CN110840901A - 溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 - Google Patents
溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 Download PDFInfo
- Publication number
- CN110840901A CN110840901A CN201911273194.1A CN201911273194A CN110840901A CN 110840901 A CN110840901 A CN 110840901A CN 201911273194 A CN201911273194 A CN 201911273194A CN 110840901 A CN110840901 A CN 110840901A
- Authority
- CN
- China
- Prior art keywords
- lysophosphatidylethanolamine
- inflammatory bowel
- bowel diseases
- relieving
- medicines
- 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.)
- Withdrawn
Links
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 title claims abstract description 35
- CWRILEGKIAOYKP-SSDOTTSWSA-M [(2r)-3-acetyloxy-2-hydroxypropyl] 2-aminoethyl phosphate Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCCN CWRILEGKIAOYKP-SSDOTTSWSA-M 0.000 title claims abstract description 35
- 208000022559 Inflammatory bowel disease Diseases 0.000 title claims abstract description 26
- 239000003814 drug Substances 0.000 title claims abstract description 10
- 229940079593 drug Drugs 0.000 title claims abstract description 6
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 206010061218 Inflammation Diseases 0.000 claims description 8
- 230000004054 inflammatory process Effects 0.000 claims description 8
- 230000000968 intestinal effect Effects 0.000 claims description 8
- 208000004232 Enteritis Diseases 0.000 abstract description 6
- 210000005026 intestinal epithelial barrier Anatomy 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 26
- 241000699670 Mus sp. Species 0.000 description 19
- 241000699666 Mus <mouse, genus> Species 0.000 description 16
- 241000588724 Escherichia coli Species 0.000 description 10
- 244000052616 bacterial pathogen Species 0.000 description 10
- 210000001072 colon Anatomy 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 8
- 210000003608 fece Anatomy 0.000 description 7
- 208000015181 infectious disease Diseases 0.000 description 6
- 230000001717 pathogenic effect Effects 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- 101150044426 blc gene Proteins 0.000 description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 description 5
- 230000007358 intestinal barrier function Effects 0.000 description 5
- 230000004807 localization Effects 0.000 description 5
- 230000009469 supplementation Effects 0.000 description 5
- 102000002029 Claudin Human genes 0.000 description 4
- 108050009302 Claudin Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 210000005027 intestinal barrier Anatomy 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000004888 barrier function Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 3
- 230000000153 supplemental effect Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 238000011814 C57BL/6N mouse Methods 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102000004162 Claudin-1 Human genes 0.000 description 2
- 108090000600 Claudin-1 Proteins 0.000 description 2
- 206010009900 Colitis ulcerative Diseases 0.000 description 2
- 102000010970 Connexin Human genes 0.000 description 2
- 108050001175 Connexin Proteins 0.000 description 2
- 208000011231 Crohn disease Diseases 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- 229930193140 Neomycin Natural products 0.000 description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 2
- 102000003940 Occludin Human genes 0.000 description 2
- 108090000304 Occludin Proteins 0.000 description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 description 2
- 108010059993 Vancomycin Proteins 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 210000000436 anus Anatomy 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 230000003870 intestinal permeability Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 229960000282 metronidazole Drugs 0.000 description 2
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 2
- 229960004927 neomycin Drugs 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000007634 remodeling Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 210000000813 small intestine Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229960003165 vancomycin Drugs 0.000 description 2
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 2
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 239000008096 xylene Substances 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010000077 Abdominal mass Diseases 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 102000000905 Cadherin Human genes 0.000 description 1
- 108050007957 Cadherin Proteins 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 206010057071 Rectal tenesmus Diseases 0.000 description 1
- 102000011154 Tight junction protein ZO-1 Human genes 0.000 description 1
- 108050001370 Tight junction protein ZO-1 Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 210000004082 barrier epithelial cell Anatomy 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000004921 distal colon Anatomy 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000004890 epithelial barrier function Effects 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 208000037817 intestinal injury Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 208000012271 tenesmus Diseases 0.000 description 1
- PBYZMCDFOULPGH-UHFFFAOYSA-N tungstate Chemical compound [O-][W]([O-])(=O)=O PBYZMCDFOULPGH-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
- A61K31/685—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols one of the hydroxy compounds having nitrogen atoms, e.g. phosphatidylserine, lecithin
-
- 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]
Landscapes
- Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于医药领域，涉及溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用。本发明公开了溶血磷脂酰乙醇胺18:1通过增加肠道上皮屏障的稳定性，以致于缓解或治疗肠炎和炎症性肠病。是一种有效的治疗炎症性肠病的药物。
Description
技术领域
本发明属于医药领域，涉及溶血磷脂酰乙醇胺18:1缓解肠道炎症的发生发展从而预防或治疗肠炎和炎症性肠病；具体涉及溶血磷脂酰乙醇胺18:1通过增加肠道上皮屏障的稳定性，以致于缓解或治疗肠炎和炎症性肠病。
背景技术
随着生产物资的日益丰富，人们的饮食也日渐精致，便随而来的是炎症性肠病(IBD)的发病率日益提高。世界范围来看，IBD在北美以及西欧的年发病率超过10/105；在国内，IBD的年发病率从1950年的0.3/105上升到2002年的1.4/105，并呈现出逐年上升的趋势。
炎症性肠病(IBD)，是一种累及结肠、小肠的特发性炎症性疾病。克罗恩病和溃疡性结肠炎是炎性肠病的主要类型。克罗恩病可影响小肠、大肠、口腔、食道、胃和肛门，为非连续性全层炎症，最常累及部位为末端回肠、结肠和肛周；溃疡性结肠炎是结肠黏膜层和黏膜下层连续性炎症，疾病通常先累及直肠，逐渐向全结肠蔓延。炎症性肠病常伴有腹泻、腹痛、里急后重、腹块、贫血、发热、营养不良等征象。
溶血磷脂酰乙醇胺(LPE)18:1为白色粉末，极难溶于水，极易溶于三氯甲烷。分子式为C23H46NO7PC23H46NO7P，化学结构式如下图(I)所示。关于溶血磷脂酰乙醇胺18:1在治疗炎症性肠病的作用目前尚无报道。
目前为止，炎症性肠病的病因并不明确以及有效的至于手段都有待进一步研究。最近的研究指出，炎症性肠病的发病与肠道菌群关系密切。肠道炎症情况下，肠杆菌科的细菌(尤其是大肠杆菌)呈现出爆发性增长；而特异性的用钨酸盐抑制大肠杆菌的增长，有益于改善肠道炎症。这些研究指出，找到有效的手段治疗炎症性肠病势在必行；而针对大肠杆菌去寻找新的炎症性肠病诊疗方法也是有意义的。
发明内容
本发明的目的是提供溶血磷脂酰乙醇胺18:1的新应用。
本发明公开了溶血磷脂酰乙醇胺18:1在制备缓解肠道炎症药物中的应用。
本发明公开了溶血磷脂酰乙醇胺18:1在制备治疗炎症性肠病药物中的应用。
通过秀丽隐杆线虫对于Keio单基因缺失大肠杆菌库的筛选以及小鼠实验，我们发现了大肠杆菌的blc基因存在多态性，其中基因blca251(蛋白BlcE84)与炎症性肠病呈现出相关性。之后的研究发现BlcE84蛋白是因为减少了与脂质溶血磷脂酰乙醇胺18:1的结合；炎症性肠病病人粪便样品也显示出，病人肠道中的溶血磷脂酰乙醇胺18:1含量显著下降。人为补充溶血磷脂酰乙醇胺18:1可以逆转表达blcE84蛋白的致病性细菌(blcE84细菌)感染导致的小鼠的肠道损伤。
人为补充溶血磷脂酰乙醇胺18:1(20微摩尔每升，200微升每只鼠)可以逆转blcE84细菌感染导致的小鼠的肠道损伤。
附图说明
图1为小鼠感染致病性大肠杆菌后，粪便中溶血磷脂酰乙醇胺18:1显著下降；
图2为炎症性肠病病人对比正常人，粪便中溶血磷脂酰乙醇胺18:1下调。
图3为补充溶血磷脂酰乙醇胺18:1可以改善因为感染致病性大肠杆菌导致的小鼠肠道通透性下降；
图4为补充溶血磷脂酰乙醇胺18:1上调肠道屏障相关细胞连接蛋白的表达量下降；
图5为图4的灰度分析统计；
图6为补充溶血磷脂酰乙醇胺18:1重塑肠道紧密连接蛋白Claudin-1的定位；
图7为补充溶血磷脂酰乙醇胺18:1重塑肠道紧密连接蛋白ZO-1的定位；
图8为补充溶血磷脂酰乙醇胺18:1重塑肠道紧密连接蛋白Occludin的定位。
*表示与感染致病菌组小鼠与感染非致病菌组小鼠比较具有统计学意义p<0.05；**表示p<0.01；****表示p<0.0001。
具体实施方式
实施例：
实验一：致病性大肠杆菌感染小鼠模型，检测肠炎小鼠、病人粪便中溶血磷脂酰乙醇胺18:1含量
I实验材料
实验动物选取6周龄雄性C57BL/6N小鼠，体重17±2克，南京大学模式动物研究所。
II试验方法和结果
2.1动物实验
(1).将16只C57BL/6N小鼠传进生物隔离仓；
(2).灌胃抗生素混合溶液(Ampicillin 5mg每只鼠，Neomycin 5mg每只鼠，Vancomycin 5mg每只鼠，Metronidazole 4mg每只鼠)，连续一周；
(3).更换饮用水为混合抗生素水一周(Ampicillin 1g每升，Neomycin 1g每升，Vancomycin 0.5g每升，Metronidazole 1g每升)；
(4).更换为正常饮用水；
(5).将小鼠随机分为4组，分别为感染非致病细菌+溶剂组(BW25113+Vehicle)，非致病细菌+溶血磷脂酰乙醇胺18:1组(BW25113+LPE)，致病细菌+溶剂组(blcE84+Vehicle)，致病细菌+溶血磷脂酰乙醇胺18:1组(blcE84+LPE)。
2.2粪便中细菌脂质提取
(1).取小鼠/人新鲜粪便0.5g每只，置于玻璃管；
(2).加入4毫升萃取液(二氯甲烷：甲醇，2:1)，匀浆成浑浊液；
(3).于4摄氏度，200rpm萃取4小时；
(4).用玻璃注射器吸取最下层液相(有机相)；
(5).于4度离心机，2500rpm离心去沉淀；
(6).氮吹仪吹干；
(7).溶于100微升萃取液；
(8).液相色谱-质谱连用检测样品中溶血磷脂酰乙醇胺18:1。
2.3实验结果
图1－2是机体出现肠道炎症的情况下，粪便中溶血磷脂酰乙醇胺18:1会下降：图1为小鼠感染致病性大肠杆菌后，伴随着肠炎的发生，粪便中溶血磷脂酰乙醇胺18:1出现显著下降；图2为患有炎症性肠病的病人对比正常人，粪便中溶血磷脂酰乙醇胺18:1出现下调。*表示与感染致病菌组小鼠与感染非致病菌组小鼠比较具有统计学意义p<0.05；**表示p<0.01。
实验二：补充溶血磷脂酰乙醇胺18:1，改善肠炎小鼠表型
I实验材料
实验动物选取6周龄雄性C57BL/6N小鼠，体重17±2克，南京大学模式动物研究所。
II试验方法和结果
2.1动物实验
同实验一；
2.2FITC-dextran检测小鼠肠道屏障完整性
(1).配置80毫克每毫升的FITC-dextran(Sigma-Aldrich，46944-500MG)PBS溶液；
(2).给每只小鼠麻醉后，通过肛门灌注100微升；
(3).给药后2小时，从小鼠眼眶静脉丛采血；
(4).取血浆，荧光酶标仪检测FITC含量。
2.3提取小鼠结肠蛋白，蛋白质免疫印迹检测小鼠结肠屏障相关细胞连接蛋白(E-cadherin,Occludin,Claudin-1)表达量(图4-图5)
(1).取小鼠远端结肠1厘米，匀浆；
(2).加入RIPA提取蛋白；
(3).于4摄氏度，12000rpm离心15min，取上清；
(4).加入适量loading buffer，95摄氏度5min使蛋白变性；
(5).进行蛋白质免疫印迹检实验；
(6).用image-J进行灰度分析。
2.4小鼠结肠石蜡包埋，免疫荧光检测小鼠结肠屏障相关紧密连接蛋白定位
(1).取小鼠远端结肠1厘米，放入4％多聚甲醛固定24小时；
(2).用PBS 5min每次清洗样品三次，按照如下程序进行脱水：30％乙醇30min，50％乙醇30min，70％乙醇60min，80％乙醇60min，95％乙醇60min，100％乙醇60min，100％乙醇60min；
(3).用二甲苯透明，之后透蜡4小时；
(4).石蜡包埋；
(5).石蜡切片机切片，厚度5微米；
(6).用二甲苯脱蜡，10min两次；
(7).按照以下程序复水：100％乙醇5min，100％乙醇10min，95％乙醇10min，80％乙醇10min，70％乙醇10min，50％乙醇10min，双蒸水10min，双蒸水10min；
(8).抗原修复60min；
(9).进行肠道屏障相关紧密连接蛋白免疫荧光检测。
2.5实验结果
图3－8是认为补充溶血磷脂酰乙醇胺18:1后，感染致病性大肠杆菌的小鼠相较于补充溶剂组小鼠出现肠炎缓解：图3为感染致病性大肠杆菌的小鼠在感染后第二天，补充溶血磷脂酰乙醇胺18:1组相较于溶剂组，肠道通透性下降，肠道屏障功能更为健全；图4－5为感染后第二天，补充溶血磷脂酰乙醇胺18:1组相较于溶剂组，肠道屏障相关紧密连接蛋白的表达量出现增加；图6－8为感染后第二天，补充溶血磷脂酰乙醇胺18:1组相较于溶剂组，肠道屏障相关紧密连接蛋白的定位更加正常。*表示与感染致病菌组小鼠与感染非致病菌组小鼠比较具有统计学意义p<0.05；**表示p<0.01；****表示p<0.0001。
Claims (2)
1.溶血磷脂酰乙醇胺18:1在制备缓解肠道炎症药物中的应用。
2.溶血磷脂酰乙醇胺18:1在制备治疗炎症性肠病药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911273194.1A CN110840901A (zh) | 2019-12-12 | 2019-12-12 | 溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911273194.1A CN110840901A (zh) | 2019-12-12 | 2019-12-12 | 溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110840901A true CN110840901A (zh) | 2020-02-28 |
Family
ID=69608874
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911273194.1A Withdrawn CN110840901A (zh) | 2019-12-12 | 2019-12-12 | 溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110840901A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114324621A (zh) * | 2020-10-11 | 2022-04-12 | 北京泰德制药股份有限公司 | 一种改进的药物制剂中溶血磷脂酰乙醇胺的检测方法 |
| CN117434277A (zh) * | 2023-12-21 | 2024-01-23 | 中国人民解放军总医院 | 用于早期诊断或筛查克罗恩病的粪便代谢标志物组合及应用 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1297329A (zh) * | 1997-11-10 | 2001-05-30 | 威斯康星校友研究基金会 | 溶血磷脂酰乙醇胺(18:1)与溶血磷脂酰肌醇对于延缓衰老和促进果实成熟的用途 |
| CN101287499A (zh) * | 2005-09-21 | 2008-10-15 | 沃尔夫冈·斯特里梅尔 | 具治疗效果的脂质体的用途以及对器官/组织有特异性并且有治疗效果的脂质体的生产方法 |
| WO2011060144A2 (en) * | 2009-11-11 | 2011-05-19 | Virginia Tech Intellectual Properties, Inc. | Compositions and methods to protect cells by blocking entry of pathogen proteins |
| US20150250812A1 (en) * | 2013-12-05 | 2015-09-10 | Buriva, LLC | Dietary supplement containing phospholipid-dha and b vitamins |
| CN107308111A (zh) * | 2017-06-30 | 2017-11-03 | 华仁药业股份有限公司 | 一种含橄榄油中/长链脂肪乳注射液及其制备方法 |
| CN107340385A (zh) * | 2016-11-14 | 2017-11-10 | 南京拓睿谱基因科技有限公司 | 生物标记用于制备腺性膀胱炎诊断试剂的用途 |
-
2019
- 2019-12-12 CN CN201911273194.1A patent/CN110840901A/zh not_active Withdrawn
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1297329A (zh) * | 1997-11-10 | 2001-05-30 | 威斯康星校友研究基金会 | 溶血磷脂酰乙醇胺(18:1)与溶血磷脂酰肌醇对于延缓衰老和促进果实成熟的用途 |
| CN101287499A (zh) * | 2005-09-21 | 2008-10-15 | 沃尔夫冈·斯特里梅尔 | 具治疗效果的脂质体的用途以及对器官/组织有特异性并且有治疗效果的脂质体的生产方法 |
| WO2011060144A2 (en) * | 2009-11-11 | 2011-05-19 | Virginia Tech Intellectual Properties, Inc. | Compositions and methods to protect cells by blocking entry of pathogen proteins |
| US20150250812A1 (en) * | 2013-12-05 | 2015-09-10 | Buriva, LLC | Dietary supplement containing phospholipid-dha and b vitamins |
| CN107340385A (zh) * | 2016-11-14 | 2017-11-10 | 南京拓睿谱基因科技有限公司 | 生物标记用于制备腺性膀胱炎诊断试剂的用途 |
| CN107308111A (zh) * | 2017-06-30 | 2017-11-03 | 华仁药业股份有限公司 | 一种含橄榄油中/长链脂肪乳注射液及其制备方法 |
Non-Patent Citations (6)
| Title |
|---|
| DAVID K. GILES ET AL: "Remodeling of the Vibrio cholerae membrane by incorporation of exogenous fatty acids from host and aquatic environments", 《MOL MICROBIOL》 * |
| DAYUAN ZOU ET AL: "A SNP of bacterial blc disturbs gut lysophospholipid homeostasis and induces inflammation through epithelial barrier disruption", 《EBIOMEDICINE》 * |
| T SAWAI ET AL.: "The Effect of Phospholipids and Fatty Acids on Tight-Junction Permeability and Bacterial Translocation", 《PEDIATR SURG INT》 * |
| WOLFGANG STREMMEL ET AL: "Phospholipase A2 of Microbiota as Pathogenetic Determinant to Induce Inflammatory States in Ulcerative Colitis: Therapeutic Implications of Phospholipase A2 Inhibitors", 《INFLAMM INTEST DIS》 * |
| 中国食品药品检定研究院: "《国家药品标准物质使用说明书》", 24 November 2014 * |
| 陈华等: "脂质组学在高脂血症的研究进展", 《药物分析杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114324621A (zh) * | 2020-10-11 | 2022-04-12 | 北京泰德制药股份有限公司 | 一种改进的药物制剂中溶血磷脂酰乙醇胺的检测方法 |
| CN117434277A (zh) * | 2023-12-21 | 2024-01-23 | 中国人民解放军总医院 | 用于早期诊断或筛查克罗恩病的粪便代谢标志物组合及应用 |
| CN117434277B (zh) * | 2023-12-21 | 2024-03-08 | 中国人民解放军总医院 | 用于早期诊断或筛查克罗恩病的粪便代谢标志物组合及应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhong et al. | Arsenic exposure induces intestinal barrier damage and consequent activation of gut-liver axis leading to inflammation and pyroptosis of liver in ducks | |
| Ma et al. | Gut microbiota translocation promotes autoimmune cholangitis | |
| Gomes et al. | Mucormycosis caused by unusual mucormycetes, non-Rhizopus,-Mucor, and-Lichtheimia species | |
| Su et al. | Fusobacterium nucleatum promotes the development of ulcerative colitis by inducing the autophagic cell death of intestinal epithelial | |
| Jiang et al. | Optimal immunosuppressor induces stable gut microbiota after liver transplantation | |
| CN110840901A (zh) | 溶血磷脂酰乙醇胺18:1在制备缓解、治疗炎症性肠病药物中的应用 | |
| Shi et al. | Nuciferine improves high-fat diet-induced obesity via reducing intestinal permeability by increasing autophagy and remodeling the gut microbiota | |
| Chen et al. | Influence of skin wounds on the intestinal inflammatory response and barrier function: Protective role of dietary Shewanella putrefaciens SpPdp11 administration to gilthead seabream (Sparus aurata L.) | |
| Han et al. | Polysaccharides from Acanthopanax senticosus enhances intestinal integrity through inhibiting TLR4/NF‐κB signaling pathways in lipopolysaccharide‐challenged mice | |
| US20140031434A1 (en) | Use of Patchouli Alcohol in Preparation of Drug Against Helicobacter Pylori | |
| CN114344325A (zh) | 脆弱拟杆菌及其两性离子荚膜多糖在制备用于防治生殖泌尿系统肿瘤的药物中的应用 | |
| Zeng et al. | Ameliorative effect of Saccharomyces cerevisiae JKSP39 on fusobacterium nucleatum and dextran sulfate sodium-induced colitis mouse model | |
| Duchaussoy et al. | Gastrointestinal granuloma due to Candida albicans in an immunocompetent cat | |
| Oshima et al. | Successful endoscopic submucosal dissection of gastric carcinoid in a patient with autoimmune gastritis and systemic lupus erythematosus | |
| CN107496438B (zh) | 桑黄多糖在制备药物及保健食品中的应用 | |
| Qiu et al. | Fungal–bacterial interactions in mice with dextran sulfate sodium (DSS)-induced acute and chronic colitis | |
| WO2023036203A1 (zh) | Cs-4发酵菌丝体杂聚多糖及其制备方法与用途 | |
| Mitchell et al. | Metal transporter SLC39A14/ZIP14 modulates regulation between the gut microbiome and host metabolism | |
| CN115624562A (zh) | 黄芩苷在制备治疗对免疫检查点抑制剂无应答/超进展的肿瘤的药物中的应用 | |
| CN116172997A (zh) | 苯基乳酸在抑制幽门螺杆菌感染中的应用 | |
| US11925634B2 (en) | Use of koumine in preparation of medicament for treatment of inflammatory bowel disease | |
| CN109276574B (zh) | 链霉素在制备用于治疗帕金森病药物方面的应用 | |
| CN1354187A (zh) | 抗幽门螺杆菌细胞毒素鸡卵黄抗体其制备及其应用 | |
| Sun et al. | The Acetic Acid Produced by Lactobacillus Species Regulates Immune Function to Alleviate PEDV Infection in Piglets | |
| Shi et al. | Dynamic gut microbiome-metabolome in c-BSA induced experimental immune-complex glomerulonephritis and effect of losartan and MMF on microbiota modulation |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20200228 |