CN112843068A - 白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 - Google Patents
白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 Download PDFInfo
- Publication number
- CN112843068A CN112843068A CN202110017015.9A CN202110017015A CN112843068A CN 112843068 A CN112843068 A CN 112843068A CN 202110017015 A CN202110017015 A CN 202110017015A CN 112843068 A CN112843068 A CN 112843068A
- Authority
- CN
- China
- Prior art keywords
- nematode
- polyphenol
- nematodes
- product
- application
- 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
- 241000244206 Nematoda Species 0.000 title claims abstract description 77
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 50
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 48
- 241000222640 Polyporus Species 0.000 title description 6
- 238000002360 preparation method Methods 0.000 title description 5
- 230000003712 anti-aging effect Effects 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims description 12
- 241001428259 Hypnea Species 0.000 claims description 9
- 241000588724 Escherichia coli Species 0.000 claims description 8
- 101100098479 Caenorhabditis elegans glp-4 gene Proteins 0.000 claims description 5
- ZRUOGCKQVBXKOD-RUDWDYDBSA-N [3-[(5s,11ar)-3,11a-bis[(2s)-butan-2-yl]-2,4-dihydroxy-5-oxido-1-oxopyrazino[1,2-b][1,4,2]benzodioxazin-5-ium-9-yl]-2-acetyloxy-4,5-dihydroxy-6-(4-hydroxyphenyl)phenyl] acetate Chemical compound O([N@@+]1([O-])C(O)=C(N(C(=O)[C@@]1([C@@H](C)CC)OC1=C2)O)[C@@H](C)CC)C1=CC=C2C(C(=C1OC(C)=O)OC(C)=O)=C(O)C(O)=C1C1=CC=C(O)C=C1 ZRUOGCKQVBXKOD-RUDWDYDBSA-N 0.000 claims description 5
- 235000003642 hunger Nutrition 0.000 claims description 4
- 230000037351 starvation Effects 0.000 claims description 4
- 241000722826 Ardisia Species 0.000 claims description 2
- 241000001493 Sarcodon aspratus Species 0.000 claims description 2
- 239000000679 carrageenan Substances 0.000 claims description 2
- 235000010418 carrageenan Nutrition 0.000 claims description 2
- 229920001525 carrageenan Polymers 0.000 claims description 2
- 229940113118 carrageenan Drugs 0.000 claims description 2
- -1 carrageenan polyphenol Chemical class 0.000 claims description 2
- 235000013373 food additive Nutrition 0.000 claims description 2
- 239000002778 food additive Substances 0.000 claims description 2
- 239000002417 nutraceutical Substances 0.000 claims description 2
- 235000021436 nutraceutical agent Nutrition 0.000 claims description 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 2
- 229940127557 pharmaceutical product Drugs 0.000 claims description 2
- HEXAABFDZKPMQT-UHFFFAOYSA-N 2-[3,11a-di(butan-2-yl)-2,4-dihydroxy-5-oxido-1-oxopyrazino[1,2-b][1,4,2]benzodioxazin-5-ium-9-yl]-3,6-dihydroxy-5-(4-hydroxyphenyl)cyclohexa-2,5-diene-1,4-dione Chemical compound C1=C2OC3(C(C)CC)C(=O)N(O)C(C(C)CC)=C(O)[N+]3([O-])OC2=CC=C1C(C(C=1O)=O)=C(O)C(=O)C=1C1=CC=C(O)C=C1 HEXAABFDZKPMQT-UHFFFAOYSA-N 0.000 claims 2
- 241001149558 Trichoderma virens Species 0.000 claims 1
- 239000000575 pesticide Substances 0.000 abstract description 2
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 25
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 10
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- 241000244203 Caenorhabditis elegans Species 0.000 description 6
- 238000001035 drying Methods 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 230000032683 aging Effects 0.000 description 5
- 235000012000 cholesterol Nutrition 0.000 description 5
- 239000000469 ethanolic extract Substances 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- QEEAPRPFLLJWCF-UHFFFAOYSA-K potassium citrate (anhydrous) Chemical compound [K+].[K+].[K+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O QEEAPRPFLLJWCF-UHFFFAOYSA-K 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 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 3
- 241000238631 Hexapoda Species 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 238000007664 blowing Methods 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
- 229910001628 calcium chloride Inorganic materials 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 241000221198 Basidiomycota Species 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- YASYEJJMZJALEJ-UHFFFAOYSA-N Citric acid monohydrate Chemical compound O.OC(=O)CC(O)(C(O)=O)CC(O)=O YASYEJJMZJALEJ-UHFFFAOYSA-N 0.000 description 2
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 2
- 241001531333 Hyphantria Species 0.000 description 2
- 241001534815 Hypsizygus marmoreus Species 0.000 description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 244000269722 Thea sinensis Species 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000007605 air drying Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007844 bleaching agent Substances 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229960002303 citric acid monohydrate Drugs 0.000 description 2
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000011565 manganese chloride Substances 0.000 description 2
- 229910052603 melanterite Inorganic materials 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 239000001508 potassium citrate Substances 0.000 description 2
- 229960002635 potassium citrate Drugs 0.000 description 2
- 229940050931 potassium citrate monohydrate Drugs 0.000 description 2
- 235000011082 potassium citrates Nutrition 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 229930182947 sarcodictyin Natural products 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000001360 synchronised effect Effects 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 235000013616 tea Nutrition 0.000 description 2
- 229910021654 trace metal Inorganic materials 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 239000000811 xylitol Substances 0.000 description 2
- 235000010447 xylitol Nutrition 0.000 description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 2
- 229960002675 xylitol Drugs 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- 239000011686 zinc sulphate Substances 0.000 description 2
- NUFKRGBSZPCGQB-FLBSXDLDSA-N (3s)-3-amino-4-oxo-4-[[(2r)-1-oxo-1-[(2,2,4,4-tetramethylthietan-3-yl)amino]propan-2-yl]amino]butanoic acid;pentahydrate Chemical compound O.O.O.O.O.OC(=O)C[C@H](N)C(=O)N[C@H](C)C(=O)NC1C(C)(C)SC1(C)C.OC(=O)C[C@H](N)C(=O)N[C@H](C)C(=O)NC1C(C)(C)SC1(C)C NUFKRGBSZPCGQB-FLBSXDLDSA-N 0.000 description 1
- SERLAGPUMNYUCK-YJOKQAJESA-N 6-O-alpha-D-glucopyranosyl-D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O SERLAGPUMNYUCK-YJOKQAJESA-N 0.000 description 1
- WBZFUFAFFUEMEI-UHFFFAOYSA-M Acesulfame k Chemical compound [K+].CC1=CC(=O)[N-]S(=O)(=O)O1 WBZFUFAFFUEMEI-UHFFFAOYSA-M 0.000 description 1
- 241000222501 Agaricaceae Species 0.000 description 1
- 241000222382 Agaricomycotina Species 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 239000004377 Alitame Substances 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000244202 Caenorhabditis Species 0.000 description 1
- 241000253115 Carpesium Species 0.000 description 1
- 241000123346 Chrysosporium Species 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 241001167795 Escherichia coli OP50 Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 239000004378 Glycyrrhizin Substances 0.000 description 1
- 241000959662 Hydnaceae Species 0.000 description 1
- 229920001908 Hydrogenated starch hydrolysate Polymers 0.000 description 1
- 241001428258 Hypnea musciformis Species 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- 102100024295 Maltase-glucoamylase Human genes 0.000 description 1
- 240000001794 Manilkara zapota Species 0.000 description 1
- 235000011339 Manilkara zapota Nutrition 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000318836 Pleurotus nebrodensis Species 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- WINXNKPZLFISPD-UHFFFAOYSA-M Saccharin sodium Chemical compound [Na+].C1=CC=C2C(=O)[N-]S(=O)(=O)C2=C1 WINXNKPZLFISPD-UHFFFAOYSA-M 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000577946 Sarcodon Species 0.000 description 1
- 241000271252 Sarcodon leucopus Species 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- OMHUCGDTACNQEX-OSHKXICASA-N Steviolbioside Natural products O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O OMHUCGDTACNQEX-OSHKXICASA-N 0.000 description 1
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 description 1
- 239000004376 Sucralose Substances 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000566952 Thelephorales Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 235000010358 acesulfame potassium Nutrition 0.000 description 1
- 229960004998 acesulfame potassium Drugs 0.000 description 1
- 239000000619 acesulfame-K Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 235000019409 alitame Nutrition 0.000 description 1
- 108010009985 alitame Proteins 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 108010028144 alpha-Glucosidases Proteins 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910052927 chalcanthite Inorganic materials 0.000 description 1
- JLPRGBMUVNVSKP-AHUXISJXSA-M chembl2368336 Chemical compound [Na+].O([C@H]1[C@@H](O)[C@H](O)[C@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C([O-])=O)[C@@H]1O[C@@H](CO)[C@@H](O)[C@H](O)[C@@H]1O JLPRGBMUVNVSKP-AHUXISJXSA-M 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000000625 cyclamic acid and its Na and Ca salt Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- QGGZBXOADPVUPN-UHFFFAOYSA-N dihydrochalcone Chemical compound C=1C=CC=CC=1C(=O)CCC1=CC=CC=C1 QGGZBXOADPVUPN-UHFFFAOYSA-N 0.000 description 1
- PXLWOFBAEVGBOA-UHFFFAOYSA-N dihydrochalcone Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC(C(=O)CC(O)C=2C=CC(O)=CC=2)=C1O PXLWOFBAEVGBOA-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000009650 gentamicin protection assay Methods 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 229960001462 sodium cyclamate Drugs 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229940013618 stevioside Drugs 0.000 description 1
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 description 1
- 235000019202 steviosides Nutrition 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 235000019408 sucralose Nutrition 0.000 description 1
- BAQAVOSOZGMPRM-QBMZZYIRSA-N sucralose Chemical compound O[C@@H]1[C@@H](O)[C@@H](Cl)[C@@H](CO)O[C@@H]1O[C@@]1(CCl)[C@@H](O)[C@H](O)[C@@H](CCl)O1 BAQAVOSOZGMPRM-QBMZZYIRSA-N 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/539—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines having two or more oxygen atoms in the same ring, e.g. dioxazines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
-
- 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
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
- A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
-
- 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
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Nutrition Science (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Physiology (AREA)
- Botany (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Birds (AREA)
- Microbiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本申请涉及中医药技术领域，尤其涉及一种白肉虎掌菌多酚在制备延长线虫寿命产品中的应用。本申请的方案以线虫为施药对象，研究了白肉虎掌菌的抗衰老功能，发现浓度为200μg/mL的白肉虎掌菌多酚可以显著延长线虫的寿命。
Description
技术领域
本申请涉及中医药技术领域，尤其涉及一种白肉虎掌菌多酚在制备延长线虫寿命产品中的应用。
背景技术
衰老是生命体的机体生理机能持续退化的一个过程，其中伴随着运动能力、环境抵御能力下降及患病率的升高。衰老自由基学说指出，衰老使机体清除自由基的能力逐渐减弱，导致体内活性氧(reactive oxygen species，ROS)积累，进而引起蛋白质、核酸及脂质过氧化，从而加速衰老。抗氧化能力能维持机体自由基产生于清除平衡，对预防衰老具有重要意义。
秀丽隐杆线虫，作为首个完成基因组测序的生物，其基因与人类基因具有60％-80％的同源性。秀丽隐杆线虫是一种身体通透、结构简单、发育快，生命周期短、易培养观察，以细菌为食且培养成本低的模式生物；生命力强，逆境条件下可进入多尔(Dauer)状态，待外界条件满足生存需求时便可直接从Dauer期进入到L4期。线虫的多种优势使得其作为模式生物被广泛的应用于人类复杂疾病的研究，并发展为药物开发的工具。
发明内容
本申请提供了一种白肉虎掌菌多酚在制备延长线虫寿命产品中的应用。
第一方面，本申请提供了白肉虎掌菌多酚在制备延长线虫寿命产品中的应用。
在一个实施例中，所述线虫为秀丽隐杆线虫glp4(bn-2)突变体。
在一个实施例中，所述白肉虎掌菌多酚包括episarcoviolin β、episarcodoninα、sarcoviolin β和sarcodonin α。
第二方面，本申请提供了白肉虎掌菌多酚在制备延缓衰老产品中的应用。
本申请提供的白肉虎掌菌多酚可以和药剂学上可接受的辅料按传统或现代生产工艺制成临床可以接受的口服剂型。如散剂、茶剂、颗粒剂、胶囊剂、片剂、浓缩丸、滴丸、膜剂、煎膏剂、口服液或口服乳等等。所用的药物辅料可以是淀粉、糊精、乳糖、微晶纤维、明胶、蔗糖、硬脂酸镁，赤藓糖醇、木糖醇、山梨糖醇、甘露糖醇、麦芽糖醇、异麦芽糖醇、氢化淀粉水解物、甜菊苷、甜菊双糖苷、二氢查耳酮、甘草甜素、木糖醇、阿斯巴甜、阿力甜、糖精、甜蜜素、安塞蜜、三氯蔗糖等。临床上需要长期用药，制成固体制剂贮存方便，患者服用便利，也更适应市场需求。优选的剂型是胶囊剂、颗粒剂、片剂。
在一个实施例中，所述白肉虎掌菌多酚包括episarcoviolin β、episarcodoninα、sarcoviolin β和sarcodonin α。
在一个实施例中，所述产品为药品、保健食品、食品添加剂中的一种或多种。
第三方面，一种延长线虫寿命的方法，所述方法包括：向线虫施予白肉虎掌菌多酚，以延长所述线虫的寿命。
在一个实施例中，所述方法包括：在线虫处于L4阶段时，向线虫施予白肉虎掌菌多酚。
在一个实施例中，所述方法包括：重悬L1阶段的线虫，使得每毫升线虫重悬液中含有300-500只线虫；向线虫重悬液中加入E.coli OP50，使E.coli OP50终浓度为每毫升1.2x109细菌；在25℃下孵育，直到线虫达到L4阶段；向线虫重悬液中添加的白肉虎掌菌多酚，使得白肉虎掌菌多酚的终浓度为200μg/mL；继续25℃培养线虫，其中，每个一周，向线虫重悬液中加入E.coli OP50，以防止线虫饥饿。
在一个实施例中，所述线虫为秀丽隐杆线虫glp4(bn-2)突变体。
本申请的方案采用线虫为施药对象，研究了白肉虎掌菌的抗衰老功能，发现浓度为200μg/mL的白肉虎掌菌多酚可以显著延长线虫的寿命。
说明书附图
图1为白肉虎掌菌多酚作用下的线虫的生存曲线。
具体实施方式
下面在具体实施例中，对本说明书提供的方案进行举例描述。
多酚是一类重要的抗氧化剂，具有清除自由基、延缓衰老等良好的生理功能。多酚在水果、茶和蘑菇中广泛存在。白肉虎掌菌(Sarcodon leucopus)隶属于担子菌门(Basidiomycota)，伞菌纲(Agaricomycetes)，革菌目(Thelephorales)，齿菌科(Bankeraceae)，肉齿菌属(Sarcodon)，是一种食药两用真菌。本申请的发明人经过实验，发现白肉虎掌菌的多酚提取物具有较好的体外抗氧化及α-葡萄糖苷酶抑制活性，可能具有抗衰老功能，将白肉虎掌菌多酚提取物作为抗衰老成分进行应用可能具有重大的现实意义。
为此，本申请的发明人以线虫为实验对象，进行了如下实验，以研究白肉虎掌菌多酚提取物的抗衰老功能。其中，为方便表述，在下文中，可以将白肉虎掌菌多酚提取物称为白肉虎掌菌多酚。
实施例1，提取白肉虎掌菌多酚
取采集的新鲜白肉虎掌菌子实体，干燥粉碎后称取200g，用无水乙醇按照料液比1:10进行超声提取30min后浸泡过夜，过滤收集滤液，重复提取3次，合并滤液，于36-40℃减压浓缩、干燥过得乙醇提取物13.8g。
使用500mL乙醇/水溶液(70:30)复溶乙醇提取物，将获得的溶液通过D-101大孔树脂(总体积为1500毫升，径高比为1:10)进行分离，滤液反复吸附4次后，分别用蒸馏水、20％的乙醇水和80％的乙醇水进行洗脱，每个洗脱体积为2L。将80％的乙醇水洗脱下来的洗脱液进行合并，并于36-40℃减压浓缩，真空干燥获得富含生物碱的多酚粗提液PAE组分3.79g，于-20℃保存，用于动物实验。
为方便表述，实施例1提取得到的白肉虎掌菌多酚也可以称为PAE。
实施例2，提取白肉虎掌菌多酚
在本实施例中，提取白肉虎掌菌多酚的方法包括以下步骤：
(1)干燥新鲜白肉虎掌菌子实体，进行干燥；粉碎干燥后的白肉虎掌菌子实体，得到药粉；
(2)按照每1g药粉添加10g无水乙醇的比例，往药粉中加入无水乙醇，得到第一混合液；
(3)对所述第一混合液进行超声，然后，静止12小时，过滤，得到滤液；其中，超声的功率为500W,温度为30℃，时间为30分钟。
(4)浓缩并干燥所述滤液，得到乙醇提取物；
(5)按照每1g所述乙醇提取物加入35-38毫升的乙醇体积分数为70％的乙醇水溶液的比例，向所述乙醇提取物加入乙醇体积分数为70％的乙醇水溶液，得到复溶溶液；
(6)将所述复溶溶液通过到填塞有D-101大孔树脂的层析柱(体积为1500毫升，径高比为1：10)，以进行挂柱；然后，依次使用2升蒸馏水、2升乙醇体积分数为20％的乙醇水溶液、2升乙醇体积分数为80％的乙醇水溶液进行洗脱；
(7)对使用乙醇体积分数为80％的乙醇水溶液进行洗脱得到的洗脱液进行浓缩、干燥，得到所述白肉虎掌菌多酚。
为方便表述，实施例2提取得到的白肉虎掌菌多酚也可以称为PAE。
实施例3，白肉虎掌菌多酚成分分析
对实施例1和实施例2提取得到的白肉虎掌菌多酚进行GC-MS分析，结果显示白肉虎掌菌多酚包括episarcoviolin β、episarcodonin α、sarcoviolin β和sarcodonin α等成分。
实施例4，白肉虎掌菌多酚的抗衰老功能鉴定
(1)本实施例采用的仪器与试剂：YW-042-L电热鼓风干燥箱(上海元沃仪器设备有限公司)、CO2培养箱(美国Thermo Fisher公司)、电子分析天平AL240(瑞士梅特勒公司、托利多仪器公司)、CX31RTSF光学显微镜(日本东京奥林巴斯公司)、二级生物安全柜(北京五洲东方科技发展有限公司)、美的微波炉(美的厨房电器制造有限公司)。
(2)本实施例采用的试剂：氯化钙(CaCl2)、硫酸镁(MgSO4)、磷酸氢二钾(K2HPO4)、磷酸二氢钾(KH2PO4)、氢氧化钠(NaOH)、次氯酸钠(NaClO)、磷酸氢二钠(Na2HPO4)、氯化钠(NaCl)、EDTA二钠、FeSO4·7H2O、MnCl2·4H2O、ZnSO4·7H2O、CuSO4·5H2O、一水合柠檬酸、一水合柠檬酸钾均购自上海阿拉丁生化科技股份有限公司，胆固醇和琼脂购自索莱宝生物科技有限公司，蛋白胨和酵母购自北京翰隆达科技发展有限公司。
实施例2制备的白肉虎掌菌多酚。
(3)实验储备液配制：
31、1mol/L CaCl2：5.55g CaCl2，加水定容至100mL，121℃，20min灭菌备用。
32、1mol/LMgSO4：12.04g MgSO4，加水定容至100mL，121℃，20min灭菌备用。
33、5mg/mL胆固醇：取0.25g胆固醇溶解于50mL无水乙醇中，使用针式滤器无菌过滤，待用。
34、M9 buffer的配制：分别取3g KH2PO4，15g Na2HPO4，5g NaCl，1mL 1mol/LMgSO4，加水至1L，高压蒸汽灭菌。
35、5mol/L NaOH：取6g NaOH溶解于24mL蒸馏水待用。
36、10％NaClO：取3mL NaClO溶液加入27mL蒸馏水待用。
37、Bleach溶液(裂解线虫)：现配现用(分别取1mL的5mol/L NaOH和2mL的10％NaClO，加入去离子水定容至10mL)。
38、S basal溶液：5.85g NaCl，1g K2HPO4，6g KH2PO4，1mL 5mg/mL胆固醇，加水至1L，高压蒸汽灭菌。
39、1mol/L柠檬酸钾缓冲液：10g一水合柠檬酸，146.75g一水合柠檬酸钾，PH调至6.0，加水至500mL，高压蒸汽灭菌。
310、微量金属溶液：1.86g EDTA(乙二胺四乙酸)二钠，0.69g FeSO4·7H2O，0.2gMnCl2·4H2O，0.29g ZnSO4·7H2O，0.025g CuSO4·5H2O，加水至1L，高压蒸汽灭菌。
311、S medium：在超净台中加入1L S basal，10mL 1mol/L柠檬酸钾缓冲液，10mL微量金属溶液，3mL 1mol/L CaCl2，3mL 1mol/L MgSO4。
(4)LB培养基配制：
41、液体培养基：分别称取胰蛋白胨10g，酵母提取物5g，NaCl 10g，加入去离子水定容至1L，121℃，20min灭菌后备用。
42、固体培养基：分别称取胰蛋白胨10g，酵母提取物5g，NaCl 10g，琼脂15g，加入去离子水定容至1L，121℃，20min灭菌后倒平板，待凝固后备用。
(5)NGM空白培养基配制：取1L锥形瓶，加入1.5g NaCl，8.5g琼脂，1.25g蛋白胨，加水至500mL。高压灭菌后，在超净台中加入灭菌的500μl1mol/L CaCl2，500μl 1mol/L MgSO4，500μl 5mg/mL胆固醇(用乙醇溶)，12.5mL1mol/L KPO4，混匀，倒入直径为90mm或者60mm的培养皿中，吹风晾干30min，紫外照射5min，平板晾干后备用。
(6)秀丽隐杆线虫的培养
实验中使用的秀丽隐杆线虫为glp4(bn-2)线虫，其为一种温度敏感25℃不孕突变体。线虫食物大肠杆菌OP50(E.coli OP50)是一种尿嘧啶缺陷型菌株。
61、E.coli OP50的培养：取出储存于-80℃的E.coli OP50，用接种环蘸取少量菌液接种到10mL LB液体培养基上，在37℃摇床中培养12h。将摇好的E.coli OP50加入到两瓶500mL LB液体培养基中，37℃摇床孵育过夜。
62、E.coli OP50的制备：将E.coli OP50转移到无菌且称重的离心管中，4000rpm离心20分钟，弃上清，将管中的液体吸净，称量管重，以确定OP50的重量。将OP50沉淀用S-complete重悬为浓度100mg/mL。100mg/mL OP50的浓度对应于每毫升2x1010细菌。制备完成的E.coli OP50储存在4℃冰箱中备用。
63、线虫的一般培养：在无菌条件下用铲子切取适当大小的有线虫的琼脂块，有线虫的一面朝下放置于含有OP50的NGM培养基上，置于20℃培养箱中培养。
64、线虫的同步化培养：用M9 Buffer将体内有卵的虫子从NGM培养基上冲洗下来，收集在15mL离心管中；静置，待虫子全部沉到管底，吸去上层液体后加入M9 Buffer，轻轻颠倒离心管进行清洗，重复2-3次，至上层液体澄清；吸出15mL离心管中上层液体，加入6mL M9Buffer，再加入6mL Bleach溶液，颠倒摇晃离心管；显微镜下观察，待大部分虫子破裂有卵溢出后，离心，4,000rpm，10sec；弃上层液体，加入12mL M9 Buffer，吹散沉淀，离心，4,000rpm，10sec，重复2-3次，弃去上层液体；加入4-6mL M9 Buffer，吹散沉淀，15℃，培养1-2天。直到线虫已经全部孵化并因为饥饿生长发育停止在L1期。将停滞于L1期的线虫在无菌操作台中，滴加至NGM空白培养基上培养。即得到可用于实验的同一代时的线虫。此阶段的线虫可以进行实验，本实验中同步化均采用此法。
(7)秀丽隐杆线虫寿命的测定：显微镜观察卵是否孵化，吸取10μL线虫溶液显微镜计数，确定管中线虫浓度，至少计数10次取平均值。使用S-complete溶液，重悬线虫溶液，使得每毫升300-500只线虫。向线虫溶液中加入OP50，使OP50终浓度为6mg/mL(每毫升1.2x109细菌)。保证线虫处于悬浮状态，向96孔板每孔加入100μl虫液，此时每孔中约30-50只线虫。将96孔板封口以避免污染和蒸发。轻轻摇晃96孔板两三分钟，在25℃下孵育，直到线虫达到L4阶段。添加50-400μg/mL的白肉虎掌菌多酚，设置三个重复并设置对照。封口，轻轻摇晃。记下每个孔中线虫总数，25℃培养。每隔一周，每孔加入5μL 100mg/mL OP50溶液，防止线虫饥饿。每天观察统计线虫死亡条数，直至最后一条线虫死亡为止。线虫死亡标志为摇晃孔板后线虫在培养液中不运动。实验组最后一条线虫死亡的时间为该组线虫的最长寿命，实验重复3次。
(8)秀丽隐杆线虫的安全性评估--存活率测定：同期化的L4期线虫给药24h后统计线虫的存活率，实验重复3次。
寿命分析：
图1为白肉虎掌菌多酚作用下线虫的生存曲线。与对照相比，200μM白肉虎掌菌多酚能延长线虫的寿命；图1示出了肉虎掌菌多酚对秀丽隐杆线虫抗衰老的影响。与对照相比200μg/mL白肉虎掌菌多酚能延长线虫的平均寿命和最长寿命，其中平均寿命延长了2.12天，延长百分比为11.78％；最长寿命延长了2.89天，延长百分比为8.50％。因此，白肉虎掌菌多酚具有抗衰老作用。
以上所述的具体实施方式，对本申请的目的、技术方案和有益效果进行了进一步详细说明，所应理解的是，以上所述仅为本申请的具体实施方式而已，并不用于限定本申请的保护范围，凡在本申请的技术方案的基础之上，所做的任何修改、等同替换、改进等，均应包括在本申请的保护范围之内。
Claims (10)
1.白肉虎掌菌多酚在制备延长线虫寿命产品中的应用。
2.根据权利要求1所述的应用，其特征在于，所述线虫为秀丽隐杆线虫glp4(bn-2)突变体。
3.根据权利要求1或2所述的应用，其特征在于，所述白肉虎掌菌多酚包括episarcoviolinβ、episarcodoninα、sarcoviolinβ和sarcodoninα。
4.白肉虎掌菌多酚在制备延缓衰老产品中的应用。
5.根据权利要求4所述的应用，其特征在于，所述白肉虎掌菌多酚包括episarcoviolinβ、episarcodoninα、sarcoviolinβ和sarcodoninα。
6.根据权利要求4所述的产品，其特征在于，所述产品为药品、保健食品、食品添加剂中的一种或多种。
7.一种延长线虫寿命的方法，其特征在于，所述方法包括：向线虫施予白肉虎掌菌多酚，以延长所述线虫的寿命。
8.根据权利要求7所述的方法，其特征在于，所述方法包括：在线虫处于L4阶段时，向线虫施予白肉虎掌菌多酚。
9.根据权利要求8所述的方法，其特征在于，所述方法包括：
重悬L1阶段的线虫，使得每毫升线虫重悬液中含有300-500只线虫；
向线虫重悬液中加入E.coli OP50，使E.coli OP50终浓度为每毫升1.2x109细菌；
在25℃下孵育，直到线虫达到L4阶段；向线虫重悬液中添加的白肉虎掌菌多酚，使得白肉虎掌菌多酚的终浓度为200μg/mL；
继续25℃培养线虫，其中，每个一周，向线虫重悬液中加入E.coliOP50，以防止线虫饥饿。
10.根据权利要求7-9任一项所述的方法，其特征在于，所述线虫为秀丽隐杆线虫glp4(bn-2)突变体。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110017015.9A CN112843068A (zh) | 2021-01-07 | 2021-01-07 | 白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110017015.9A CN112843068A (zh) | 2021-01-07 | 2021-01-07 | 白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112843068A true CN112843068A (zh) | 2021-05-28 |
Family
ID=76004659
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110017015.9A Pending CN112843068A (zh) | 2021-01-07 | 2021-01-07 | 白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112843068A (zh) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102692493A (zh) * | 2012-01-15 | 2012-09-26 | 河南科技大学 | 一种具有抗细胞畸变植物提取物的快速筛选方法 |
| CN103804392A (zh) * | 2014-02-19 | 2014-05-21 | 中国科学院微生物研究所 | 两种三联苯并二恶嗪衍生物及其应用 |
| CN105309387A (zh) * | 2014-06-04 | 2016-02-10 | 兰州红菌生物技术有限责任公司 | 一种筛选抗衰老药物的试剂盒及其使用方法 |
| CN111635417A (zh) * | 2020-06-12 | 2020-09-08 | 西藏自治区高原生物研究所 | 用于调节肥胖人群肠道菌群的白肉虎掌菌多酚制备方法及其应用 |
| CN111759837A (zh) * | 2020-07-08 | 2020-10-13 | 南昌大学 | 野菊花内酯作为制备秀丽隐杆线虫抗应激药物或延长寿命药物中的应用 |
-
2021
- 2021-01-07 CN CN202110017015.9A patent/CN112843068A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102692493A (zh) * | 2012-01-15 | 2012-09-26 | 河南科技大学 | 一种具有抗细胞畸变植物提取物的快速筛选方法 |
| CN103804392A (zh) * | 2014-02-19 | 2014-05-21 | 中国科学院微生物研究所 | 两种三联苯并二恶嗪衍生物及其应用 |
| CN105309387A (zh) * | 2014-06-04 | 2016-02-10 | 兰州红菌生物技术有限责任公司 | 一种筛选抗衰老药物的试剂盒及其使用方法 |
| CN111635417A (zh) * | 2020-06-12 | 2020-09-08 | 西藏自治区高原生物研究所 | 用于调节肥胖人群肠道菌群的白肉虎掌菌多酚制备方法及其应用 |
| CN111759837A (zh) * | 2020-07-08 | 2020-10-13 | 南昌大学 | 野菊花内酯作为制备秀丽隐杆线虫抗应激药物或延长寿命药物中的应用 |
Non-Patent Citations (7)
| Title |
|---|
| DAVID W LIN 等: "Synthesis-guided structure revision of the sarcodonin, sarcoviolin, and hydnellin natural product family", 《J ORG CHEM》 * |
| KE MA 等: "Two sarcoviolins with antioxidative and α-glucosidase inhibitory activity from the edible mushroom Sarcodon leucopus collected in Tibet", 《J NAT PROD》 * |
| 吴补领: "《老年口腔医学》", 31 January 2019, 西安交通大学出版社 * |
| 孟思进: "《骨骼肌细胞分子生物学研究进展》", 31 December 2010, 武汉理工大学出版社 * |
| 江建勋: "《生物医学》", 31 January 2013, 长春出版社 * |
| 海春旭: "《自由基医学》", 31 December 2006, 第四军医大学出版社 * |
| 王开贞: "《现代女性合理用药指南》", 31 October 2001, 人民卫生出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20180263871A1 (en) | Lightening composition comprising ginseng-derived exosome-like vesicles | |
| WO2017133468A1 (zh) | 白头翁皂苷类化合物作为ev71病毒抑制剂的用途 | |
| CN102008528A (zh) | 复方海参制剂及其制备方法 | |
| JP2007526768A (ja) | イシクラゲ（Ｎｏｓｔｏｃｃｏｍｍｕｎｅ）のコロニー、可食イシクラゲを培養するための方法、可食イシクラゲ製剤および健康を促進するためのそれらの使用 | |
| CN111803533A (zh) | 降血糖、降血脂组合物、制备方法及其应用 | |
| CN103360452B (zh) | 甜瓜蒂四环三萜葫芦素类化合物的制备和应用 | |
| CN116948901B (zh) | 食窦魏斯氏菌d-2胞外多糖在抑制结肠癌细胞中的应用 | |
| CN111419829B (zh) | 和厚朴酚在抑制猪链球菌或其生物被膜中的用途 | |
| CN112843068A (zh) | 白肉虎掌菌多酚在制备延长线虫寿命产品中的应用 | |
| CN108823118B (zh) | 一株能抑制α-葡萄糖苷酶活性的勒克菌及其应用 | |
| CN101723997A (zh) | 葛根素糖基化衍生物、其药物组合物、及其制备方法和用途 | |
| CN102526157B (zh) | 红花提取物在预防或治疗神经退行性疾病中的用途 | |
| CN106511528B (zh) | 一种皱皮木瓜提取物、组合物及用途 | |
| CN113521262A (zh) | 一种具有抗炎效果的溶菌酶制剂 | |
| CN108771070B (zh) | 冠突散囊菌发酵芹菜汁的复合功能饮品及制备方法和用途 | |
| CN102961443B (zh) | 北豆根抗肿瘤提取物、制备方法及用途 | |
| CN104906146B (zh) | 蝙蝠蛾拟青霉菌株ph40在抗抑郁症中的医用用途 | |
| CN111920833A (zh) | 茶藨子叶状层菌发酵液在制备治疗免疫因子风暴药物上的应用 | |
| CN111803528A (zh) | 一种冠突散囊菌制品及其应用 | |
| Tauber et al. | Preparation and some properties of Neisseria gonorrhoeae endotoxin. | |
| CN110664834A (zh) | 苦荞麦多糖制备治疗非酒精性脂肪肝药物中的应用 | |
| WO2006053487A1 (fr) | Utilisation de derives vegetaux de l’anthraquinone et de polysaccharides vegetaux destines au traitement du virus de l’immunodeficience humaine (vih) | |
| KR101835457B1 (ko) | 미세조류 추출물을 포함하는 항암, 항염증 또는 세포 보호용 조성물 | |
| CN109953999B (zh) | 由灵芝多糖、猪苓多糖组成的具有免疫增强作用的组合物 | |
| CN107252454B (zh) | 一种含有黑茶提取物的药品组合物在治疗脓毒症中的应用 |
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 |