CN111973585A - 银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的用途 - Google Patents
银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的用途 Download PDFInfo
- Publication number
- CN111973585A CN111973585A CN202010935058.0A CN202010935058A CN111973585A CN 111973585 A CN111973585 A CN 111973585A CN 202010935058 A CN202010935058 A CN 202010935058A CN 111973585 A CN111973585 A CN 111973585A
- Authority
- CN
- China
- Prior art keywords
- myeloid
- ginkgetin
- health product
- resisting
- cells
- 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.)
- Granted
Links
- 239000003814 drug Substances 0.000 title claims abstract description 26
- SQGLUEWZRKIEGS-UHFFFAOYSA-N Ginkgetin Natural products C1=CC(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(OC)C(C=3C(=CC=C(C=3)C=3OC4=CC(O)=CC(O)=C4C(=O)C=3)O)=C2O1 SQGLUEWZRKIEGS-UHFFFAOYSA-N 0.000 title claims abstract description 20
- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 title claims abstract description 20
- AIFCFBUSLAEIBR-UHFFFAOYSA-N ginkgetin Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C(C=1)=CC=C(OC)C=1C1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 AIFCFBUSLAEIBR-UHFFFAOYSA-N 0.000 title claims abstract description 20
- 230000036541 health Effects 0.000 title claims abstract description 18
- 210000004985 myeloid-derived suppressor cell Anatomy 0.000 title abstract description 31
- 238000002360 preparation method Methods 0.000 title abstract description 4
- 229940079593 drug Drugs 0.000 claims abstract description 8
- 206010028980 Neoplasm Diseases 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 9
- 230000004054 inflammatory process Effects 0.000 claims description 8
- 208000023275 Autoimmune disease Diseases 0.000 claims description 7
- 208000037581 Persistent Infection Diseases 0.000 claims description 7
- 210000004479 myeloid suppressor cell Anatomy 0.000 claims description 5
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 4
- 208000037976 chronic inflammation Diseases 0.000 claims description 4
- 206010017758 gastric cancer Diseases 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 201000011549 stomach cancer Diseases 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 239000002417 nutraceutical Substances 0.000 claims 1
- 235000021436 nutraceutical agent Nutrition 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 24
- 230000002401 inhibitory effect Effects 0.000 abstract description 6
- 239000000047 product Substances 0.000 description 13
- 238000001565 modulated differential scanning calorimetry Methods 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 239000006228 supernatant Substances 0.000 description 8
- 238000002156 mixing Methods 0.000 description 7
- 241000218628 Ginkgo Species 0.000 description 6
- 235000011201 Ginkgo Nutrition 0.000 description 6
- 235000008100 Ginkgo biloba Nutrition 0.000 description 6
- 241000282414 Homo sapiens Species 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 239000012980 RPMI-1640 medium Substances 0.000 description 5
- 210000004443 dendritic cell Anatomy 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 210000001185 bone marrow Anatomy 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 210000005259 peripheral blood Anatomy 0.000 description 4
- 239000011886 peripheral blood Substances 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- UFJORDZSBNSRQT-UHFFFAOYSA-N 3-(4-oxo-2-phenylchromen-3-yl)-2-phenylchromen-4-one Chemical compound O1C2=CC=CC=C2C(=O)C(C=2C(C3=CC=CC=C3OC=2C=2C=CC=CC=2)=O)=C1C1=CC=CC=C1 UFJORDZSBNSRQT-UHFFFAOYSA-N 0.000 description 3
- 210000002798 bone marrow cell Anatomy 0.000 description 3
- 230000002490 cerebral effect Effects 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 210000003714 granulocyte Anatomy 0.000 description 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 210000005087 mononuclear cell Anatomy 0.000 description 3
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000002303 tibia Anatomy 0.000 description 3
- 206010002383 Angina Pectoris Diseases 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 2
- 102100022338 Integrin alpha-M Human genes 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000006450 immune cell response Effects 0.000 description 2
- 210000002414 leg Anatomy 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 206010065559 Cerebral arteriosclerosis Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000218791 Ginkgoaceae Species 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 101000746373 Homo sapiens Granulocyte-macrophage colony-stimulating factor Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 208000032984 Intraoperative Complications Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 210000004394 hip joint Anatomy 0.000 description 1
- 102000046157 human CSF2 Human genes 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 201000005851 intracranial arteriosclerosis Diseases 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000066 myeloid cell Anatomy 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 210000001694 thigh bone Anatomy 0.000 description 1
- 230000008733 trauma Effects 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- 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]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- 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)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Transplantation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的用途。本申请发明人经过研究发现，银杏双黄酮能明显抑制髓系抑制性细胞的产生，可用于制备抗髓系抑制性细胞的药物和/或保健品。本发明提供了银杏双黄酮的药物新用途，同时为制备抗髓系抑制性细胞的药物或保健品提供了新的选择。
Description
技术领域
本发明属于生物医药技术领域，具体涉及银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的用途。
背景技术
髓系抑制性细胞(Myeloid-derived suppressor cells，MDSCs)是骨髓来源的一群异质性细胞，是树突状细胞(dendritic cells，DCs)、巨噬细胞和(或)粒细胞的前体，具有显著抑制免疫细胞应答的能力。MDSCs来源于骨髓祖细胞和未成熟髓细胞(immaturemyeloid cells，IMCs)。正常情况下，是树突状细胞(DC)、巨噬细胞和粒细胞的前体，能迅速地分化为成熟的粒细胞、DCs和巨噬细胞，并进入相应的器官、组织，发挥正常免疫功能，IMCs占外周血单个核细胞的0.5％左右。在肿瘤、感染、炎症、败血症、外科损伤等其它病理条件下，受细胞因子的作用，这些髓系来源的前体细胞成熟受阻，因而停留在各个分化阶段，成为具有免疫抑制功能的MDSCs。它们并在细胞因子的作用下被募集、迁移、扩增，使得其在外周血中的数量和比例增加十倍左右，约占患者外周血单个核细胞(PBMC)的10％，贯穿疾病发生的整个过程。MDSCs主要分为两个亚群：粒细胞PMN-MDSCs和单核M-MDSCs。在大多数类型的癌症中，PMN-MDSCs占全部MDSCs的70-80％，而M-MDSCs通常不超过20％。
银杏双黄酮，银杏科植物银杏的干燥叶提取物，现有研究表明可增加脑血管流量，降低脑血管阻力，改善脑血管循环功能，保护脑细胞，免受缺血损害，扩张冠状动脉，防止心绞痛及心肌梗塞，抑制血小板聚集，防止血栓形成，清除有害的氧化自由基，提高免疫能力，具有防癌抗衰功能。对治疗冠心病、心绞痛、脑动脉硬化、老年性痴呆、高血压等病有很好的疗效。
目前的研究表明MDSCs与肿瘤、自身免疫性疾病、慢性感染、创伤和炎症等多种疾病的发生发展密切相关。
发明内容
基于此，本发明提供了银杏双黄酮的药物新用途，银杏双黄酮能有效抑制髓系抑制性细胞的产生，可用于制备抗髓系抑制性细胞的药物和/或保健品。
为实现上述目的，本发明采用的技术方案为：
银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的用途。
优选地，所述抗髓系抑制性细胞的药物和/或保健品可防治肿瘤、自身免疫性疾病、慢性感染或炎症。
优选地，所述肿瘤为胃癌。
本发明还提供了一种抗髓系抑制性细胞的药物和/或保健品。
为实现上述目的，本发明采用的技术方案为：
一种抗髓系抑制性细胞的药物和/或保健品，所述药物和/或保健品的活性成分包含银杏双黄酮。
银杏双黄酮作为活性成分，可与药物学可接受的载体一起，制备得到抗髓系抑制性细胞的药物和/或保健品。
优选地，所述药物和/或保健品可防治肿瘤、自身免疫性疾病、慢性感染或炎症。
优选地，所述肿瘤为胃癌。
优选地，所述药物和/或保健品的剂型为片剂、颗粒剂、溶液剂、粉剂、胶囊剂、注射液或粉针剂。
相对于现有技术，本发明的有益效果为：发明人经过大量的研究发现，银杏双黄酮能显著抑制小鼠以及人MDSCs的产生，可用于制备抗MDSCs的药物和/或保健品。MDSCs是一种具有显著抑制免疫细胞应答能力的细胞，与多种疾病的发生和发展密切相关，包括肿瘤、自身免疫性疾病、慢性感染、创伤和炎症等疾病。因此，抑制MDSCs的药物或保健品可用于防治肿瘤、自身免疫性疾病、慢性感染、创伤和炎症等疾病。
附图说明
图1为银杏双黄酮抑制小鼠MDSCs的流式检测结果图。
图2为银杏双黄酮抑制人MDSCs的流式检测结果图。
具体实施方式
为了便于理解本发明，下面将参照实施例对本发明进行更全面的描述，以下给出了本发明的较佳实施例。但是，本发明可以以许多不同的形式来实现，并不限于本文所描述的实施例。提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。应理解，下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。实施例中所用到的各种常用试剂，均为市售产品。
除非另有定义，本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的，不是旨在于限制本发明。
实施例1银杏双黄酮抑制小鼠MDSCs
1、实验方法
(1)从小鼠骨髓中分离骨髓细胞：
1)颈椎脱位法处死8周龄C57小鼠，将小鼠浸泡在75％酒精中5min；取出小鼠，用眼科剪剪开小鼠双侧腿部皮肤，自髋关节剪下双侧腿骨；小心剪去股骨与胫骨的附着肌肉，勿伤及关节面，忌暴露骨髓腔；
2)将剥离出的股骨、胫骨浸泡于75％酒精中3min后转移至预冷的RPMI1640培养基中；
3)用眼科剪轻轻剪开股骨或胫骨两端，然后用5ml无菌注射器吸取RPMI1640培养基，对准50ml离心管冲出骨髓腔中骨髓组织；
4)3000rpm/min，室温离心5min，弃上清，加入5ml红细胞裂解液ACK，吹打混匀后裂解5min，然后加入30ml预冷的PBS终止裂解；室温3000rpm/min，离心5min以获取细胞沉淀；
5)弃上清后再加入10ml预冷的RPMI 1640，吹打重悬细胞，显微镜细胞计数板计数，备用。
(2)对步骤(1)获得的骨髓细胞进行体外培养：
1)细胞计数后，用RPMI1640+10％FBS+100U/ml青霉素+0.1mg/ml链霉素+50uMβ-ME的完全培养基将细胞密度调整至1×106cells/ml；
2)培养体系中加入重组小鼠源GM-CSF细胞因子(20ng/ml)，充分混匀后，将细胞悬液按每孔1ml铺于48孔板中。
(3)银杏双黄酮处理骨髓细胞
向步骤(2)所述培养体系中加入银杏双黄酮(1.5umol/ml)处理，继续进行体外培养6天，DMSO处理组作为对照。
(4)流式细胞术检测：
体外培养6天后，收集细胞进行流式细胞术检测：
1)取含0.5million的单细胞悬液，加入5ml PBS混匀，3000rpm/min离心5min，弃尽上清；
2)用于分析小鼠MDSC的流式抗体染色组合为：PMN-MDSC(CD11b+Gr1+Ly6C-Ly6Ghi)和M-MDSC(CD11b+Gr1+Ly6ChigLy6G-)；
3)按照抗体说明书，抗体1:200使用PBS稀释，每个样本用100μl抗体稀释液。将流式管于振荡器上振荡，充分混匀；于4℃冰箱避光染色30min。加入4ml PBS，3000rpm/min离心5min，弃去上清；加入500μl PBS重悬细胞后，流式上机检测，获取数据并用FlowJo10软件进行分析。
2、实验结果
结果如图1所示，银杏双黄酮能显著抑制小鼠MDSCs的产生，使培养体系中MDSCs的比例下降，PMN-MDSCs和M-MDSC两种亚型均受到抑制。
实施例2银杏双黄酮抑制人MDSCs
1、实验方法
(1)从人外周血中分离单核细胞：
取健康志愿者外周血5ml，分离得到单个核细胞：
1)用含EDTA抗凝采血管收集健康志愿者外周血，然后加入等3倍体积的PBS溶液，轻轻混匀；
2)向50ml无菌离心管中加入3ml淋巴细胞分离液；然后用吸管将血稀释液沿着管壁缓慢的加至淋巴细胞分离液的上层，于18℃下600g(升5，降0)，离心22min；
3)离心后，用移液枪将中间白膜层吸出，放于一干净的15ml离心管中，加入10ml预冷的PBS溶液，混匀；室温下，600g离心10min，弃尽上清；
4)加入1ml ACK红细胞裂解液进行裂红，3min后，加入10ml预冷的PBS进行终止；室温下，600g离心10min，弃尽上清；
5)用5ml PBS进行重悬细胞沉淀，进行细胞计数，放冰上备用。
(2)对步骤(1)获得的单核细胞进行体外培养：
1)细胞计数后，用RPMI1640+10％FBS+100U/ml青霉素+0.1mg/ml链霉素+50uMβ-ME的完全完全培养基将细胞密度调整至1×106cells/ml；
2)培养体系中加入重组人源GM-CSF细胞因子(20ng/ml)，充分混匀后，将细胞悬液按每孔1ml铺于48孔板中；
(3)银杏双黄酮处理单核细胞
向步骤(2)所述培养体系中加入银杏双黄酮(1.5umol/ml)处理，续进行体外培养6天，DMSO处理组作为对照。
(4)流式细胞术检测：
体外培养6天后，收集细胞进行流式细胞术检测：
1)取含0.5million的单细胞悬液，加入5ml PBS混匀，3000rpm/min离心5min，弃尽上清；
2)用于分析人MDSC的流式抗体染色组合为：M-MDSC(HLA-DR-/low CD11bhiCD33hi)和PMN-MDSC(HLA-DR-/lowCD11bmedCD33med)。
3)按照抗体说明书，抗体1:200使用PBS稀释，每个样本用100μl抗体稀释液。将流式管于振荡器上振荡，充分混匀；于4℃冰箱避光染色30min。加入4ml PBS，3000rpm/min离心5min，弃去上清；加入500μl PBS重悬细胞后，流式上机检测，获取数据并用FlowJo10软件进行分析。
2、实验结果
结果如图2所示，银杏双黄酮能显著抑制人MDSCs的产生，使培养体系中MDSCs的比例下降，尤其是使PMN-MDSCs受到抑制。
上述结果表明，银杏双黄酮能显著抑制MDSCs的产生，可用于制备抗MDSCs的药物或保健品。
最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。
Claims (7)
1.银杏双黄酮在制备抗髓系抑制性细胞的药物和/或保健品中的应用。
2.根据权利要求1所述的应用，其特征在于，所述抗髓系抑制性细胞的药物和/或保健品可防治肿瘤、自身免疫性疾病、慢性感染或炎症。
3.根据权利要求2所述的应用，其特征在于，所述肿瘤为胃癌。
4.一种抗髓系抑制性细胞的药物和/或保健品，其特征在于，所述药物和/或保健品的活性成分包含银杏双黄酮。
5.根据权利要求4所述的药物和/或保健品，其特征在于，所述药物和/或保健品可防治肿瘤、自身免疫性疾病、慢性感染或炎症。
6.根据权利要求5所述的药物和/或保健品，其特征在于，所述肿瘤为胃癌。
7.根据权利要求4～6任一项所述的药物和/或保健品，其特征在于，所述药物和/或保健品的剂型为片剂、颗粒剂、溶液剂、粉剂、胶囊剂、注射液或粉针剂。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010935058.0A CN111973585B (zh) | 2020-09-08 | 2020-09-08 | 银杏双黄酮在制备抗髓系抑制性细胞的药物中的用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010935058.0A CN111973585B (zh) | 2020-09-08 | 2020-09-08 | 银杏双黄酮在制备抗髓系抑制性细胞的药物中的用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111973585A true CN111973585A (zh) | 2020-11-24 |
| CN111973585B CN111973585B (zh) | 2022-04-08 |
Family
ID=73447736
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010935058.0A Active CN111973585B (zh) | 2020-09-08 | 2020-09-08 | 银杏双黄酮在制备抗髓系抑制性细胞的药物中的用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111973585B (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104940184A (zh) * | 2014-03-25 | 2015-09-30 | 上海医药工业研究院 | 一种银杏黄素类化合物的应用 |
| CN105193784A (zh) * | 2015-10-20 | 2015-12-30 | 吉林大学 | 银杏素在制备治疗猪链球菌感染药物中的应用 |
-
2020
- 2020-09-08 CN CN202010935058.0A patent/CN111973585B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104940184A (zh) * | 2014-03-25 | 2015-09-30 | 上海医药工业研究院 | 一种银杏黄素类化合物的应用 |
| CN105193784A (zh) * | 2015-10-20 | 2015-12-30 | 吉林大学 | 银杏素在制备治疗猪链球菌感染药物中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| YOON JUNG JEON 等: "Ginkgetin inhibits the growth of DU_145 prostate cancer cells through inhibition of signal transducer and activator of transcription 3 activity", 《CANCER SCI》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111973585B (zh) | 2022-04-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2759508C1 (ru) | Композиция, включающая экзосому, полученную из стволовых клеток, для индукции адипогенной дифференцировки, регенерации жировой ткани, отбеливания кожи или коррекции морщин | |
| Liang et al. | Mesenchymal stem cell transplantation for diffuse alveolar hemorrhage in SLE | |
| US20190276803A1 (en) | Method of culturing immune cells, kit for thereof, immune cell cultured medium obtained by same method, cosmetic composition and pharmaceutical composition comprising thereof | |
| Billingham et al. | Quantitative studies on the induction of tolerance of skin homografts and on runt disease in neonatal rats | |
| CN113456645A (zh) | Dmxaa在制备防治骨质疏松症药物中的应用 | |
| CN111184711A (zh) | 银杏内酯a的新应用 | |
| CN111773211B (zh) | 血筒素在制备抗类风湿关节炎药物中的应用 | |
| Ballas | Lymphokine-activated killer (LAK) cells. I. Differential recovery of LAK, natural killer cells, and cytotoxic T lymphocytes after a sublethal dose of cyclophosphamide. | |
| EP4349348A1 (en) | Cancer cachexia ameliorating agent and cancer cachexia amelioration method | |
| WO2022016098A1 (en) | Compositions and methods for treatment of inflammatory and thromboinflammatory disorders using modified exosomes | |
| CN111973585B (zh) | 银杏双黄酮在制备抗髓系抑制性细胞的药物中的用途 | |
| Lv et al. | Functional distinction of rat liver natural killer cells from spleen natural killer cells under normal and acidic conditions in vitro | |
| KR20190074720A (ko) | 골수이식 후 생착 촉진용 조성물 | |
| CN111991403B (zh) | 丹参酮ⅰ在制备抗髓系抑制性细胞的药物中的用途 | |
| WO2019235789A1 (ko) | 편도 유래 중간엽 줄기세포의 조정 배지를 포함하는 이식편대숙주 질환 예방 혹은 치료용 조성물 | |
| BG62239B1 (bg) | Приложение на лизозимен димер | |
| CN111821312A (zh) | 异槲皮苷在制备抗髓系抑制性细胞的药物和/或保健品中的用途 | |
| CN111197029A (zh) | 一种尿酸钠诱导产生髓系抑制性细胞的方法 | |
| WO2022194094A1 (zh) | 无细胞脂肪提取物用于治疗脊髓损伤 | |
| KR20210002205A (ko) | 면역세포 배양액 함유 조성물의 제조방법 및 기능성 화장품 조성물 | |
| CN118291375A (zh) | 一种细胞修复蛋白提取物及其制备方法和其应用 | |
| CN108853129B (zh) | 升麻素苷在制备髓源性抑制细胞抑制药物中的应用 | |
| JPWO2016195086A1 (ja) | アレルギー性疾患の治療薬 | |
| CN110229786A (zh) | 一种间充质干细胞的制备方法及其应用 | |
| CN116925186B (zh) | 一种新生儿肺发育不良的间充质干细胞治疗方法 |
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 |