CN116549551A - Application of aloe whole leaf powder or active ingredient thereof in preparation of medicine for treating chronic atrophic gastritis - Google Patents
Application of aloe whole leaf powder or active ingredient thereof in preparation of medicine for treating chronic atrophic gastritis Download PDFInfo
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- aloe
- polysaccharide
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- 241001116389 Aloe Species 0.000 title claims abstract description 82
- 235000011399 aloe vera Nutrition 0.000 title claims abstract description 80
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- 208000016644 chronic atrophic gastritis Diseases 0.000 title claims abstract description 46
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- 238000002360 preparation method Methods 0.000 title claims abstract description 11
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- 235000002961 Aloe barbadensis Nutrition 0.000 description 1
- 244000186892 Aloe vera Species 0.000 description 1
- 208000004300 Atrophic Gastritis Diseases 0.000 description 1
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
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- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/886—Aloeaceae (Aloe family), e.g. aloe vera
-
- 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
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the field of traditional Chinese medicines, and discloses application of aloe whole leaf powder in preparation of a medicine for treating chronic atrophic gastritis. The aloe whole leaf powder can remarkably improve the phenotype of reduced cell number, gland expansion, neck mucus cell increase and main cell decrease of the mouse stomach wall induced by tamoxifen, and has the efficacy of treating chronic atrophic gastritis. The invention discloses application of at least one of aloe polysaccharide, aloe polyphenol or aloe flavone in preparing a medicament for treating chronic atrophic gastritis. The polysaccharide, flavone and polyphenol in the aloe whole leaf powder have the efficacy of treating chronic atrophic gastritis, and the polysaccharide is more than the polyphenol is more than the flavone. The invention discloses a composition for treating chronic atrophic gastritis, which is prepared from polysaccharide, polyphenol and flavone according to the mass ratio of (11.247-65.32) (0.373-4.551) (0.452-7.182), and has the advantages of remarkably improving the efficacy of chronic atrophic gastritis and having a synergistic effect.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicines, and relates to application of aloe whole leaf powder or active ingredients thereof in preparing a medicine for treating chronic atrophic gastritis, and application of a composition of aloe polysaccharide, aloe polyphenol and aloe flavone prepared from aloe whole leaf powder in preparing the medicine for treating chronic atrophic gastritis.
Technical Field
Chronic atrophic gastritis (Chronic atrophic gastritis, CAG) is a common digestive disorder in clinic, and is mainly manifested by cytopenia of gastric mucosa wall, wider gastric gland inner cavity, cytopenia of gastric gland, and is closely related to helicobacter pylori infection, age, smoking, drinking and other factors, and the incidence and detection rate of the chronic atrophic gastritis increase with age. Modern medical treatment CAG mainly improves and alleviates patient symptoms by eradicating helicobacter pylori (Helicobacter Pylori, hp) infection, strengthening barrier function, reducing bile reflux, promoting epithelial growth, surgical treatment and the like, and the treatment method has a certain effect but is not ideal and has higher recurrence rate. Chronic atrophic gastritis is also a common disease of premalignant lesions. Therefore, the development of a practical and effective medicament for treating chronic atrophic gastritis has important scientific research significance and application value.
Aloe is evergreen succulent herb plant of Aloe genus of Liliaceae, and is a medicinal plant integrating edible, medicinal, skin caring and health promoting effects. Wherein, aloe vera is the most widely used aloe species, about 98.5% -99.5% of the aloe species is water, the rest of the aloe species is very complex, and about more than 200 chemical components exist, and mainly comprise carbohydrates, amino acids, vitamins, minerals, organic acids, lignin, phenolic compounds, anthraquinone, phytosterol and the like. The natural active ingredients are abundant, so that the composition has the effects of improving intestinal absorption, promoting wound healing, regulating organism immunity, resisting cancer, resisting virus, resisting bacteria, resisting inflammation, resisting oxidation, whitening, moisturizing and the like. At present, no research shows that aloe has the function of treating chronic atrophic gastritis, and no relevant report exists on active substance groups for resisting chronic atrophic gastritis.
Disclosure of Invention
The inventor utilizes tamoxifen to construct a model of the chronic atrophic gastritis of the mice, and the model has the phenotype of wall cell disappearance, gland expansion and neck mucus cell increase and main cytopenia, and is matched with the pathological characteristics of clinical chronic atrophic gastritis patients.
The inventor screens the activity of aloe whole leaf powder for treating chronic atrophic by using the model, and the result shows that the aloe whole leaf powder can remarkably improve the phenotype of reduced cell number, gland expansion, neck mucus cell increase and main cell reduction of mice stomach wall induced by tamoxifen, and proves that the aloe whole leaf powder has the efficacy of treating chronic atrophic gastritis. In order to further determine the active ingredients for treating the chronic atrophic gastritis in the aloe whole leaf powder, the inventor takes the aloe whole leaf powder as a raw material to obtain high-purity polysaccharide, flavone and polyphenol, and utilizes a tamoxifen-induced mouse chronic atrophic gastritis model to perform activity detection on the ingredients, and the result shows that the polysaccharide, the flavone and the polyphenol in the aloe whole leaf powder have the effect of treating the chronic atrophic gastritis, and the polysaccharide is more than the polyphenol is more than the flavone.
Based on the traditional Chinese medicine compatibility theory, the different medicinal materials can fully exert the synergistic effect according to the proportion of 'monarch, minister, assistant and guide', and the component traditional Chinese medicine theory considers that the ingredients in the traditional Chinese medicinal materials can be mainly synergistic to improve the medicinal effect according to the principle of 'monarch, minister, assistant and guide'. The inventor combines the components to screen out an active composition with the efficacy of obviously improving the chronic atrophic gastritis, and the activities of the active composition are obviously higher than those of single components, so that the active composition has a synergistic effect.
The invention aims to provide application of aloe whole leaf powder in preparing a medicine for treating chronic atrophic gastritis, wherein the aloe whole leaf powder is solid powder obtained by dehydrating aloe whole leaves.
The aloe whole leaf powder contains polysaccharide, flavone and polyphenol.
It is another object of the present invention to provide the use of at least one of aloe polysaccharides, aloe polyphenols or aloe flavonoids in the manufacture of a medicament for treating chronic atrophic gastritis.
The aloe polysaccharide is prepared by the following steps: adding aloe whole leaf powder into 80% ethanol according to the mass-volume ratio of aloe whole leaf powder to 80% ethanol of 1:8kg/L, standing overnight at 4deg.C, centrifuging, discarding supernatant, precipitating, and vacuum drying to obtain crude polysaccharide I; dissolving the crude polysaccharide I in distilled water, adding papain accounting for 0.1-0.2% of the mass of the crude polysaccharide I, heating to 60 ℃, stirring for 6h, centrifuging, and taking supernatant to obtain deproteinized crude polysaccharide II; and D101 macroporous adsorption resin wet packing, preparing the crude polysaccharide II into a sample solution with the mass fraction of 20% by using distilled water, loading the sample solution on the D101 macroporous adsorption resin column, eluting by using 50% ethanol with the volume equal to that of the sample solution, recovering the ethanol from the eluent until no alcohol smell exists, concentrating in a water bath, and drying in vacuum to obtain aloe polysaccharide.
The aloe polyphenol is prepared by the following steps: dissolving aloe whole leaf powder in 60% ethanol at a mass-volume ratio of 1:20kg/L, ultrasonic treating at 60deg.C for 45min, filtering to obtain extractive solution, and concentrating the extractive solution into solid powder; dissolving the solid powder in 60% ethanol according to the mass-volume ratio of 1:2kg/L of the solid powder to 60% ethanol, and adding a proper amount of CaCl 2 And (3) collecting the precipitate until no precipitate is separated out, adding hydrochloric acid until the precipitate is completely dissolved, extracting with ethyl acetate, collecting an organic phase, and concentrating to obtain aloe polyphenol.
The aloe flavone is prepared by the following method: adding aloe whole leaf powder into 80% ethanol according to the mass volume ratio of 1:8kg/L, centrifuging, collecting supernatant, loading to D101 macroporous adsorbent resin column, eluting with distilled water 2 times the volume of the loading liquid to remove impurities, eluting with 80% ethanol until no flavone reaction occurs, collecting 80% ethanol eluate, recovering ethanol from 80% ethanol eluate until no ethanol smell exists, concentrating in water bath, and vacuum drying to obtain total flavone.
It is another object of the present invention to provide a composition for treating chronic atrophic gastritis, wherein the composition is a composition comprising polysaccharide, flavone and polyphenol in the mass ratio of (11.247-65.32): 0.373-4.551): 0.452-7.182.
Preferably, the mass ratio of polysaccharide, flavone and polyphenol is 11.247:0.373:0.452, 65.32:4.551:7.182.
It is another object of the present invention to provide the use of said composition for the preparation of a medicament for the treatment of chronic atrophic gastritis.
The invention also aims to provide a medicine for treating chronic atrophic gastritis, which is prepared from the composition serving as a main active ingredient and a pharmaceutically acceptable carrier.
The preparation forms are tablets and capsules.
Description of the drawings:
FIG. 1 shows the results of treatment of tamoxifen-induced chronic atrophic gastritis in mice with aloe whole leaf powder.
Figure 2 shows the results of treatment of tamoxifen induced chronic atrophic gastritis in mice with polysaccharides, polyphenols and flavonoids isolated from aloe whole leaf powder.
Fig. 3 shows the treatment results of different proportions of polysaccharide, flavone and polyphenol separated from aloe whole leaf powder on tamoxifen-induced chronic atrophic gastritis of mice.
FIG. 4 is a graph showing the results of a mathematical model for predicting the treatment of atrophic gastritis with different proportions of polysaccharides, polyphenols and flavonoids; wherein, the graph A is a contour diagram of the mathematical model, and the graph B is a 3D response surface of the mathematical model.
Detailed Description
The following examples further illustrate the technical aspects of the invention. The examples are illustrative and should not be construed as limiting the invention.
The aloe whole leaf powder used in the examples was decolorized aloe whole leaf powder (Hainan bell morning bioengineering Co., ltd.) and was dehydrated aloe whole leaf solid powder.
Example 1
The aloe whole leaf powder is administrated to a tamoxifen-induced chronic atrophic gastritis mouse model by stomach irrigation, and the change of parietal cells, main cell number and gland number in stomach histopathological section is observed to judge whether the aloe whole leaf powder has the activity of treating chronic atrophic gastritis.
Experiment design: after 7 days of adaptive feeding, the C57/BL mice were randomly placed in cages and divided into 3 groups of mice: control, model, whole leaf powder, 2 cages per group, 4 per cage. The Model group and the whole leaf powder group mice are respectively prepared according to 250mg/kg of gastric-irrigation tamoxifen (the mixed solvent of absolute ethyl alcohol and sunflower seed oil in the volume ratio of 1:9, the same applies to the preparation), and the time interval between the administration and the molding is not less than 4 hours for 15 days after the 4 th day, the Model group and the whole leaf powder group mice are respectively prepared according to 62.5mg/kg (initial dose of 1/4) of gastric-irrigation tamoxifen, and the whole leaf powder group mice are simultaneously administered with aloe whole leaf powder (3.9 g/kg of crude drug, the mixed solvent of absolute ethyl alcohol and sunflower seed oil in the volume ratio of 1:9); control mice were perfused daily with a blank solution (mixed solvent of absolute ethanol and sunflower seed oil at a volume ratio of 1:9). During the experiment, the body weight and the food intake of the mice were recorded every day. After the end of the administration, the stomach tissue was sectioned in paraffin and stained for H & E.
Experimental results: the pathological results are shown in fig. 1, and compared with the Control group, the Model group and the whole leaf powder group mice have rough and uneven gastric mucosal surfaces, granular appearance and wall cell disappearance, and the gastric mucosal wall cells are obviously reduced, and glands are expanded. Compared with the Model group, after the aloe whole leaf powder is given, the cells of the gastric mucosa wall of the mice are obviously recovered, and the atrophic glands in the visual field range are obviously reduced, and the gland atrophy is not obviously improved although the mucosal gland cavity is wider, but the aloe whole leaf powder has the efficacy of improving the chronic atrophic gastritis as a whole.
Example 2
The aloe whole leaf powder is used as raw materials to extract high-purity aloe polysaccharide, aloe flavone and aloe polyphenol, and the polysaccharide, the flavone and the polyphenol are respectively acted on a tamoxifen-induced chronic atrophic gastritis mouse model to observe the change of the number of parietal cells and main cells and the change of the number of glands in pathological sections of stomach tissues, so as to determine whether the polysaccharide, the flavone and the polyphenol in the aloe whole leaf powder have the efficacy of treating the chronic atrophic gastritis.
Preparation of aloe flavone: adding aloe whole leaf powder into 80% ethanol according to the mass volume ratio of 1:8kg/L, centrifuging for 10min at 1350r/min, collecting supernatant, loading to D101 macroporous adsorbent resin column, eluting with distilled water with volume 2 times of the volume of the loading solution (i.e. supernatant) to remove impurities, eluting with 80% ethanol until no flavone reaction, collecting 80% ethanol eluate, recovering ethanol from 80% ethanol eluate until no ethanol smell exists, concentrating in water bath, and vacuum drying to obtain total flavone.
Content determination of aloe flavone:
precisely weighing 1.0g of total flavone (dried to constant weight at 120 ℃), and adding methanol to prepare 1g/mL of mother liquor of the test sample.
Preparation of a color reagent: precisely weighing AlCl 3 1.34 g, methanol is fixed to 100mL to prepare AlCl with the concentration of 0.1mol/L 3 A solution. Accurate weighing CH 3 COONa·3H 2 O2.72 g, double distilled water was dissolved to 100mL to prepare a sodium acetate solution having a concentration of 0.2 mol/L. 1.15mL of glacial acetic acid is precisely measured, and double distilled water is diluted to 100mL to prepare acetic acid solution test solution with the concentration of 0.2 mol/L.
Precisely measuring 1mL of mother solution of the sample, placing in a 10mL volumetric flask, adding 2mL of 0.1mol/L AlCl 3 The solution was fixed in volume with methanol and developed well for 20min, and absorbance was recorded at 270 nm. Preparing standard solution (standard solution is 0.025g rutin is dissolved in methanol to constant volume of 50 mL) by taking rutin as a standard substance, absorbing different amounts of standard solution, measuring absorbance value through optimized experimental steps, and performing linear fitting by using a least square method to obtain a relation between rutin concentration C and absorbance value A; the absorbance values are in a better linear relationship in the range of 0.1 to 0.7A. The aloe flavone content was measured to be 90.2%.
Preparation of aloe polysaccharide:
adding aloe whole leaf powder into 80% ethanol according to the mass volume ratio of aloe whole leaf powder to 80% ethanol of 1:8kg/L, standing overnight at 4deg.C, centrifuging for 10min at 1350r/min, discarding supernatant, precipitating, and vacuum drying to obtain crude polysaccharide I; adding distilled water into the crude polysaccharide I to dissolve completely, adding papain with mass of 0.1% of that of the crude polysaccharide I, heating to 60deg.C, stirring for 6 hr, centrifuging, and collecting supernatant to obtain deproteinized crude polysaccharide, which is denoted as crude polysaccharide II. Loading the D101 macroporous adsorption resin in a wet method, weighing crude polysaccharide II, preparing a sample solution with the mass fraction of 20% by using distilled water, loading the sample solution into the D101 macroporous adsorption resin column, eluting with 50% ethanol with the volume equal to that of the sample solution, the flow rate being 1/2 of that of the sample solution, recovering ethanol from the eluent until no alcohol smell exists, concentrating in a water bath, and drying in vacuum to obtain aloe polysaccharide.
Content determination of aloe polysaccharide:
accurately weighing 50mg of glucose (dried to constant weight at 105 ℃), placing in a 500mL volumetric flask, preparing glucose standard solutions with concentrations of 0.02, 0.04, 0.06, 0.08 and 0.1mg/mL by using distilled water, respectively taking 1mL of distilled water and 1mL of glucose standard solutions with different concentrations, respectively adding 1mL of 5% phenol solution, rapidly adding 5mL of concentrated sulfuric acid, standing for 10min, shaking, standing at 30 ℃ for 30min, measuring an OD value at 490nm, taking the glucose content as an abscissa and the OD value as an ordinate, and preparing a standard curve.
Taking aloe polysaccharide, preparing a sample solution with the concentration of 0.1mg/mL by adopting distilled water, sucking 1.0mL of the sample solution, measuring OD 490nm according to the operation of the steps, substituting a standard curve to calculate the total sugar content of the sample, and measuring the aloe polysaccharide content to be 86.4%.
Extraction of aloe polyphenol:
dissolving aloe whole leaf powder in 60% ethanol at a mass-volume ratio of 1:20kg/L, ultrasonic treating at 60deg.C for 45min, filtering to obtain extractive solution, and concentrating the extractive solution into solid powder; dissolving the solid powder in 60% ethanol according to the mass-volume ratio of 1:2kg/L of the solid powder to 60% ethanol, and adding a proper amount of CaCl 2 And (3) collecting the precipitate until no precipitate is separated out, slowly adding hydrochloric acid with the concentration of 1M until the precipitate is completely dissolved, extracting with ethyl acetate, collecting an organic phase, and concentrating to obtain aloe polyphenol.
Content determination of aloe polyphenol:
precisely weighing 5.00mg of gallic acid standard substance, placing into a 50mL volumetric flask, fixing volume to scale with 60% ethanol, and shaking to obtain gallic acid standard solution with concentration of 0.1 mg/mL. Precisely removing gallic acid standard solution 0mL, 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL, and 0.7mL respectively in 10mL volumetric flask, adding 0.5mL Fu Lin Fen reagent, and then adding 1.5mL Na with concentration of 1M 2 CO 3 The solution was fixed in volume with distilled water, heated in a 75℃water bath for 10min, naturally cooled, and scanned at full wavelength with maximum absorption at 760 nm. Drawing a standard curve by taking the gallic acid concentration (c) as an abscissa and the absorbance (A) at 760nm as an ordinate, and fitting to obtain a linear regression equation: a=121c-0.0562, r 2 = 0.9996. The aloe polyphenol content was found to be 89.9%.
Experiment design: after 7 days of adaptive feeding, the C57/BL mice were randomly placed in cages and divided into 5 groups of mice: control (Control), model, polysaccharide (3.9 g/kg crude drug dose), huang Tongzu (3.9 g/kg crude drug dose), polyphenol (3.9 g/kg crude drug dose), 2 cages per group, 4 cages per group. The Model group, the polysaccharide group, the flavone group and the polyphenol group are respectively used for pouring the stomach tamoxifen according to 250mg/kg in the first 3 days, the Model group and the 3 administration groups are respectively used for pouring the stomach tamoxifen according to 62.5mg/kg (initial dose 1/4) from the 4 th day, and the 3 administration groups are respectively provided with corresponding extraction parts (prepared by adopting mixed solvents of absolute ethyl alcohol and sunflower seed oil in a volume ratio of 1:9), and the time interval between the administration and the molding is not less than 4 hours and lasts for 15 days; the Control group was filled with a blank solution (prepared from a mixed solvent of absolute ethanol and sunflower seed oil in a volume ratio of 1:9) daily. During the experiment, the body weight and the food intake of the mice were recorded every day. After the end of the administration, the stomach tissue was sectioned in paraffin and stained for H & E.
Experimental results: the pathological results are shown in fig. 2, and after 15 days of administration, compared with the Model group, the gastric mucosal wall cells of the mice in the polysaccharide group and the polyphenol group are completely restored, and the gastric mucosal wall cells of the mice in the flavone group are completely restored under the condition of no glandular atrophy in the visual field, and the gastric mucosal wall cells of the mice in the flavone group are obviously improved although the glandular atrophy exists in the visual field. The polysaccharide, flavone and polyphenol prepared from aloe whole leaf powder serving as raw materials can be used for remarkably improving the chronic atrophic gastritis induced by tamoxifen in mice, and the polysaccharide efficacy is greater than the polyphenol efficacy and greater than the flavone efficacy.
Example 3
The effect of the combination of aloe polysaccharides, aloe flavonoids and aloe polyphenols prepared in example 2 (compatibility ratio 1, mass ratio of polysaccharides, flavonoids and polyphenols=11.247:0.373:0.452, i.e. 93.17:3.090:3.744; compatibility ratio 2, mass ratio of polysaccharides, flavonoids and polyphenols=65.32:4.551:7.182, i.e. 84.77:5.906:9.321) on tamoxifen-induced chronic atrophic gastritis mouse model was examined.
Experiment design: after 7 days of adaptive feeding, the C57/BL mice were randomly placed in cages and divided into 4 groups of mice: control group, model group, compatibility proportion 1 group, compatibility proportion 2 group, 2 cages of each group, 4 cages of each group. The Model group, the compatibility proportion 1 group and the compatibility proportion 2 group mice are filled with the tamoxifen according to 250mg/kg in the first 3 days, and the Model group and the two administration groups are filled with the tamoxifen according to 62.5mg/kg (initial dose of 1/4) from the 4 th day, and simultaneously the two administration groups are given with corresponding compositions (the total amount of the compositions is consistent with the administration dose of aloe whole leaf powder in the embodiment 1, and the administration and molding time interval is not less than 4 hours and lasts for 15 days, wherein the total amount of the compositions is prepared by adopting a mixed solvent of absolute ethyl alcohol and sunflower seed oil in a volume ratio of 1:9). The Control group was filled with a blank solution (prepared from a mixed solvent of absolute ethanol and sunflower seed oil in a volume ratio of 1:9) daily. During the experiment, the body weight and the food intake of the mice were recorded every day. After the end of the administration, the stomach tissue was sectioned in paraffin and stained for H & E.
Experimental results: the pathological results are shown in fig. 3, and after 15 days of administration, compared with the Model group, the pathological section results of the compatibility proportion 1 group and the compatibility proportion 2 group show that parietal cells are basically recovered, atrophic glands are basically disappeared in the visual field range, and the two compositions have stronger efficacy of improving chronic atrophic gastritis and have higher activity than single components.
Example 4
The aloe polysaccharide, aloe flavone and aloe polyphenol prepared in the example 2 are taken as compatibility units, and the polysaccharide, the flavone and the polyphenol are blended into corresponding mass ratios, and are acted on a mouse model induced by tamoxifen, and the detection is carried out only by taking the change number of gastric mucosa wall cells as an index. In the embodiment, a 3D model is constructed by using a D-optimal design method and a response surface curve method on the premise of keeping the total amount consistent, so that the optimal drug effect compatibility ratio is obtained.
Experiment design: after 7 days of adaptive rearing, the C57/BL mice were randomly placed in cages, and were divided into 13 groups (see Table 1), 2 cages per group, and 4 per cage. The 13 groups are named Control group, model group, compatibility ratio 3 group and compatibility ratio 13 group. The Model group and the first 3 days of the compatibility proportion of 1-11 groups are respectively filled with tamoxifen according to 250mg/kg, the Model group and 11 administration groups are respectively filled with the stomach according to 62.5mg/kg (initial dose of 1/4) from the 4 th day, meanwhile, the 11 administration groups are respectively provided with corresponding compositions (the total amount of the compositions is consistent with the administration dose of aloe whole leaf powder in the embodiment 1, and the compositions are prepared by adopting a mixed solvent with anhydrous ethanol and sunflower seed oil in a volume ratio of 1:9), and the time interval between the administration and the molding is not less than 4 hours and lasts for 15 days. The Control group was filled with a blank solution (prepared from a mixed solvent of absolute ethanol and sunflower seed oil in a volume ratio of 1:9) daily. During the experiment, the body weight and the food intake of the mice were recorded every day. After the end of the administration, the stomach tissue was sectioned in paraffin and stained for H & E.
TABLE 1 different mass ratios of polysaccharide, flavone and polyphenol
Experimental results: the results are shown in figure 4, and the difference of different compatibility ratios on the cell number of the stomach wall of the mice induced by tamoxifen is found that the model has a synergistic effect when the mass ratio of polysaccharide to flavone to polyphenol is 11.247-65.32:0.373-4.551:0.452-7.182.
Claims (9)
1. The application of aloe whole leaf powder in preparing medicine for treating chronic atrophic gastritis is disclosed.
2. The use according to claim 1, characterized in that: the aloe whole leaf powder contains polysaccharide, flavone and polyphenol.
3. Application of at least one of aloe polysaccharide, aloe polyphenol or aloe flavone in preparing medicine for treating chronic atrophic gastritis is provided.
4. A use according to claim 3, characterized in that: the aloe polysaccharide is prepared by the following steps: adding aloe whole leaf powder into 80% ethanol according to the mass-volume ratio of aloe whole leaf powder to 80% ethanol of 1:8kg/L, standing overnight at 4deg.C, centrifuging, vacuum drying the precipitate to obtain crude polysaccharide I; dissolving the crude polysaccharide I in distilled water, adding papain accounting for 0.1-0.2% of the mass of the crude polysaccharide I, heating to 60 ℃, stirring for 6h, centrifuging, and taking supernatant to obtain crude polysaccharide II; loading the D101 macroporous adsorption resin in a wet method, preparing the crude polysaccharide II into a sample solution with the mass fraction of 20% by using distilled water, loading the sample solution on the D101 macroporous adsorption resin column, eluting with 50% ethanol with the volume equal to that of the sample solution, recovering ethanol from the eluent until no alcohol smell exists, concentrating in a water bath, and drying in vacuum to obtain aloe polysaccharide;
the aloe polyphenol is prepared by the following steps: dissolving aloe whole leaf powder in 60% ethanol at a mass-volume ratio of 1:20kg/L, ultrasonic treating at 60deg.C for 45min, filtering to obtain extractive solution, and concentrating the extractive solution into solid powder; dissolving the solid powder in 60% ethanol according to the mass-volume ratio of 1:2kg/L of the solid powder to 60% ethanol, and adding a proper amount of CaCl 2 Collecting precipitate until no precipitate is separated out, adding hydrochloric acid until precipitate is completely dissolved, extracting with ethyl acetate, collecting organic phase, and concentrating to obtain aloe polyphenol;
the aloe flavone is prepared by the following method: adding aloe whole leaf powder into 80% ethanol according to the mass volume ratio of 1:8kg/L, centrifuging, collecting supernatant, loading to D101 macroporous adsorbent resin column, eluting with distilled water 2 times the volume of the loading liquid to remove impurities, eluting with 80% ethanol until no flavone reaction occurs, collecting 80% ethanol eluate, recovering ethanol from 80% ethanol eluate until no ethanol smell exists, concentrating in water bath, and vacuum drying to obtain total flavone.
5. A composition for treating chronic atrophic gastritis, characterized in that: the composition is a composition of polysaccharide, flavone and polyphenol according to the mass ratio of (11.247-65.32) (0.373-4.551) (0.452-7.182).
6. The composition of claim 5, wherein: the mass ratio of the polysaccharide to the flavone to the polyphenol is 11.247:0.373:0.452, 65.32:4.551:7.182.
7. Use of the composition of claim 5 for the preparation of a medicament for the treatment of chronic atrophic gastritis.
8. A medicament for treating chronic atrophic gastritis, which is characterized in that: the medicine is prepared from the composition of claim 5 serving as a main active ingredient and a pharmaceutically acceptable carrier.
9. A medicament according to claim 8, characterized in that: the preparation forms are tablets and capsules.
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