CN117224625A - Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis - Google Patents
Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis Download PDFInfo
- Publication number
- CN117224625A CN117224625A CN202311241662.3A CN202311241662A CN117224625A CN 117224625 A CN117224625 A CN 117224625A CN 202311241662 A CN202311241662 A CN 202311241662A CN 117224625 A CN117224625 A CN 117224625A
- Authority
- CN
- China
- Prior art keywords
- fan palm
- chinese fan
- palm seed
- extract
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 240000007163 Livistona chinensis Species 0.000 title claims abstract description 143
- 239000000284 extract Substances 0.000 title claims abstract description 86
- 230000001684 chronic effect Effects 0.000 title claims abstract description 41
- 206010009887 colitis Diseases 0.000 title claims abstract description 40
- 239000003814 drug Substances 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims description 12
- 150000004676 glycans Chemical class 0.000 claims abstract description 17
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 17
- 239000005017 polysaccharide Substances 0.000 claims abstract description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 86
- 239000000287 crude extract Substances 0.000 claims description 39
- 235000019441 ethanol Nutrition 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 18
- OAABHEHWRQAHEJ-UHFFFAOYSA-N butan-1-ol;chloroform Chemical compound ClC(Cl)Cl.CCCCO OAABHEHWRQAHEJ-UHFFFAOYSA-N 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 15
- 102000004169 proteins and genes Human genes 0.000 claims description 15
- 108090000623 proteins and genes Proteins 0.000 claims description 15
- 239000011259 mixed solution Substances 0.000 claims description 14
- 210000000582 semen Anatomy 0.000 claims description 14
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 9
- 239000004952 Polyamide Substances 0.000 claims description 9
- 229920002647 polyamide Polymers 0.000 claims description 9
- 238000000502 dialysis Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 230000037396 body weight Effects 0.000 claims description 5
- 238000005238 degreasing Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000002386 leaching Methods 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 239000000443 aerosol Substances 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000006210 lotion Substances 0.000 claims description 2
- 239000003094 microcapsule Substances 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 240000008243 Corypha umbraculifera Species 0.000 claims 2
- 229940098465 tincture Drugs 0.000 claims 1
- 210000001072 colon Anatomy 0.000 abstract description 23
- 230000008595 infiltration Effects 0.000 abstract description 12
- 238000001764 infiltration Methods 0.000 abstract description 12
- 210000002175 goblet cell Anatomy 0.000 abstract description 10
- 210000004969 inflammatory cell Anatomy 0.000 abstract description 9
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 7
- 241001465754 Metazoa Species 0.000 abstract description 6
- 238000010171 animal model Methods 0.000 abstract description 6
- 238000004904 shortening Methods 0.000 abstract description 5
- 230000006727 cell loss Effects 0.000 abstract description 4
- 230000006378 damage Effects 0.000 abstract description 4
- 208000027418 Wounds and injury Diseases 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract description 3
- 208000014674 injury Diseases 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 description 35
- 230000000694 effects Effects 0.000 description 11
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 8
- 229920003045 dextran sodium sulfate Polymers 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 239000003651 drinking water Substances 0.000 description 6
- 235000020188 drinking water Nutrition 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 208000025865 Ulcer Diseases 0.000 description 5
- 210000004400 mucous membrane Anatomy 0.000 description 5
- 231100000397 ulcer Toxicity 0.000 description 5
- 210000004185 liver Anatomy 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000009266 disease activity Effects 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 210000004876 tela submucosa Anatomy 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 2
- 206010023799 Large intestinal ulcer Diseases 0.000 description 2
- 241000202661 Livistona Species 0.000 description 2
- 206010030113 Oedema Diseases 0.000 description 2
- 230000008378 epithelial damage Effects 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 230000003628 erosive effect Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- -1 oral preparations Substances 0.000 description 2
- 230000000306 recurrent effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 241000233788 Arecaceae Species 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 208000004262 Food Hypersensitivity Diseases 0.000 description 1
- 206010016946 Food allergy Diseases 0.000 description 1
- 206010018691 Granuloma Diseases 0.000 description 1
- 206010019851 Hepatotoxicity Diseases 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 206010022941 Iridocyclitis Diseases 0.000 description 1
- 206010025476 Malabsorption Diseases 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 201000004612 anterior uveitis Diseases 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000019902 chronic diarrheal disease Diseases 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000020932 food allergy Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 230000007686 hepatotoxicity Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000004968 inflammatory condition Effects 0.000 description 1
- 208000002551 irritable bowel syndrome Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000005976 liver dysfunction Effects 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007119 pathological manifestation Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Classifications
-
- 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
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The application relates to an application of a Chinese fan palm seed extract in preparing a medicament for preventing and/or treating chronic colitis, and belongs to the technical field of biological medicines. According to the application, the Chinese fan palm seed extract with the polysaccharide content of more than 60% is proved at the animal level for the first time, so that the phenomenon of colon shortening can be relieved, inflammatory cell infiltration, surface epithelial injury and goblet cell loss of intestinal tracts can be reduced, and DAI score and histological double-blind score of an animal model can be reduced, thereby playing a role in treating chronic colitis.
Description
Technical Field
The application relates to the technical field of biological medicines, in particular to application of a Chinese fan palm seed extract in preparation of a medicament for preventing and/or treating chronic colitis.
Background
Chronic colitis is a chronic, recurrent, multiple inflammatory edema of the intestinal tract, ulcers, hemorrhagic lesions, clinically manifested as chronic diarrhea, recurrent abdominal pain, concomitant other inflammatory conditions such as arthritis, iridocyclitis, liver dysfunction, etc. due to various pathogenic causes. Chronic colitis has a complex etiology, and common etiology is non-specific enteritis, such as irritable bowel syndrome, inflammatory bowel disease, intestinal dysbacteriosis, small intestinal malabsorption, etc. Chronic colitis is generally considered to be associated with infection, immune inheritance, environmental, food allergy, and prevention of dysfunction and mental factors.
The semen Livistonae chinensis is seed of R.Br. of Livistona chinensis Livistona chinensis (Jacq.) of family Palmae, has sweet and bitter taste, is classified into lung, liver, kidney and stomach channels, has effects of promoting blood circulation for removing blood stasis, softening and resolving hard mass, and is used for treating chronic hepatitis. Modern medical research discovers that Chinese fan palm seed has the functions of resisting oxidization, protecting liver and resisting nasopharyngeal carcinoma, but no related report on the prevention and treatment effects of Chinese fan palm seed extracts mainly comprising Chinese fan palm seed polysaccharide on chronic colitis exists.
Disclosure of Invention
The application aims to overcome the defects of the prior art and provide the application of the Chinese fan palm seed extract in preparing medicines for preventing and/or treating chronic colitis.
In order to achieve the above purpose, the technical scheme adopted by the application is as follows:
in a first aspect, the application provides an application of a Chinese fan palm seed extract in preparing a medicament for preventing and/or treating chronic colitis; the Chinese fan palm seed extract comprises Chinese fan palm seed polysaccharide.
According to the application, the Chinese fan palm seed extract can be found to be effective in treating chronic colitis on an animal experiment level for the first time, and the Chinese fan palm seed extract can be found to be effective in reducing colon shortening, inflammatory cell infiltration, surface epithelial damage and goblet cell loss caused by chronic colitis by adopting the Chinese fan palm seed extract to treat a chronic colitis animal model, and can reduce DAI score and histological double-blind score of the animal model.
As a preferred embodiment of the application of the application, the mass percentage of the Chinese fan palm seed polysaccharide in the Chinese fan palm seed extract is more than 60%, and the relative molecular mass of the Chinese fan palm seed polysaccharide is 8000-14000kDa.
As a preferred embodiment of the application of the present application, the dosage form of the medicament for preventing and/or treating chronic colitis is at least one of capsules, tablets, oral preparations, microcapsule preparations, injections, suppositories, sprays, ointments, gels, solutions, powders, lotions, tinctures, oils, creams and aerosols.
As a preferred embodiment of the use according to the application, the medicament for preventing and/or treating chronic colitis is for oral administration.
As a preferred embodiment of the use according to the application, the medicament for preventing and/or treating chronic colitis is administered orally in a dose of 50-100mg per 1kg body weight of the organism. In the experimental process, 50-100mg of the Chinese fan palm seed extract can obviously reduce the DAI score of a chronic colitis animal model, and the oral administration dosage for treating the chronic colitis is 50-100mg of the Chinese fan palm seed extract per 1kg body weight of an organism, so that the symptoms of the chronic colitis can be effectively relieved.
As a preferred embodiment of the application of the application, the Chinese fan palm seed extract can reduce infiltration of inflammatory cells in the intestinal tract, and relieve epithelial damage on the surface of the intestinal tract and loss of goblet cells.
In a second aspect, the application provides a preparation method of a Chinese fan palm seed extract, which comprises the following steps:
(1) Adding the Chinese fan palm seed powder into ethanol for degreasing, and volatilizing the ethanol to obtain degreased Chinese fan palm seeds;
(2) Mixing the defatted Chinese fan palm seeds obtained in the step (1) with water, leaching, and filtering to obtain Chinese fan palm seed extract;
(3) Concentrating the Chinese fan palm seed extract obtained in the step (2), adding ethanol for precipitation, filtering, washing the obtained filter residues, and drying to obtain a Chinese fan palm seed crude extract;
(4) Decolorizing the crude extract of the Chinese fan palm seeds obtained in the step (3) to obtain the decolorized crude extract of the Chinese fan palm seeds;
(5) Removing protein in the decolorized crude extract of the Chinese fan palm seeds obtained in the step (4), dialyzing, and drying the dialyzed solution to obtain the Chinese fan palm seed extract.
The application successfully extracts and purifies the Chinese fan palm seed extract with the polysaccharide content of more than 60 percent through degreasing, leaching, alcohol precipitation, decoloring, deproteinizing and dialyzing, and can be applied to the prevention and treatment of chronic colitis.
As a preferred embodiment of the preparation method of the present application, in the step (1), the degreasing method is as follows: mixing semen Livistonae chinensis powder with ethanol, transferring into reflux device, refluxing in water bath at 80deg.C for 2 hr twice;
in the step (3), the washing is carried out twice by adopting absolute ethyl alcohol, diethyl ether and acetone;
in the step (4), the decoloring method comprises the following steps: adding polyamide into the crude extract water solution of Chinese fan palm, stirring and decoloring for 30min at 50 ℃; the mass ratio of the crude extract of the Chinese fan palm seeds to the polyamide in the aqueous solution of the crude extract of the Chinese fan palm seeds is as follows: polyamide=1:4;
in step (5), the method of dialysis is: transferring the crude extract of semen Livistonae chinensis with protein removed into dialysis bag, dialyzing at room temperature under running water for 48 hr, and dialyzing with distilled water for 24 hr; the specification of the dialysis bag is 8000-14000kDa.
As a preferred embodiment of the preparation method, in the step (1), the Chinese fan palm seed powder is obtained by sieving with a 60-mesh sieve and drying at 40 ℃ for 3-4 hours;
in the step (2), the method for mixing the defatted Chinese fan palm seeds with water comprises the following steps: ultrasonic mixing at 30deg.C for 20 min;
in step (2), the leaching method comprises the following steps: reflux-extracting in water bath at 90deg.C for 2 hr;
in step (3), the concentration method comprises the following steps: evaporating and concentrating under reduced pressure at 50deg.C until the volume of concentrated semen Livistonae chinensis extractive solution is one fifth of the volume before concentration.
As a preferred embodiment of the application of the present application, in the step (1), the ratio of the Chinese fan palm seed powder to the ethanol is the Chinese fan palm seed powder: ethanol = 1g:10ml of ethanol, wherein the ethanol is absolute ethanol;
in the step (2), the ratio of the defatted Chinese fan palm seeds to the water is the defatted Chinese fan palm seeds: water = 1g:20ml;
in the step (3), the ethanol is added to the mixture of the Chinese fan palm seed extract and the ethanol, wherein the ethanol content is 80%, and the ethanol is absolute ethanol.
As a preferred embodiment of the preparation method of the present application, in the step (5), the method for removing proteins in the decolorized crude extract of Chinese fan palm seed comprises: dissolving the decolorized crude extract of semen Livistonae chinensis into crude extract solution of semen Livistonae chinensis, adding chloroform-n-butanol mixed solution, oscillating for 30min, standing for layering, centrifuging, adding chloroform-n-butanol mixed solution into supernatant, oscillating, repeating for 8-12 times, and removing protein from crude extract of semen Livistonae chinensis.
The application adopts chloroform-n-butanol mixed solution to destroy the protein structure in the crude extract of Chinese fan palm seed, so that the protein can be removed by filtering after losing the solubility of the protein, and the protein in the crude extract of Chinese fan palm seed can completely react with the chloroform-n-butanol mixed solution after standing for multiple times and layering and shaking, thereby improving the removal rate of the protein. The chloroform-n-butanol mixed solution has poor solubility on polysaccharide, and does not influence the structure of polysaccharide and cause precipitation of polysaccharide, so that the chloroform-n-butanol mixed solution is used for removing protein in the crude extract of Chinese fan palm seed.
As a preferred embodiment of the preparation method of the application, the volume ratio of chloroform to n-butanol in the chloroform-n-butanol mixed solution is chloroform: n-butanol=4:1, and the volume ratio of the crude extract solution of Chinese fan palm seed to the chloroform-n-butanol mixed solution is that: chloroform-n-butanol mixture=5:1.
In a third aspect, the application provides a medicament for treating chronic colitis, the medicament comprising a Livistona chinensis extract.
As a preferred embodiment of the medicament according to the application, the medicament further comprises a pharmaceutically acceptable carrier.
Compared with the prior art, the application has the beneficial effects that:
according to the application, the Chinese fan palm seed extract with the polysaccharide content of more than 60% is proved at the animal level for the first time, so that the phenomenon of colon shortening can be relieved, inflammatory cell infiltration, surface epithelial injury and goblet cell loss of intestinal tracts can be reduced, and DAI score and histological double-blind score of an animal model can be reduced, thereby playing a role in treating chronic colitis.
Drawings
FIG. 1 is a graph showing the results of weight analysis of mice in the different treatment groups of Experimental example 3 on day 45 after administration, wherein the graph shows that the model group has significant differences, and P < 0.01, compared with the control group;
FIG. 2 is a graph showing the results of Disease Activity Index (DAI) analysis of mice in the different treatment groups in experimental example 3, wherein "#" in the graph shows that the model group has significant differences, P < 0.05, compared with the control group; ", is that there was a significant difference between the low dose group of chinese fan palm seed extract or the high dose group of chinese fan palm seed extract compared to the model group; p < 0.05, "/x" is that there is a significant difference between the low dose group of livistona chinensis extract or the high dose group of livistona chinensis extract compared to the model group, P < 0.01;
FIG. 3 is a graph showing the comparison of colon length of mice in the different treatment groups of Experimental example 3 after 45 days of administration;
FIG. 4 is a graph showing the statistical results of colon length of mice in the different treatment groups of experimental example 3 on day 45 after administration, wherein "# # #" is a significant difference between the model group and the control group, and P < 0.0001; ", is that there was a significant difference between the low dose group of chinese fan palm seed extract or the high dose group of chinese fan palm seed extract compared to the model group;
FIG. 5 shows the results of colon HE staining of mice in the different treatment groups of Experimental example 3 after 45 days of administration;
FIG. 6 is a graph showing the histopathological scores of mice in the different treatment groups of Experimental example 3 on day 45 after dosing, "# #", with significant differences in model groups compared to control groups, P < 0.0001; "" is that the low dose group of the Chinese fan palm seed extract or the high dose group of the Chinese fan palm seed extract has significant difference compared with the model group, and P is less than 0.0001;
FIG. 7 shows the results of hepatotoxicity index measurement of mice in the different treatment groups of Experimental example 3 at 45 days after administration.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present application, the present application will be further described with reference to the following specific examples.
The reagents and consumables used in the following examples were all commercially available without any particular explanation.
Experimental example 1 extraction and identification of Livistona seed extract
1. Extracting semen Livistonae chinensis extract.
S1, drying Chinese fan palm seeds, crushing by a crusher, sieving by a 60-mesh sieve, and drying at 40 ℃ for 3-4 hours to obtain Chinese fan palm seed powder;
s2, adding the Chinese fan palm seed powder obtained in the step S1 into absolute ethyl alcohol, transferring into a reflux device, refluxing in a water bath at 80 ℃ for 2 hours each time, and obtaining defatted Chinese fan palm seeds; the ratio of the Chinese fan palm seed powder to the absolute ethyl alcohol is the Chinese fan palm seed powder: absolute ethanol = 1g:10ml;
s3, mixing the defatted Chinese fan palm seeds obtained in the step S2 with water according to the defatted Chinese fan palm seeds: water = 1g: mixing 20ml of the Chinese fanpalm seed extract at 30 ℃ for 20min by ultrasonic treatment, transferring the mixture into a reflux device, carrying out water bath reflux extraction for 2h at 90 ℃, and filtering to obtain the Chinese fanpalm seed extract;
s4, decompressing, evaporating and concentrating the Chinese fan palm seed extract obtained in the step S4 at 50 ℃ until the volume is one fifth of the volume before concentration, adding absolute ethyl alcohol until the alcohol content in the mixture of the Chinese fan palm seed extract and the alcohol is 80%, precipitating at 4 ℃ overnight, filtering, washing the obtained filter residues twice by adopting absolute ethyl alcohol, diethyl ether and acetone, and drying to obtain the Chinese fan palm seed crude extract;
s5, preparing the crude extract of the Chinese fan palm seeds obtained in the step S4 into an aqueous solution of the crude extract of the Chinese fan palm seeds, adding polyamide, and stirring and decoloring for 30min at 50 ℃ to obtain a decolored crude extract of the Chinese fan palm seeds; the mass ratio of the crude extract of the Chinese fan palm seeds to the polyamide in the aqueous solution of the crude extract of the Chinese fan palm seeds is as follows: polyamide=1:4;
s6, dissolving the decolored crude extract of the Chinese fan palm seeds obtained in the step S5 into a crude extract solution of the Chinese fan palm seeds, adding chloroform-n-butyl alcohol mixed solution (chloroform: n-butyl alcohol=4:1, v/v), oscillating for 30min, standing for layering, centrifuging, taking supernatant, adding the chloroform-n-butyl alcohol mixed solution again, oscillating, repeating for 8-12 times, removing proteins in the crude extract of the Chinese fan palm seeds, transferring into a dialysis bag, dialyzing for 48h at room temperature under running water, and dialyzing for 24h by adopting distilled water to obtain the Chinese fan palm seed extract; the volume ratio of the crude extract solution of the Chinese fan palm seeds to the chloroform-n-butanol mixed solution is that: chloroform-n-butanol mixture = 5:1; the specification of the dialysis bag is 8000-14000kDa.
2. And (5) identifying the Chinese fan palm seed extract.
The polysaccharide content in the extracted Chinese fan palm seed extract is measured by a sulfuric acid-phenol method, and the measurement result shows that the polysaccharide content in the Chinese fan palm seed extract extracted by the experimental example is 62.17%, so that the Chinese fan palm seed extract with the polysaccharide content of more than 60% is proved to be successfully extracted by the application.
Experimental example 2 Effect of Livistona seed extract on mice with chronic colitis
In order to verify whether the Chinese fan palm seed extract has the effect of treating the chronic colitis or not, the effect of the Chinese fan palm seed extract on treating the chronic colitis is evaluated through an animal test, and the specific scheme is as follows:
1. establishment of mouse chronic colitis model
SPF grade C57BL/6J male mice (8 weeks old, 20-22g BW) were used as subjects and weighed into a model group and an untreated group.
On day 0, 1-3% dextran sulfate sodium salt (DSS) was given to model group mice, and normal water was given to untreated group mice;
on experiment day 2, the model-building mice were replaced with 1.5% DSS drinking water;
on experiment day 4, the model group mice were replaced with 1.5% DSS drinking water;
on experiment day 7, the model mice were replaced with DSS-free drinking water;
repeating the operation on days 0-4 from day 17-21;
repeating the operation on the 31 st day to 35 th day and the 0 th day to 4 th day to increase the concentration of DSS to 3%;
on day 38, the model mice were replaced with DSS-free drinking water.
The weight of the mice is obviously reduced, and the feces are in a dilute liquid form and severe occult blood, which proves that the chronic colitis mouse model is successfully established.
2. Effect of Livistona chinensis seed extract on mouse body weight and DAI score
The following treatment groups were set up for this experiment: a control group, a model group, a low dose group of Chinese fan palm seed extract and a high dose group of Chinese fan palm seed extract, wherein mice in the control group are given drinking water without DSS, and the mice in the other treatment groups are given drinking water with DSS according to a method for establishing a chronic colitis animal model; the low dose group of the Chinese fan palm seed extract is used for administering the Chinese fan palm seed extract to mice according to the dosage of 50mg/kg, and the high dose group of the Chinese fan palm seed extract is used for administering the Chinese fan palm seed extract to the mice according to the dosage of 100 mg/kg.
The model group, the low dose group of the Chinese fan palm seed extract and the high dose group of the Chinese fan palm seed extract are subjected to gastric lavage administration according to the following period from day 0, the liquid medicine volume is 0.1ml/g, and the administration is continued for 45 days. The administration of the mice was carried out in the first stage of 7-17 days, the administration of 24-31 days, the administration of the mice was carried out in the second stage of 38-45 days, the disease activity index (disease activity index, DAI) of the mice in the three stages was evaluated, the DAI scoring criteria were shown in Table 1, one item of fecal occult blood in the DAI scoring criteria was measured by a test paper method, the body weight results of the mice after the administration of the mice in the first stage of 7 days and 17 days were shown in FIG. 1, and the DAI scoring results were shown in FIG. 2.
TABLE 1DAI scoring criteria
As shown in fig. 1, the weight of mice in the model group, the low dose group of the Chinese fan palm seed extract and the high dose group of the Chinese fan palm seed extract are lower than those in the control group, wherein the evaluation weight of the high dose group of the Chinese fan palm seed extract is highest, the low dose group of the Chinese fan palm seed extract is inferior to that of the model group, and the Chinese fan palm seed extract can relieve the weight loss caused by chronic colitis.
As shown in fig. 2, in the DAI scores at 3 stages, the control mice were completely normal, while the scores of the model group were significantly increased. The scores of the low-dose group and the high-dose group of the Chinese fan palm seed extract are reduced to different degrees, and in the stage 3, the DAI scores of the low-dose group and the high-dose group of the Chinese fan palm seed extract are respectively reduced by 35.56 percent and 44.45 percent compared with the model group, which shows that the Chinese fan palm seed extract can obviously reduce the DAI scores of mice with chronic colitis, namely the Chinese fan palm seed extract has the effects of relieving and treating the chronic colitis.
3. Effect of Livistona chinensis seed extract on colon length, spleen weight and histopathology in mice
Mice from each treatment group in experiment three were randomly selected, sacrificed and dissected at day 45 of the experiment, and colon, spleen and liver tissues were removed. Colon length of mice was measured, colon tissues were HE and PAS stained and histologically double blind scored (scoring criteria see table 2); ALT, AST and AST/ALT indices of mouse livers were determined and the results are shown in FIGS. 3-7.
TABLE 2 animal colon tissue apparent pathology score criteria (histological double blind score)
| Scoring of | Integrity of epithelium | Inflammatory cell infiltration |
| 0 | Normal morphology | No infiltration |
| 1 | Goblet cell disappearing | Infiltration around the crypt |
| 2 | Most of the goblet cells disappeared | Infiltrating mucosal layer |
| 3 | Crypt vanishes | Massive infiltration of mucosal layer |
| 4 | Most of the crypt disappears | Submucosa infiltration |
As shown in fig. 3, the colon tissue of the mouse taken out is observed with naked eyes, the inner wall of the colon of the mouse in the control group is complete, the folds are regular, the vein textures are clear, no macroscopic erosion, ulcer or granuloma exists, and slight congestion is rarely visible; the model group can be used for shortening large intestine, thinning intestinal wall, remarkably filling and edema of mucous membrane, scattered erosion or ulcer with bleeding, large-area ulcer, and remarkably lower colon length than the control group; the colon of the mice in the low and high dose groups can also be damaged pathologically to different degrees, but the length of the colon and the thinning condition of the intestinal wall are improved to different degrees.
As shown in fig. 4, the average colon length of mice in the high dose group of the Chinese fan palm seed extract is significantly increased, and the average colon length of mice in the low dose group of the Chinese fan palm seed extract is not significantly changed, compared with the model group, which indicates that the high dose of the Chinese fan palm seed extract can effectively improve the colon length reduction caused by chronic colitis.
As shown in fig. 5, the control group of mice had colon mucosal epithelial cells aligned, no atrophy, deformation, necrosis and inflammatory cell infiltration were seen, goblet cells were abundant, and capillaries and a small amount of inflammatory cells were seen in the lamina propria. The pathological section of the DSS model group has typical acute stage pathological manifestations, multiple colonic ulcers, reduction or disappearance of goblet cells, crypt destruction and abscess occurrence, complete absence of epithelium or only a few residues, and inflammatory cells infiltrating the mucous membrane and submucosa. The colon surface mucous membrane of the mice in the group with low and high dosage of the Chinese fanpalm seed extract is basically recovered, the ulcer healing is better, the goblet cells are increased, the crypt inflammation is obviously reduced, the mucous membrane and the submucosa neutrophil are obviously reduced, but the interstitial mass still has a small amount of lymphocyte infiltration.
As shown in fig. 6, the histological double-blind score of the control group was extremely low, the score of the model group was significantly increased compared to the control group, the score of the high dose group of the livistona chinensis extract was significantly decreased compared to the model group, but the score of the low dose group of the livistona chinensis extract was not significantly different from the model group.
The data indicate that the Chinese fan palm seed extract can relieve chronic colitis by healing colonic ulcers and reducing inflammation.
As shown in fig. 7, the ALT and AST indices of the liver of mice were not significantly affected by the different treatment groups, indicating that the extract of chinese fan palm seed of the present application did not affect liver function of animals.
In conclusion, the experiment shows that the Chinese fan palm seed extract can obviously relieve the colon length reduction caused by chronic colitis, and the treatment effect of the Chinese fan palm seed extract is mainly aimed at intestinal tracts; further researches show that the Chinese fan palm seed extract with the polysaccharide content of more than 60 percent has the effect of repairing inflammatory intestinal tracts, and is particularly characterized by obviously relieving colon shortening, reducing inflammatory cell infiltration, surface epithelial injury and goblet cell loss, and the Chinese fan palm seed extract has the effect of preventing and/or treating chronic colitis.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present application and not for limiting the scope of the present application, and although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present application may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present application.
Claims (10)
1. Application of semen Livistonae chinensis extract in preparing medicine for preventing and/or treating chronic colitis is provided; the Chinese fan palm seed extract comprises Chinese fan palm seed polysaccharide.
2. The use according to claim 1, wherein the fraction of fan palm polysaccharide in the fan palm extract is greater than 60% by mass; the relative molecular mass of the Chinese fan palm seed polysaccharide is 8000-14000kDa.
3. The use according to claim 1, wherein the medicament for preventing and/or treating chronic colitis is in the form of at least one of a capsule, a tablet, an oral preparation, a microcapsule, an injection, a suppository, a spray, an ointment, a gel, a solution, a powder, a lotion, a tincture, an oil, a cream and an aerosol.
4. The use according to claim 1, wherein the medicament for preventing and/or treating chronic colitis is for oral administration.
5. The use according to claim 4, wherein the medicament for preventing and/or treating chronic colitis is administered orally in a dose of 50-100mg per 1kg body weight of the organism.
6. The preparation method of the Chinese fan palm seed extract is characterized by comprising the following steps:
(1) Adding the Chinese fan palm seed powder into ethanol for degreasing, and volatilizing the ethanol to obtain degreased Chinese fan palm seeds;
(2) Mixing the defatted Chinese fan palm seeds obtained in the step (1) with water, leaching, and filtering to obtain Chinese fan palm seed extract;
(3) Concentrating the Chinese fan palm seed extract obtained in the step (2), adding ethanol for precipitation, filtering, washing the obtained filter residues, and drying to obtain a Chinese fan palm seed crude extract;
(4) Decolorizing the crude extract of the Chinese fan palm seeds obtained in the step (3) to obtain the decolorized crude extract of the Chinese fan palm seeds;
(5) Removing protein in the decolorized crude extract of the Chinese fan palm seeds obtained in the step (4), dialyzing, and drying the dialyzed solution to obtain the Chinese fan palm seed extract.
7. The method of claim 6, wherein in step (1), the degreasing method comprises: mixing semen Livistonae chinensis powder with ethanol, transferring into reflux device, refluxing in water bath at 80deg.C for 2 hr twice;
in the step (3), the washing is carried out twice by adopting absolute ethyl alcohol, diethyl ether and acetone;
in the step (4), the decoloring method comprises the following steps: adding polyamide into the crude extract water solution of Chinese fan palm, stirring and decoloring for 30min at 50 ℃; the mass ratio of the crude extract of the Chinese fan palm seeds to the polyamide in the aqueous solution of the crude extract of the Chinese fan palm seeds is as follows: polyamide=1:4;
in step (5), the method of dialysis is: transferring the crude extract of semen Livistonae chinensis with protein removed into dialysis bag, dialyzing at room temperature under running water for 48 hr, and dialyzing with distilled water for 24 hr; the specification of the dialysis bag is 8000-14000kDa.
8. The method of claim 6, wherein in step (1), the ratio of the Chinese fan palm seed powder to the ethanol is the Chinese fan palm seed powder: ethanol = 1g:10ml of ethanol, wherein the ethanol is absolute ethanol;
in the step (2), the ratio of the defatted Chinese fan palm seeds to the water is the defatted Chinese fan palm seeds: water = 1g:20ml;
in the step (3), the ethanol is added to the ethanol content of 80% in the mixture of the extract of Chinese fan palm and the ethanol, and the ethanol is absolute ethanol.
9. The method of claim 6, wherein in step (5), the method for removing proteins in the decolorized crude extract of Chinese fan palm seed comprises: dissolving the decolorized crude extract of semen Livistonae chinensis into crude extract solution of semen Livistonae chinensis, adding chloroform-n-butanol mixed solution, oscillating for 30min, standing for layering, centrifuging, adding chloroform-n-butanol solution into supernatant, oscillating, repeating for 8-12 times, and removing protein from crude extract of semen Livistonae chinensis.
10. The method according to claim 9, wherein the volume ratio of chloroform to n-butanol in the chloroform-n-butanol mixed solution is chloroform: n-butanol=4:1, and the volume ratio of the crude extract solution of Chinese fan palm seed to the chloroform-n-butanol mixed solution is that: chloroform-n-butanol mixture=5:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311241662.3A CN117224625A (en) | 2023-09-25 | 2023-09-25 | Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311241662.3A CN117224625A (en) | 2023-09-25 | 2023-09-25 | Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117224625A true CN117224625A (en) | 2023-12-15 |
Family
ID=89096467
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311241662.3A Pending CN117224625A (en) | 2023-09-25 | 2023-09-25 | Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117224625A (en) |
-
2023
- 2023-09-25 CN CN202311241662.3A patent/CN117224625A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR100671432B1 (en) | Herb composition for asthma maintenance therapy | |
| US7947283B2 (en) | Compositions and methods for treating psoriasis by Ganoderma lucidum (Reishi) polysaccharides | |
| CN107184702B (en) | Traditional Chinese medicine composition for preventing and treating proventriculitis and myogastritis of poultry and preparation method thereof | |
| CN117224625A (en) | Application of Chinese fan palm seed extract in preparation of medicine for preventing and/or treating chronic colitis | |
| CN115025112B (en) | Application of radix angelicae polysaccharide in preparation of medicine for preventing and treating ulcerative colitis | |
| CN113940945A (en) | Application of houttuynia polysaccharide in preparation of medicine for preventing and treating inflammatory bowel disease | |
| CN116617249A (en) | Application of dendrobium candidum stem/leaf polysaccharide | |
| CN114404462B (en) | Composition and application thereof in preparation of medicine for treating gastric injury and/or gastric ulcer | |
| CN101214337B (en) | Chinese medicinal composition for treating stomach and duodenal ulcer and preparation | |
| CN110812396B (en) | Traditional Chinese medicine composition for treating ulcerative colitis as well as preparation method and application thereof | |
| CN104352539A (en) | Extraction method for achyranthes aspera extract | |
| JP3157728B2 (en) | Diabetes treatment | |
| CN117323402B (en) | Pharmaceutical composition for treating enteritis and preparation method thereof | |
| CN116763870B (en) | Extract for improving inflammatory bowel disease and application thereof | |
| CN115998779B (en) | Artemisia princeps polyphenol extract and preparation method and application thereof | |
| CN111297887B (en) | Preparation method and application of liver-protecting active component of Yunnan ginseng | |
| CN110917198B (en) | Application of dehydroevodiamine in preparation of medicine for treating acute gastritis | |
| CN116211993B (en) | Composition for treating senile chronic disease and its preparation method | |
| CN111297886A (en) | Application of gentian polysaccharide in preparation of medicine for preventing or treating liver injury diseases | |
| CN115969877A (en) | Oligomeric ginseng pectin nano selenium compound and preparation method and application thereof | |
| CN115671131A (en) | Application of trifoliate acanthopanax polysaccharide in preparation of medicine for treating hyperuricemia | |
| CN114344368A (en) | Application of nauclea officinalis extract in preparation of medicine for preventing and treating oral ulcer | |
| CN116549555A (en) | Cough relieving and phlegm reducing traditional Chinese medicine for treating cough caused by liver-lung yin deficiency | |
| CN117695319A (en) | Application of pharmaceutical composition in preparation of medicine for preventing and/or treating rheumatoid arthritis | |
| CN117731702A (en) | Application of herba Sonchi arvensis water extract in preventing and treating ulcerative colitis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |