TSAB NTAWV

Lub hom phiaj:Maclura pomifera (Rafin.) Schneider yog ib hom kab mob thoob plaws ntiaj teb, uas yog nquag siv los ua ornamental. Cov kev tshawb fawb yav dhau los tau qhia tias M. pomifera txiv hmab txiv ntoo yog nplua nuj nyob rau hauv prenylated isoflavonoids, nthuav tawm cov kev ua ub no tseem ceeb, thiab muaj txiaj ntsig zoo, tshwj xeeb tshaj yog thaum siv tshuaj pleev ib ce. Xav tias phenolic tebchaw yog qhov tseem ceeb hauv kev txhim kho cov khoom siv tshuaj pleev ib ce, qhov kev tshawb fawb no tau tshawb xyuas qhov kev tiv thaiv kev laus ntawm M. pomifera 80 feem pua ​​​​methanolic extract (MPM) los ntawm kev ntsuam xyuas antioxidant thiab extracellular matrix-degrading enzymes inhibiting kev ua si.

Glycoside ntawm cistanche tuaj yeem ua rau muaj kev ua haujlwm ntawm SOD hauv plawv thiab daim siab cov ntaub so ntswg, thiab txo cov ntsiab lus ntawm lipofuscin thiab MDA hauv txhua cov ntaub so ntswg, tshem tawm ntau yam reactive oxygen radicals (OH-, H₂O₂, thiab lwm yam) thiab tiv thaiv DNA puas. los ntawm OH-radicals. Cistanche phenylethanoid glycosides muaj peev xwm tshem tawm cov dawb radicals, muaj peev xwm txo tau ntau dua li cov vitamin C, txhim kho cov haujlwm ntawm SOD hauv cov phev ncua, txo cov ntsiab lus ntawm MDA, thiab muaj qee yam kev tiv thaiv ntawm cov phev ua haujlwm. Cistanche polysaccharides tuaj yeem txhim kho kev ua haujlwm ntawm SOD thiab GSH-Px hauv erythrocytes thiab ntsws cov ntaub so ntswg ntawm cov nas senescent sim los ntawm D-galactose, nrog rau txo cov ntsiab lus ntawm MDA thiab collagen hauv lub ntsws thiab ntshav, thiab nce cov ntsiab lus ntawm elastin, muaj Cov nyhuv scavenging zoo ntawm DPPH, ncua lub sijhawm ntawm hypoxia hauv cov nas senescent, txhim kho cov haujlwm ntawm SOD hauv cov ntshav, thiab ncua lub physiological degeneration ntawm lub ntsws hauv kev sim cov nas nrog cellular morphological degeneration, kev sim tau pom tias Cistanche muaj peev xwm antioxidant zoo. thiab muaj peev xwm los ua ib qho tshuaj tiv thaiv thiab kho cov kab mob ntawm daim tawv nqaij laus. Nyob rau tib lub sijhawm, echinacoside hauv Cistanche muaj lub peev xwm tseem ceeb txhawm rau txhawm rau DPPH dawb radicals thiab muaj peev xwm tshem tawm cov pa oxygen reactive thiab tiv thaiv dawb radical-induced collagen degradation, thiab kuj muaj kev kho zoo ntawm thymine dawb radical anion puas.

Nyem rau ntawm Cistanche Tubulosa Ntxiv

【Yog xav paub ntxiv:george.deng@wecistanche.com / WhatApp:86 13632399501 】

Cov ntaub ntawv thiab cov txheej txheem:Rau txoj kev tshawb no, qhov inhibitory muaj peev xwm ntawm 80 feem pua ​​MPM tiv thaiv cov enzymes sib txawv cuam tshuam nrog cov txheej txheem kev laus raug soj ntsuam. Muab lub luag haujlwm tsis sib xws ntawm oxidative kev nyuaj siab hauv kev laus, hauv vitro antioxidant kev sim tau ua haujlwm ib yam. Ntxiv mus, osajin tau txiav txim siab tias yog qhov tseem ceeb bioactive isoflavonoid ntawm cov qauv los ntawm kev ua haujlwm siab nyias-txheej chromatography tsom.

Cov txiaj ntsig:Cov txiaj ntsig ntawm kev sib txawv ntawm cov tshuaj tua kab mob antioxidant tau nthuav tawm qhov tseem ceeb antioxidant peev xwm ntawm cov extract. Qhov inhibition muaj peev xwm ntawm MPM tawm tsam hyaluronidase, collagenase, thiab elastase enzymes, uas ncaj qha txuas mus rau qhov nrawm ntawm cov txheej txheem kev laus, tau tshawb xyuas thiab cov txiaj ntsig tau qhia tias MPM inhibited cov enzymes saum toj no meej. MPM muaj cov ntsiab lus phenolic thiab flavonoid tshwj xeeb; 113.92 ± 2.26 mg gallic acid sib npaug / g thiab 66.41 ± 0.74 mg QE / g, feem. Thaum tag nrho cov tshuaj tiv thaiv kab mob antioxidant tau raug txiav txim siab, nws muaj peev xwm hais tias MPM yog tus neeg sawv cev tiv thaiv kev laus.

Xaus:Yog li ntawd, txoj kev tshawb no nthuav tawm tias cov txiv hmab txiv ntoo ntawm M. pomifera muaj peev xwm tiv thaiv kev laus thiab tuaj yeem siv rau lub hom phiaj no.

Ntsiab lus:Maclura pomifera, anti-aging, antioxidants, HPTLC, osajin

Taw qhia

Ib yam li ntawd, txhua yam hauv nruab nrog cev, suav nrog tib neeg cov tawv nqaij tau dhau los ua ntau yam kev hloov pauv ntawm lub cev nrog lub hnub nyoog.1 Ob chav kawm ntawm kev laus muaj nyob: kev laus hauv nruab nrab yog tswj los ntawm noob caj noob ces thiab extrinsic aging yog lub ntuj tsim los ntawm kev hloov kho lub cev vim kev puas tsuaj ntawm ib puag ncig yam xws li ultraviolet (UV. ) hluav taws xob, tshuaj lom neeg, thiab kev haus luam yeeb.1,2 Cov kab mob ntawm cov hlab ntsha thiab cov qog nqaij hlav degradation, fibrous cov ntaub so ntswg poob, thiab txo cov cell regeneration yog qhov tseem ceeb ntawm kev laus, 3 uas ua rau nce hauv cov ntaub so ntswg degeneration, wrinkles, thiab decrement ntawm extracellular matrix (ECM) ).4

Raws li lub cev loj tshaj plaws, daim tawv nqaij muaj ntau lub luag haujlwm xws li kev tiv thaiv, kev tswj hwm ntawm lub cev kub, thiab kev paub txog kev hnov ​​​​qab.5 Cov tawv nqaij muaj cov epidermis, dermis, thiab cov ntaub so ntswg subcutaneous thiab yog thawj qhov thaiv ntawm tib neeg lub cev thiab sab nrauv. ib puag ncig.6,7 ECM yog lub tsev loj tshaj plaws ntawm cov dermis thiab txhawb kev loj hlob thiab elasticity los ntawm kev nthuav qhia cov qauv scaffold.8 Collagen, elastin, thiab fibronectin, uas yog tsim los ntawm dermal fibroblasts, ua ECM thiab fused nrog proteoglycans.5 Collagen yog ib qho tseem ceeb protein uas muaj kwv yees li ntawm 25-35 feem pua ​​​​ntawm tag nrho cov ntsiab lus ntawm cov protein nyob hauv lub cev thiab pom nyob rau hauv lub extracellular qhov chaw ntawm ntau hom tsiaj connective cov ntaub so ntswg.3 Ib qho ntawm cov ua rau ntawm daim tawv nqaij laus thiab wrinkle tsim yog kev hloov ntawm collagen. Structure.1 Elastin yog cov protein, uas muaj qhov tshwj xeeb physiological elasticity rau ntawm daim tawv nqaij thiab muaj nyob rau hauv ob peb cov ntaub so ntswg.6 daim tawv nqaij hydration, thiab ua kom daim tawv nqaij du, moist, thiab lubricated yog ib qho tseem ceeb uas tiv thaiv daim tawv nqaij laus, thiab ib tug loj. glycosaminoglycan (GAG), hyaluronic acid, plays lub luag haujlwm tseem ceeb hauv cov dej num no.8 Cov ntsiab lus tseem ceeb no yog degraded los ntawm hyaluronidase, collagenase, thiab elastase enzymes, yog li, ua rau kom nrawm ntawm daim tawv nqaij laus. Tsis tas li ntawd, raug cov kab mob, kev ua qias tuaj, hluav taws xob ionizing, tshuaj lom neeg, thiab co toxins ua rau tsim cov pa oxygen reactive hom (ROS) thiab nws yog qhov tsim kev puas tsuaj rau daim tawv nqaij laus.9 ROS tuaj yeem pib txoj hauv kev nyuaj molecular thiab vim li ntawd, collagenase, elastase. , thiab hyaluronidase kev ua haujlwm tuaj yeem nce ntxiv, ua rau pom ECM kev puas tsuaj thiab kev hloov kho ntawm daim tawv nqaij.10 Rau cov laj thawj tau hais, cov neeg ua haujlwm tshiab tshiab, uas txo qis ROS tsim thiab inhibit ECM degrading proteases, tuaj yeem ncua kev laus ntawm daim tawv nqaij.11

Maclura pomifera (Rafin.) Schneider belongs rau Moraceae los yog mulberry tsev neeg thiab tseem hu ua osage txiv kab ntxwv tsob ntoo, cultivated yuav luag thoob ntiaj teb.12 M. pomifera muaj ntau yam kab mob xws li tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, cytotoxic, antitumor, estrogenic, thiab antimalarial13 vim nws cov isoflavones prenylated, piv txwv li osajin thiab pomiferin, uas yog suav hais tias yog cov metabolites loj ntawm cov txiv hmab txiv ntoo.14 Nyob rau hauv anti-aging cosmetic ntau lawm, phenolic tebchaw yog ib qho tseem ceeb ntawm ntuj tsim. Yog li, muaj kev txaus siab rau kev kawm phenolic compound-nplua nuj nroj tsuag xws li M. pomifera rau cov dej num no. Cov kev tshawb fawb yav dhau los tau pom tias isoflavones ntawm Maclura nce qib kev qhia ntawm collagen, elastin, thiab fibrillin piv lossis zoo dua rau qhov sib npaug ntawm cov retinol. Li no, nws yuav xav tias Maclura isoflavones yog cov muaj zog ECM protein stimulants.15 Xav txog cov ntaub ntawv no, txoj kev tshawb no tsom mus soj ntsuam cov kev tiv thaiv kev laus ntawm M. pomifera 80 feem pua ​​​​methanolic extract (MPM) los ntawm kev tshawb nrhiav nws lub peev xwm rau antioxidant bioactivity thiab inhibiting. ECM degrading enzymes. Tsis tas li ntawd, kev soj ntsuam ntau ntawm cov khoom siv bioactive tseem ceeb ntawm cov extract, osajin tau ntsuas los ntawm kev ua haujlwm siab nyias txheej chromatography (HPTLC), thiab tag nrho cov ntsiab lus phenolic thiab tag nrho cov ntsiab lus flavonoid kev soj ntsuam tau ua rau kev nkag siab ntau dua ntawm phenolic profile. Cov txiaj ntsig tau pom tias M. pomifera tej zaum yuav yog ib qho tseem ceeb ntawm cov khoom siv los tiv thaiv kev laus.

Cov khoom siv thiab cov txheej txheem

Tshuaj

Sigma Chemical Co. (St. Louis, MO, USA) tau them tag nrho cov enzymes, tshuaj, thiab cov ntaub ntawv pov thawj ua haujlwm hauv kev sim. Qhov zoo ntawm tag nrho cov tshuaj yog ntawm analytical qib.

Cov khoom cog

Cov txiv hmab txiv ntoo ntawm M. pomifera tau sau los ntawm Uşak, Türkiye, thaum lub Tsib Hlis 2020. Dr. Hilal Bardakcı tau ua cov txheej txheem kev txheeb xyuas cov nroj tsuag ntawm cov qauv cog. Ib daim ntawv pov thawj ntawm cov nroj tsuag tau muab tso rau hauv Acıbadem University, Herbarium ntawm Kws Qhia Tshuaj (ACUPH 00002).

Kev npaj ntawm extracts

Cov txiv hmab txiv ntoo tau muab cais rau hauv me me pieces thiab dhau los ntawm ib tug blender. Cov txiv hmab txiv ntoo (6.45 kg) tau macerated nrog 3125 mL ntawm 80 feem pua ​​​​methanol (MeOH) los ntawm kev siv lub tshuab co co rau peb hnub ntawm chav tsev kub hauv qhov chaw tsaus. Lub macerate tau lim thiab cov txheej txheem no tau rov ua dua ob zaug. Cov filtrates tau muab sib sau ua ke thiab, tom qab ntawd, methanol tau evaporated nyob rau hauv lub rotary evaporator. Cov kua nplaum methanolic extract tau lyophilized (cov qoob loo yog 204.37 g, 3.16 feem pua) thiab khaws cia ntawm -20 degree (MPM).

Cov txheej txheem Quantification ntawm cov tshuaj bioactive loj los ntawm HPTLC

Tag nrho cov tshuaj thiab cov tshuaj reagents siv tau yog qib ntsuas. Chloroform (CHCl3) thiab ethyl acetate (EtOAc) tau yuav los ntawm Sigma-Aldrich. Ib qho kev lag luam muaj qauv ntawm osajin tau yuav los ntawm Sigma-Aldrich (SMB {{10}}}0092). Kev tshuaj xyuas HPTLC tau ua tiav ntawm 20 cm × 10 cm iav HPTLC silica gel 60 F254 daim hlau (Merck, Darmstadt, Lub teb chaws Yelemees). Osajin cov ntsiab lus hauv MPM tau txiav txim siab los ntawm CAMAG HPTLC analytical system. Lub sijhawm txawb siv nyob rau hauv txoj kev tshawb fawb tam sim no tau piav qhia yav dhau los los ntawm Bozkurt et al.16 thaum lub sijhawm kev sib cais ntawm cov ntsiab lus tseem ceeb ntawm M. pomifera. 10 mg / mL MeOH extract tau siv los ua cov tshuaj ntsuam xyuas tshuaj. Cov txheej txheem kev daws teeb meem (0.5 mg / mL) ntawm osajin tau npaj siv 2 mL ntawm acetone. Ib qho kev daws teeb meem ua haujlwm nrog 50 µg / mL concentration ntawm cov qauv sib xyaw tau npaj los ntawm dilution nrog acetone los ntawm cov tshuaj hauv Tshuag. Txhua tus qauv raug lim los ntawm 0.45 µm syringe lim. 10 μL ntawm cov extract nrog rau tsawg kawg tsib qhov sib txawv ntawm cov txheej txheem kev daws teeb meem (3.3-4.7 μL) tau siv rau hauv daim ntawv bands ntawm 8 hli ntev ntawm silica gel iav HPTLC daim hlau 60 F254 nrog CAMAG Tsis Siv Neeg TLC Sampler IV. Kev txhim kho tau ua tiav hauv CAMAG Automatic Developing Chamber-2 (ADC- 2) thiab theem mobile yog CHCl3: EtOAc [8:2 (v/v)]. Chamber tau saturated rau 10 min thiab lub phaj yog preconditioned rau 5 min ua ntej txoj kev loj hlob. Cov av noo tau tswj los ntawm ADC-2 siv MgCl2 (33 feem pua ​​RH) rau 10 feeb. Kev ntsuas densitometric tau ua tiav siv CAMAG TLC Scanner IV hauv hom fluorescence. Qhov loj me me tau khaws cia ntawm 5 × 0.2 hli, micro, thiab qhov ntsuas ceev tau teeb tsa ntawm 20 mm / s. Cov ntsiab lus txheem tau txais los ntawm kev sib piv cov cheeb tsam hauv qab lub receiver operating yam ntxwv nkhaus (AUCs) nrog rau calibration nkhaus ntawm cov qauv ntawm 280 nm. Lub xub ntiag ntawm cov qauv hauv cov extract tau paub tseeb los ntawm kev sib piv ntawm ob qho tib si khaws cia yam tseem ceeb (Rf) thiab overlaying UV spectra ntawm txhua cov extract thiab cov qauv. Qhov ntau ntawm osajin tau txiav txim siab los ntawm kev sib piv ntawm qhov siv ntawm diffusely reflected lub teeb los ntawm cov extract thiab feem ntau nrog cov qauv compound.

Osajin cov ntsiab lus nyob rau hauv crude cog extract tau ntsuas siv HPTLC-densitometry. Tus nqi Rf ntawm tus qauv osajin tau pom tias yog 0.556. Qhov tshwm sim ntawm osajin hauv cov qauv kuaj tau txheeb xyuas los ntawm kev sib piv ntawm lawv cov txiaj ntsig Rf thiab sib tshooj lawv cov UV spectra (Daim duab 1). Quantification tau them los ntawm kev sib piv cov AUCs ntawm cov qauv nrog cov calibration nkhaus tau siv cov qauv compound osajin. calibration muaj nuj nqi yog y=2.268*10-8x. Qhov sib txheeb coefficient (R) thiab qhov sib txawv ntawm qhov sib txawv ntawm qhov kev ua haujlwm calibration yog 0.998 feem pua ​​​​thiab 1.06 feem pua, raws li. HPTLC tsom xam pom tias MPM muaj 0.22 feem pua ​​(w/w) osajin. Cov txiaj ntsig ntawm HPTLC txoj kev tshawb fawb tau muab rau hauv Table 1.

Phenolic profile soj ntsuam

Tag nrho cov ntsiab lus phenolic xeem

Qhov kev ntsuam xyuas tau ua los ntsuas tag nrho cov ntsiab lus phenolic ntawm cov qauv los ntawm Folin-Ciocalteu txoj kev, raws li yav dhau los siv los ntawm Kurt-Celep li al. ), thiab 100 μL FCR (Folin-Ciocalteu reagent) diluted nrog H2 O (1: 9). Tom qab incubation rau 30 min ntawm 45 degree, lub absorbance ntawm cov mixs tau nyeem spectrophotometrically ntawm 765 nm. Cov txiaj ntsig tau qhia tias yog mg gallic acid sib npaug (GAE) ib g ntawm extract.

Kev ntsuam xyuas tag nrho cov ntsiab lus flavonoid

Kev ntsuas ntawm tag nrho cov ntsiab lus flavonoid ntawm cov feem yog ua raws li ib txoj kev uas yav tas los qhia los ntawm Bardakci li al.18 Ncua tseg, freshly npaj 1 M CH3 COONa thiab 10 feem pua ​​AlCl3 tau tov nrog cov qauv. Tom qab ntawd, 30 min ntawm incubation ntawm mixs yog ua nyob rau hauv chav tsev kub thiab nyob rau hauv qhov tsaus ntuj nti. Tom qab cov txheej txheem incubation, lub absorbance yog xam ntawm 415 nm. Cov txiaj ntsig tau lees paub tias yog mg quercetin sib npaug (QE) hauv 1 g ntawm cov qauv.

Kev txiav txim siab ntawm kev ua haujlwm hauv vitro antioxidant

2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity test

Txhawm rau txiav txim siab DPPH radical scavenging kev ua ub no, kev sib xyaw ua ke ntawm cov qauv tshiab diluted cov qauv kev daws teeb meem (ntau ntau yam npaj los ntawm 1 mg / mL cov tshuaj hauv Tshuag) thiab cov tshuaj methanolic DPPH (100 mM) tau ua tiav. Tom qab lub sij hawm incubation ntawm chav tsev kub rau 45 min, lub absorbance tau nyeem ntawm 517 nm. Butylated hydroxytoluene (BHT) tau siv los ua ib qho kev sib txuas kom tau txais kev ntsuas nkhaus. IC50 qhov tseem ceeb ntawm cov txiaj ntsig tau teev tseg raws li µg/mL.19

Ferric txo antioxidant zog (FRAP) kuaj

Yuav kom tau txais FRAP reagent, 25 mL ntawm 300 mM acetate buffer (pH 3.6), 2.5 mL TPTZ [2,4,6-tris (2-}pyridyl)-s-triazine] , thiab 2.5 mL ntawm FeCl3 .6H2 O (20 mM) tau sib xyaw. Tom qab ntawd, 10 mL ntawm cov qauv yog ntxiv rau 260 mL ntawm FRAP reagent thiab diluted rau 300 mL nrog dej distilled nyob rau hauv 96 zoo-phaj. Tom qab incubating rau 30 min ntawm 37 degree, qhov ntsuas ntawm absorbance tau ua ntawm 593 nm. BHT tau siv los ua qhov sib xyaw ua ke. Ferrous chloride tov (0.252 mM) tau siv kom tau ib qho qauv nkhaus thiab cov txiaj ntsig tau muab raws li mM FeSO4 hauv 1 g qhuav extract.20

Cupric-txo antioxidant muaj peev xwm (CUPRAC) kuaj

Kev ntsuas CUPRAC tau kwv yees raws li txoj kev tau piav qhia ua ntej los ntawm Barak li al.21 Qhov sib npaug ntawm 10 mM CuSO4, neocupraine, thiab ammonium acetate buffer (85 mL) tau muab sib xyaw rau hauv 96- zoo phaj. Tom qab ntawd, 51 mL ntawm distilled dej thiab 43 mL ntawm cov qauv kev daws tau ntxiv rau qhov sib tov, raws li. Tom qab incubation rau 20 min, lub absorbance tau nyeem ntawm 450 nm. Cov txiaj ntsig tau hais tias yog mg ascorbic acid sib npaug hauv 1 g qhuav extract.

Kev txiav txim siab ntawm tag nrho cov khoom muaj peev xwm antioxidant (TOAC).

Tag nrho cov tshuaj antioxidant muaj peev xwm raug xam raws li txoj kev phosphomolybdenum piav qhia ua ntej los ntawm Barak li al.22 Ua ntej, kom tau txais TOAC tov; 28 mM sodium phosphate monobasic, 4 mM ammonium molybdate thiab 600 mM H2 SO4 tau sib xyaw. Tom qab ntawd, 300 µL ntawm TOAC tov tau tov nrog 30 µL ntawm cov qauv kev daws teeb meem hauv 96 lub phaj noj qab haus huv. Tom qab lub sij hawm incubation ntawm 95 degree rau 90 min, lub absorbance tau nyeem ntawm 695 nm. Ascorbic acid tau siv kom tau txais tus qauv nkhaus thiab cov txiaj ntsig tau suav ua mg Trolox sib npaug hauv 1 g qhuav extract.

Inhibitory kev ua ntawm daim tawv nqaij laus-txog enzymes

Anti-collagenase kev ua haujlwm

Txhawm rau ntsuas qhov kev ua haujlwm los tiv thaiv collagenase ntawm MPM, 50 mM tricine buffer tov (pH: 7.5) tau npaj (400 mM NaCl thiab 10 mM CaCl2 ). Clostridium histolyticum (ChC - EC. 3.4.23.3) tau siv los ua lub hauv paus ntawm collagenase, uas tau yaj hauv 50 mM tricine buffer tov kom ua tiav qhov pib concentration ntawm 0.8 U / mL. 2 mM ntawm N-[3-(2-furyl) acryloyl]-Leu-Gly-Pro-Ala (FALGPA) yaj nyob rau hauv tricine tsis yog siv los ua substrate. Cov extracts tau incubated nrog collagenase enzyme nyob rau hauv ib tug tsis haum tshuaj rau 15 min ua ntej ntxiv substrate los pib cov tshuaj tiv thaiv. Qhov kawg cov tshuaj tiv thaiv sib tov muaj tag nrho cov ntim ntawm 150 µL; tricine tsis, 0.8 mM FALGPA, 0.1 units ntawm ChC, thiab 25 μL MPM. Cov dej tau siv rau cov txiaj ntsig dawb. Tom qab qhov sib ntxiv ntawm substrate, qhov ntsuas ntawm absorbance tau ua tam sim ntawd. Kev tswj tau zoo ua tau epigallocatechin gallate (EGCG).23

Anti-elastase kev ua haujlwm

Kev ntsuam xyuas ntawm MPM rau kev ua haujlwm los tiv thaiv elastase yog ua los ntawm kev siv 0.2 mM Tris-HCl buffer solution (pH: 8.0). Ib qho kev daws teeb meem ntawm elastase (PE, EC 3.4.21.36) tau los ntawm porcine pancreas tau npaj nrog dej distilled ntawm qhov concentration ntawm 3.33 mg / mL. N-Succinyl-Ala-Ala-p-nitroanilide (AAAPVN) siv los ua cov substrate tau yaj hauv cov tshuaj tsis haum (1.6 mM). MPM extract yog incubated nrog 1 µg / mL ntawm PE rau 15 min ntawm 37 degree ua ntej substrate ntxiv. Thaum kawg ntawm 15 min pre-incubation, 0.8 mM AAAPVN substrate tau ntxiv rau cov enzyme sib tov uas muaj 1 mg / mL cog extract, thiab incubation tau ua dua rau 15 min ntawm 37 degree . Thaum siv 0.25 mg/mL EGCG raws li kev tswj xyuas qhov zoo, cov qauv kuaj no muaj tib lub ntim ntawm EGCG es tsis txhob MPM, thiab qhov kev ntsuam xyuas tau rov ua dua. Tom qab lub sijhawm incubation, kev ntsuas tau coj los ntawm 4 lub sijhawm sib txawv rau 5 mus rau 30 feeb los ntawm Thermo Scientific Multiskan SkyHigh Microplate Spectrophotometer ntawm 365 nm excitation thiab 410 nm emission.24, 25

Anti-hyaluronidase kev ua haujlwm

Cov kev ua los tiv thaiv hyaluronidase tau ua los ntawm kev hloov kho txoj kev piav qhia los ntawm Kolayli li al.26 thiab Lee et al.3 Ua ntej, cov khoom lag luam hyaluronidase (EC 3.2.1.35, Sigma-Aldrich) tau yaj hauv 0. 02 M phosphate buffer (pH: 3.5) muaj NaCl thiab bovine serum albumin. Tom qab ntawd, hyaluronic acid, ib qho tsim nyog substrate ntawm enzyme, tau npaj rau hauv acetate tsis ({21}}.1 M, pH: 3.5) thiab npaj rau kev siv. Cov tshuaj sib xyaw uas muaj 20 µL ntawm MPM ntawm qhov concentration ntawm 1 mg / mL, 10 µL ntawm hyaluronidase, thiab 60 µL ntawm 0.1 M acetate tsis yog pre-incubated rau 20 min ntawm 37 degree. Tom qab lub sij hawm incubation, 10 µL ntawm hyaluronic acid tau ntxiv rau qhov sib tov thiab incubated dua ntawm 37 degree rau 20 min. Thaum kawg ntawm tag nrho lub sij hawm incubation, kev ntsuas tau ua nyob rau lub sij hawm sib txawv cov ntsiab lus los ntawm Thermo Scientific Multiskan SkyHigh Microplate Spectrophotometer ntawm 600 nm. Cov pab pawg dawb huv tsis muaj cov enzymes hauv kev sim teeb tsa, thaum pawg tswj hwm tsis muaj cov extract. Qhov feem pua ​​​​ntawm cov kev ua los tiv thaiv hyaluronidase tau suav nrog kev sib npaug hauv qab no:

Kev tiv thaiv kev laus (%)= [(Abs of control - Abs of sample)/ Abs of control] × 100

Kev txheeb cais

Anti-elastase, anti-collagenase, thiab anti-hyaluronidase kev ua ub no suav nrog hauv txoj kev tshawb no tau rov ua peb zaug ntawm nws tus kheej. Qhov sib txawv ntawm kev txheeb cais tau txheeb xyuas siv t-test ntawm GraphPad Prism 8 software (p Tsawg dua lossis sib npaug rau 0.05).

TSEEM CEEB THIAB PAUB

Kev txiav txim siab ntawm qhov muaj peev xwm tiv thaiv kev laus

Elastin, collagen, thiab hyaluronic acid yog paub cov ntsiab lus ntawm ECM, uas muaj lub luag haujlwm tseem ceeb hauv cov hluas ntawm daim tawv nqaij. Elastin yog ib qho tseem ceeb ntawm cov protein kom tswj cov elasticity ntawm daim tawv nqaij, yog li ntawd, kev txo qis ntawm elastin hauv ECM ua rau kom nrawm rau hauv cov txheej txheem kev laus.27 Cov ntaub ntawv yav dhau los qhia txog kev sib txuas ncaj qha ntawm qhov tawv nqaij thiab kev laus ntawm daim tawv nqaij nrog qis qis ntawm elastin.28 Hyaluronic acid yog hydrophobic GAG molecule, uas yog depolymerized ntawm hyaluronidase. Hyaluronic acid yog qhov tseem ceeb los ua kom cov tawv nqaij smoothness thiab noo noo ruaj khov; Yog li ntawd, nws tau pom tias kev puas tsuaj ntau dhau ua rau cov tawv nqaij qhuav thiab tawv nqaij.29 Los ntawm cov txheej txheem kev laus nrog lub sijhawm, cov qib collagen txo qis ua rau nyias nyias ntawm daim tawv nqaij, uas yog suav tias yog ib qho kev qhia tshwj xeeb hauv kev tshuaj xyuas microscopic.3{{23 }} Nws tau raug qhia meej tias kev ncua sijhawm ntawm kev tawg ntawm collagen los ntawm collagenase inhibitors thiaj li cuam tshuam qhov wrinkling thiab aging ntawm daim tawv nqaij qauv.5 Xav txog cov ntaub ntawv no, cov khoom uas inhibit elastase, collagenase, thiab hyaluronidase muaj peev xwm tseem ceeb rau cov khoom tiv thaiv kev laus. Cov kev tshawb fawb yav dhau los tau pom tias ntau yam isoflavonoids nthuav tawm qhov tseem ceeb inhibitory bioactivity tiv thaiv cov enzymes uas tau hais tseg. Addotey thiab al.31 tau qhia tias plaub qhov sib txawv isoflavonoids inhibit hyaluronidase txog 61.2 feem pua. Kim et al.32 tau qhia tias ib qho isoflavonoid cais tawm los ntawm Glycyrrhiza uralensis Fisch., licoricidine, muaj cov elastase inhibitory tseem ceeb. IC50 tus nqi ntawm licorice raug xam raws li 61.2 ± 4.2 µM thaum oleanolic acid raug xam raws li 131.4 ± 11.4 raws li cov ntaub ntawv siv. Cov txiaj ntsig tau qhia tias isoflavonoids tuaj yeem cuam tshuam elastase enzymes. Raws li txoj kev tshawb no, Kim et al.33 tau kawm txog cuaj qhov sib txawv prenylated isoflavonoids, muaj feem cuam tshuam nrog osajin, cais los ntawm cov hauv paus hniav ntawm Flemingia philippinensis Merr. & Rolfe. Cov kws tshawb fawb tau tshaj tawm tias tsib ntawm cov isoflavones prenylated muaj zog tiv thaiv kev tiv thaiv neutrophil elastase, IC50 qhov tseem ceeb sib txawv ntawm 1.9-12.0 µM, thaum IC50 tus nqi ntawm oleanolic acid yog 28.4 µM. Hauv lwm txoj kev tshawb fawb, Ergene Öz li al. 34 tshawb xyuas hauv vitro inhibitory kev ua ntawm tsib isoflavonoids cais los ntawm cov hauv paus hniav ntawm Ononis spinoza L. tiv thaiv hyaluronidase, collagenase, thiab elastase. Hyaluronidase inhibitory kev ua ntawm cov isoflavones tau tshaj tawm tias nyob nruab nrab ntawm 22.08-45.58 feem pua, thaum, ntawm tib qhov concentration, tannic acid pom 88.32 feem pua ​​inhibition. Collagenase inhibition cov txiaj ntsig tau suav nrog ntawm 20.41- 28.49 feem pua ​​thiab elastase inhibition tau ntsuas li 20.47-46.88 feem pua ​​. EGCG tau siv los ua ib qho kev siv rau ob qho tib si kev soj ntsuam thiab kev ua haujlwm inhibition ntawm tib qhov concentration tau ntsuas li 41.18 feem pua ​​thiab 84.64 feem pua. Lwm txoj kev tshawb fawb tau tshawb xyuas cov tshuaj kho mob ntawm pomiferin ncaj qha cais los ntawm M. pomifera. 15 Pomiferin yog ib qho prenylated isoflavonoid, uas tuaj yeem pom hauv cov txiv hmab txiv ntoo M. pomifera thiab nws cov qauv molecular zoo ib yam li osajin. Cov kws tshawb fawb tau tshaj tawm tias pomiferin tau nthuav tawm ECM cov protein stimulation muaj zog los ntawm kev nce collagen thiab elastin uas zoo dua lossis sib npaug rau cov ntaub ntawv siv, retinol. Tag nrho cov kev tshawb fawb tau hais qhia tias isoflavones yog nruab nrab rau cov muaj zog inhibitors ntawm cov enzymes thiab muaj peev xwm tseem ceeb raws li cov khoom siv los tiv thaiv kev laus.

Hauv txoj kev tshawb no, hauv vitro hyaluronidase, collagenase, thiab elastase inhibitory kev ua ub no ntawm MPM tau tshawb xyuas rau kev txiav txim siab ntawm kev tiv thaiv kev laus. Kev sib piv kev sib piv tau ua rau kev ntsuam xyuas collagenase inhibition ntawm ob lub sijhawm, xws li 20 thiab 40 min, rau MPM thiab cov ntaub ntawv siv, EGCG. Cov txiaj ntsig tau pom tias collagenase inhibition kev ua haujlwm tau nce nrog lub sijhawm. 1 mg/mL MPM pom 84.55 ± 1.99 feem pua ​​inhibition, thaum 25{{40}} µg/mL EGCG pom 84.66 ± 1.83 feem pua ​​tom qab 20 min incubation. Inhibitory bioactivity tau nthuav dav los ntawm lub sijhawm, tom qab 40 min MPM thiab EGCG inhibited collagenase 94.68 ± 2.42 feem pua ​​thiab 94.98 ± 2.81 feem pua, feem. Raws li cov ntaub ntawv, MPM tau nthuav tawm cov haujlwm tseem ceeb inhibition tiv thaiv elastase. Cov txiaj ntsig tau ntsuas rau plaub lub sijhawm (5, 10, 20, thiab 30 min) thiab ua kom pom kev nce ntxiv hauv lub sijhawm (Daim duab 2). EGCG (250 µg/mL) tau siv los ua kev siv thiab inhibition kev ua haujlwm tau nce txhua lub sijhawm (44.07 ± 0.00 feem pua ​​, 52.19 ± 0.00 feem pua ​​, 64.69 ± 0.{{41 }} feem pua ​​, thiab 86.21 ± 0.00 feem pua ​​, feem). Meanwhile MPM nyob rau hauv 1 mg / mL concentration qhia ntau dua li thiab inhibition kev ua si, uas tsa los ntawm 34.70 ± 0.57 feem pua ​​​​rau 97.40 ± 1.04 feem pua ​​​​ntawm 5 min mus rau 30 min. Ib yam li ntawd, 1 mg / mL MPM inhibited hyaluronidase enzyme tseem ceeb tom qab 40 min incubation. 83.91 ± 2.36 feem pua ​​​​ inhibition tau ntsuas tom qab 40 min, tom qab ntawd 80 min ntawm incubation, tus nqi inhibition amplified rau 97.19 ± 0.45 feem pua. Thaum tag nrho cov enzyme inhibition kev soj ntsuam tau raug txiav txim siab, cov txiaj ntsig pom tau pom tias MPM yuav yog ib qho tseem ceeb ntawm kev tiv thaiv kev laus thiab tuaj yeem siv rau hauv cov ntsiab lus ntawm ntau yam khoom tiv thaiv kev laus; yog li ntawd, M. pomifera tej zaum yuav tau txais nyiaj txiag tseem ceeb ntxiv.

Kev txiav txim siab ntawm qhov muaj peev xwm antioxidant

Ntau yam exogenous thiab endogenous yam ua rau daim tawv nqaij laus ntawm ntau yam mechanisms. Feem ntau ntawm cov xwm txheej no raug cuam tshuam ncaj qha lossis cuam tshuam los ntawm ROS tsim nyob rau hauv ECM ntawm daim tawv nqaij.35 Txij li thaum daim tawv nqaij npog sab nrauv ntawm peb lub cev, nws ntsib ntau yam ntawm UV irradiation hauv lub neej niaj hnub. Yog li, feem ntau ntawm cov teeb meem ntawm daim tawv nqaij xws li sunburn, hyperpigmentation, thiab daim tawv nqaij carcinogenesis los yog muaj feem xyuam rau cov teebmeem ntawm UV hluav taws xob. Ib yam li ntawd, kev yees duab yog ib qho txiaj ntsig ntxiv ntawm nws cov khoom muaj kev phom sij.36 Ntxiv mus, UV teeb tsim ROS tsim thiab, yog li ntawd, oxidative stress nyob rau hauv daim tawv nqaij, uas yog ib qho tseem ceeb mechanisms uas ua rau photoaging.37 Nws yog hypothesized tias txij li thaum ntau dhau lawm. ROS tsim ua rau cov tawv nqaij laus ntxov ntxov, cov neeg ua haujlwm uas muaj peev xwm tiv thaiv oxidative tseem ceeb yuav yog cov cuab yeej muaj txiaj ntsig tiv thaiv kev phom sij ntawm UV hluav taws xob. Raws li, cov kev tshawb fawb soj ntsuam qhia tau hais tias cov tshuaj tua kab mob antioxidant muaj kev tiv thaiv ntawm daim tawv nqaij.38

Tom qab kev sib txuas ntawm kev siv tshuaj tua kab mob antioxidant thiab ncua kev laus ntawm daim tawv nqaij, uas tau tsim muaj nyob rau hauv cov ntaub ntawv dhau los, kev tshuaj xyuas hauv vitro antioxidant peev xwm ntawm MPM muab cov ntaub ntawv tseem ceeb rau nws muaj peev xwm tiv thaiv kev laus thaum siv tshuaj pleev xim. Cov ntaub ntawv yav dhau los qhia tau hais tias cov extracts nrog ntau cov isoflavonoids tej zaum yuav muaj txiaj ntsig antioxidants. Hauv kev tshawb fawb yav dhau los, ib qho isoflavonoid-nplua nuj extract ntawm F. macrophylla extract txo UVB-induced tawv nqaij puas tsuaj los ntawm scavenging ROS.39 Santos thiab Silva4{{10}} qhia tias prenylated isoflavonoids muaj peev xwm antioxidant tseem ceeb vim lawv. flavonoid moiety thiab ntxiv cov nyhuv ntawm prenyl sidechain. Lub peev xwm antioxidant ntawm cov extracts thiab isoflavonoids ntawm M. pomifera tau soj ntsuam hauv kev tshawb fawb yav dhau los. Cov txiaj ntsig tau pom tias hydroalcoholic extract thiab ntshiab osajin tau pom cov haujlwm tseem ceeb ntawm DPPH, FRAP, thiab TOAC kev soj ntsuam, txawm tias pomiferin thiab ethyl acetate feem ntau pom muaj kev ua haujlwm siab dua.41 Rau txoj kev tshawb no, DPPH radical scavenging kev ua, FRAP, CUPRAC, thiab TOAC kev soj ntsuam tau ua rau kev txiav txim siab dav dav hauv vitro antioxidant peev xwm ntawm MPM (Table 2). MPM tau nthuav tawm cov haujlwm tseem ceeb ntawm DPPH radical scavenging, qhov twg IC50 tus nqi raug ntsuas ntawm 1998.86 ± {{20}}.02. FRAP thiab TOAC kev soj ntsuam kuj ua rau muaj qhov tseem ceeb ntawm kev txo cov hlau, 0.191 ± 0.01 mM FeSO4 / DE thiab 114.43 ± 0.02 AAE / g DE, feem. Cov kev tshawb pom no tau ua raws li txoj kev tshawb fawb yav dhau los luam tawm los ntawm Orhan et al.41 CUPRAC kev soj ntsuam tau ua rau M. pomifera txiv hmab txiv ntoo rho tawm thawj zaug rau peb qhov kev paub. Ua ke, MPM tau pom qhov tseem ceeb ntawm kev txo cov tooj liab hauv CUPRAC kev soj ntsuam, qhov txiaj ntsig tau ntsuas li 73.928 ± 0.01 AAE / g DE. Thaum tag nrho cov tshuaj tiv thaiv kab mob antioxidant tau raug txiav txim siab, nws muaj peev xwm hais tias MPM yog tus neeg sawv cev tiv thaiv kev laus.

Phenolic profile thiab HPTLC tsom xam

Isoflavonoids yog cov tshuaj phenolic, uas yog lub npe hu ua cov nroj tsuag muaj lub luag haujlwm rau ntau yam kev ua ub ua no tseem ceeb xws li antioxidant, anticancer, thiab tiv thaiv cov teeb meem gynecological.42 Cov kev tshawb fawb yav dhau los tau pom tias prenylated isoflavonoids yog cov tshuaj phenolic loj hauv M. pomifera txiv hmab txiv ntoo.43 Ntau cov kev tshawb fawb pom osajin. thiab pomiferin raws li cov khoom xyaw tseem ceeb ntawm M. pomifera txiv hmab txiv ntoo, uas yog feem ntau lav rau nws yog biological kev ua ub no.12 Osajin thiab pomiferin yog heev zoo sib xws prenylated isoflavonoids uas tsuas yog txawv nrog ib pawg hydroxyl.44 Yav dhau los cov ntaub ntawv qhia tau pom kev tsis sib haum xeeb tshwm sim rau osajin thiab pomiferin ntsiab lus. ntawm M. pomifera txiv hmab txiv ntoo. Kartal et al.45 tau tsim ib txoj kev LC-MS rau kev txiav txim siab ntawm osajin thiab pomiferin hauv M. pomifera, uas tau sau los ntawm Ankara xeev ntawm Türkiye. Cov txiaj ntsig tau pom tias pomiferin cov ntsiab lus siab dua me ntsis ntawm osajin cov ntsiab lus hauv ntau qhov sib txawv ntawm cov qauv txiv hmab txiv ntoo. Hauv lwm txoj kev tshawb nrhiav, M. pomifera cov qauv txiv hmab txiv ntoo tau sau los ntawm ntau thaj tsam ntawm Midwest thiab Southern United States thiab osajin thiab pomiferin cov ntsiab lus tau ntsuas los ntawm HPLC tshiab txoj kev tsom xam. Cov txiaj ntsig tau pom tias qhov sib txawv ntawm thaj chaw ua rau muaj kev hloov pauv tseem ceeb ntawm cov nyiaj isoflavonoid hauv cov qauv.46 Tsao et al.47 txiav txim siab cov ntsiab lus osajin thiab pomiferin ntawm cov txiv hmab txiv ntoo sau los ntawm Canada thiab pom tias pomiferin cov ntsiab lus siab dua li cov ntsiab lus osajin. Hauv qhov sib piv, Hwang et al.48 tau sau tseg ntau qhov kev tshawb fawb uas pom tias osajin ntau dua li pomiferin ntau npaum li cas hauv ntau cov extracts. Nws tuaj yeem thov tias osajin thiab pomiferin cov ntsiab lus ntawm M. pomifera txiv hmab txiv ntoo yog qhov sib txawv tshwj xeeb nrog thaj chaw sib txawv thiab cov txheej txheem rho tawm vim lawv cov qauv tshuaj lom neeg txiav txim siab. Rau txoj kev tshawb no, tus nqi ntawm MPM tau ntsuas los ntawm HPTLC tsom xam, thawj zaug rau peb qhov kev paub. Cov txiaj ntsig ntawm kev txheeb xyuas pom tias osajin yog cov khoom tseem ceeb ntawm MPM uas tau sau los ntawm Uşak xeev, 0.22 feem pua ​​​​ntawm cov qauv muaj osajin (Table 1). Tsis tas li ntawd, txhawm rau ua tiav kev soj ntsuam ntxiv ntawm phenolic profile ntawm MPM, tag nrho phenolic thiab tag nrho cov ntsiab lus flavonoid tau soj ntsuam. Cov txiaj ntsig tau pom tias MPM muaj cov ntsiab lus tseem ceeb phenolic thiab flavonoid raws li hauv qab no; 113.92 ± 2.26 mg GAE/g thiab 66.41 ± 0.74 mg QE/g, feem. Cov txiaj ntsig ntawm kev ntsuam xyuas phenolic profile tau pom tias MPM yuav yog tus neeg sib tw tseem ceeb ua tus neeg sawv cev tshiab los tiv thaiv kev laus.

TEEB MEEM

Txawm hais tias kev tshawb fawb soj ntsuam kev siv tshuaj pleev xim rau M. pomifera txiv hmab txiv ntoo yog qhov tshiab, kev saib xyuas hauv qhov no tau nce ntxiv nrog cov lus ceeb toom txhawb nqa. Yog li ntawd, txoj kev tshawb no tsom mus piav qhia txog kev ntsuam xyuas ntawm qhov ua tau los tiv thaiv kev laus ntawm M. pomifera txiv hmab txiv ntoo extract. Kev tsom xam HPTLC tau siv rau M. pomifera txiv hmab txiv ntoo rau kev txiav txim siab ntawm cov ntsiab lus isoflavonoid thawj zaug rau peb qhov kev paub, nrog rau kev tshawb fawb hauv vitro rau kev txiav txim siab tag nrho cov phenolic profile. Cov txiaj ntsig tau pom tias osajin yog cov khoom tseem ceeb ntawm cov qauv. Tsis tas li ntawd, nyob rau hauv vitro antioxidant muaj peev xwm ntawm cov extract tau soj ntsuam nrog plaub qhov kev ntsuam xyuas sib txawv thiab cov txiaj ntsig tau pom tias muaj peev xwm antioxidant tseem ceeb ntawm MPM. Tsis tas li ntawd, kev ua haujlwm inhibitory tiv thaiv cov enzymes, uas cuam tshuam nrog cov txheej txheem kev laus, tau ntsuas thiab pom tias MPM muaj qhov tseem ceeb ntawm kev ua haujlwm enzyme inhibition. Hauv kev xaus, qhov kev tshawb fawb no muab cov ntaub ntawv, uas yuav ua rau kev tsim cov khoom siv tawv nqaij tshiab.

Kev ncaj ncees

Ethics Committee Pom Zoo:Pawg neeg saib xyuas kev ncaj ncees tsis tas yuav tsum muaj kev pom zoo rau txoj kev kawm.

Qhia Kev Pom Zoo:Tsis tsim nyog.

Peer-review:Externally peer-saib xyuas.

Authorship Kev Pabcuam

Concept: THB, TBŞ., HB, Design: THB, İ.KC, EC, Data Collection or processing: THB, İ.KC, HB, Analysis or interpretation: THB, İ.KC, Literature Search: THB, TBŞ., Sau: THB, EC

Kev tsis sib haum xeeb ntawm kev txaus siab:Tsis muaj teeb meem ntawm kev txaus siab tau tshaj tawm los ntawm cov kws sau ntawv.

Kev Qhia Txog Nyiaj Txiag:Txoj kev tshawb no tau txais kev txhawb nqa los ntawm Acıbadem Mehmet Ali Aydınlar University Commission of Scientific Projects (no: 2020/02/07).

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