CN111773334A - 一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品 - Google Patents
一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品 Download PDFInfo
- Publication number
- CN111773334A CN111773334A CN202010787755.6A CN202010787755A CN111773334A CN 111773334 A CN111773334 A CN 111773334A CN 202010787755 A CN202010787755 A CN 202010787755A CN 111773334 A CN111773334 A CN 111773334A
- Authority
- CN
- China
- Prior art keywords
- dss
- aom
- millet
- group
- food
- 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
- 206010009944 Colon cancer Diseases 0.000 title claims abstract description 31
- 208000001333 Colorectal Neoplasms Diseases 0.000 title claims abstract description 25
- 235000013305 food Nutrition 0.000 title claims abstract description 22
- 208000022559 Inflammatory bowel disease Diseases 0.000 title claims abstract description 18
- 235000013361 beverage Nutrition 0.000 title description 2
- 244000062793 Sorghum vulgare Species 0.000 claims abstract description 68
- 235000019713 millet Nutrition 0.000 claims abstract description 68
- 235000005911 diet Nutrition 0.000 claims description 14
- 230000037213 diet Effects 0.000 claims description 14
- 206010009887 colitis Diseases 0.000 claims description 12
- 241000282414 Homo sapiens Species 0.000 claims description 7
- 230000003203 everyday effect Effects 0.000 claims 1
- 210000001072 colon Anatomy 0.000 abstract description 27
- 210000002966 serum Anatomy 0.000 abstract description 14
- 238000011161 development Methods 0.000 abstract description 9
- 206010061218 Inflammation Diseases 0.000 abstract description 8
- 230000004054 inflammatory process Effects 0.000 abstract description 8
- 102100021943 C-C motif chemokine 2 Human genes 0.000 abstract description 7
- 101710155857 C-C motif chemokine 2 Proteins 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 3
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 79
- 229920003045 dextran sodium sulfate Polymers 0.000 description 78
- 102100039019 Nuclear receptor subfamily 0 group B member 1 Human genes 0.000 description 77
- 241000209094 Oryza Species 0.000 description 50
- 235000007164 Oryza sativa Nutrition 0.000 description 50
- 235000009566 rice Nutrition 0.000 description 50
- 241000699670 Mus sp. Species 0.000 description 38
- 230000014509 gene expression Effects 0.000 description 23
- 210000001519 tissue Anatomy 0.000 description 20
- 238000000034 method Methods 0.000 description 18
- 230000000968 intestinal effect Effects 0.000 description 16
- 230000002757 inflammatory effect Effects 0.000 description 13
- 206010028980 Neoplasm Diseases 0.000 description 12
- 230000002829 reductive effect Effects 0.000 description 12
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 11
- 108020004999 messenger RNA Proteins 0.000 description 11
- 210000003608 fece Anatomy 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 150000004666 short chain fatty acids Chemical class 0.000 description 9
- GOLXRNDWAUTYKT-UHFFFAOYSA-N 3-(1H-indol-3-yl)propanoic acid Chemical compound C1=CC=C2C(CCC(=O)O)=CNC2=C1 GOLXRNDWAUTYKT-UHFFFAOYSA-N 0.000 description 8
- 208000029742 colonic neoplasm Diseases 0.000 description 8
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 8
- 230000004083 survival effect Effects 0.000 description 8
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 210000001035 gastrointestinal tract Anatomy 0.000 description 7
- 102000005962 receptors Human genes 0.000 description 7
- 108020003175 receptors Proteins 0.000 description 7
- 102000003814 Interleukin-10 Human genes 0.000 description 6
- 108090000174 Interleukin-10 Proteins 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 5
- 108090000695 Cytokines Proteins 0.000 description 5
- 101000713169 Homo sapiens Solute carrier family 52, riboflavin transporter, member 2 Proteins 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 description 5
- 108010017324 STAT3 Transcription Factor Proteins 0.000 description 5
- 102100024040 Signal transducer and activator of transcription 3 Human genes 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000002207 metabolite Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 235000021391 short chain fatty acids Nutrition 0.000 description 5
- PLVPPLCLBIEYEA-WAYWQWQTSA-N (z)-3-(1h-indol-3-yl)prop-2-enoic acid Chemical compound C1=CC=C2C(\C=C/C(=O)O)=CNC2=C1 PLVPPLCLBIEYEA-WAYWQWQTSA-N 0.000 description 4
- 208000003200 Adenoma Diseases 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 102100040133 Free fatty acid receptor 2 Human genes 0.000 description 4
- 101100227054 Homo sapiens FFAR2 gene Proteins 0.000 description 4
- 108090001005 Interleukin-6 Proteins 0.000 description 4
- 102000004889 Interleukin-6 Human genes 0.000 description 4
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 description 4
- 238000011529 RT qPCR Methods 0.000 description 4
- 102100036862 Solute carrier family 52, riboflavin transporter, member 2 Human genes 0.000 description 4
- 230000003110 anti-inflammatory effect Effects 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 4
- 239000002158 endotoxin Substances 0.000 description 4
- 239000003290 indole 3-propionic acid Substances 0.000 description 4
- 239000003617 indole-3-acetic acid Substances 0.000 description 4
- PLVPPLCLBIEYEA-UHFFFAOYSA-N indoleacrylic acid Natural products C1=CC=C2C(C=CC(=O)O)=CNC2=C1 PLVPPLCLBIEYEA-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- ZFRKQXVRDFCRJG-UHFFFAOYSA-N skatole Chemical compound C1=CC=C2C(C)=CNC2=C1 ZFRKQXVRDFCRJG-UHFFFAOYSA-N 0.000 description 4
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 102100027581 Forkhead box protein P3 Human genes 0.000 description 3
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 3
- 101000861452 Homo sapiens Forkhead box protein P3 Proteins 0.000 description 3
- 102100037850 Interferon gamma Human genes 0.000 description 3
- 108010074328 Interferon-gamma Proteins 0.000 description 3
- 102000003777 Interleukin-1 beta Human genes 0.000 description 3
- 108090000193 Interleukin-1 beta Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 3
- 229940054051 antipsychotic indole derivative Drugs 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 231100000517 death Toxicity 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 235000018823 dietary intake Nutrition 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 3
- 229960002986 dinoprostone Drugs 0.000 description 3
- 230000009266 disease activity Effects 0.000 description 3
- 230000002550 fecal effect Effects 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 208000035861 hematochezia Diseases 0.000 description 3
- 150000002475 indoles Chemical class 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- 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 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- XGILAAMKEQUXLS-UHFFFAOYSA-N 3-(indol-3-yl)lactic acid Chemical compound C1=CC=C2C(CC(O)C(O)=O)=CNC2=C1 XGILAAMKEQUXLS-UHFFFAOYSA-N 0.000 description 2
- 206010001233 Adenoma benign Diseases 0.000 description 2
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000692822 Bacteroidales Species 0.000 description 2
- 206010009900 Colitis ulcerative Diseases 0.000 description 2
- 208000011231 Crohn disease Diseases 0.000 description 2
- 108050006400 Cyclin Proteins 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 2
- 102000013691 Interleukin-17 Human genes 0.000 description 2
- 108050003558 Interleukin-17 Proteins 0.000 description 2
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 2
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 2
- 102000003940 Occludin Human genes 0.000 description 2
- 108090000304 Occludin Proteins 0.000 description 2
- 102000009339 Proliferating Cell Nuclear Antigen Human genes 0.000 description 2
- 238000010162 Tukey test Methods 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004534 cecum Anatomy 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 230000008951 colonic inflammation Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 235000013325 dietary fiber Nutrition 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000007358 intestinal barrier function Effects 0.000 description 2
- 230000007413 intestinal health Effects 0.000 description 2
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 235000019260 propionic acid Nutrition 0.000 description 2
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 208000016261 weight loss Diseases 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- DGAKHGXRMXWHBX-ONEGZZNKSA-N Azoxymethane Chemical compound C\N=[N+](/C)[O-] DGAKHGXRMXWHBX-ONEGZZNKSA-N 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 108010075254 C-Peptide Proteins 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 208000018458 Colitis-Associated Neoplasms Diseases 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 108090000045 G-Protein-Coupled Receptors Proteins 0.000 description 1
- 102000003688 G-Protein-Coupled Receptors Human genes 0.000 description 1
- 101000725401 Homo sapiens Cytochrome c oxidase subunit 2 Proteins 0.000 description 1
- 101000605127 Homo sapiens Prostaglandin G/H synthase 2 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 241000239218 Limulus Species 0.000 description 1
- 206010028124 Mucosal ulceration Diseases 0.000 description 1
- 101000972289 Mus musculus Mucin-2 Proteins 0.000 description 1
- 206010061309 Neoplasm progression Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- -1 P-STAT3 Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 206010038063 Rectal haemorrhage Diseases 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 239000006180 TBST buffer Substances 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 101150116184 abi gene Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- ZRYCZAWRXHAAPZ-UHFFFAOYSA-N alpha,alpha-dimethyl valeric acid Chemical compound CCCC(C)(C)C(O)=O ZRYCZAWRXHAAPZ-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000007234 antiinflammatory process Effects 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 235000021196 dietary intervention Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 230000036732 histological change Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000005027 intestinal barrier Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000013077 scoring method Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000005751 tumor progression Effects 0.000 description 1
- 238000007492 two-way ANOVA Methods 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 239000013585 weight reducing agent Substances 0.000 description 1
- 235000020985 whole grains Nutrition 0.000 description 1
Images
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)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
-
- 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
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明创造提供了一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,所述饮食品中包含小米。本发明创造所述的含有小米的饮食品对炎症性肠病及相关结直肠癌的发展有改善作用;所述饮食品可降低MPO、MCP‑1、C‑P等炎症标志物在结肠或血清中的含量。
Description
技术领域
本发明创造属于医药领域,尤其是涉及一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品。
背景技术
最新数据显示,癌症是全球人口死亡的第一大疾病,其中结直肠癌的死亡率位列癌症的第二位,严重威胁人类健康。近年来,一些发展中国家的结直肠癌发病率和死亡率仍在上升。仅2015年中国新增结直肠癌病例37.63万例,相关死亡19.1万人。炎症性肠病(IBD)是一种由异常免疫介导,有复发倾向的胃肠道炎症(GIT),包括溃疡性结肠炎(UC)和克罗恩病(CD)。流行病学研究表明,IBD患者的结直肠癌发病率增加,并且患癌率与IBD患者的病龄成正相关,与IBD相关的促炎细胞因子释放和信号传导增强在与结肠炎相关的结直肠癌(CAC)发展中起关键作用。流行病学研究表明,全谷物或粗粮的摄入与结肠癌的发病率呈负相关。但干预性实验中,特定谷物的摄入是否能够降低结肠癌发病率仍缺乏研究,且机制不清。
小米,禾本科植物,它耐干旱耐盐碱,生命力顽强,主要分布于干旱地区和半干旱地区,是中国的传统农作物之一。小米营养价值丰富,常见的烹饪方式是将其熬制成小米粥或是掺进大米中蒸熟。小米中营养成分的结构与大米,碳水化合物含量为60.9%,低于大米(76%),主要成分是淀粉,其中直链淀粉占比约为25%;小米中的纤维含量约为8%,约比大米高出7%。小米中富含色氨酸,色氨酸是主要依靠外源性的饮食摄入来补充的人类必需氨基酸,在肠道菌群作用下产生吲哚衍生物,通过AHR受体介导抗炎反应。除此之外,小米中维生素、矿物质和多酚等活性物质也可对肠道菌群起到调节作用。大米和小米均为在中国乃至亚洲地区被广泛接纳的主食来源,两者营养成分之间存在诸多差异,人体摄入后肠道代谢产物也发生相应变化,继而也会影响到肠道菌群结构,长期累积是否会对结肠炎或结直肠癌的发展造成不同的影响引起了学界的关注。
发明内容
有鉴于此,本发明创造旨在克服现有技术中的缺陷,提出一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品。
为达到上述目的,本发明创造的技术方案是这样实现的:
一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,所述饮食品中包含小米。
优选地,所述的结直肠癌为结肠炎诱导的结直肠癌。
优选地,所述饮食品提供每日人体正常摄入的全部热量,其中,所述饮食品中小米的能量占比为38%-39%。
优选地,所述饮食品提供每日人体摄入的全部食物,其中,所述饮食品中小米的总量为180-330g。
相对于现有技术,本发明创造具有以下优势:
本发明创造所述的饮食品中的精小米对结肠炎及相关结直肠癌的发展有改善作用;所述饮食品可降低MPO、MCP-1、C-P等炎症标志物在结肠或血清中的含量;所述饮食品可以降低多种炎症通路蛋白的表达水平;所述饮食品可以抑制肿瘤进展关进通路STAT3的磷酸化,降低与肿瘤细胞生长、增值和存活相关的多种蛋白的表达;所述饮食品可增加肠道SCFA和色氨酸代谢物含量,增加肠道受体GPCR41、GPCR43和AHR的基因表达;所述饮食品可显著增加菌群α-多样性、回复β-多样性、显著增加Bacteroidales S24-7这一产短链脂肪酸微生物的相对丰度,降低条件致病菌Alistipes的相对丰度。
附图说明
图1为AOM/DSS诱导小鼠结肠炎相关性癌模型;
图2为对NM、AOM/DSS、AOM/DSS+millet和AOM/DSS+rice组中小鼠的表观观察。(A)生存率;(B)体重;(C)病情活动指数;(D)肿瘤数目(每只小鼠);(E)肿瘤负荷(每只小鼠总肿瘤体积,mm3)。(A)生存率采用Log-rank(Mantel-Cox)test的分析方法。(B,C)采用Two-wayANOVA和Tukey post hoc test分析整个研究过程中某段时间的意义(在实验的最后,每组样本数n=4-10)。括号表示在一段时间上各组之间的比较。在图2C中,*p<0.05用于AOM/DSSvs AOM/DSS+millet;#p<0.05用于AOM/DSS+millet vs AOM/DSS+rice;&p<0.05用于AOM/DSS vs AOM/DSS+rice。(D,E)One-way ANOVA后进行Tukey post-hoc test。数据表示为为平均值±方差。不同字母的平均值差异显著(p<0.05)。n=4-10/组;
图3为结肠炎症的宏观和组织学评价。结肠组织(A)MPO和(B)MCP-1;血清(C)C-P,(D)IFN-γ,(E)IL-1β,和(F)LPS.(G)具有代表性的福尔马林固定结肠组织H&E染色切片。黑色的椭圆表示炎性浸润;蓝色箭头表示隐窝缺损;红色曲线表示隐窝变形。放大倍数:×100倍。数据表示为为平均值±方差。带有不同字母的平均值间具有显著性差异(p<0.05)。NM,AOM/DSS,AOM/DSS+millet和AOM/DSS+rice组中的样本数分别为10,6,7,4;
图4为结肠组织中(A)TNF-α,(B)IL-6,(C)IL-17,(D)COX-2,(E)iNOS,(F)FOXP3,(G)IL-10,(H)IL-22,(I)ZO-1和(J)occludin在mRNA水平上的表达情况。血清中(K)TNF-α和(L)IL-10中的表达情况。结肠组织中(M)COX-2和(N)PGE2的表达情况。数据表示为为平均值±方差。带有不同字母的平均值间具有显著性差异(p<0.05)。NM,AOM/DSS,AOM/DSS+millet和AOM/DSS+rice组中的样本数分别为10,6,7,4;
图5为(A)结肠组织中Bcl-2、PCNA、P-STAT3和STAT3的蛋白免疫印迹分析。(B)结肠组织中VEGF在mRNA水平上的表达情况。数据表示为为平均值±方差。带有不同字母的平均值间具有显著性差异(p<0.05)。NM,AOM/DSS,AOM/DSS+millet和AOM/DSS+rice组中的样本数分别为10,6,7,4;
图6为粪便中(A)短链脂肪酸和(B)色氨酸代谢物的浓度。结肠组织中(C)GPCR43,(D)GPCR41,和(E)AHR在mRNA水平上的表达情况。数据表示为为平均值±方差。带有不同字母的平均值间具有显著性差异(p<0.05)。NM,AOM/DSS,AOM/DSS+millet和AOM/DSS+rice组中的样本数分别为10,6,7,4;
图7为小米和大米对肠道微生物多样性和组成的影响。(A)索布斯指数和稀释曲线。(B)香农指数和稀释曲线。(C)OTU水平肠道微生物宏基因组的主成分分析。(D)AOM/DSS+millet组与AOM/DSS+rice组间微生物群落的差异。*p<0.05,**p<0.001。分析方法采用Kruskal-Wallis分析和Dunn’s test事后检验。NM,AOM/DSS,AOM/DSS+millet和AOM/DSS+rice组中的样本数分别为10,6,7,4;
具体实施方式
除有定义外,以下实施例中所用的技术术语具有与本发明创造所属领域技术人员普遍理解的相同含义。以下实施例中所用的试验试剂,如无特殊说明,均为常规生化试剂;所述实验方法,如无特殊说明,均为常规方法。
下面结合实施例来详细说明本发明创造。
1.材料
1.1主要试剂
本实验例中的AOM购于美国Sigma公司;硫酸葡聚糖钠DSS购于美国MPBiomedicals公司,MW:36000-50000。TRIzolTM Reagent(Thermo Fisher Sientific);LunaScriptTM SuperMix Kit(New England BioLabs);SYBR qPCR Master Mix(ChamQTMUniversal)
1.2动物
40只4周龄SPF级雄性BALB/c小鼠,购自斯贝福(北京)实验动物科技有限公司。小鼠饲养于中国医学科学院放射医学研究所动物中心,并维持在标准实验室条件下饲养:25±2℃,相对湿度50±5%,12小时循环光照,自由取水取食,相关实验操作符合动物实验伦理学要求。普通饲料喂养1周适应环境,实验从第二周正式开始。
1.3结肠炎及相关结肠癌模型的建立
本实验例采用AOM/DSS法构建结肠炎及相关结肠癌小鼠模型。参照图1,一周适应期后,将40只小鼠随机分成四组(NM组,CON组,RM组和RR组),每组10只。CON组、RM组和RR组的小鼠腹腔注射AOM(10mg/kg),NM组不注射AOM。注射AOM后1周,给予CON组、RM组和RR组的小鼠含2%DSS的饮水1周,NM组饮用水中不含DSS,之后恢复正常饮水2周,重复上述步骤3次。另外,在注射AOM后第1天NM、CON组的小鼠饲料为AIN-93G,RM、RR组的小鼠饲料分别为AIN-93G-精小米、AIN-93G-精大米。AIN-93G-精小米、AIN-93G-精大米均以AIN-93G配方为基础,纤维素添加量为3%,玉米淀粉分别替换为100%精小米粉、100%精大米粉。
表1.实验中三组饲料的组成成分
实验过程中,每周记录一次体重和摄食量,开始饮用DSS水溶液后,每周观察并记录小鼠的活动情况、大便形态、肛周潜血、脱肛现象和存活状况。所有小鼠在第十周末进行眼眶取血,随后脱颈处死(处死前一天采集粪便样品,禁食一晚),小鼠解剖后取肝脏、脾脏、盲肠及盲肠末端至肛门处整段结直肠并称重。测量结肠长度后,纵向剖开整段肠组织并将内容物清理干净,记录结肠上可见的肿瘤数目(肿瘤直径以3mm为分界线,分别计数)。取结肠组织末端5mm,浸泡于10%福尔马林中固定24h,其余样品用液氮速冻后保存于-80℃。
2实验方法
2.1病理情况打分方法
在结直肠炎动物模型中,病情活动指数(DAI)常用于评估疾病状态。小鼠的结肠炎症状主要表现为粪便黏稠度和是否有便血。DAI评分采用3分制计算:(1)0分:大便稠度正常,无便血;(2)1分:大便柔软,粪便带有血迹;(3)2分:腹泻,直肠出血;(4)3分:病情严重至死亡。
2.2细胞因子测定方法
血清中IL-1β,IL-4,IL-6,IL-10,IFN-γ,TNF-α和血清C肽含量的测定按ELISA试剂盒的方法进行。结肠组织中的MCP-1,MPO,COX2和PGE2含量的测定根据ELISA试剂盒的方法进行。血清中内毒素的含量测定根据鲎试剂盒方法进行。
2.3组织病理学评估方法
对福尔马林固定的结肠组织进行石蜡包埋、切片、苏木精和伊红染色(H&E)或1%阿尔新蓝染色,在显微镜下观察炎症浸润、隐窝损伤、粘膜溃疡、有无水肿等组织学改变。
2.4粪便中短链脂肪酸测定方法
取50mg小鼠粪便,加入0.4mL 50%甲醇溶液(含0.1%甲酸),剪碎粪便并充分混悬。混悬液13200g于4℃离心20min,取上清,重复该操作一次。通过DB-FFAP capillarycolumn(30m×0.25mm i.d.,0.25μm,Agilent)色谱柱分离。气相色谱条件:80℃,2分钟;80-180℃,10分钟;180℃,5分钟。以二甲基戊酸为外标,绘制乙酸、丙酸、正丁酸、异丁酸、正戊酸标准曲线。数据处理采用Agilent’s MSD ChemStation(E.02.00.493)软件。
2.5粪便中色氨酸代谢物测定方法
采用LC-MS法测定粪便氨基酸代谢物含量。
2.6蛋白质印迹分析方法
取100mg结肠组织样本,加入1mLRAPI裂解液(含1%的蛋白酶抑制剂PMSF),冰上静止5min,然后超声均质。取得蛋白匀浆12000rpm,4℃,离心10min,收集上清液。用BCA蛋白浓度测定试剂盒测定上清液总蛋白浓度。用5×Loading Buffer和PBS稀释蛋白样品,在沸水浴中煮沸5min,制成上样液备用。计算后移取蛋白上样量40μg,进行SDS-PAGE电泳,转膜后5%脱脂牛奶室温封闭1h,一抗4℃过夜孵育,二抗37℃孵育45min。TBST洗膜后,利用化学发光凝胶成像系统进行曝光显色。将胶片进行扫描,用凝胶图象处理系统(Gel-Pro-Analyzer软件)分析目标条带的光密度值。
2.7RT-qPCR定量分析方法
使用TRIzolTM Reagent(Thermo Fisher Sientific)提取结肠组织中总RNA,具体操作按照说明书进行。提取后用超微量分光光度计(Implen,Germany)于260/280吸光度测定浓度,适当稀释后取1μg RNA逆转录为cDNA,逆转录酶为LunaScriptTM SuperMix Kit(NewEngland BioLabs)。采用SYBR qPCR Master Mix(ChamQTM Universal)进行扩增,反应在Bio-Rad CFX ConnectTM Real-Time System中进行。引物序列如下表所示,β-actin为内参。
表2引物序列[sequence 5’-3’].
2.8DNA提取和高通量测序方法
粪便微生物DNA提取按试剂盒方法进行,溶解DNA前残留乙醇需彻底挥干。用1%琼脂糖凝胶电泳检测DNA提取质量。细菌16S rRNA V3-V4可变区通过338F(5’-ACTCCTACGGGAGGCAGCAG-3’)和806R(5’-GGACTACHVGGGTWTCTAAT-3’)引物扩增,扩增程序为:95℃预变性3min,27个循环(95℃变性30s,55℃退火30s,72℃延伸30s),最后72℃延伸10min(PCR仪:ABI Gene9700型)。扩增体系为20μL,4μL5*FastPfu缓冲液,2μL 2.5mMdNTPs,0.8μL引物(5μM),0.4μL FastPfu聚合酶;10ng DNA模板。
使用2%琼脂糖凝胶回收PCR产物,利用AxyPrep DNA Gel Extraction Kit(Axygen Biosciences,Union City,CA,USA)进行纯化,Tris-HCl洗脱,2%琼脂糖电泳检测。利用QuantiFluorTM-ST(Promega,USA)进行检测定量。根据Illumina MiSeq平台(Illumina,San Diego,USA)标准操作规程将纯化后的扩增片段构建PE2*300的文库。构建文库步骤:(1)连接“Y”字形接头;(2)使用磁珠筛选去除接头自连片段;(3)利用PCR扩增进行文库模板的富集;(4)氢氧化钠变性,产生单链DNA片段。利用Illumina公司的MiseqPE300平台进行测序(上海美吉生物医药科技有限公司)。原始数据上传至NCBI数据库中(序列号:SRP239677)。
3.结果分析
3.1改善AOM/DSS诱导结直肠炎相关癌症小鼠的生存状态,延缓病情的发展
采用AOM/DSS法构建炎症相关的结直肠癌小鼠模型,共干预9周,精小米组与精大米组相比可显著增加结直肠癌小鼠的存活率(参考图2A)、抑制体重降低(图2b)、降低病变指数(参考图2c),降低腺瘤发生率(图2D)和肿瘤负担(图2E)。在造模过程中,四组小鼠体重变化差异较为明显。NM组小鼠体重稳定增加;DSS组、RM组及RR组小鼠前四周体重呈增加趋势,第二次DSS处理后小鼠出现体重减轻、腹泻和便血等病症,同时病情活动指数升高。DSS停药后的两周恢复期,体重有回升迹象,病情也有改善。9周干预结束时,AOM/DSS+millet组的存活率低于NM组,但高于AOM/DSS+rice组(图2A)。AOM/DSS+rice组小鼠的死亡率最高,DSS组次之。另外,AOM/DSS+millet组小鼠的体重显著高于AOM/DSS+rice组,AOM/DSS组小鼠体重与AOM/DSS+rice组无显著差异(图2B)。造模期间AOM/DSS+millet组小鼠的病情相较于AOM/DSS组和AOM/DSS+rice组也有明显减轻(图2C)。AOM/DSS+millet组中平均每只小鼠结肠组织上的腺瘤数(直径<3mm)相比AOM/DSS组明显减少(图2D);与AOM/DSS+millet组相比,直径<3mm和>3mm的腺瘤数均显著降低。最后,AOM/DSS+millet组小鼠的肿瘤负荷较AOM/DSS组和AOM/DSS+rice组也显著降低(图2E)。以上数据表明小米可以改善结肠炎,抑制结肠炎相关腺瘤的形成。
3.2改善炎症组织病理学
炎症标志分子测定结果和H&E染色组织学结果如图3所示。由图3A-E可以看出,与大米组相比,小米可降低MPO、MCP-1、C-P等炎症标志物在结肠或血清中的含量。AOM/DSS处理后结肠组织中MPO和MCP-1的水平显著提高,分别表示中性粒细胞和巨噬细胞募集。与AOM/DSS组和AOM/DSS+rice组相比,小米干预后小鼠结肠中MPO和MCP-1水平降低。另外AOM/DSS+millet组小鼠血清中的C-P和IFN-γ的水平与AOM/DSS+rice组相比也显著降低,但血清中IL-1β的含量在两组间无显著差异。同时,AOM/DSS+rice组小鼠血清中的C-P和IFN-γ水平也显著高于AOM/DSS组。此外,血清中LPS(图3F)的浓度在AOM/DSS组中明显高于NM组。与AOM/DSS组和AOMM/DSS+rice组相比,小米组中LPS浓度显著降低。
由图3G(a-d)得知,小米可以改善AOM/DSS处理对小鼠结肠组织造成的验证损伤。NM组小鼠的结肠切片组织结构正常,无炎症反应(图3G(a))。在AOM/DSS组的小鼠结肠组织中,观察到炎性浸润,隐窝缺损和扭曲变形的现象(图3G(b))。另外,与AOM/DSS组和AOM/DSS+rice组相比,小米组小鼠组织病理损伤得到缓解,炎性浸润、隐窝缺损和扭曲现象均有明显改善,病理学评分与AOM/DSS组和AOM/DSS+rice组相比也显著降低(图3G(c-e))。
3.3降低多种炎症通路蛋白的表达水平
结肠炎及相关结肠癌的发展伴随着细胞因子表达水平的变化与AOM/DSS组和AOM/DSS+rice组相比,小米膳食干预可以显著抑制结肠炎症因子TNF-α,IL-6,IL-17,COX-2和iNOS在mRNA水平上的表达(图4A-E)。另外,小米组中的抗炎因子FOXP3,IL-10和IL-22在mRNA水平上的表达显著高于AOM/DSS和AOM/DSS+rice组(图4F-H)。同时,小米组中与肠屏障健康相关的指标ZO-1和Occludin的mRNA表达量也显著上调(图4I-J)。
以上RT-qPCR的实验结果通过ELISA进一步得到证实(图K-N)。AOM/DSS+rice组小鼠血清中炎症因子TNF-α的含量显著高于AOM/DSS+millet组。同时,AOM/DSS+millet组血清中抗炎因子IL-10的表达显著高于AOM/DSS组。另一方面,膳食补充小米的小鼠中结肠组织中炎症因子COX-2和PGE2显著低于AOM/DSS和AOM/DSS+rice组。
总之,与大米相比,小米能够调节与结肠炎症相关的关键细胞因子的表达,并改善肠屏障功能。
3.4抑制肿瘤的发生
饮食中补充小米可以通过抑制STAT3磷酸化和相关下游炎症蛋白的表达抑制结直肠癌的发展。由蛋白印迹分析结果可知(图5),与AOM/DSS组和AOM/DSS+rice组相比,小米干预可以显著抑制STAT3的磷酸化,同时抑制与肿瘤细胞存活(Bcl-2)、增殖(PCNA)相关炎症蛋白的表达。另外,与AOM/DSS和AOM/DSS+rice组相比,AOM/DSS+millet组小鼠结肠中与肿瘤生长相关的指标VEGF在mRNA水平上的表达也显著下调。
3.5增加肠道受体和菌群代谢物
与大米相比,小米中含有更丰富的色氨酸和膳食纤维,这些营养成分经肠道菌群作用产生的代谢产物(吲哚衍生物和短链脂肪酸)能激活肠道受体,从而促进肠道健康。为了探讨小米影响结肠健康的机制,我们测定了谷子粪便中色氨酸和膳食纤维代谢物的含量(图6A-B),并测定了特异性肠道受体的mRNA表达水平(图6C-E)。
各组粪便中短链脂肪酸的含量如图6A所示。与NM组相比,AOM/DSS组粪便中的乙酸、丙酸和丁酸均显著减少,这可能由于产短链脂肪酸的细菌数量减少导致的。与大米组相比,膳食摄入小米使粪便中乙酸、丙酸和丁酸的含量显著增加。小米组与AOM/DSS组相比,乙酸和丁酸的含量无显著差异。
另外,各组粪便中色氨酸代谢物的含量如图6B所示。在已测定的色氨酸代谢产物中,吲哚-3-丙酸(IPA)、3-甲基吲哚(3ML)、吲哚乳酸(ILA)、吲哚乙酸(IAA)和吲哚丙烯酸(IA)是AHR的特异性配体。与NM组相比,AOM/DSS组的IPA、IAA和IA含量显著降低。与AOM/DSS组和AOM/DSS+rice组相比,小米显著提高了IPA、IAA和IA的含量。此外,AOM/DSS+millet组中具有AHR激活性质的总色氨酸代谢物含量增加。
最后,我们检测了AHR、GPCR41和GPCR43在mRNA水平上的表达情况(图6C-E)。与色氨酸代谢产物和短链脂肪酸的减少一致,AOM/DSS处理使AHR、GPCR43和GPCR41的mRNA表达水平相对于NM组显著下调。AOM/DSS+millet组中AHR的表达明显高于AOM/DSS组和AOM/DSS+rice组。同时,AOM/DSS+millet组中小鼠的两种短链脂肪酸受体的mRNA表达显著高于大米处理小鼠。
以上结果表明,膳食摄入小米可以增加色氨酸代谢产物和短链脂肪酸的产生,并激活相关的肠道受体AHR,GPCR41和GPCR43。一方面,这些肠道受体的激活通过下调包括TNF-α和IL-6在内的抗炎细胞因子,抑制STAT3的磷酸化及相关炎症蛋白表达,从而抑制结肠炎及相关结肠癌发展。另一方面,也可以上调FOXP3、IL-10和IL-22等抗炎因子表达,从而共同促进结肠粘蛋白的分泌,起到维护肠道健康的作用。
3.6增加肠道菌群的多样性
宏基因组测序测定粪便菌群16S rRNA V3-V4可变区序列,研究各组小鼠肠道菌群的变化情况。索布斯(图7A)和香农(图7B)稀释曲线可反映菌群α-多样性,AOM/DSS处理与NM组相比,菌群α-多样性显著降低,小米组与AOM/DSS组相比,索布斯指数显著增加,大米组与AOM/DSS组相比,索布斯指数无显著差异,提示小米膳食可增加菌群多样性。
基于Unweighted-unifrac算法的PCoA分析中(图7C),AOM/DSS组菌群与NM组相比分布存在显著差异,小米膳食可显著回调AOM/DSS处理导致的肠道菌群紊乱,大米组与AOM/DSS组相比无显著差异。
对大米组和小米组的菌群丰度进行差异分析(图7D),小米膳食可显著增加肠道产短链脂肪酸微生物Bacteroidales S24-7科的相对丰度,显著增加益生菌Bifidobacterium的相对丰度,显著增加可导致肠炎致病菌Alistipes的相对丰度。
肠道菌群紊乱伴随的有益菌丰度降低和条件致病菌丰度增加是炎性肠病和结肠癌的重要诱因。以上结果提示,相对于大米膳食和对照组,小米膳食可显著调节肠道菌群紊乱,增加有益微生物相对丰度,降低有害微生物相对丰度,从而发挥改善炎性肠病和结肠癌的作用。
以上所述仅为本发明创造的较佳实施例而已,并不用以限制本发明创造,凡在本发明创造的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明创造的保护范围之内。
Claims (4)
1.一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,其特征在于:所述饮食品中包含小米。
2.根据权利要求1所述的用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,其特征在于:所述的结直肠癌为结肠炎诱导的结直肠癌。
3.根据权利要求1所述的用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,其特征在于:所述饮食品提供每日人体正常摄入的全部热量,其中,所述饮食品中小米的能量占比为38%-39%。
4.根据权利要求1所述的用于预防和/或治疗炎症性肠病或结直肠癌的饮食品,其特征在于:所述饮食品提供每日人体摄入的全部食物,其中,所述饮食品中小米的总量为180-330g。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010787755.6A CN111773334A (zh) | 2020-08-07 | 2020-08-07 | 一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010787755.6A CN111773334A (zh) | 2020-08-07 | 2020-08-07 | 一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN111773334A true CN111773334A (zh) | 2020-10-16 |
Family
ID=72766004
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010787755.6A Pending CN111773334A (zh) | 2020-08-07 | 2020-08-07 | 一种用于预防和/或治疗炎症性肠病或结直肠癌的饮食品 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111773334A (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004089140A (ja) * | 2002-09-03 | 2004-03-25 | Tabeyonomori Toyama:Kk | すし飯 |
CN102617718A (zh) * | 2012-03-27 | 2012-08-01 | 山西大学 | 一种谷糠抗肿瘤活性蛋白及其制备方法和应用 |
CN108813299A (zh) * | 2018-05-16 | 2018-11-16 | 浙江省中医药研究院 | 一种恢复溃疡性结肠炎患者肠道的速食粥及其制备方法 |
-
2020
- 2020-08-07 CN CN202010787755.6A patent/CN111773334A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004089140A (ja) * | 2002-09-03 | 2004-03-25 | Tabeyonomori Toyama:Kk | すし飯 |
CN102617718A (zh) * | 2012-03-27 | 2012-08-01 | 山西大学 | 一种谷糠抗肿瘤活性蛋白及其制备方法和应用 |
CN108813299A (zh) * | 2018-05-16 | 2018-11-16 | 浙江省中医药研究院 | 一种恢复溃疡性结肠炎患者肠道的速食粥及其制备方法 |
Non-Patent Citations (7)
Title |
---|
NALAN GULSXEN UNAL ET AL.: "Anti-Inflammatory Effect of Crude Momordica charantia L. Extract on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis Model in Rat and the Bioaccessibility of its Carotenoid Content", 《JOURNAL OF MEDICINAL FOOD》 * |
RUIPENG YANG等: "Inhibitory Effects of Bound Polyphenol From Foxtail Millet Bran on Colitis-Associated Carcinogenesis by the Restoration of Gut Microbiota in a Mice Model", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
刘宝琴: "《百姓健康专家谈》", 31 May 2019, 陕西科学技术出版社 * |
单树花等: "小米米糠中抗癌细胞增殖活性蛋白的分离纯化 ", 《食品科学》 * |
张时报: "食物抗癌力分等级", 《烹调知识》 * |
徐峰: "《大医生系列 防癌抗癌饮食真经》", 30 November 2015, 广东科技出版社 * |
李辉: "自制粗粮馒头营养又美味 ", 《家庭医药.快乐养生》 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Xue et al. | Probiotics may delay the progression of nonalcoholic fatty liver disease by restoring the gut microbiota structure and improving intestinal endotoxemia | |
Kumar et al. | A novel role of SLC26A3 in the maintenance of intestinal epithelial barrier integrity | |
Gaudier et al. | Butyrate specifically modulates MUC gene expression in intestinal epithelial goblet cells deprived of glucose | |
Ge et al. | Effect of industrial trans-fatty acids-enriched diet on gut microbiota of C57BL/6 mice | |
van Rhijn et al. | Proton pump inhibitors partially restore mucosal integrity in patients with proton pump inhibitor–responsive esophageal eosinophilia but not eosinophilic esophagitis | |
Støy et al. | Bovine colostrum improves intestinal function following formula-induced gut inflammation in preterm pigs | |
Pang et al. | Vitamin A supplementation ameliorates ulcerative colitis in gut microbiota–dependent manner | |
Kotlo et al. | The olfactory G protein-coupled receptor (Olfr-78/OR51E2) modulates the intestinal response to colitis | |
Suriano et al. | Particle size determines the anti-inflammatory effect of wheat bran in a model of fructose over-consumption: Implication of the gut microbiota | |
Jena et al. | Influence of gut microbiota on inflammation and pathogenesis of sugar rich diet induced diabetes | |
Pouille et al. | Chicory root flour–A functional food with potential multiple health benefits evaluated in a mice model | |
Lin et al. | Dietary Lactobacillus reuteri prevent from inflammation mediated apoptosis of liver via improving intestinal microbiota and bile acid metabolism | |
Wan et al. | Dicaffeoylquinic acids from Ilex kudingcha attenuate dextran sulfate sodium-induced colitis in C57BL/6 mice in association with the modulation of gut microbiota | |
EP3134543B1 (en) | Method for diagnosing hepatic fibrosis | |
Liu et al. | Early life Lactobacillus rhamnosus GG colonisation inhibits intestinal tumour formation | |
Xue et al. | The effects of live and pasteurized Akkermansia muciniphila on DSS-induced ulcerative colitis, gut microbiota, and metabolomics in mice | |
Bialkowski et al. | Effects of microencapsulated blend of organic acids and botanicals on growth performance, intestinal barrier function, inflammatory cytokines, and endocannabinoid system gene expression in broiler chickens | |
Liao et al. | Hylocereus undatus flower extract suppresses OVA-induced allergic asthma in BALb/c mice by reducing airway inflammation and modulating gut microbiota | |
Li et al. | Probiotic effects of Lacticaseibacillus rhamnosus 1155 and Limosilactobacillus fermentum 2644 on hyperuricemic rats | |
Park et al. | Heat-inactivated Lactobacillus plantarum nF1 promotes intestinal health in Loperamide-induced constipation rats | |
Hu et al. | Chicory fibre improves reproductive performance of pregnant rats involving in altering intestinal microbiota composition | |
Peng et al. | Early life administration of Bifidobacterium bifidum BD-1 alleviates long-term colitis by remodeling the gut microbiota and promoting intestinal barrier development | |
Pan et al. | Rapid gut adaptation to preterm birth involves feeding-related DNA methylation reprogramming of intestinal genes in pigs | |
Xu et al. | Retinoid acid induced 16 deficiency aggravates colitis and colitis-associated tumorigenesis in mice | |
Zhou et al. | Miao sour soup influences serum lipid via regulation of high‐fat diet‐induced intestinal flora in obese rats |
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 | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201016 |