Antioxidant, Anti-aging thiab lub cev tiv thaiv los ntawm tag nrho Saponins los ntawm Cistanche

Xaus: TSAT yog cov khoom lag luam zoo tshaj plaws nrog cov tshuaj tua kab mob antioxidant thiab tiv thaiv kev laus uas tuaj yeem txo D-gal-induced aging puas hauv nas.

Ntsiab lus:Cistanche tag nrho saponinsD-galactose, antioxidant,anti-aging

Nyem qhov no kom tau txais Cistanche Ntxiv rau Kev Tiv Thaiv Kev Laus

Taw qhia

Txij li thaum lub xyoo pua 21st, nrog rau lub ntiaj teb txoj kev laus ntawm kev laus thiab kev txhim kho ntawm kev ua neej nyob thoob ntiaj teb, cov teeb meem kev noj qab haus huv los ntawm kev laus thiab cov kab mob ntawm cov laus tau nce zuj zus.1 Yog li ntawd, yuav ua li cas tswj kev noj qab haus huv ntawm cov neeg laus thiab ncua sijhawm. Qhov tshwm sim ntawm kev laus thiab cov kab mob uas muaj feem rau kev laus tau dhau los ua ib lub ntsiab lus tshawb fawb nrov. Kev laus yog ib qho irreversible thiab complex degenerative physiological txheej txheem uas tshwm sim nyob rau hauv tag nrho cov qhov chaw ntawm lub cev, ua rau lub structural thiab functional deterioration ntawm cov ntaub so ntswg thiab lub cev thiab ua rau muaj kev pheej hmoo ntawm ntau yam kab mob, xws li kub siab, ntshav qab zib, kab mob neurodegenerative, thiab daim tawv nqaij melanosis. .2 Ntau cov ntawv tshaj tawm tau qhia tias txoj kev xav dawb radical ntawm kev laus, ib qho ntawm cov kev xav nrov tshaj plaws, tuaj yeem piav qhia txog kev puas tsuaj rau lub cev los ntawm cov txheej txheem kev laus.3 Thaum lub sij hawm aerobic metabolism, reactive oxygen hom (ROS) yog tsim nyob rau hauv cov hlwb, xws li singlet oxygen (1 O2), superoxide anions (•O2-), hydroxyl radicals (HO•), thiab hydrogen peroxide (H2O2).4,5 Thaum cov dawb radicals, uas muaj unpaired thiab reactive electrons, yog tsim nyob rau hauv ntau, lawv ncaj qha ua rau oxidative kev nyuaj siab thiab proinflammatory cov lus teb, ua raws li redox tsis txaus, ua rau cov protein thiab lipid peroxidation, nrog rau mitochondrial thiab DNA puas, thaum kawg ua rau hloov cell thiab cov ntaub so ntswg muaj nuj nqi thiab pathological tej yam kev mob kom ceev aging. .6,7 Interestingly, natural endogenous antioxidant systems muaj nyob rau hauv tib neeg / mammalian system, nrog rau cov koom nrog SOD, CAT thiab GSH-Px, nrog rau nonenzymatic antioxidants xws li ascorbic acid thiab carotenoids, tsim kom muaj kev sib npaug ntawm cov dawb radicals. thiab antioxidants.8,9 Txawm li cas los xij, cov tshuaj tiv thaiv antioxidant tsis txaus los tiv thaiv oxidative puas tsuaj los ntawm cov dawb radicals ntau dhau, thiab ntxiv nrog cov tshuaj antioxidants tuaj yeem txo qhov kev laus thiab cuam tshuam oxidative puas tsuaj.10 Piv nrog ntuj antioxidants, hluavtaws antioxidants (piv txwv li, BHT thiab BHA) tau pom tias ua rau muaj kev cuam tshuam endocrine, kev muaj menyuam tsis taus thiab txawm tias carcinogenesis hauv kev siv mus ntev.11,12 Yog li ntawd, kev tshawb nrhiav ntuj, kev nyab xeeb thiab zoo dawb radical scavengers yog qhov tseem ceeb heev rau tib neeg kev noj qab haus huv. kev tiv thaiv.

Cistanche yog ib hom nroj tsuag ntawm tib genus li Aralia elata (Miq.) Seem.13 Nws yog dav faib nyob rau hauv lub Qinba Roob sab hnub poob Tuam Tshoj thiab yog ib tug tshuaj thiab edible nroj tsuag. Cistanche paus bark extract tau ntev tau siv nyob rau hauv Tuam Tshoj raws li ib tug pej xeem tshuaj rau kev kho mob ntawm cov ntshav qab zib, thiab nws lub ntsiab active ingredient yog triterpene saponin.14 Nws tau pom tias muaj antioxidant, antihyperglycaemic thiab antihyperlipidaemic teebmeem.14-16 Ntau cov kev tshawb fawb tau pom. uas triterpene saponins xws li asiaticoside B, madecassoside thiab parkside A tuaj yeem hnov ​​​​mob dawb radicals thiab tiv thaiv ntxiv kev loj hlob ntawm cov dawb radical chain reactions, yog li exerting anti-oxidant teebmeem.17,18 Cov ntaub ntawv dhau los tau piav qhia tias TSAT nthuav tawm superior DPPH radical scavenging kev ua rau ib leeg. saponins, raws li zoo raws li antioxidant muaj peev xwm tiv thaiv lipid peroxidation nyob rau hauv nas daim siab microsomes, thiab tau pom tias muaj superior antioxidant teebmeem rau ginseng, Schisandra thiab lwm yam saponins.14,15 Tsis tas li ntawd, TSAT kuj attenuated D-gal-induced daim siab puas siab puas ntsws hauv kev laus. nas los ntawm nws cov nyhuv antioxidant.19 Kev tshawb fawb ntxiv-cov txiaj ntsig kev sib raug zoo tau pom tias qhov sib txawv ntawm cov qauv xws li qeb thiab qhov sib lawv liag ntawm cov saw oligosaccharide ntawm C-3 hauv cov saponins no ua lub luag haujlwm tseem ceeb hauv cov ntsiab lus ntawm cov ntshav qab zib mellitus. antioxidant thiab antiglycation kev ua ub no.20

D-gal yog ib qho kev sim siv rau kev laus inducer. Cov pov thawj ntau ntxiv qhia tias cov phenotypes cuam tshuam txog kev laus xws li kev tsim cov dawb radical ntau dhau, kev paub tsis zoo, kev tiv thaiv kab mob tsis zoo thiab kev raug mob ntawm lub cev tuaj yeem raug soj ntsuam raws li D-gal stimulation ntev. thiab paub txog kev nco hauv nas, raws li pom los ntawm kev ua haujlwm tsis zoo ntawm cov teeb meem hauv kev nrhiav lub hom phiaj platform thiab txo tus naj npawb ntawm lub sij hawm hla lub hom phiaj platforms hauv Morris dej maze sim.22 Yog li ntawd, kev siv lub sijhawm ntev D-gal stimulation tau siv los kawm txog kev laus ntawm lub cev. thiab aging-related nco poob, redox imbalance thiab lub cev raug mob.

Raws li cov kev tshawb fawb yav dhau los ntawm TSAT, muaj qhov laj thawj los ntseeg tias cov nroj tsuag tshuaj thiab cov khoom noj muaj txiaj ntsig zoo rau kev siv hauv kev tiv thaiv kev laus. Txawm li cas los xij, tsuas muaj ib txoj kev tshawb fawb txog kev tshem tawm cov dawb radicals hauv vitro los ntawm qhov triterpenoid saponin extract, thiab lwm yam physiologically cuam tshuam tus yam ntxwv ntawm dawb radical scavenging tsis muaj. Tsis tas li ntawd, kev paub txog kev tiv thaiv los ntawm TSAT tiv thaiv kev nco, redox tsis txaus thiab kev raug mob ntawm lub cev hauv D-gal-induced subacute aging qauv yog txwv. Yog li, qhov kev tshawb fawb no tau sib sau ua ke cov khoom antioxidant ntawm TSAT hauv vitro los ntawm kev tshem tawm DPPH, ABTS, HO•, •O2- thiab inhibiting tyrosinase kev ua. Ntxiv mus, kev sim coj tus cwj pwm, ntsuas qhov ntsuas ntawm oxidative kev ntxhov siab, thiab cov kab mob histopathological ntawm cov kab mob loj hauv cov nas tau raug soj ntsuam los muab cov ntaub ntawv ntxiv ntawm TSAT lub peev xwm los ameliorate D-gal-induced subacute oxidative thiab kev laus puas tsuaj, sib sau ua ke nws cov khoom siv tshuaj.

Cov ntaub ntawv thiab cov txheej txheem

Lub hauv paus bark ntawm Cistanche tau sau los ntawm Qinba Toj siab, Shaanxi Province, Tuam Tshoj, thiab tau botanically txheeb xyuas los ntawm Dr. Jitao Wang (Shaanxi University of suav tshuaj). Daim ntawv pov thawj (SUCM, No. 20201003) tau muab tso rau hauv Herbarium ntawm Shaanxi University of Chinese Medicine. Tarasaponin IV

DPPH (Sigma-Aldrich, St. Louis, MO, USA), ABTS (Maclin Biochemical Co., Ltd, Shanghai, China), nceb tyrosinase (Yuan Ye Biotechnology Co., Ltd, Shanghai, Tuam Tshoj), L{{1} }, 4-dihydroxyphenylalanine (L-DOPA) thiab D-gal (Yuan Ye Biotechnology Co., Ltd, Shanghai, Tuam Tshoj) tau yuav. Superoxide anion (Khoom naj npawb: A052-1-1), Hydroxyl dawb radical (khoom naj npawb: A018-1-1), BCA (tus naj npawb: A045-4-2), SOD ( Yam khoom: A{{ 8}}), GSH-Px (Khoom naj npawb: A005-1-1), CAT (tus lej khoom: A007-1-1), MDA (tus lej khoom: A003-1-2), T-AOC ( Yam khoom naj npawb: A015-2-1), AST ( Yam khoom naj npawb: C010-2-1) thiab ALT (Khoom naj npawb: C009-2-1) cov khoom siv tshuaj ntsuam xyuas tau txais los ntawm Nanjing JianCheng Bioengineering Institute (Nanjing, Tuam Tshoj) . Tag nrho cov tshuaj thiab lwm yam reagents siv yog cov qhab nia ntsuas.

Cistanche paus bark yog ntxuav nrog ob distilled dej, qhuav thiab crushed ntawm 60 degree. Ua ntej qhov kev sim, cov hmoov tau muab tso rau hauv qhov chaw txias thiab qhuav. Cov hmoov (3 kg) tau muab rho tawm nrog 70 feem pua ​​​​ethanol rau peb zaug nyob rau hauv reflux. Cov kev daws teeb meem tau ua ke thiab cov supernatant tau txais los ntawm kev kub ceev centrifugation thiab pom. Lub supernatant tau huv si los ntawm kev thauj khoom rau hauv HPD300 macroporous resin kem thiab tom qab ntawd eluting nrog 70 feem pua ​​​​ethanol. Cov tshuaj uas eluted nrog 70 feem pua ​​​​ethanol tau evaporated kom yield ib residue (356 g) tom qab hnyav tshem tawm hauv vacuo. Cov txiaj ntsig ntawm TSAT yog 11.87 feem pua ​​(w/w) thiab tau khaws cia rau hauv lub tub yees rau kev siv ntxiv.

duab kab (4.6 ID × 250 mm). Cov theem ntawm lub xov tooj ntawm tes muaj xws li acetonitrile (cov kuab tshuaj A) thiab 0.1 feem pua ​​​​phosphoric acid aqueous tov (V/V) (hloov B). Qhov kev pab cuam gradient tau raws li hauv qab no: 0–10 min, 5–20 feem pua ​​A; 10–25 feeb 20–28 feem pua ​​A; 25–35 feeb, 28–33 feem pua ​​A; 35–45 feeb, 33–38 feem pua ​​A; 45–55 feeb, 38–46 feem pua ​​A; 55–60 feeb, 46–60 feem pua ​​A; 60-70 feeb. 60-75% A; 70–80 feeb, 75–45 feem pua ​​A; 80–90 min, 45–5 feem pua ​​​​A. Cov kev ua haujlwm ntawm qhov ntsuas yog qhov kub thiab txias ntawm 30 degree thiab tus nqi ntws ntawm 0.8 mL / min. Qhov ntim ntawm cov qauv thiab cov qauv yog 10 μL. Kev kuaj pom wavelength yog 203 nm. Ua ntej txhaj tshuaj, tag nrho cov qauv tau dhau los ntawm 0.22 μm lim. Tom qab sib piv nrog tus qauv, lub ncov yog txiav txim siab los ntawm lub sijhawm khaws cia. Lub linear calibration nkhaus ntawm tus qauv yog siv rau quantification.

Antioxidant Kev Ua Haujlwm hauv vitro

DPPH Assay Qhov kev ntsuam xyuas DPPH tau hloov kho me ntsis los ntawm daim ntawv tshaj tawm dhau los.23 Ib qho sib xyaw ntawm anhydrous ethanol (100} μL), DPPH tov (100 μL, 0.2 mM) thiab cov qauv (100 μL, 0.01 - 1 mg / L) tau shaken ceev thiab khaws cia (25 degree, 30 min) hauv qhov tsaus ntuj. VC tau siv los ua qhov zoo tswj Lub absorbance (517 nm) tau ntsuas. Lub DPPH radi

cal scavenging tus nqi tau xam raws li hauv qab no:

qhov twg A0 sawv cev rau kev nqus ntawm kev tswj tsis zoo (DPPH plus absolute ethanol); A1 sawv cev rau qhov kev xeem uas muaj cov qauv (DPPH plus TSAT); thiab A2 yog qhov nqus ntawm qhov khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob. 50 feem pua ​​​​ inhibition concentration tus nqi (IC50) ntawm DPPH dawb radical scavenging tus nqi raug xam, uas sawv cev rau lub peev xwm antioxidant ntawm TSAT. Tag nrho cov kev khiav hauj lwm yog ua nyob rau hauv parallel peb zaug.

ABTS plus Assay ABTS tau txiav txim siab siv txoj kev ntawm Rafique li al nrog kev hloov kho me ntsis.24 Ua ntej, cov txheej txheem TSAT (0.01– 1 mg/mL) cov kev daws teeb meem tau npaj. Kev daws ABTS ntawm 7 mM thiab K2S2O8 daws ntawm 2.45 mM tau npaj. Tom qab muab sib xyaw rau hauv tib lub ntim, ABTS ntxiv tau txais los ntawm reacting (12-16 h) hauv qhov tsaus ntuj. Cov npaj ABTS ntxiv tau diluted nrog ob chav distilled dej mus rau ib qho absorbance ntawm 0.7 ± 0.05 ntawm 734 nm. Tom qab ntawd, qhov sib xyaw ntawm 180 μL diluted ABTS ntxiv rau kev daws thiab 20 μL cov qauv tau tshwm sim (25 degree, 10 min) hauv qhov tsaus ntuj. VC tau siv los ua kev tswj xyuas zoo. Lub absorbance (734 nm) tau ntsuas. Lub peev xwm ABTS ntxiv rau kev tshem tawm tau suav los ntawm cov qauv hauv qab no:

qhov twg A0 sawv cev rau kev nqus ntawm kev tswj tsis zoo (ABTS ntxiv rau ob chav dej distilled); A1 sawv cev rau qhov kev xeem uas muaj cov qauv (ABTS plus plus TSAT); thiab A2 yog qhov nqus ntawm qhov khoob khoob (TSAT ntxiv rau ob chav dej distilled). Tus nqi IC50 ntawm ABTS ntxiv rau qhov kev tshem tawm tau raug xam. Tag nrho cov haujlwm tau ua nyob rau hauv parallel peb zaug.

Kev txiav txim siab ntawm Mushroom Tyrosinase Inhibition Activity Lub tyrosinase inhibitory kev ua haujlwm tau txiav txim siab raws li kev txhim kho me ntsis ntawm cov txheej txheem yav dhau los qhia los ntawm Morais li al.25 A series ntawm 50 μL TSAT (0.01–1 mg / mL, hauv 50 feem pua ​​DMSO) cov kev daws teeb meem tau ntxiv rau cov tshuaj tiv thaiv uas muaj 100 μL ntawm PBS tsis (0.1 M, pH 6.8) thiab 50 μL tyrosinase (100 U / mL), feem, thiab hnov ​​​​mob ntawm 25 degree rau 15 min. . Tom qab ntawd, 50 μL L L-DOPA (3.5 mM) tov tau ntxiv rau cov tshuaj tiv thaiv kab mob thiab incubated ntawm 37 degree rau 10 min. Lub absorbance (475 nm) tau txiav txim siab. VC tau siv los ua kev tswj xyuas zoo. Cov kev ua haujlwm inhibition iof TSAT raug xamlos ntawm cov qauv hauv qab no:

qhia txog tyrosinase inhibitory kev ua ntawm TSAT. Tag nrho cov haujlwm tau ua nyob rau hauv parallel peb zaug.