CN113116920A - 枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 - Google Patents
枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 Download PDFInfo
- Publication number
- CN113116920A CN113116920A CN202110512216.6A CN202110512216A CN113116920A CN 113116920 A CN113116920 A CN 113116920A CN 202110512216 A CN202110512216 A CN 202110512216A CN 113116920 A CN113116920 A CN 113116920A
- Authority
- CN
- China
- Prior art keywords
- lycium barbarum
- barbarum polysaccharide
- distilled water
- centrifuging
- fatigue
- 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
- 239000008518 lycium barbarum polysaccharide Substances 0.000 title claims abstract description 27
- 239000003814 drug Substances 0.000 title claims abstract description 22
- 230000002929 anti-fatigue Effects 0.000 title claims abstract description 14
- 235000013305 food Nutrition 0.000 title claims abstract description 14
- 238000002360 preparation method Methods 0.000 title abstract description 6
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 102000019197 Superoxide Dismutase Human genes 0.000 claims abstract description 13
- 108010012715 Superoxide dismutase Proteins 0.000 claims abstract description 13
- 229940118019 malondialdehyde Drugs 0.000 claims abstract description 13
- 210000004369 blood Anatomy 0.000 claims abstract description 7
- 239000008280 blood Substances 0.000 claims abstract description 7
- 229940079593 drug Drugs 0.000 claims abstract description 7
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 210000002966 serum Anatomy 0.000 claims abstract description 6
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 claims abstract description 4
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 3
- 239000004310 lactic acid Substances 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 239000012153 distilled water Substances 0.000 claims description 19
- 150000004676 glycans Chemical class 0.000 claims description 19
- 229920001282 polysaccharide Polymers 0.000 claims description 18
- 239000005017 polysaccharide Substances 0.000 claims description 18
- 235000019441 ethanol Nutrition 0.000 claims description 14
- 239000002244 precipitate Substances 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 238000004108 freeze drying Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 229920005654 Sephadex Polymers 0.000 claims description 5
- 239000012507 Sephadex™ Substances 0.000 claims description 5
- 238000005303 weighing Methods 0.000 claims description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- 238000005238 degreasing Methods 0.000 claims description 4
- 238000000502 dialysis Methods 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 238000001704 evaporation Methods 0.000 claims description 3
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 239000012465 retentate Substances 0.000 claims description 3
- 238000004440 column chromatography Methods 0.000 claims description 2
- 239000006185 dispersion Substances 0.000 claims description 2
- 239000003480 eluent Substances 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000012544 monitoring process Methods 0.000 claims description 2
- 230000010355 oscillation Effects 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 18
- 229920002527 Glycogen Polymers 0.000 abstract description 17
- 229940096919 glycogen Drugs 0.000 abstract description 17
- 210000004185 liver Anatomy 0.000 abstract description 9
- 210000003205 muscle Anatomy 0.000 abstract description 9
- 238000003860 storage Methods 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 21
- 230000009182 swimming Effects 0.000 description 17
- 101100048228 Homo sapiens UBP1 gene Proteins 0.000 description 14
- 244000241838 Lycium barbarum Species 0.000 description 14
- 235000015459 Lycium barbarum Nutrition 0.000 description 14
- 101100117629 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) LCB3 gene Proteins 0.000 description 14
- 102100040065 Upstream-binding protein 1 Human genes 0.000 description 14
- 206010016256 fatigue Diseases 0.000 description 9
- 101100500049 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) YSR3 gene Proteins 0.000 description 8
- 101100446025 Caenorhabditis elegans lbp-3 gene Proteins 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 5
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 5
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000002035 prolonged effect Effects 0.000 description 3
- 244000241872 Lycium chinense Species 0.000 description 2
- 235000015468 Lycium chinense Nutrition 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000012869 ethanol precipitation Methods 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 102000006587 Glutathione peroxidase Human genes 0.000 description 1
- 108700016172 Glutathione peroxidases Proteins 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000005373 Panax quinquefolius Species 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 206010062519 Poor quality sleep Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000208292 Solanaceae Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000037080 exercise endurance Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- PWAOOJDMFUQOKB-WCZZMFLVSA-N ginsenoside Re Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@H]3C(C)(C)[C@@H](O)CC[C@]3(C)[C@@H]3[C@@]([C@@]4(CC[C@@H]([C@H]4[C@H](O)C3)[C@](C)(CCC=C(C)C)O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C)(C)C2)O[C@H](CO)[C@@H](O)[C@@H]1O PWAOOJDMFUQOKB-WCZZMFLVSA-N 0.000 description 1
- AOGZLQUEBLOQCI-UHFFFAOYSA-N ginsenoside-Re Natural products CC1OC(OCC2OC(OC3CC4(C)C(CC(O)C5C(CCC45C)C(C)(CCC=C(C)C)OC6OC(CO)C(O)C(O)C6O)C7(C)CCC(O)C(C)(C)C37)C(O)C(O)C2O)C(O)C(O)C1O AOGZLQUEBLOQCI-UHFFFAOYSA-N 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 230000004112 neuroprotection Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 210000004279 orbit Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000037081 physical activity Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 230000003860 sleep quality Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 230000009385 viral infection 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- 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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- 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)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Diabetes (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Hematology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Obesity (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Toxicology (AREA)
- Materials Engineering (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Sustainable Development (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了枸杞多糖在制备抗疲劳药物、保健品或食品中的应用。分子量为92.4kDa的枸杞多糖能够明显增加肝糖原和肌糖原储备，提高超氧化物歧化酶活力，显著降低血清尿素氮、血乳酸、丙二醛水平，相较于其他分子量的枸杞多糖而言，表现出更为优异的抗疲劳作用，适用于制备抗疲劳药物、保健品或食品。
Description
技术领域
本发明属于药物技术领域，涉及枸杞多糖在制备抗疲劳药物、保健品或食品中的应用。
背景技术
疲劳是机体在从事脑力活动或体力活动时由于消耗过大，达到一定程度时出现的一种正常的生理现象。疲劳通常会导致一系列继发性问题，例如抑郁和焦虑，同时还与认知障碍、睡眠质量差、身体机能障碍和能量失衡有关。长期疲劳会增加罹患癌症、多发性硬化症、人类免疫缺陷病毒感染和帕金森氏病的风险。消除和缓解疲劳一直以来备受运动医学、军事医学和航天医学等领域的关注。疲劳是一种复杂而普遍的生理现象，没有明确的病因，可能需要长期服药，而长期用药可能会导致一些不良反应或毒副作用。因此，找到具有明确功效且较少副作用的抗疲劳潜在药物或制剂是目前关注的重点。
枸杞为茄科植物枸杞的成熟果实，广泛分布于中国西北部、欧洲东南部和地中海干旱半干旱地区。从枸杞提取物中分离得到的枸杞多糖(L.barbarum polysaccharide，LBP)是枸杞中含量最丰富的成分之一，占干果的5％～8％，被认为是发挥生物学功效的重要组成成分。枸杞多糖因具有抗衰老、抗氧化、抗糖尿病、抗癌、细胞保护、神经保护和免疫调节等保健作用，常被作为药物和功能性食品使用。然而天然多糖分子量与其在体内的吸收和运输密切相关，从而影响生物活性。
发明内容
本发明的目的在于提供一种具有特定分子量的枸杞多糖在制备抗疲劳药物、保健品或食品中的应用。
本发明中，所述的枸杞多糖的分子量为92.4kDa。
本发明中，所述的枸杞多糖通过以下步骤制备：
(1)将枸杞粉末在80℃下用95％乙醇回流脱脂，重复数次，收集滤渣并干燥；
(2)将干燥后的滤渣按照料液比1:30加入蒸馏水，在90℃下提取，重复数次，5℃下离心收集上清液，浓缩，加入95％乙醇分散，4℃下静置过夜，5℃下离心收集沉淀；
(3)将沉淀用蒸馏水溶解，再按体积比5:1加入Sevage试剂，充分振荡，然后在5℃下离心收集上清液，重复操作至中间无变性蛋白质层出现；
(4)将上清液减压浓缩蒸发除去残留的Sevage试剂，并在4℃下透析，将截留液冷冻干燥获得粗制枸杞多糖：
(5)然后称取粗制枸杞多糖溶于蒸馏水中，缓慢加入无水乙醇，使最终溶液的乙醇浓度为30％，于4℃静置过夜，然后5℃下离心，收集沉淀，冷冻干燥得到30％的醇沉组分LBP-Ⅰ；
(6)将LBP-Ⅰ溶于蒸馏水中，经葡聚糖凝胶Sephadex G-100柱层析纯化，用蒸馏水进行洗脱，收集洗脱液，通过苯酚-硫酸法跟踪监测多糖，合并馏分，浓缩并冻干，得到纯化的分子量为92.4kDa的枸杞多糖。
优选地，步骤(1)中，脱脂时间为2h，重复次数为3次。
优选地，步骤(2)具体为：将干燥后的滤渣按照料液比1:30加入蒸馏水，在90℃下提取2h，重复2次，4000r/min、5℃下离心10min，合并上清液，浓缩至原来体积的四分之一，加入4倍体积的95％乙醇，4℃下静置过夜，4000r/min、5℃下离心10min，收集沉淀。
优选地，步骤(3)中，振荡时间为20min，离心转速为4000r/min，离心时间为10min。
优选地，步骤(4)中，透析袋的截留分子量为3500Da，透析时间为48h。
优选地，步骤(5)中，离心转速为8000r/min，离心时间为15min。
优选地，步骤(6)中，蒸馏水的洗脱速度为0.3mL/min。
本发明中，所述的药物、保健品或食品为缓解运动疲劳的药物、保健品或食品。
进一步地，所述的药物是血清尿素氮(BUN)、乳酸(BLA)和丙二醛(MDA)含量，提高血糖、超氧化物歧化酶(SOD)水平的药物、保健品或食品。
本发明首次发现不同分子量的枸杞多糖的抗疲劳提升作用存在明显差异，其中分子量为92.4kDa的抗疲劳提升效果最为显著，可以显著延长小鼠负重游泳时间，有效降低疲劳小鼠BLA和BUN以及MDA含量，提高SOD、肝糖原和肌糖原水平，与其他分子量的枸杞多糖相比较，表现出更为优异的抗疲劳作用。
附图说明
图1为不同分子量枸杞多糖对小鼠肝糖原(A)和肌糖原(B)的影响。其中，*表示存在显著性差异(p<0.05)；**表示存在极显著性差异(p<0.01)。
图2为不同分子量枸杞多糖对小鼠BUN(A)、BLA(B)、MDA(C)和SOD(D)水平的影响。其中，*表示存在显著性差异(p<0.05)；**表示存在极显著性差异(p<0.01)。
具体实施方式
下面结合具体实施例和附图对本发明做进一步详细描述。
实施例1
1.实验材料
1.1材料与试剂
糖原、尿素氮(BUN)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)以及丙二醛(MDA)测定试剂盒(南京建成生物工程研究所)；枸杞子(甘肃省定西市漳县金钟镇三农中药材种植农民专业合作社)、西洋参口服液(每100mL含：总皂甙(以人参皂甙Re计)230mg，福建省力菲克药业有限公司)
1.2实验仪器
紫外可见分光光度计(Lambda 35，美国珀金埃尔默股份有限公司)；多功能酶标仪(Tecan infinite 200pro，Tecan Austria GmbH公司)
1.3实验动物
雄性ICR小鼠(20±2g)购自扬州大学比较医学中心，动物合格证号为SCXK(苏)2017-0007。使小鼠在25℃的环境中适应一周，每天光/暗(12/12h)循环，并提供足够的饲料和饮用水
2.实验方法
2.1枸杞多糖的制备
将枸杞粉末在80℃下用95％乙醇回流脱脂2h，重复3次，收集滤渣并干燥。称取干燥后的滤渣按照料液比1:30加入蒸馏水，在90℃下提取2h，重复2次，4000r/min、5℃下离心10min，合并上清液，浓缩至原来体积的四分之一，加入4倍体积的95％乙醇，4℃下静置过夜，4000r/min、5℃下离心10min，收集沉淀。将所得沉淀用蒸馏水溶解，再按体积比5:1加入Sevage试剂，充分振荡20min，在4000r/min、5℃下离心10min后收集上清液，重复操作至中间无变性蛋白质层出现。将上清液减压浓缩蒸发除去残留的Sevage试剂，并在4℃下透析48h(Mw 3500Da)，将截留液冷冻干燥获得粗制枸杞多糖(LBP)。然后称取LBP溶于蒸馏水中，缓缓加入无水乙醇，使最终溶液的乙醇浓度为30％，于4℃静置过夜，然后离心(8000r/min，15min，5℃)，收集沉淀，冷冻干燥得到30％的醇沉组分LBP-Ⅰ；在上清液中继续缓缓加入乙醇，使最终溶液的乙醇浓度为50％，4℃下静置过夜后，离心(8000r/min，15min，5℃)，收集沉淀，冷冻干燥得到50％的醇沉组分LBP-Ⅱ；按相同的操作，使最终溶液的乙醇浓度为70％时，得到70％的醇沉组分LBP-Ⅲ。最后称取LBP-Ⅰ、LBP-Ⅱ和LBP-Ⅲ各40mg溶于5mL蒸馏水中，经葡聚糖凝胶Sephadex G-100柱(1.5cm×60cm)层析纯化，用蒸馏水以0.3mL/min速度进行洗脱，自动收集每管3mL。通过苯酚-硫酸法跟踪监测多糖，合并馏分，浓缩并冻干，得纯化多糖LBP-Ⅰ-1、LBP-Ⅱ-1和LBP-Ⅲ-1，命名为LBP1、LBP2和LBP3。
LBP1、LBP2和LBP3纯度分别为80.1％、87.3％和73.4％，分子量分别为92,441Da、7714Da和3188Da。采用红外光谱法、气相色谱法、1H NMR谱和35C NMR谱等方法对多糖结构进行表征。结果表明3种多糖结构基本相似，均不含有β构型的糖残基。LBP1由阿拉伯糖(24.0％)、木糖(25.0％)、葡萄糖(20.2％)和半乳糖(23.1％)组成；LBP2由阿拉伯糖(28.3％)、木糖(17.8％)、葡萄糖(13.4％)和半乳糖(23.4％)组成；LBP3由阿拉伯糖(26.3％)和葡萄糖(30.3％)组成。
2.2实验分组
将小鼠随机分成10组：空白组(给予等体积生理盐水)、阳性对照组(给予西洋参口服液，剂量参考说明书中推荐每日摄入量为6.7mg/kg/d)、LBP1、LBP2和LBP3的低、中、高剂量组(低、中、高剂量组分别灌胃5、10、20mg/kg/d枸杞多糖溶液)，每组20只，连续灌胃30天。
2.3测定负重游泳时间
最后一次给药1h后，从每组中随机选取10只小鼠并按体重的7％在尾部附加铅片置于游泳箱(50cm×50cm×40cm)中，游泳箱中水深不低于30cm，水温维持在30±1℃。以小鼠头部沉没水中8s以上作为试验终点，记录开始游泳至力竭的时间，为负重力竭游泳时间。
2.4测定肝糖原、肌糖原、BUN、BLA、SOD和MDA水平
每组中另外10只小鼠进行无负重游泳30min，结束后立即采用眼眶取血，室温静置15min使血液凝固，并以3000r/min，4℃离心10min收集血清，置于冰箱中冷藏备用，按照试剂盒中的方法测定血清中BUN、BLA、SOD和MDA水平。同时，收集肝脏和后腿肌肉，生理盐水漂洗后，用滤纸吸干，分别称取肝脏100mg，肌肉500mg置于试管中，按照试剂盒中的方法分析肝糖原和肌糖原含量。
2.5统计学处理
实验所得数据采用平均值±标准偏差(SD)表示，使用SPSS 23.0软件进行单因素方差分析多重比较，p<0.05表示具有显著性差异，p<0.01表示具有极显著差异。
3.实验结果
3.1不同分子量枸杞多糖对小鼠负重游泳时间的影响
如表1所示，经过30天的灌胃各组小鼠体征正常，未出现死亡情况，体重正常增长。与空白组相比，灌胃不同剂量的LBP1、LBP2和LBP3小鼠负重游泳时间均有所延长，说明不同分子量的枸杞多糖均能够有效缓解小鼠运动疲劳。其中，高剂量的LBP1组中小鼠负重游泳时间较阳性对照组有所提升，但不存在显著性差异。当灌胃剂量相同时，LBP1实验组小鼠负重游泳时间较LBP2和LBP3两组有所延长，其中高剂量的LBP1组游泳时间显著长于高剂量的LBP3。说明LBP1对延长小鼠负重游泳时间的效果最为显著，更能够提高其抗疲劳能力。
表1
3.2不同分子量枸杞多糖对小鼠肝糖原、肌糖原含量的影响
图1显示了各组小鼠在不负重游泳30min之后肝糖原和肌糖原的含量。结果表明LBP1和LBP2各剂量组中肝糖原和肌糖原含量均极显著高于空白组(p<0.01)，表明LBP1和LBP2具有增强糖原储备，为身体提供能量，缓解身体疲劳和增强运动耐力的能力。
3.3不同分子量枸杞多糖对小鼠BUN、BLA、SOD和MDA水平的影响
图2显示了各组小鼠不负重游泳30min后血液中SOD活性以及BUN、BLA和MDA的水平。结果显示，经不同分子量的枸杞多糖治疗后BUN、BLA和MDA含量较空白组明显降低，同时SOD活力显著提高，并且LBP1组治疗效果更明显。
综上所述，经小鼠游泳模型发现，不同分子量的枸杞多糖均能有效缓解运动疲劳症状，特别是分子量为92.4kDa的LBP1，能够显著延长小鼠负重游泳时间，并对运动后小鼠血液生化指标有改善作用，具有良好的抗疲劳作用。
Claims (10)
1.枸杞多糖在制备抗疲劳药物、保健品或食品中的应用，其特征在于，所述的枸杞多糖的分子量为92.4kDa。
2.根据权利要求1所述的应用，其特征在于，所述的枸杞多糖通过以下步骤制备：
(1)将枸杞粉末在80℃下用95％乙醇回流脱脂，重复数次，收集滤渣并干燥；
(2)将干燥后的滤渣按照料液比1:30加入蒸馏水，在90℃下提取，重复数次，5℃下离心收集上清液，浓缩，加入95％乙醇分散，4℃下静置过夜，5℃下离心收集沉淀；
(3)将沉淀用蒸馏水溶解，再按体积比5:1加入Sevage试剂，充分振荡，然后在5℃下离心收集上清液，重复操作至中间无变性蛋白质层出现；
(4)将上清液减压浓缩蒸发除去残留的Sevage试剂，并在4℃下透析，将截留液冷冻干燥获得粗制枸杞多糖：
(5)然后称取粗制枸杞多糖溶于蒸馏水中，缓慢加入无水乙醇，使最终溶液的乙醇浓度为30％，于4℃静置过夜，然后5℃下离心，收集沉淀，冷冻干燥得到30％的醇沉组分LBP-Ⅰ；
(6)将LBP-Ⅰ溶于蒸馏水中，经葡聚糖凝胶Sephadex G-100柱层析纯化，用蒸馏水进行洗脱，收集洗脱液，通过苯酚-硫酸法跟踪监测多糖，合并馏分，浓缩并冻干，得到纯化的分子量为92.4kDa的枸杞多糖。
3.根据权利要求2所述的应用，其特征在于，步骤(1)中，脱脂时间为2h，重复次数为3次。
4.根据权利要求2所述的应用，其特征在于，步骤(2)具体为：将干燥后的滤渣按照料液比1:30加入蒸馏水，在90℃下提取2h，重复2次，4000r/min、5℃下离心10min，合并上清液，浓缩至原来体积的四分之一，加入4倍体积的95％乙醇，4℃下静置过夜，4000r/min、5℃下离心10min，收集沉淀。
5.根据权利要求2所述的应用，其特征在于，步骤(3)中，振荡时间为20min，离心转速为4000r/min，离心时间为10min。
6.根据权利要求2所述的应用，其特征在于，步骤(4)中，透析袋的截留分子量为3500Da，透析时间为48h。
7.根据权利要求2所述的应用，其特征在于，步骤(5)中，离心转速为8000r/min，离心时间为15min。
8.根据权利要求2所述的应用，其特征在于，步骤(6)中，蒸馏水的洗脱速度为0.3mL/min。
9.根据权利要求1所述的应用，其特征在于，所述的药物、保健品或食品为缓解运动疲劳的药物、保健品或食品。
10.根据权利要求1所述的应用，其特征在于，所述的药物、保健品或食品为降低血清尿素氮、乳酸和丙二醛含量，提高血糖、超氧化物歧化酶水平的药物、保健品或食品。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110512216.6A CN113116920A (zh) | 2021-05-11 | 2021-05-11 | 枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110512216.6A CN113116920A (zh) | 2021-05-11 | 2021-05-11 | 枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113116920A true CN113116920A (zh) | 2021-07-16 |
Family
ID=76781586
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110512216.6A Pending CN113116920A (zh) | 2021-05-11 | 2021-05-11 | 枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113116920A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114470174A (zh) * | 2022-03-09 | 2022-05-13 | 扬州大学 | 预防及治疗骨质疏松的组合物 |
| CN116195700A (zh) * | 2023-02-01 | 2023-06-02 | 青海大学 | 一种抗疲劳的枸杞多糖泡腾片、制备方法及应用 |
-
2021
- 2021-05-11 CN CN202110512216.6A patent/CN113116920A/zh active Pending
Non-Patent Citations (1)
| Title |
|---|
| GOUYAN LIU ET AL: "Synthesis, stability and anti-fatigue activity of selenium nanoparticles stabilized by Lycium barbarum polysaccharides", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MAROMOLECULES》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114470174A (zh) * | 2022-03-09 | 2022-05-13 | 扬州大学 | 预防及治疗骨质疏松的组合物 |
| CN114470174B (zh) * | 2022-03-09 | 2023-11-10 | 扬州大学 | 预防及治疗骨质疏松的组合物 |
| CN116195700A (zh) * | 2023-02-01 | 2023-06-02 | 青海大学 | 一种抗疲劳的枸杞多糖泡腾片、制备方法及应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113116920A (zh) | 枸杞多糖在制备抗疲劳药物、保健品或食品中的应用 | |
| CN111035649B (zh) | 一种nmn+glp复配营养补充剂及其制备方法和应用 | |
| CN109846940A (zh) | 一种黄精多糖提取物及其提取方法和用途 | |
| US20180193373A1 (en) | Method for Preparing Linseed Polysaccharide Having Antiviral Activity and Immunological Activity, and Use of the Linseed Polysaccharide | |
| CN102349934A (zh) | 一种虫草菌丝体有效成分的提取工艺 | |
| CN104997883A (zh) | 一种桦褐孔菌树莓刺莓组合物、桦褐孔菌树莓刺莓复合口服液及其制备方法和应用 | |
| CN113061195A (zh) | 一种具有降血脂作用的复合多糖及其制备方法和应用 | |
| CN108014150B (zh) | 天然药物组合物在制备抗缺氧和抗辐射的药物或食品中的应用 | |
| CN1235601C (zh) | 一种海洋星虫提取物及制备方法和应用 | |
| CN106214847A (zh) | 一种可增强免疫力的口服液及其制备方法 | |
| CN107412254B (zh) | 金线莲多糖提取物的应用 | |
| CN108743669A (zh) | 用于治疗痛风病的中药组合物及其制备方法和应用 | |
| CN101755953A (zh) | 具有降血糖作用的绿茶提取物及其制备方法 | |
| CN115844937B (zh) | 一种解酒护脑功能的苦苣菜提取物的制备方法及应用 | |
| CN111870627A (zh) | 一种解酒制剂及其制备方法和用途 | |
| CN100509856C (zh) | 一种隐孔菌多糖及其制备与应用 | |
| CN106727902B (zh) | 含冬虫夏草的解酒护肝中药组合物及其应用 | |
| CN113354748B (zh) | 一种铁皮石斛叶多糖及其制备和应用 | |
| CN110483657B (zh) | 一种半边莲均一多糖及其制备方法和应用 | |
| CN107087795A (zh) | 一种抗疲劳耐缺氧保健品组合物 | |
| CN107669795B (zh) | 一种减轻酒精所致化学性肝损伤的组合物 | |
| CN110898180A (zh) | 一种活血降脂减肥组合物及其制备方法 | |
| CN115624565B (zh) | 一种具有减肥及降脂作用的青稞嫩叶水提多糖及其应用 | |
| CN112641829B (zh) | 金果榄水提取物及其新应用 | |
| CN113637089B (zh) | 一种醇水两溶性党参葡果聚糖、制备工艺及抗肿瘤应用 |
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 |
Application publication date: 20210716 |
|
| RJ01 | Rejection of invention patent application after publication |