CN116942784A - Exosome preparation for relieving liver injury and preparation method and application thereof - Google Patents
Exosome preparation for relieving liver injury and preparation method and application thereof Download PDFInfo
- Publication number
- CN116942784A CN116942784A CN202311018167.6A CN202311018167A CN116942784A CN 116942784 A CN116942784 A CN 116942784A CN 202311018167 A CN202311018167 A CN 202311018167A CN 116942784 A CN116942784 A CN 116942784A
- Authority
- CN
- China
- Prior art keywords
- exosome
- preparation
- parts
- liver injury
- liver
- 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
- 210000001808 exosome Anatomy 0.000 title claims abstract description 157
- 238000002360 preparation method Methods 0.000 title claims abstract description 86
- 206010067125 Liver injury Diseases 0.000 title claims abstract description 54
- 231100000753 hepatic injury Toxicity 0.000 title claims abstract description 52
- 241000196324 Embryophyta Species 0.000 claims abstract description 34
- 235000008708 Morus alba Nutrition 0.000 claims abstract description 28
- 240000000249 Morus alba Species 0.000 claims abstract description 25
- 239000003814 drug Substances 0.000 claims abstract description 24
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 22
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 16
- 241000405414 Rehmannia Species 0.000 claims abstract description 15
- 208000026594 alcoholic fatty liver disease Diseases 0.000 claims abstract description 15
- 208000010706 fatty liver disease Diseases 0.000 claims abstract description 15
- 239000002994 raw material Substances 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 240000000233 Melia azedarach Species 0.000 claims abstract description 13
- 240000002948 Ophiopogon intermedius Species 0.000 claims abstract description 13
- 108010024636 Glutathione Proteins 0.000 claims abstract description 11
- 102000010445 Lactoferrin Human genes 0.000 claims abstract description 11
- 108010063045 Lactoferrin Proteins 0.000 claims abstract description 11
- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 claims abstract description 11
- 229940107187 fructooligosaccharide Drugs 0.000 claims abstract description 11
- 229960003180 glutathione Drugs 0.000 claims abstract description 11
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 claims abstract description 11
- 229940078795 lactoferrin Drugs 0.000 claims abstract description 11
- 235000021242 lactoferrin Nutrition 0.000 claims abstract description 11
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims abstract description 11
- 239000004299 sodium benzoate Substances 0.000 claims abstract description 11
- 235000010234 sodium benzoate Nutrition 0.000 claims abstract description 11
- 239000006188 syrup Substances 0.000 claims abstract description 11
- 235000020357 syrup Nutrition 0.000 claims abstract description 11
- 208000007082 Alcoholic Fatty Liver Diseases 0.000 claims abstract description 10
- 206010016262 Fatty liver alcoholic Diseases 0.000 claims abstract description 10
- 235000003969 glutathione Nutrition 0.000 claims abstract description 4
- 229960003885 sodium benzoate Drugs 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 26
- 239000006228 supernatant Substances 0.000 claims description 26
- 239000000843 powder Substances 0.000 claims description 17
- 238000001914 filtration Methods 0.000 claims description 15
- 239000000706 filtrate Substances 0.000 claims description 12
- 238000009472 formulation Methods 0.000 claims description 12
- 239000008215 water for injection Substances 0.000 claims description 12
- 241000007126 Codonopsis pilosula Species 0.000 claims description 11
- 235000014360 Punica granatum Nutrition 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 239000008213 purified water Substances 0.000 claims description 11
- 241000382455 Angelica sinensis Species 0.000 claims description 10
- 241000207199 Citrus Species 0.000 claims description 10
- 235000020971 citrus fruits Nutrition 0.000 claims description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 10
- 235000006886 Zingiber officinale Nutrition 0.000 claims description 9
- 235000008397 ginger Nutrition 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 150000002772 monosaccharides Chemical class 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 241000218231 Moraceae Species 0.000 claims description 4
- 239000013049 sediment Substances 0.000 claims description 4
- 239000004677 Nylon Substances 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 229920001778 nylon Polymers 0.000 claims description 3
- 244000294611 Punica granatum Species 0.000 claims 2
- 244000273928 Zingiber officinale Species 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 239000004615 ingredient Substances 0.000 claims 1
- 235000012054 meals Nutrition 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 25
- 210000004185 liver Anatomy 0.000 abstract description 25
- 239000008280 blood Substances 0.000 abstract description 11
- 210000004369 blood Anatomy 0.000 abstract description 10
- 235000001287 Guettarda speciosa Nutrition 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 4
- 238000005728 strengthening Methods 0.000 abstract description 4
- 210000000952 spleen Anatomy 0.000 abstract description 2
- 230000001502 supplementing effect Effects 0.000 abstract description 2
- 230000007812 deficiency Effects 0.000 abstract 3
- 240000002045 Guettarda speciosa Species 0.000 abstract 1
- 230000017531 blood circulation Effects 0.000 abstract 1
- 210000003734 kidney Anatomy 0.000 abstract 1
- 230000001737 promoting effect Effects 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 48
- 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 24
- 210000002966 serum Anatomy 0.000 description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 238000012360 testing method Methods 0.000 description 15
- 239000000047 product Substances 0.000 description 14
- 235000012000 cholesterol Nutrition 0.000 description 11
- 210000005228 liver tissue Anatomy 0.000 description 11
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 9
- 108010082126 Alanine transaminase Proteins 0.000 description 9
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 9
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 9
- 241000219991 Lythraceae Species 0.000 description 9
- 230000037396 body weight Effects 0.000 description 9
- 239000000725 suspension Substances 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 241000234314 Zingiber Species 0.000 description 7
- 230000002757 inflammatory effect Effects 0.000 description 7
- 150000002632 lipids Chemical class 0.000 description 7
- 230000003442 weekly effect Effects 0.000 description 7
- 241000287828 Gallus gallus Species 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- 239000008118 PEG 6000 Substances 0.000 description 6
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 description 6
- 229960000583 acetic acid Drugs 0.000 description 6
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 6
- 239000012266 salt solution Substances 0.000 description 6
- 229920006395 saturated elastomer Polymers 0.000 description 6
- 210000000941 bile Anatomy 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 210000005229 liver cell Anatomy 0.000 description 5
- 230000003908 liver function Effects 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 231100000240 steatosis hepatitis Toxicity 0.000 description 5
- 241000125175 Angelica Species 0.000 description 4
- 108010028554 LDL Cholesterol Proteins 0.000 description 4
- 230000001476 alcoholic effect Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000037406 food intake Effects 0.000 description 4
- 235000012631 food intake Nutrition 0.000 description 4
- 235000021588 free fatty acids Nutrition 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 3
- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004889 Interleukin-6 Human genes 0.000 description 3
- 108090001005 Interleukin-6 Proteins 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 241000411851 herbal medicine Species 0.000 description 3
- 208000019423 liver disease Diseases 0.000 description 3
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 230000002633 protecting effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 229940087562 sodium acetate trihydrate Drugs 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 230000007863 steatosis Effects 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000756943 Codonopsis Species 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 208000004930 Fatty Liver Diseases 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 108010023302 HDL Cholesterol Proteins 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 241000711549 Hepacivirus C Species 0.000 description 2
- 206010019708 Hepatic steatosis Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000036528 appetite Effects 0.000 description 2
- 235000019789 appetite Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 208000019425 cirrhosis of liver Diseases 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 231100000234 hepatic damage Toxicity 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000037356 lipid metabolism Effects 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 230000008818 liver damage Effects 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 2
- 230000035807 sensation Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- 210000003934 vacuole Anatomy 0.000 description 2
- 241000208173 Apiaceae Species 0.000 description 1
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 1
- 241000208671 Campanulaceae Species 0.000 description 1
- 102000011727 Caspases Human genes 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 206010019837 Hepatocellular injury Diseases 0.000 description 1
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 241000158728 Meliaceae Species 0.000 description 1
- 241000218213 Morus <angiosperm> Species 0.000 description 1
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 1
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 1
- 235000014676 Phragmites communis Nutrition 0.000 description 1
- IIXHQGSINFQLRR-UHFFFAOYSA-N Piceatannol Natural products Oc1ccc(C=Cc2c(O)c(O)c3CCCCc3c2O)cc1O IIXHQGSINFQLRR-UHFFFAOYSA-N 0.000 description 1
- 241000013557 Plantaginaceae Species 0.000 description 1
- 108090000340 Transaminases Proteins 0.000 description 1
- 102000003929 Transaminases Human genes 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 231100000354 acute hepatitis Toxicity 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 108010081577 aldehyde dehydrogenase (NAD(P)+) Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 102000055102 bcl-2-Associated X Human genes 0.000 description 1
- 108700000707 bcl-2-Associated X Proteins 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 210000004766 cell nucleus structure Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 230000003090 exacerbative effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000031891 intestinal absorption Effects 0.000 description 1
- 210000005027 intestinal barrier Anatomy 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000006372 lipid accumulation Effects 0.000 description 1
- 231100000849 liver cell damage Toxicity 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 108091070501 miRNA Proteins 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- CDRPUGZCRXZLFL-OWOJBTEDSA-N piceatannol Chemical compound OC1=CC(O)=CC(\C=C\C=2C=C(O)C(O)=CC=2)=C1 CDRPUGZCRXZLFL-OWOJBTEDSA-N 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/34—Campanulaceae (Bellflower family)
- A61K36/344—Codonopsis
-
- 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)
-
- 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)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
-
- 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)
- A61K36/58—Meliaceae (Chinaberry or Mahogany family), e.g. Azadirachta (neem)
-
- 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)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
- A61K36/605—Morus (mulberry)
-
- 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)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
-
- 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)
- A61K36/80—Scrophulariaceae (Figwort family)
- A61K36/804—Rehmannia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8968—Ophiopogon (Lilyturf)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9068—Zingiber, e.g. garden ginger
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- 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)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of medicines, and particularly relates to an exosome preparation for relieving liver injury, and a preparation method and application thereof. The exosome preparation for relieving liver injury is mainly prepared from raw materials such as dangshen, angelica, dwarf lilyturf tuber, mulberry, prepared rehmannia root, szechwan chinaberry fruit, plant exosome extract, glutathione, lactoferrin, sodium benzoate, fructo-oligosaccharide, single syrup and the like. The prepared exosome preparation has the effects of strengthening spleen, supplementing qi, strengthening exterior, harmonizing ying and wei, promoting blood circulation, nourishing blood, tonifying liver and kidney, and the like. The exocrine liquid preparation provided by the invention has good treatment effects on liver injury diseases such as spleen-kidney deficiency, qi-blood deficiency, liver-kidney deficiency and the like, particularly has obvious treatment effects on alcoholic fatty liver, and is a novel medicament for treating liver injury with small side effect and obvious treatment effects.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an exosome preparation for relieving liver injury, and a preparation method and application thereof.
Background
According to the global disease burden project, the number of the death of diseases such as acute hepatitis, liver cirrhosis, liver cancer and the like accounts for about 4% of the total number of the death worldwide. It is estimated that in china, about one fifth of the population suffers from some form of liver disease, including Hepatitis B Virus (HBV), hepatitis C Virus (HCV), cirrhosis, liver cancer, non-alcoholic fatty liver disease (NAFLD), alcoholic Liver Disease (ALD), drug-induced liver damage, and the like. Despite the development of vaccine and antiviral drugs that can be related, liver disease still severely affects human health.
At present, research shows that exosomes can have the effect of relieving and treating liver injury. Patent document CN114107179A discloses application of chicken bile exosome in injection medicine for treating chicken liver injury and medicine, and the research shows that the chicken bile exosome has excellent protecting effect on LPS induced liver cell injury, can obviously reduce ALT and AST levels of cell culture supernatant caused by LPS, reduce IL-6, TNF-alpha and iNOS, caspase, bax expression levels and improve Bcl-2 expression levels. The effect of intraperitoneal injection of chicken bile exosomes on the liver injury induced by LPS is observed by using an LPS-induced liver injury animal model, so that the effect of the injection of the chicken bile exosomes on the liver injury induced by LPS can be obviously reduced after one-time administration, and the treatment effect of the chicken bile exosomes on the liver injury is proved.
Patent document CN113171379a discloses an application of mesenchymal stem cell exosomes in preparing a medicament for treating fatty liver disease, wherein the fatty liver disease is alcoholic fatty liver disease. Experiments show that the mesenchymal stem cell exosome has obvious effect in treating fatty liver disease, especially alcoholic fatty liver disease; the combination of mesenchymal stem cell exosomes and piceatannol can remarkably improve the enzyme activity of acetaldehyde dehydrogenase, and delay the occurrence of alcoholic fatty liver disease and alcohol-related cancer.
As research proceeds, researchers have found that plant exosomes have unique advantages over animal exosomes, including undetected by the immune system, higher bioavailability and innocuity. Plant exosomes exhibit better bioavailability than mirnas that are free or bound to proteins. Plant exosomes have been demonstrated to have stability in the gastrointestinal tract, and many studies have shown that plant exosomes can be used in therapeutic applications for oral or intranasal administration. Plant exosomes can target specific organs with higher solubility, higher barrier permeability, faster intestinal absorption and fewer side effects than natural products. Meanwhile, plant-derived exosomes have further advantages in clinical applications: (1) The plant source exosome has the functions of repairing tissue injury and regulating immunity. (2) The plant exosome has stronger adaptability to freeze thawing damage and high in vivo stability. (3) Plant-derived exosomes do not replicate themselves, without potential risk of neoplasia. (4) The exosomes are smaller in cell volume and the membrane surface tissue antigens are low/non-expressed and not prone to elicit an immune response. (5) plant-derived exosomes can penetrate the blood-brain barrier. The plant exosome biomembrane structure can be fused with the organism with high efficiency, and can be used as a carrier of medicines for oral administration of intravenous injection medicines.
However, the utilization and development of plant exosomes are still deficient at present, and research and development of functional products based on the plant exosomes for liver injury have great significance.
Disclosure of Invention
The invention aims to provide an exosome preparation for relieving liver injury, a preparation method and application thereof, and the exosome preparation for relieving liver injury is an oral preparation which is prepared by combining plant exosome and Chinese herbal medicines and can effectively improve liver function and achieve the effects of protecting liver, relieving alcoholic liver and fatty liver. The inventor has the effect of enhancing and improving liver function by optimizing the combination and synergy of the Chinese herbal medicine formula and the plant exosomes, and has remarkable treatment effect on alcoholic fatty liver.
The invention provides an exosome preparation for relieving liver injury, which comprises the following components in parts by weight:
15 to 25 parts of pilose asiabell root, 15 to 25 parts of Chinese angelica, 25 to 35 parts of dwarf lilyturf tuber, 15 to 25 parts of mulberry, 10 to 20 parts of prepared rehmannia root, 0.2 to 0.6 part of szechwan chinaberry fruit, 10 to 20 parts of plant exosome extract, 0.5 to 0.9 part of glutathione, 0.1 to 0.5 part of lactoferrin, 0.01 to 0.05 part of sodium benzoate, 1 to 5 parts of fructo-oligosaccharide and 5 to 15 parts of single syrup.
Further, the exosome preparation for relieving liver injury comprises the following components in parts by weight:
20 parts of codonopsis pilosula, 20 parts of angelica sinensis, 30 parts of dwarf lilyturf tuber, 20 parts of mulberry, 14 parts of prepared rehmannia root, 0.4 part of szechwan chinaberry fruit, 14 parts of plant exosome extract, 0.8 part of glutathione, 0.2 part of lactoferrin, 0.01 part of sodium benzoate, 2 parts of fructo-oligosaccharide and 10 parts of monosaccharide syrup.
Further, the plant exosome extract is formed by mixing (2-4): 3-5): 1-3 by mass ratio of the pomegranate exosome extract, the ginger exosome extract and the citrus exosome extract.
Further, the preparation method of the pomegranate exosome extract comprises the following steps:
a: peeling fresh pomegranate, taking out soft seeds, squeezing juice, centrifuging the squeezed juice supernatant at 4 ℃ and 4000g for 30min, removing residual soft seeds, meat fragments and proteins secreted by the pomegranate seeds in a large amount, filtering the centrifuged supernatant by a 0.45um filter, and removing impurity proteins above 450nm to obtain supernatant;
b: weighing 6000240g of food-grade PEG, 41g of food-grade NaCl29.25g of sodium acetate trihydrate, dissolving in water for injection, adding water for injection to a volume of 1L, uniformly mixing, completely dissolving, refrigerating at 4 ℃ and standing for 24 hours, filtering and sterilizing with a filter membrane with a pore diameter of 0.22um and a filter to prepare 0.5mol of 24% PEG6000 saturated salt solution;
c: the supernatant fluid prepared in the step A and the 24% PEG6000 saturated salt solution prepared in the step B are respectively packaged in 500ml centrifuge tubes according to the volume ratio of 1:1, the pH value is regulated by glacial acetic acid or diluted acetic acid to 4.7-5.0, a tube cover is covered, the mixture is inverted and mixed uniformly, the mixture is transferred into a refrigerator at the temperature of 4 ℃, and the mixture is stood and settled for 12 hours to obtain suspension;
d: directly transferring the suspension obtained in the step C into a centrifuge, centrifuging at 4 ℃ for 30min at 5000g, removing supernatant from the centrifuged suspension, covering a cover, standing upside down for 5min, and completely draining the redundant supernatant; each 100ml of supernatant was resuspended in 2ml of physiological saline, and the resuspended exosome pellet was sealed with parafilm and stored in a-80℃refrigerator.
Further, the preparation method of the ginger exosome extract comprises the following steps:
a: squeezing fresh raw Jiang Baopi, centrifuging at 4deg.C and 4000g for 30min, filtering the supernatant with 0.45um filter to remove impurity proteins above 450nm to obtain supernatant;
b: weighing 6000240g of food-grade PEG, 41g of food-grade NaCl29.25g of sodium acetate trihydrate, dissolving in water for injection, adding water for injection to a volume of 1L, uniformly mixing, completely dissolving, refrigerating at 4 ℃ and standing for 24 hours, filtering and sterilizing with a filter membrane with a pore diameter of 0.22um and a filter to prepare 0.5mol of 24% PEG6000 saturated salt solution;
c: the supernatant fluid prepared in the step A and the 24% PEG6000 saturated salt solution prepared in the step B are respectively packaged in 500ml centrifuge tubes according to the volume ratio of 1:1, the pH value is regulated by glacial acetic acid or diluted acetic acid to 4.7-5.0, a tube cover is covered, the mixture is inverted and mixed uniformly, the mixture is transferred into a refrigerator at the temperature of 4 ℃, and the mixture is stood and settled for 12 hours to obtain suspension;
d: directly transferring the suspension obtained in the step C into a centrifuge, centrifuging at 4 ℃ for 30min at 5000g, removing supernatant from the centrifuged suspension, covering a cover, standing upside down for 5min, and completely draining the redundant supernatant; each 100ml of supernatant was resuspended in 2ml of physiological saline, and the resuspended exosome pellet was sealed with parafilm and stored in a-80℃refrigerator.
Further, the preparation method of the citrus exosome extract comprises the following steps:
a: peeling fresh citrus, taking out soft seeds, squeezing juice, centrifuging 4000g of the squeezed juice supernatant at 4 ℃ for 30min, and filtering the centrifuged supernatant by a 0.45um filter to remove impurity proteins above 450nm to obtain supernatant;
b: weighing 6000240g of food-grade PEG, 41g of food-grade NaCl29.25g of sodium acetate trihydrate, dissolving in water for injection, adding water for injection to a volume of 1L, uniformly mixing, completely dissolving, refrigerating at 4 ℃ and standing for 24 hours, filtering and sterilizing with a filter membrane with a pore diameter of 0.22um and a filter to prepare 0.5mol of 24% PEG6000 saturated salt solution;
c: the supernatant fluid prepared in the step A and the 24% PEG6000 saturated salt solution prepared in the step B are respectively packaged in 500ml centrifuge tubes according to the volume ratio of 1:1, the pH value is regulated by glacial acetic acid or diluted acetic acid to 4.7-5.0, a tube cover is covered, the mixture is inverted and mixed uniformly, the mixture is transferred into a refrigerator at the temperature of 4 ℃, and the mixture is stood and settled for 12 hours to obtain suspension;
d: directly transferring the suspension obtained in the step C into a centrifuge, centrifuging at 4 ℃ for 30min at 5000g, removing supernatant from the centrifuged suspension, covering a cover, standing upside down for 5min, and completely draining the redundant supernatant; each 100ml of supernatant was resuspended in 2ml of physiological saline, and the resuspended exosome pellet was sealed with parafilm and stored in a-80℃refrigerator.
Further, the plant exosome extract is formed by mixing a pomegranate exosome extract, a ginger exosome extract and a citrus exosome extract according to a mass ratio of 3:4:2.
Meanwhile, the invention also provides a preparation method of the exosome preparation for relieving liver injury, which comprises the following steps:
s1, crushing codonopsis pilosula, chinese angelica, dwarf lilyturf tuber, prepared rehmannia root and szechwan chinaberry fruit into coarse powder to obtain raw material coarse powder, adding purified water, heating to 70-80 ℃ for soaking for 5-7 h, filtering, collecting filtrate, concentrating the volume of the filtrate to 1/4 by vacuum freeze concentration, centrifuging, and removing sediment to obtain a traditional Chinese medicine concentrated solution;
s2, drying and crushing mulberries to obtain mulberry powder, then putting the mulberry powder into a stirring tank, adding purified water, heating to 65-70 ℃ for soaking for 3-4 hours, taking supernatant, filtering with 400-mesh, 200-mesh, 100-mesh and 80-mesh nylon gauze to obtain filtrate, and concentrating the filtrate volume to 1/10 to obtain mulberry juice;
and S3, uniformly mixing the traditional Chinese medicine concentrated solution prepared in the step S1 with the mulberry juice prepared in the step S2, then adding the plant exosome extract, glutathione, lactoferrin, sodium benzoate, fructo-oligosaccharide and monosaccharide syrup, uniformly stirring to obtain a raw material liquid, then adding water for injection, wherein the volume ratio of the water for injection to the raw material liquid is 100:1, uniformly stirring to obtain a mixed liquid, then adjusting the pH, filtering by using a 0.22um filter membrane, and filling to obtain the traditional Chinese medicine.
Further, the feed liquid ratio of the raw material coarse powder to the purified water in the step S1 is 1 g:20-30 mL.
Further, in the step S2, the feed liquid ratio of the mulberry powder to the purified water is 1g:50mL.
Further, the pH of the mixed solution in the step S3 is 5.7-7.4.
In addition, the invention also provides the application of the exosome preparation for relieving liver injury or the exosome preparation prepared by the method for relieving liver injury in preparing medicaments for treating liver injury.
Further, the liver injury is alcoholic fatty liver.
The legal standard of each medicine taste in the traditional Chinese medicine composition provided by the invention is from the part 2020 of Chinese pharmacopoeia, and is specifically as follows:
radix codonopsis pilosulae: the product is dried root of Codonopsis pilosula (frank.) Nannf. Of Campanulaceae, codonopsis pilosula (French.) Nannf. Var. Modesta (Nannf.) L.T. shen or Codonopsis pilosula (Chuan) Codonopsis tangshenOliv.). Collected in autumn, washed and dried in the sun. The product meets the relevant regulations under the 293 pages of pilose asiabell root item in the edition 2020 of Chinese pharmacopoeia.
Chinese angelica root: the product is dry root of Angelica sinensis Diels (Oliv.) belonging to Umbelliferae. Digging in the late autumn, removing fibrous roots and silt, bundling into small bundles after water is slightly evaporated, and slowly smoking with smoke and fire. The product meets the relevant regulations under the item of Chinese pharmacopoeia 2020 edition one 139 pages of Chinese angelica.
Radix Ophiopogonis: the product is a dried root tuber of Ker-Gawl. Collected in summer, washed, repeatedly exposed to the sun, piled up until the roots are dried, removed and dried. The product meets the relevant regulations of the first 162 pages of dwarf lilyturf tuber in the 2020 edition of Chinese pharmacopoeia.
Mulberry: the product is dry ear of Morus morusalbal belonging to Moraceae. Harvesting and sun-drying the fruits after the fruits turn red for 4 to 6 months, or sun-drying the fruits after slightly steaming. The product meets the relevant regulations under the 313 pages mulberry item in the first edition of Chinese pharmacopoeia 2020.
Prepared rehmannia root: the product is fresh or dried root tuber of rehmannia Rehmanniaglutinososis Libosch of Scrophulariaceae. Digging in autumn, removing reed heads, fibrous roots and sediment, and fresh using; or baking rehmanniae radix slowly until it is about eight times dry. The preparation method comprises stewing radix rehmanniae under alcohol (general rule 0213) until the alcohol is absorbed, taking out, air drying until the skin mucus is slightly dry, slicing into thick slices or blocks, and drying. The product meets the relevant regulations of 130 pages of prepared rehmannia root in the first edition of Chinese pharmacopoeia 2020.
Fructus Toosendan: the product is obtained by collecting dried mature fruit of Melia toosendan of Meliaceae, removing impurities, and drying. The product meets the relevant regulations under the 44 pages of Chuanzhi in the first edition of Chinese pharmacopoeia 2020.
The exosome preparation provided by the invention is prepared by matching plant exosome with Chinese herbal medicines, and can relieve symptoms of alcoholic liver and fatty liver. The inventor selects the pomegranate exosome extract, the ginger exosome extract and the citrus exosome extract to be compounded according to a certain proportion, and then cooperates with traditional Chinese medicine compositions such as dangshen, angelica, dwarf lilyturf tuber, mulberry, prepared rehmannia root, szechwan chinaberry fruit and the like to play roles in strengthening spleen, supplementing qi and strengthening exterior, harmonizing ying and wei, activating blood and nourishing blood and protecting liver, has better treatment effect on liver injury, and particularly has remarkable treatment effect on alcoholic fatty liver. Meanwhile, the exosome preparation can also solve the problems that active substances or nutrient substances are difficult to be absorbed by liver cells and the like.
Compared with the prior art, the exosome preparation for relieving liver injury has the advantages of good absorption, small side effect, rich raw materials and remarkable effect of relieving liver injury, and is an ideal medicine for relieving liver injury.
Description of the drawings:
FIG. 1 is a graph showing weekly changes in body weight of mice after each group of administration;
FIG. 2 is a graph showing the weekly change in average food intake of mice after each group of administration;
FIG. 3 is a graph showing blood lipid and 5 index plots of mice after each group of administration;
FIG. 4 is a graph showing the concentration of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase in serum of mice of each group after administration;
FIG. 5 is a graph showing the effect of serum inflammatory factors in mice after each group of administration;
FIG. 6 is a graph of liver tissue of mice after each group of doses;
FIG. 7 is a tissue map of liver sections of mice after each group of dosing;
FIG. 8 is a graph of liver slice staining of mice after each group of dosing;
FIG. 9 is a diagram of a stock solution of an exosome preparation for alleviating liver damage prepared in example 2.
Detailed Description
The invention is further illustrated by the following description of specific embodiments, which are not intended to be limiting, and various modifications or improvements can be made by those skilled in the art in light of the basic idea of the invention, but are within the scope of the invention without departing from the basic idea of the invention. The materials and reagents involved in the present invention are all available commercially or by means conventional in the art.
Example 1 an exosome preparation for alleviating liver injury
The exosome preparation for relieving liver injury consists of the following components in parts by weight:
15g of codonopsis pilosula, 15g of angelica, 25g of dwarf lilyturf tuber, 15g of mulberry, 10g of prepared rehmannia root, 0.2g of szechwan chinaberry fruit, 10g of plant exosome extract, 0.5g of glutathione, 0.1g of lactoferrin, 0.01g of sodium benzoate, 2g of fructo-oligosaccharide and 6g of monosaccharide syrup; the plant exosome extract is formed by mixing a pomegranate exosome extract, a ginger exosome extract and a citrus exosome extract according to a mass ratio of 2:5:1.
The preparation method comprises the following steps:
s1, crushing codonopsis pilosula, chinese angelica, dwarf lilyturf tuber, prepared rehmannia root and szechwan chinaberry fruit into coarse powder to obtain raw material coarse powder, adding purified water into the raw material coarse powder, wherein the feed liquid ratio of the raw material coarse powder to the purified water is 1g to 25mL, heating to 75 ℃, soaking for 6 hours, filtering, collecting filtrate, concentrating the filtrate to 1/4 of the volume by vacuum freezing and concentrating, centrifuging, and removing sediment to obtain a traditional Chinese medicine concentrated solution;
s2, drying and crushing mulberries to obtain mulberry powder, then putting the mulberry powder into a stirring tank, adding purified water, heating the mixture to 65 ℃ for 3 hours, taking supernatant, filtering with 400-mesh, 200-mesh, 100-mesh and 80-mesh nylon gauze to obtain filtrate, and concentrating the volume of the filtrate to 1/10 to obtain mulberry juice;
and S3, uniformly mixing the traditional Chinese medicine concentrated solution prepared in the step S1 with the mulberry juice prepared in the step S2, then adding the plant exosome extract, glutathione, lactoferrin, sodium benzoate, fructo-oligosaccharide and monosaccharide syrup, uniformly stirring to obtain a raw material liquid, then adding water for injection, wherein the volume ratio of the water for injection to the raw material liquid is 100:1, uniformly stirring to obtain a mixed solution, adjusting the pH value to 5.7-7.4, filtering by using a 0.22um filter membrane, and filling.
Example 2 exosome preparation for alleviating liver injury
The exosome preparation for relieving liver injury consists of the following components in parts by weight:
20g of codonopsis pilosula, 20g of angelica, 30g of dwarf lilyturf tuber, 20g of mulberry, 14g of prepared rehmannia root, 0.4g of szechwan chinaberry fruit, 14g of plant exosome extract, 0.8g of glutathione, 0.2g of lactoferrin, 0.01g of sodium benzoate, 2 parts of fructo-oligosaccharide and 10g of monosaccharide syrup; the plant exosome extract is formed by mixing a pomegranate exosome extract, a ginger exosome extract and a citrus exosome extract according to a mass ratio of 3:4:2.
The preparation is similar to that of example 1.
Example 3 exosome preparation for alleviating liver injury
The exosome preparation for relieving liver injury consists of the following components in parts by weight:
25g of codonopsis pilosula, 25g of angelica, 35g of dwarf lilyturf tuber, 25g of mulberry, 20g of prepared rehmannia root, 0.6g of szechwan chinaberry fruit, 18g of plant exosome extract, 0.9g of glutathione, 0.3g of lactoferrin, 0.02g of sodium benzoate, 3g of fructo-oligosaccharide and 12g of monosaccharide syrup; the plant exosome extract is formed by mixing a pomegranate exosome extract, a ginger exosome extract and a citrus exosome extract according to a mass ratio of 4:3:3.
The preparation is similar to that of example 1.
Test example one efficacy test of exosome preparation for alleviating liver injury
1. The test raw materials:
1.1, use instrumentation and reagents: electronic balance, ten-thousandth balance, autoclave, electrothermal constant temperature blast drying oven, water purifier, ultra-low temperature refrigerator, high speed refrigerated centrifuge, enzyme-labeled instrument, inverted microscope, ELISA kit (Shanghai enzyme-linked organism), hematoxylin and eosin dye solution (Shanghai Song Van organism)
1.2, dose selection: the human body of the exosome preparation for relieving liver injury is recommended to be 100ml/d, and the test sets 3 dosage groups of the exosome preparation, namely, low dosage group, medium dosage group and high dosage group, wherein 0.2ml, 0.4ml and 0.8ml are equivalent to 0.5, 1 and 2 times of the same dosage of the human body.
1.3, test animals: 40C 57BL/6J male mice, 20-25 g, SPF grade, offered by Huateng biological medicine technologies, inc. of Guangzhou.
1.4, feeding environment: the ambient temperature of the animal house in the test period is 24.0+/-2.0 ℃, the relative humidity is 54-65%, the ventilation times are 15 times/hour, the illumination is 12 hours/darkness is 12 hours, and the brightness is alternate.
2. The test method comprises the following steps:
2.1, establishing an alcoholic fatty liver model mouse model and verifying liver-protecting oral treatment effect:
the exosome preparation for alleviating liver injury prepared in example 2 was selected, and the dosage was low: 0.2ml medium dose: 0.4ml, high dose: 0.8ml, each group was lavaged with 0.4ml, corresponding to 100ml taken daily by an adult.
2.2, administration period and mice modeling:
50 SPF-class 7-8-week-old C57 male mice (20-25 g) are selected, and fed for 7 days in an SPF-class environment in an adaptive manner, and animals eat and drink water freely to feed common feed. Mice were then randomly divided into 2 groups. Blank 10, normal feed; and (5) feeding 40 models by using common feed and alcohol for gastric lavage. The model group is fed for 10 weeks to form an alcoholic fatty liver model, mice in the model group are randomly divided into 5 test groups for test, and the test groups are respectively a blank group, a control group, an exosome preparation low-dose group, an exosome preparation medium-dose group and an exosome preparation high-dose group, 10 mice in each group are numbered and weighed. The corresponding doses of the exosome preparation sample solution for alleviating liver injury prepared in example 2 were subjected to intragastric administration on a daily basis in low, medium and high dose groups, and the blank group and the control group were subjected to intragastric administration with equal volumes of distilled water for 10 weeks. The body weight of the mice was recorded weekly and the amount of lavage was adjusted according to the change in body weight of the mice. After the end of the test, the test animals were fasted for 12 hours. The mice are anesthetized by diethyl ether, then the eyeballs are picked up for blood sampling, liver tissues are rapidly taken out, the middle tissue of the left lobe of the liver is placed in 4% formalin for fixation for histopathological examination, and part of the liver tissues are frozen by liquid nitrogen and then placed in-80 ℃ for freezing for later detection of various biochemical indexes.
2.3, statistical treatment:
the statistical spss method is adopted to carry out the comparison analysis of the significance difference among groups, and the P value is smaller than 0.05, which indicates that the statistical difference exists.
2.4, detection indexes:
2.4.1, body weight and food intake:
each group of mice was weighed at weekly timing. The residual amount of the feed of each group of mice is weighed at regular intervals every week, and the daily administration amount of each group of mice is obtained by subtracting the residual amount from the current day of the week, so that the weekly food intake of each group of mice is obtained.
2.4.2 content of serum inflammatory factors in mice
Blood was collected, centrifuged at 800g/min in a high-speed centrifuge at 4℃for 15min. The supernatant was taken and assayed for mouse serum IL-1. Beta., TNF-alpha., IL-6 and IL-10 levels using ELISA kit (Shanghai ELISA).
2.4.3 ALT and AST content in serum
Blood was collected, centrifuged at 800g/min in a high-speed centrifuge at 4℃for 15min. The supernatant was taken and the levels of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase in the serum of mice were determined using a common kit.
2.4.4 total cholesterol and total triglyceride content in liver homogenates.
Approximately 0.2g-0.5g of liver tissue was weighed using a thousandth analytical balance and homogenized in a sample mill. After centrifugation, the supernatant was taken and the total cholesterol and total triglyceride levels in the mouse liver were determined using the kit.
2.4.5 liver histopathological examination
Dissecting mice, cutting liver tissue with proper size, embedding in embedding box, fixing with formaldehyde, dewatering, embedding, pre-cooling at-20deg.C, and slicing with slicer. The cut pieces were stained with hematoxylin-eosin. Finally, the fiber staining in the liver lobule was observed using an electron microscope, and a photograph was taken using a digital microscope camera.
3. Test results:
3.1, influence of exosome preparation on mice feeding and body weight:
no significant differences were observed between the weekly weights of the blank and the low, medium, and high dose groups with the exosome formulation prior to dosing. The differences between the body weight groups of the mice in each group after the end of the test were statistically significant (0.01 < p < 0.05). The exosome preparation provided by the invention has obvious influence on animal weight under the test condition. The results are shown in Table 1 and FIG. 1. Table 2 and fig. 2 are average weekly feed intake per mouse in each group of exosome formulations under the experimental conditions. The mice in each group given the alcohol lavage had no significant difference in food intake, and were lower than the mice in the control group. The long-term alcoholic diet reduces appetite of mice, while the exosome preparation does not affect appetite of mice with alcoholic fatty liver.
Table 1 exosome formulation change in body weight of mice before and after administration (n=10)
| Group of | Average body weight before administration (g) | Average body weight (g) after administration | 0.05<P<0.01 effective |
| Blank group | 26.08 | 27.46 | P*<0.12 |
| Control group | 30.12 | 32.83 | P**<0.37 |
| Low dose group | 32.11 | 31.69** | P***<0.06 |
| Medium dose group | 35.32 | 34.87* | P****<0.04** |
| High dose group | 33.53 | 32.289*** | P*****<0.07 |
Note that: p <0.06, P <0.04, P <0.07 compared to control group
TABLE 2 weight changes in mice before and after administration of exosome formulations
3.2, influence of exosome preparation on the content of four blood lipids and free fatty acids in mouse serum:
the four levels of blood lipids are usually representative of the change of body lipid metabolism, and free fatty acids are important bioactive substances in the human body, which participate in various physiological functions, have high metabolic activity and are susceptible to fat metabolism. Compared with a blank group, the total cholesterol and total triglyceride content of the control group is obviously increased, the difference has statistical significance, and the alcoholic fatty liver model is established. The serum total cholesterol content of the low and medium dosage groups of the exosome preparation is obviously reduced compared with the control group; the total serum triglyceride content of the exosome preparation group is reduced, and the significance is low. The low-density lipoprotein cholesterol content of each dosage group of the exosome preparation is obviously reduced, and the high-density lipoprotein cholesterol content of the low, medium and high dosage groups of the exosome preparation is obviously increased. The content of free fatty acid in the exosome preparation is obviously reduced in the high-dose group. Indicating that the four index results of total cholesterol, total triglyceride, high density lipoprotein cholesterol, low density lipoprotein cholesterol and free fatty acid are all obviously improved. The results are shown in Table 3 and FIG. 3.
TABLE 3 blood lipid and five indicators of mice after administration
3.3 effect of exosome preparation on total cholesterol and total triglyceride content in mouse liver homogenate:
compared with the control group, the total triglyceride content of liver tissues of each dosage group of the exosome preparation is obviously reduced; the total cholesterol content of liver tissues in the dosage group in the exosome preparation is significantly reduced. I.e. the dosage group in the exosome formulation significantly improved the total cholesterol and total triglyceride content in liver tissue, the results are shown in table 4.
TABLE 4 Total cholesterol and Total Triglycerides levels in mice liver after dosing
| Group of | Total cholesterol (mmol/L) | Total triglycerides (mmol/L) |
| Blank group | 0.0143 | 0.0092 |
| Control group | 0.0238 | 0.0147 |
| Low dose group | 0.0154 | 0.0108 |
| Medium dose group | 0.0149 | 0.0098 |
| High dose group | 0.0167 | 0.0101 |
3.4, influence of exosome preparation on glutamic pyruvic transaminase and glutamic oxaloacetic transaminase content in mouse serum:
glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase are main indexes of liver functions, and compared with a control group, the concentrations of glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase in serum are obviously reduced in the exosome preparation with low dosage, medium dosage and high dosage, so that the increase of the glutamic-pyruvic transaminase and the glutamic-oxaloacetic transaminase induced by alcohol can be reduced in the exosome preparation administration group. The results are shown in Table 5 and FIG. 4.
TABLE 5 glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase concentrations in serum of mice after administration
| Group of | Glutamic pyruvic transaminase (U/L) | Glutamic-oxaloacetic transaminase (U/L) |
| Blank group | 44.18 | 33.11 |
| Control group | 68.42 | 66.28 |
| Low dose group | 49.62 | 43.77 |
| Medium dose group | 45.33 | 38.79 |
| High dose group | 48.47 | 40.15 |
3.5 influence of exosome preparation on mouse serum inflammatory factor:
mice induced by long-term alcoholic diet further cause liver injury due to lipid accumulation, which can form massive inflammatory aggregates, exacerbating liver injury. As shown in table 6 and fig. 5, the high dose of exosome formulation and the dose group in exosome formulation significantly reduced the content of IL-1 β in the serum of mice compared to the blank group. Exosome formulations may protect the liver from secondary injury by reducing inflammatory infiltration in the late stage of alcohol-induced liver injury in mice.
TABLE 6 influence of serum inflammatory factors in mice after administration
| Group of | IL-1β(pg/ml) | IL-6(pg/ml) | TNF-α(pg/ml) | IL-10(pg/ml) |
| Blank group | 98.47 | 100.56 | 550.64 | 384.66 |
| Control group | 131.28 | 106.33 | 590.41 | 355.23 |
| Low dose group | 106.44 | 101.28 | 568.33 | 362.47 |
| Medium dose group | 101.94 | 99.61 | 561.43 | 369.58 |
| High dose group | 105.72 | 102.36 | 574.92 | 371.62 |
3.6 influence of exosome preparation on liver histopathology in mice:
as can be seen from the observation of the overall morphology of the liver of the mice, the liver of the mice in the control group is yellowish, has obvious granular sensation, and the liver of the mice in the blank group and the administration group is ruddy and glossy, and has no granular sensation on the surface. Fig. 7 is a graph of liver section of a mouse, and fig. 8 is a graph of liver section staining of a mouse, wherein as can be seen from fig. 7, liver tissue cells of a normal mouse are orderly arranged, the cell nucleus structure and the cell boundary are clear, and the cell arrangement is orderly without steatosis necrosis; the liver cells of the control group are arranged in disorder, the intercellular boundary is fuzzy, fat vacuoles with different sizes appear in the cells, and the fat degeneration appears; the dosage group in the exosome preparation has lower degree of steatosis than that of the control group, and has obvious improvement; the liver cell structure and the hepatic cable arrangement of the low and medium dose groups of the exosome preparation are basically close to normal, only a small amount of fat vacuoles appear, and the liver disease condition is obviously improved.
Summarizing: the test shows that the low, medium and high doses of the exosome preparation are respectively administered to the mice by oral gavage for 10 weeks, and the doses are respectively 0.2ml, 0.4ml and 0.8ml which are equivalent to 0.5, 1 and 2 times of the same dosage of the human body. Compared with a control group, the exosome preparation is found to be capable of remarkably reducing the total triglyceride content in the serum of the mice in both low and medium dosage groups; the dosage groups of the exosome preparation can obviously reduce the content of the low-density lipoprotein cholesterol in serum, and the dosage groups of the exosome preparation with low and medium dosage can obviously raise the content of the low-density lipoprotein cholesterol in serum. In addition, the total triglyceride and total cholesterol content of liver tissues were significantly reduced after the exosome formulation was administered, indicating that the exosome formulation was able to improve serum and liver lipid levels. Meanwhile, each dosage group of the exosome preparation can obviously reduce the content of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase in the serum of the mice, which indicates that the exosome preparation can protect liver cells in early stage of liver cell damage caused by liver lipid deposition and improve liver functions. The liver tissue sections of the mice were observed by a microscope, and it was found that the hepatic cell steatosis of each dose group of the exosome preparation was reduced, wherein the effect of the medium dose group was most remarkable. The intestinal pathological section is observed through HE staining, the exosome preparation is effectively improved, and the intestinal barrier of the mice with alcoholic fatty liver disease is damaged, wherein the effect of the medium dose group is most obvious.
In a word, four indexes of serum and blood lipid, liver, total triglyceride and total cholesterol, liver function index glutamic oxaloacetic transaminase and serum inflammatory factor are obviously improved after the exosome preparation is given. It can be determined that the exosome preparation has a relieving effect on alcoholic fatty liver through liver lipid metabolism. The development of exosome preparation into products for treating alcoholic fatty liver disease has important significance.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (10)
1. An exosome preparation for relieving liver injury is characterized by comprising the following components in parts by weight:
15 to 25 parts of pilose asiabell root, 15 to 25 parts of Chinese angelica, 25 to 35 parts of dwarf lilyturf tuber, 15 to 25 parts of mulberry, 10 to 20 parts of prepared rehmannia root, 0.2 to 0.6 part of szechwan chinaberry fruit, 10 to 20 parts of plant exosome extract, 0.5 to 0.9 part of glutathione, 0.1 to 0.5 part of lactoferrin, 0.01 to 0.05 part of sodium benzoate, 1 to 5 parts of fructo-oligosaccharide and 5 to 15 parts of single syrup.
2. The exosome formulation for alleviating liver injury of claim 1, comprising the following ingredients in parts by weight:
20 parts of codonopsis pilosula, 20 parts of angelica sinensis, 30 parts of dwarf lilyturf tuber, 20 parts of mulberry, 14 parts of prepared rehmannia root, 0.4 part of szechwan chinaberry fruit, 14 parts of plant exosome extract, 0.8 part of glutathione, 0.2 part of lactoferrin, 0.01 part of sodium benzoate, 2 parts of fructo-oligosaccharide and 10 parts of monosaccharide syrup.
3. The exosome preparation for alleviating liver injury according to claim 1, wherein the plant exosome extract is composed of a mixture of (2-4): (3-5): (1-3) by mass ratio of pomegranate exosome extract, ginger exosome extract and citrus exosome extract.
4. An exosome formulation for alleviating liver injury according to claim 3, wherein said plant exosome extract is comprised of a mixture of pomegranate exosome extract, ginger exosome extract and citrus exosome extract in a mass ratio of 3:4:2.
5. The method for preparing an exosome preparation for alleviating liver injury according to any one of claims 1 to 4, comprising the steps of:
s1, crushing codonopsis pilosula, chinese angelica, dwarf lilyturf tuber, prepared rehmannia root and szechwan chinaberry fruit into coarse powder to obtain raw material coarse powder, adding purified water, heating to 70-80 ℃ for soaking for 5-7 h, filtering, collecting filtrate, concentrating the volume of the filtrate to 1/4 by vacuum freeze concentration, centrifuging, and removing sediment to obtain a traditional Chinese medicine concentrated solution;
s2, drying and crushing mulberries to obtain mulberry powder, then putting the mulberry powder into a stirring tank, adding purified water, heating to 65-70 ℃ for soaking for 3-4 hours, taking supernatant, filtering with 400-mesh, 200-mesh, 100-mesh and 80-mesh nylon gauze to obtain filtrate, and concentrating the filtrate volume to 1/10 to obtain mulberry juice;
and S3, uniformly mixing the traditional Chinese medicine concentrated solution prepared in the step S1 with the mulberry juice prepared in the step S2, then adding the plant exosome extract, glutathione, lactoferrin, sodium benzoate, fructo-oligosaccharide and monosaccharide syrup, uniformly stirring to obtain a raw material liquid, then adding water for injection, wherein the volume ratio of the water for injection to the raw material liquid is 100:1, uniformly stirring to obtain a mixed liquid, then adjusting the pH, filtering by using a 0.22um filter membrane, and filling to obtain the traditional Chinese medicine.
6. The method for producing an exosome preparation for alleviating liver injury according to claim 5, wherein a feed liquid ratio of raw meal to purified water in the step S1 is 1 g:20-30 mL.
7. The method for preparing an exosome preparation for alleviating liver injury according to claim 5, wherein the ratio of mulberry powder to purified water in step S2 is 1g:50ml.
8. The method for producing an exosome preparation for alleviating liver injury according to claim 5, wherein the pH of the mixed solution in step S3 is 5.7 to 7.4.
9. Use of an exosome preparation for alleviating liver injury according to any one of claims 1 to 4 or an exosome preparation prepared by a method for preparing an exosome preparation for alleviating liver injury according to any one of claims 5 to 8 in the preparation of a medicament for treating liver injury.
10. The use according to claim 9, wherein the liver injury is alcoholic fatty liver.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311018167.6A CN116942784A (en) | 2023-08-11 | 2023-08-11 | Exosome preparation for relieving liver injury and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311018167.6A CN116942784A (en) | 2023-08-11 | 2023-08-11 | Exosome preparation for relieving liver injury and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116942784A true CN116942784A (en) | 2023-10-27 |
Family
ID=88446237
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311018167.6A Pending CN116942784A (en) | 2023-08-11 | 2023-08-11 | Exosome preparation for relieving liver injury and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116942784A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1843456A (en) * | 2005-04-07 | 2006-10-11 | 孙毅 | Method for extracting polysaccharide from Chinese formula of 'Yi Guan Jian' |
| CN102366008A (en) * | 2011-06-22 | 2012-03-07 | 王学文 | Life-prolonging and health-preserving tea |
| US20210353747A1 (en) * | 2018-09-06 | 2021-11-18 | Yeditepe Universitesi | Use of plant exosomes for showing modulating effects on immune system cells |
| CN114507634A (en) * | 2022-03-18 | 2022-05-17 | 中南大学湘雅医院 | Method for extracting ginger exosomes |
| CN115120695A (en) * | 2022-08-31 | 2022-09-30 | 健码制药（广东）有限公司 | Licorice probiotic compound preparation CLPP-9 and application thereof in preventing or treating non-alcoholic fatty liver disease |
-
2023
- 2023-08-11 CN CN202311018167.6A patent/CN116942784A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1843456A (en) * | 2005-04-07 | 2006-10-11 | 孙毅 | Method for extracting polysaccharide from Chinese formula of 'Yi Guan Jian' |
| CN102366008A (en) * | 2011-06-22 | 2012-03-07 | 王学文 | Life-prolonging and health-preserving tea |
| US20210353747A1 (en) * | 2018-09-06 | 2021-11-18 | Yeditepe Universitesi | Use of plant exosomes for showing modulating effects on immune system cells |
| CN114507634A (en) * | 2022-03-18 | 2022-05-17 | 中南大学湘雅医院 | Method for extracting ginger exosomes |
| CN115120695A (en) * | 2022-08-31 | 2022-09-30 | 健码制药（广东）有限公司 | Licorice probiotic compound preparation CLPP-9 and application thereof in preventing or treating non-alcoholic fatty liver disease |
Non-Patent Citations (3)
| Title |
|---|
| XIAOYING ZHUANG，等: "Ginger-derived nanoparticles protect against alcohol-induced liver damage", JOURNAL OF EXTRACELLULAR VESICLES, no. 04, pages 1 - 18 * |
| 宁煜: "中西医结合治疗酒精性肝病12 例", 国中西医结合杂志, no. 01, pages 300 * |
| 王昆，等: "加味一贯煎抗脂肪肝的实验性研究", 新中医, vol. 41, no. 12, pages 96 - 98 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103141712B (en) | Compound feed for laying hens in egg producing period and preparation method of feed | |
| CN105053637A (en) | Pig intestine membrane protein-containing pigling mixed feed and preparation method thereof | |
| CN106386694A (en) | Culture method of stocking yellow cattle | |
| WO2016202184A1 (en) | Formula feed for postpartum sows and preparation method therefor | |
| CN103141682B (en) | Composite duck feed containing Chinese medicinal herbs | |
| CN104256168A (en) | Pig functional feed and preparation method thereof | |
| CN104982751A (en) | Rice bran type piglet mixed feed and preparation method thereof | |
| CN106173548A (en) | One carp feed in autumn | |
| CN107349385A (en) | One group is used for fermented tcm composition that live pig nonreactive cultivates and preparation method thereof | |
| CN105360646A (en) | Compound feed for weaned piggies and preparation method of compound feed | |
| CN105410346A (en) | Cultured penaeus vannamei survival rate improving traditional Chinese medicine preparation and preparation method thereof | |
| CN106306533A (en) | Traditional Chinese medicine feed for preventing and treating infectious bronchitis and preparation method thereof | |
| CN106578477A (en) | Special feed for cattle and preparation method of special feed | |
| CN105360597B (en) | A kind of functional pharmaceutical chemistry and preparation method thereof and feeding method promoting soft-shelled turtle quality | |
| CN108813501B (en) | Health-preserving honey paste with functions of clearing heat, moistening lung, relieving cough, reducing phlegm, relieving asthma and regulating human body functions | |
| EP3881685A1 (en) | Ceratonia siliqua fruit composition and preparation method therefor and use thereof | |
| CN112807348A (en) | Traditional Chinese medicine compound for resisting diseases and promoting growth as well as preparation and application thereof | |
| CN104489172A (en) | Acanthopanax health tea for improving immunity and preparation method thereof | |
| KR20150051597A (en) | Food Composition for improving liver function containing extract of Arctiumlappa L. | |
| CN103920140B (en) | A kind of people is with blood sugar lowering Weight-reducing and lipid-lowering compound preparation | |
| CN104161207B (en) | The feed additive of a kind of anti-duck bacterial disease, preparation method and application | |
| CN105833075A (en) | Traditional Chinese medicine preparation for preventing rabbit haemorrhagic diseases | |
| CN116942784A (en) | Exosome preparation for relieving liver injury and preparation method and application thereof | |
| CN105053638A (en) | Meat duck mixed feed for meat ducks of 0-21 days old and 22-45 days old and preparation method thereof | |
| CN106173240A (en) | A kind of liver protecting function and service plant extract feed additive and preparation technology |
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 |