CN113549660B - Preparation method of broccoli sprout extract and anti-inflammatory activity thereof - Google Patents
Preparation method of broccoli sprout extract and anti-inflammatory activity thereof Download PDFInfo
- Publication number
- CN113549660B CN113549660B CN202110648434.2A CN202110648434A CN113549660B CN 113549660 B CN113549660 B CN 113549660B CN 202110648434 A CN202110648434 A CN 202110648434A CN 113549660 B CN113549660 B CN 113549660B
- Authority
- CN
- China
- Prior art keywords
- broccoli
- extract
- miao
- ganfen
- broccoli sprout
- 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.)
- Active
Links
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 title claims abstract description 73
- 235000017647 Brassica oleracea var italica Nutrition 0.000 title claims abstract description 73
- 240000003259 Brassica oleracea var. botrytis Species 0.000 title claims abstract description 71
- 239000000284 extract Substances 0.000 title claims abstract description 40
- 230000003110 anti-inflammatory effect Effects 0.000 title abstract description 15
- 238000002360 preparation method Methods 0.000 title abstract description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000000605 extraction Methods 0.000 claims abstract description 9
- 238000005238 degreasing Methods 0.000 claims abstract description 8
- 238000002390 rotary evaporation Methods 0.000 claims abstract description 7
- 230000003301 hydrolyzing effect Effects 0.000 claims abstract description 4
- 238000002386 leaching Methods 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000012153 distilled water Substances 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000002274 desiccant Substances 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 12
- 230000007062 hydrolysis Effects 0.000 claims description 11
- 238000006460 hydrolysis reaction Methods 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 235000012054 meals Nutrition 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 5
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 5
- 230000002378 acidificating effect Effects 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000003960 organic solvent Substances 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000010298 pulverizing process Methods 0.000 claims 1
- 239000004480 active ingredient Substances 0.000 abstract description 4
- 239000013543 active substance Substances 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract description 4
- 238000001035 drying Methods 0.000 abstract description 2
- 206010061218 Inflammation Diseases 0.000 description 14
- 230000004054 inflammatory process Effects 0.000 description 14
- SUVMJBTUFCVSAD-UHFFFAOYSA-N sulforaphane Chemical compound CS(=O)CCCCN=C=S SUVMJBTUFCVSAD-UHFFFAOYSA-N 0.000 description 14
- RUQCCAGSFPUGSZ-OBWQKADXSA-N Glucoraphanin Natural products C[S@](=O)CCCCC(=NS(=O)(=O)O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RUQCCAGSFPUGSZ-OBWQKADXSA-N 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 8
- GMMLNKINDDUDCF-JRWRFYLSSA-N [(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] (1e)-5-[(r)-methylsulfinyl]-n-sulfooxypentanimidothioate Chemical compound C[S@@](=O)CCCC\C(=N/OS(O)(=O)=O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GMMLNKINDDUDCF-JRWRFYLSSA-N 0.000 description 8
- SUVMJBTUFCVSAD-JTQLQIEISA-N 4-Methylsulfinylbutyl isothiocyanate Natural products C[S@](=O)CCCCN=C=S SUVMJBTUFCVSAD-JTQLQIEISA-N 0.000 description 7
- 235000015487 sulforaphane Nutrition 0.000 description 7
- 229960005559 sulforaphane Drugs 0.000 description 7
- 235000013311 vegetables Nutrition 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- 239000011734 sodium Substances 0.000 description 6
- 239000008399 tap water Substances 0.000 description 6
- 235000020679 tap water Nutrition 0.000 description 6
- 108010058651 thioglucosidase Proteins 0.000 description 6
- 239000000843 powder Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 206010009887 colitis Diseases 0.000 description 4
- 210000001072 colon Anatomy 0.000 description 4
- 230000014509 gene expression Effects 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 238000007605 air drying Methods 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 210000004969 inflammatory cell Anatomy 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 230000028709 inflammatory response Effects 0.000 description 3
- 150000002540 isothiocyanates Chemical class 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 244000308180 Brassica oleracea var. italica Species 0.000 description 2
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 108090000193 Interleukin-1 beta Proteins 0.000 description 2
- QKGJFQMGPDVOQE-UHFFFAOYSA-N Sulforaphen Natural products CS(=O)C=CCCN=C=S QKGJFQMGPDVOQE-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003517 fume Substances 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 208000035861 hematochezia Diseases 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000004877 mucosa Anatomy 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- QKGJFQMGPDVOQE-HWKANZROSA-N raphanin Chemical compound CS(=O)\C=C\CCN=C=S QKGJFQMGPDVOQE-HWKANZROSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 235000011331 Brassica Nutrition 0.000 description 1
- 241000219198 Brassica Species 0.000 description 1
- 235000011302 Brassica oleracea Nutrition 0.000 description 1
- 240000007124 Brassica oleracea Species 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 102100028999 High mobility group protein HMGI-C Human genes 0.000 description 1
- 101000986379 Homo sapiens High mobility group protein HMGI-C Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 1
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 206010038063 Rectal haemorrhage Diseases 0.000 description 1
- 229930182475 S-glycoside Natural products 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 229960000633 dextran sulfate Drugs 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000009266 disease activity Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 210000002175 goblet cell Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 150000003569 thioglycosides Chemical class 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 208000016261 weight loss Diseases 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C331/00—Derivatives of thiocyanic acid or of isothiocyanic acid
- C07C331/16—Isothiocyanates
- C07C331/18—Isothiocyanates having isothiocyanate groups bound to acyclic carbon atoms
- C07C331/20—Isothiocyanates having isothiocyanate groups bound to acyclic carbon atoms of a saturated carbon skeleton
Abstract
The invention relates to the field of food and the field of natural active substance extraction, in particular to a preparation method of a broccoli sprout extract, which comprises the steps of drying, crushing, degreasing, hydrolyzing, ultrasonic leaching and rotary evaporation of broccoli sprouts; wherein dichloromethane is adopted to extract the active ingredients of the broccoli sprouts; the broccoli sprout extract extracted by the preparation method shows better anti-inflammatory activity and has high application value.
Description
Technical Field
The invention relates to the field of foods and the field of natural active substance extraction, in particular to a preparation method and anti-inflammatory activity of broccoli sprout extracts.
Background
Broccoli (Brassica oleracea l.var.itica), also known as broccoli, is an annual plant of the brassica genus of the cruciferae family. The broccoli contains abundant natural active ingredients such as thioglucoside (GLS for short), polyphenol, anthocyanin, etc., and the nutritional ingredients are contained in plant perk. When thioglycoside is contacted with Myrosinase (MYR), isothiocyanates (ITCs) having remarkable anticancer effects can be produced by hydrolysis. The most abundant thioloside in broccoli is Glucoraphanin (GRA), the hydrolysate of which, glucoraphanin (Sulforaphane), is the natural ingredient with the strongest anticancer effect to date.
The sprouting vegetable is characterized by that it utilizes plant seed or other nutrient storage organ to directly grow out tender bud, sprout, bud ball, young shoot or young stem for eating under the condition of darkness or illumination. During the long-term growth of plants, the seedlings accumulate more phytochemicals, so that the plant active substances in the sprouting vegetables are 2-10 times that of the mature vegetables, and the content of glucoraphanin and glucoraphanin in the broccoli sprouts is 10-100 times that of the mature broccoli sprouts. Along with the improvement of the quality and variety requirements of consumers on vegetables, the sprouting vegetable is taken as a green food with rich nutrition and high quality, is more and more favored by people due to the unique flavor and the health care effect of homology of medicine and food, and the sprouting vegetable industry becomes an emerging vegetable industry with great development prospect.
Inflammation is the immune response of the body to the stimulation of pathogens. In general, immune cells can release various cytokines to regulate internal environmental signals during the inflammation process, promoting tissue repair and inflammation elimination. However, in the development process of long-term inflammation, inflammatory cells such as macrophages, mast cells, neutrophils, eosinophils and the like attack tissues, release various inflammatory factors such as IL-1 beta, IL-6, TNF-alpha and the like and a large amount of active oxygen, cause tissue injury, change the microenvironment of the organism, cause cellular gene mutation, further cause inflammation deterioration and finally promote tumor occurrence. Therefore, suppression of the occurrence and development of inflammation is one of the problems that are currently of practical significance. However, the current research on the antioxidant, antibacterial and anticancer activities of broccoli has been focused much, and the research on anti-inflammatory activity is very little.
Disclosure of Invention
The applicant has studied the biological activity of the broccoli sprouts in detail, and found that the broccoli sprout active substances extracted by a specific extraction method can provide good anti-inflammatory activity, and further studied the influence of different culture mediums on the anti-inflammatory activity of the broccoli sprout extracts. In fresh broccoli sprouts, the precursor substance of raphanin is present in different compartments than MYR. However, in the subsequent grinding process, glucoraphanin and MYR are released and are in the same reaction space. In the hydrolysis process, MYR is activated so as to exert physiological functions and promote the conversion of glucoraphanin into glucoraphanin. Therefore, without hydrolysis, MYR cannot be activated, i.e., raphanin cannot be produced. The extraction process of the broccoli sprouts has the characteristics of few steps, no need of other reagents in the hydrolysis process to regulate the hydrolysis environment, convenience in operation and the like.
In addition, anti-inflammatory activity is a more general concept that, if embodied, can be subdivided into various diseases. Currently, anti-inflammatory activity with respect to sulforaphane is mainly focused on inflammation of organs or tissues such as kidneys, livers, stomach, muscles, etc. And for the prevention and treatment of ulcerative colitis, few studies involving ITCs and sulforaphane have been conducted. Thus, although sulforaphane has some anti-inflammatory activity, the anti-inflammatory effect against specific diseases such as ulcerative colitis is not completely understood.
In one aspect, the present application provides a method for preparing a broccoli sprout extract, which is characterized in that: the method comprises the steps of drying, crushing, degreasing, hydrolyzing, ultrasonic leaching and rotary evaporating of broccoli sprouts; wherein dichloromethane is used for extracting the active ingredients of the broccoli sprouts.
Further, the method comprises the steps of:
grinding and sieving fresh broccoli sprouts Miao Donggan to obtain broccoli sprouts Miao Ganfen;
degreasing dry powder with n-hexane, and air-drying the organic solvent;
adding distilled water into the obtained residue, and hydrolyzing under dark condition to obtain wet meal;
adding a drying agent into the obtained wet meal, and then adding dichloromethane for extraction;
and (3) performing rotary evaporation on the extract liquid to obtain the broccoli sprout extract.
Further, wherein the broccoli sprouts are crushed to 80 mesh.
Further, the ratio of the broccoli sprouts Miao Ganfen to the n-hexane is 1:8g/mL, the degreasing temperature is room temperature, and the degreasing time is 1h.
Further, the feed liquid ratio of the broccoli sprouts Miao Ganfen to the distilled water is 1:4g/mL, the hydrolysis temperature is room temperature, and the hydrolysis time is 8 hours; the desiccant consists of NaCl and Na in a mass ratio of 4:3 2 SO 4 The mass ratio of the desiccant to the distilled water is 2:1; the feed liquid ratio of the broccoli sprouts Miao Ganfen to the dichloromethane is 1:10g/mL; dichloromethane extraction was performed 3 times.
Further, the temperature of the rotary evaporation is 30-35 ℃.
Further, the broccoli sprouts are fresh broccoli sprouts cultivated by slightly acidic electrolyzed water.
In another aspect, the present application provides broccoli sprout extracts prepared by the above preparation method.
In another aspect, the present application provides the use of the broccoli sprout extract as described above in anti-inflammatory aspects.
Further, the inflammation is ulcerative colitis.
Preferably, the inflammation is cell inflammation caused by LPS/IFN-gamma and ulcerative colitis caused by DSS.
The broccoli sprouts in the present application are not limited to specific varieties, and although the activities may be different, the broccoli sprouts of each variety may be used.
Inflammation in the present application is not limited to colitis, and inflammatory reactions of various organs on human or animal bodies can be controlled by the extract of the present application through proper verification; the inflammatory response is also not limited to cellular inflammatory responses and colitis symptoms in the examples, and inflammatory responses and symptoms conventionally recognized in the art can be controlled with the extracts of the present application.
The beneficial effects are that:
the invention provides a preparation method of a broccoli sprout extract, which greatly reserves characteristic active ingredients in broccoli sprouts, and the broccoli sprout extract prepared by the invention has better anti-inflammatory activity and high application value.
Drawings
FIG. 1 is the effect of broccoli extract on BMDMs inflammatory gene expression;
FIG. 2 shows the change in body weight, stool shape, hematochezia and DAI of mice; (Ctrl group represents Control, DSS group represents 2.5% DSS, br (TW) +dss group represents 20mg/kg·bw tw+2.5% DSS, br (sea) +dss group represents 20mg/kg·bw saiw+2.5% DSS);
FIG. 3 is a plot of colon tissue HE staining sections, histological scores, and colon length of mice; (Ctrl group represents Control, DSS group represents 2.5% DSS, BR (TW)/DSS group represents 20 mg/kg. Bw TW+2.5% DSS, BR (SEAW)/DSS group represents 20 mg/kg. Bw SAEW+2.5% DSS).
FIG. 4 shows the content of sulforaphane in different extracts, and the content of sulforaphane in the crude extract is calculated to be 19.1mg/100mg (FIG. 4A, slightly acidic water culture group) and 16.0mg/100mg (FIG. 4B, tap water culture group) respectively by comparison with a standard spectrogram, so that a significant difference exists. Fig. 4C is a map of the sulforaphane standard.
Detailed Description
The invention is further illustrated below in connection with specific examples.
Example 1
1) Fresh broccoli sprouts cultivated by slightly acidic electrolyzed water (Slightly Acidic Electrolyzed Water, SAEW) are harvested, freeze-dried and crushed to 80-mesh sieve to obtain broccoli sprout dry powder.
2) Adding n-hexane into the dry powder obtained in the step 1) according to a feed-liquid ratio of 1:8, wherein the unit is g/mL; shaking for 1h at room temperature, centrifuging at 5000r/min for 10min to obtain residue, and air drying the organic reagent in a fume hood.
3) Taking the residue obtained in the step 2), adding distilled water, and placing in a dark environment for hydrolysis at room temperature for 8 hours; the feed liquid ratio of the broccoli sprouts Miao Ganfen to distilled water is 1:4, and the unit is g/mL.
4) Adding NaCl and Na into the wet meal in the step 3) 2 SO 4 A desiccant comprising NaCl and Na 2 SO 4 The mass ratio of the added drying agent to the distilled water in the step 3) is 4:3, and the mass ratio of the added drying agent to the distilled water in the step 3) is 2:1. Adding dichloromethane, mixing, ultrasonic extracting for 30min, centrifuging at 5000r/min for 10min, and collecting extractive solution; the feed liquid ratio of the broccoli sprouts Miao Ganfen to the dichloromethane is 1:10, and the unit is g/mL. Repeating the extraction process for three times, and combining the obtained extracts.
5) And (3) performing rotary evaporation on the extract liquid obtained in the step (4) at the temperature of 30-35 ℃ to evaporate dichloromethane so as to obtain the broccoli extract.
Example 2
1) Fresh broccoli sprouts cultivated by Tap Water (TW) are harvested, freeze-dried and crushed to 80 mesh sieve to obtain broccoli sprout dry powder.
2) Adding n-hexane into the dry powder obtained in the step 1) according to a feed-liquid ratio of 1:8, wherein the unit is g/mL; shaking for 1h at room temperature, centrifuging at 5000r/min for 10min to obtain residue, and air drying the organic reagent in a fume hood.
3) Taking the residue obtained in the step 2), adding distilled water, and placing in a dark environment for hydrolysis at room temperature for 8 hours; the feed liquid ratio of the broccoli sprouts Miao Ganfen to distilled water is 1:4, and the unit is g/mL.
4) Adding NaCl and Na into the wet meal in the step 3) 2 SO 4 A desiccant comprising NaCl and Na 2 SO 4 The mass ratio of the added drying agent to the distilled water in the step 3) is 4:3, and the mass ratio of the added drying agent to the distilled water in the step 3) is 2:1. Adding dichloromethane, mixing, ultrasonic extracting for 30min, centrifuging at 5000r/min for 10min, and collecting extractive solution; the feed liquid ratio of the broccoli sprouts Miao Ganfen to the dichloromethane is 1:10, and the unit is g/mL. Repeating the extraction process for three times, and combining the obtained extracts.
5) And (3) performing rotary evaporation on the extract liquid obtained in the step (4) at the temperature of 30-35 ℃ to evaporate dichloromethane so as to obtain the broccoli extract.
Example 3
The beneficial effects of the invention are further illustrated by the following protective effect of the broccoli sprout extract prepared by the invention on inflammatory cells and colitis mice:
test data one: primary cells were extracted from BABL/C mouse Bone Marrow and induced with M-CSF to give Bone Marrow primary macrophages (Bone Marrow-derived Macrophages, BMDMs). During the induction of inflammation by LPS/IFN-gamma, the broccoli sprout extracts (Broccoli sprouts extract, BR) of example 1 or example 2 were added at different concentrations to the cell culture broth and incubated for 24h. For comparison, cells that had not been treated with LPS and that had not been treated with drug were used as a control group. After the induction, the anti-inflammatory effect of the broccoli sprout extract was evaluated by the expression of the cell inflammation marker gene.
Measurement results: as shown in FIG. 1, the broccoli sprout extract of example 1 or example 2 can significantly inhibit the expression of inflammatory genes IL-1 beta and iNOS, and the inhibition rate of the broccoli sprout extract on the expression of the two genes can reach 60.9% and 62.3% at most. From this, it can be seen that the broccoli sprout extract has a good anti-inflammatory effect.
Test data two: the C57BL/6Jnifdc mice were induced for 7 days with DSS (sodium dextran sulfate) while the broccoli sprout extracts prepared in example 1 or example 2 were perfused daily for a fixed time. As a comparison, mice not subjected to DSS induction while not being perfused with broccoli sprout extract were used as a control group. The mice were recorded for body weight changes, fecal character, rectal bleeding, and disease activity index during induction. The colon tissue of the mice was taken on the next day of final administration, and the morphology was observed.
Measurement results: as shown in fig. 2 and 3, the broccoli sprout extract of example 1 or example 2 has a certain degree of alleviation effect on loose stool, hematochezia and weight loss of the colitis mice caused by DSS, and has remarkable effect in the late treatment period (4-7 days). Meanwhile, the intervention of the broccoli sprout extract can relieve pathological features such as mucosa collapse, mucosa edema, goblet cell disappearance, crypt deformation, inflammatory cell infiltration and the like, and reduce colon shrinkage. Therefore, the broccoli sprout extract has better anti-colonitis effect.
Finally, it should be noted that: the foregoing description of the invention has been presented for purposes of illustration and description, and is not intended to be limiting. It will be apparent to those skilled in the art that several simple deductions, modifications or substitutions can be made according to the idea of the invention, and these simple modifications and combinations should also be regarded as the disclosure of the invention, which falls within the scope of the invention.
Claims (3)
1. Use of a broccoli sprout extract in the manufacture of a medicament for the treatment of ulcerative colitis, a method of preparing the broccoli sprout extract, the broccoli sprout being a fresh broccoli sprout cultivated with slightly acidic electrolyzed water, the method comprising the steps of:
(1) Freeze drying fresh broccoli sprout, and pulverizing to obtain broccoli sprout Miao Ganfen;
(2) Adding n-hexane into broccoli bud Miao Ganfen for degreasing, and airing an organic solvent, wherein the ratio of the broccoli bud Miao Ganfen to the n-hexane is 1:8g/mL, the degreasing temperature is room temperature, and the degreasing time is 1h;
(3) Adding distilled water into the residue obtained in the step (2), and hydrolyzing under dark condition to obtain wet meal, wherein the feed liquid ratio of broccoli bud Miao Ganfen to distilled water is 1:4g/mL, the hydrolysis temperature is room temperature, and the hydrolysis time is 8h;
(4) Adding a drying agent into the wet meal obtained in the step (3), adding dichloromethane, carrying out vortex mixing, carrying out ultrasonic leaching extraction for 30 minutes, and centrifugally collecting an extract, wherein the drying agent consists of NaCl and Na in a mass ratio of 4:3 2 SO 4 The mass ratio of the desiccant to the distilled water is 2:1; the feed liquid ratio of the broccoli sprouts Miao Ganfen to the dichloromethane is 1:10g/mL; extracting with dichloromethane for 3 times;
(5) And (3) performing rotary evaporation on the extract liquid to obtain the broccoli sprout extract.
2. The use according to claim 1, wherein (1) the freeze-dried fresh broccoli sprouts are crushed to 80 mesh.
3. Use according to claim 1, wherein (5) the temperature of rotary evaporation is 30-35 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110648434.2A CN113549660B (en) | 2021-06-10 | 2021-06-10 | Preparation method of broccoli sprout extract and anti-inflammatory activity thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110648434.2A CN113549660B (en) | 2021-06-10 | 2021-06-10 | Preparation method of broccoli sprout extract and anti-inflammatory activity thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113549660A CN113549660A (en) | 2021-10-26 |
| CN113549660B true CN113549660B (en) | 2023-12-29 |
Family
ID=78130427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110648434.2A Active CN113549660B (en) | 2021-06-10 | 2021-06-10 | Preparation method of broccoli sprout extract and anti-inflammatory activity thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113549660B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229211A (en) * | 2008-01-31 | 2008-07-30 | 广东药学院 | Broccoli seed extractive and preparing method thereof |
| CN101514174A (en) * | 2009-02-24 | 2009-08-26 | 黑龙江八一农垦大学 | Method for extracting multifunctional sulforaphane from broccoli sprouting vegetable |
| CN104086469A (en) * | 2014-05-22 | 2014-10-08 | 苏州泽达兴邦医药科技有限公司 | Method for extracting and purifying sulforaphane from broccoli seeds |
-
2021
- 2021-06-10 CN CN202110648434.2A patent/CN113549660B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229211A (en) * | 2008-01-31 | 2008-07-30 | 广东药学院 | Broccoli seed extractive and preparing method thereof |
| CN101514174A (en) * | 2009-02-24 | 2009-08-26 | 黑龙江八一农垦大学 | Method for extracting multifunctional sulforaphane from broccoli sprouting vegetable |
| CN104086469A (en) * | 2014-05-22 | 2014-10-08 | 苏州泽达兴邦医药科技有限公司 | Method for extracting and purifying sulforaphane from broccoli seeds |
Non-Patent Citations (3)
| Title |
|---|
| MgSO_4处理对西兰花芽苗菜生理活性物质和抗氧化能力的影响;初婷;彭畅;郭丽萍;;食品科学(第11期);全文 * |
| 王天珩.西兰花芽苗提取物提取工艺优化研究.《医学理论与实践》.2018,第31卷(第11期),1561-1564. * |
| 西兰花芽苗提取物提取工艺优化研究;王天珩;《医学理论与实践》;第31卷(第11期);1561-1564 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113549660A (en) | 2021-10-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ramírez-Rodríguez et al. | Ethnobotanical, nutritional and medicinal properties of Mexican drylands Cactaceae Fruits: Recent findings and research opportunities | |
| Jimoh et al. | Comparison of the nutritive value, antioxidant and antibacterial activities of Sonchus asper and Sonchus oleraceus | |
| CN104855738A (en) | Compound feed for pig and preparation method of compound feed | |
| CN107156828A (en) | A kind of full nutrition formula food suitable for patients with inflammatory bowel | |
| KR100939906B1 (en) | Oriental medicine extract for farming fish of the same | |
| Tabin et al. | Medical and medicinal importance of Rheum spp. collected from different altitudes of the Kashmir Himalayan range | |
| CN106942133A (en) | A kind of cultural method of cockfighting | |
| KR20140009717A (en) | Healthy food composition having anti-obesity and anti-diabetic activities | |
| Hussain et al. | Fig (Ficus Carica)—Morphology, Taxonomy, Composition and Health Benefits | |
| Yasmeen et al. | Phytochemical and biological investigation of Armoracia rusticana | |
| CN113549660B (en) | Preparation method of broccoli sprout extract and anti-inflammatory activity thereof | |
| KR20100021040A (en) | Composition for feed for poultry comprising oriental herb | |
| Subba et al. | A comprehensive review on common plants with remarkable medicinal properties: Urtica dioica | |
| CN107416919A (en) | Efficiently remove the composition of Polychlorinated biphenyls in water body | |
| WO2019033527A1 (en) | Plant extract for controlling diseases of cucumber and improving content of nutrient components of cucumber | |
| KR20110091959A (en) | Anhydrous lactobacillus fermentation health function food composition with cancer curing improvements and its preparation method | |
| JP4673220B2 (en) | Herbal composition for digestive disorders | |
| CN107213175B (en) | Wild jasmine extract and preparation method and application thereof | |
| Irshad et al. | Jujube | |
| Badole et al. | Hongay oil tree (Pongamia pinnata Linn.) seeds in health and disease benefits | |
| Ulrikh et al. | Substantiation and Selection of Promising Medicinal plants for prevention of diseases of Springer cows | |
| CN115531449B (en) | Composition for treating diarrhea of Tibetan pigs and piglets caused by escherichia coli and preparation method of composition | |
| CN109043527A (en) | It adjusts human-body sub-health and improves the composition and preparation process of immunity | |
| Nayak et al. | Traditional uses and nutritional properties of mahua (Madhuca longifolia): A multipurpose tropical tree grown by tribals of Odisha | |
| KR20030005127A (en) | Healthy foods on earth and alder |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |