is 10mg/mL, FEO is 26mg/mL, FWEO is 5, 10 and 20mg/mL respectively, corresponding medicines are pre-administered according to 10mL/kg for 4 days, equivalent amount of 0.5wt% Tween-80 solution is administered by gastric lavage of a normal group and a model group, physiological saline is administered from the 6 th day, 85% alcohol is administered by gastric lavage of 10mL/kg after each group is administered for 1 hour, and alcohol is continuously administered for 3 days, so that an acute alcoholic gastric mucosal injury mouse model is formed. After the last wine feeding for 1 hour, the eyeballs are picked up to take blood, the mice are killed by cervical dislocation, gastric tissues are separated and taken out, and the mice are weighed; shearing the appetizing cavity along the greater curvature of the stomach, cleaning with ice physiological saline, flattening on a clean flat plate, observing the general change, evaluating the damage degree of the stomach tissue, and calculating the damage index and the ulcer inhibition rate; determination of mouse serum PEG using biochemical kit 2 Levels and activity of MPO, IL-1 beta and TNF-alpha in the stomach tissue of mice.
Experimental results:
in the test process, the normal control group mice have good growth condition, liveness, vigorous appetite and luster of hair; the model group mice have symptoms of gait instability, activity reduction, food consumption reduction, emaciation, somnolence and the like after being subjected to alcohol gastric lavage, and individual mice also have sounds of oral-labial cyanosis and large-mouth wheezing. The body weight changes of the mice in each group are not statistically different, the results are shown in Table 1 and are shown in figure 1, the stomach tissue of the mice in the normal group is normal in macroscopic morphology, no obvious hemorrhagic lesions appear, and the stomach tissue of the mice in the model group is obvious in hyperemia, swelling and hemorrhagic lesions, and the ulcers are most serious. FWEO (100, 200 mg/kg) significantly ameliorates these symptoms and significantly reduces alcohol resulting in an increase in gastric index (P < 0.05).
TABLE 1 influence of FWEO on body weight and gastric index of AAGI mice
Remarks: # P＜0.05， ## p < 0.01, compared to the normal group; * P <0.05, P < 0.01, compared to model group; Δ P<0.05， ΔΔ P<0.01, compared to FWE group; ○ P<0.05， ○○ P<0.01 compared to FEO group; the stomach index is an organ index, and means the weight/body weight of the stomach×100%.
Further referring to the Guth standard pair for calculation of ulcer index, the activity of FWEO to improve gastric mucosal injury was evaluated, as shown in Table 2, and compared with the model group (18.72), FWEO was able to significantly reduce the ulcer index (16.42, 13.28, 10.63, P < 0.05) of alcohol-induced gastric mucosal injury mice, wherein the high dose (100, 200mg/kg, P < 0.05) activity was superior to FWE (100 mg/kg,16.83, P < 0.05), the high dose (200 mg/kg) activity was superior to FEO (260 mg/kg, 13.09), the efficacy was almost the same as that of omeprazole (26 mg/kg, 10.83), and the inhibition rate was 43.23%.
TABLE 2 influence of FWEO on AAGI mouse gastric ulcer index
Remarks: the different letters in the same column represent significance of the difference according to the Duncan multiple range test, the difference being at P <0.05.
PGE 2 Is an important anti-inflammatory marker for gastric tissues, and can play a role in protecting gastric mucosa by promoting secretion of gastric mucus, strengthening gastric mucosa barrier, improving gastric mucosa circulation and the like. As shown in Table 3, excessive alcohol intake resulted in PGE in serum of mice in the model group 2 The content is obviously reduced by 63.96 percent compared with the normal group, which indicates that the gastric mucosa of the mice in the model group is damaged to a certain extent, and the continuous 7-day gastric lavage FWE, FEO, FWEO has certain prevention and treatment effects on the damage, wherein the FWEO effect is obvious. FWEO (200 mg/kg) significantly increased mice serum PGE compared to the model group 2 The content (P < 0.01) is better than that of FEO (260 mg/kg) and FWE (100 mg/kg), and the difference is significant (P < 0.01). MPO is a peroxidase expressed and secreted in large quantities by activated neutrophils, and the infiltration of white blood cells in the stomach tissue of mice can be judged by MPO activity. The serum MPO activity of the mice in the model group is obviously increased (P is less than 0.01) compared with that of the mice in the normal group, which indicates that inflammatory cells invade submucosa tissues after the gastric mucosa of the mice is damaged by alcohol. FWEO (100, 200 mg/kg) can significantly reduce the MPO activity of mice with elevated alcohol stimulation, and the effect is significantly better than that of FEO (260 mg/kg) and FWE (100 mg/kg).
TABLE 3FWEO versus AAGI mouse serum PGE 2 And influence of MPO level
Remarks: # P＜0.05， ## p < 0.01, compared to the normal group; * P <0.05, P < 0.01, compared to model group; Δ P<0.05， ΔΔ P<0.01, compared to FWE group; ○ P<0.05， ○○ P<0.01, compared to FEO group.
TNF-alpha and IL-1 beta are common inflammatory factors in the body and play an important role in the inflammatory reaction of the body. As shown in table 4, the alcohol lavage resulted in significant increases in TNF- α, IL-1β content (P < 0.05) in the stomach tissue of mice, FWEO (50, 100, 200 mg/kg), FEO (260 mg/kg) and FWE (100 mg/kg) were able to significantly alleviate the increase in TNF- α, IL-1β content (P < 0.05) caused by alcohol intake, with high doses of FWEO acting significantly better than FEO and FWE, and high doses of inhibition effect close to positive control OEM. The results demonstrate that FWEO can exert a protective effect on gastric mucosal lesions by inhibiting the expression of pro-inflammatory cytokines.
TABLE 4FWEO vs. AAGI mouse stomach tissue TNF-alpha, IL-1 beta levels
Remarks: # P＜0.05， ## p < 0.01, compared to the normal group; * P <0.05, P < 0.01, compared to model group; Δ P<0.05， ΔΔ P<0.01, compared to FWE group; ○ P<0.05， ○○ P<0.01, compared to FEO group.
FIG. 2 is a graph of the effect of FWEO on the histopathological changes in the gastric mucosa of AAGI mice (x 200-fold), wherein A is the normal control group; b is a model group; c is OEM group (26 mg/kg); d is FWE group (100 mg/kg); e is FEO group (260 mg/kg); f is FWEO group (50 mg/kg); g is FWEO group (100 mg/kg); h is FWEO group (200 mg/kg). HE staining results showed that the gastric mucosa layers of the mice in the blank group had clear and complete structures, and no bleeding and inflammatory cell infiltration were seen (a in fig. 2). The gastric mucosa of the mice in the model group was severely damaged, the myometrium cells were broken and shed, various cells were severely damaged, a local area was necrotized, and a large amount of inflammatory cell infiltration was seen (B in fig. 2). The FWE group (100 mg/kg) and FWEO group (50 mg/kg) mice had a reduced gastric mucosal lesion compared to the model group, but the mucosal epithelial cell shedding was severe, and congestion and inflammatory cell infiltration were present (D in fig. 2 and F in fig. 2). The degree of gastric mucosal injury of mice in FEO group (260 mg/kg) and FWEO group (100 mg/kg) is greatly reduced, each layer of stomach tissue is clear in structure, mucosal epithelium and glands are not obviously damaged, part of surfaces are damaged, and a small amount of congestion and inflammatory cell infiltration exists locally (E in figure 2 and G in figure 2). The gastric mucosa of mice in the omeprazole positive group (26 mg/kg) and the FWEO (260 mg/kg) group were not significantly damaged, the structures of the respective layers were clear and complete, and inflammatory cell infiltration was not significant (C in FIG. 2 and H in FIG. 2).
The results show that the FEO and the FWE have better prevention and treatment effects on gastric mucosal injury caused by alcohol, the FEO and the FWE have obvious synergistic effects, the FWEO group has obvious prevention and treatment effects on gastric mucosal injury caused by alcohol stimulation (the fennel volatile oil component FEO and the fennel water-soluble component FWE are combined according to the weight ratio of 2:3), and the pharmacodynamics test result is obviously superior to that of the FEO group and the FWE group which are singly used.
While the foregoing embodiments have been described in some, but not all embodiments of the invention, other embodiments of the invention can be made and still fall within the scope of the invention without undue effort.
Claims (10)
1. A method for preparing a fennel extract composition, comprising the steps of:
mixing fructus Foeniculi powder with the first water, and steam distilling to obtain volatile oil component, first extractive solution and first residue;
mixing the first residues with the second water, and decocting and extracting to obtain a second extract;
mixing and concentrating the first extract and the second extract, and purifying by a macroporous resin column to obtain a water-soluble component of fennel;
mixing the fennel volatile oil component and the fennel water-soluble component to obtain the fennel extract composition; the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 1-3:2-4.
2. The preparation method according to claim 1, wherein the weight ratio of the fennel volatile oil component to the fennel water-soluble component is 2:3.
3. The preparation method according to claim 1, wherein the weight ratio of the fennel powder to the first water is 1:8-12.
4. A production method according to claim 1 or 3, wherein the time for the steam distillation extraction is 4 to 6 hours.
5. The method of claim 1, wherein the weight ratio of the first residue to the second water is 1:10-16.
6. The method according to claim 1 or 5, wherein the number of times of the extraction is 1 to 3, and the time of each extraction is 1 hour.
7. The method according to claim 1, wherein the specific gravity of the extract obtained by mixing and concentrating the first extract and the second extract is 1.05 to 1.20.
8. The method according to claim 1, wherein the macroporous resin column is a D101 macroporous adsorption resin column.
9. The fennel extract composition obtained by the production method of any one of claims 1 to 8, wherein the fennel extract composition comprises a fennel volatile oil component and a fennel water-soluble component;
the weight percentage of trans-anethole in the common fennel volatile oil component is 75.0-90.0%;
the weight percentage of total flavonoids in the water-soluble components of the fennel is 13.0-20.0%, the weight percentage of syringin is 5.0-8.0%, and the weight percentage of quercetin 3-O-beta-D glucuronide is 4.0-6.0%.
10. Use of a fennel extract composition of claim 9 in the manufacture of a medicament for the treatment and/or prevention of gastric mucosal lesions.
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