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Txhim kho Antimelanogenesis Thiab Antioxidant Los Ntawm Polysaccharide Los Ntawm Cuscuta Chinensis Lam Noob Tom Qab Enzymatic Hydrolysis

Mar 30, 2023

Abstract

Cuscuta chinensis polysaccharide (CPS) tau muab rho tawm siv dej kub thiab enzymatically hydrolyzed C. chinensis polysaccharide (ECPS) yog tsim los ntawm mannanase enzymatic hydrolysis txheej txheem. Lub hom phiaj ntawm qhov kev tshawb fawb no yog los tshawb xyuas cov tshuaj tiv thaiv melanogenic ntawm ECPS thiab CPS hauv B16F10 melanoma hlwb. Kev ua haujlwm hauv vitro antioxidant tau soj ntsuam los ntawm lawv cov ferric hlau txo lub zog thiab DPPH dawb radical scavenging kev ua ub no. Kev faib tawm molecular loj ntawm polysaccharides tau txiav txim siab siv SEC-MALLS-RI. CPS tau ua tiav enzymatically degraded siv mannose thiab qhov hnyav nruab nrab molecular hnyav ntawm CPS thiab ECPS yog 434.6 kDa thiab 211.7 kDa. Cov txiaj ntsig ntawm kev soj ntsuam lom neeg tau pom tias cov enzymatically hydrolyzed polysaccharide muaj kev ua haujlwm zoo tshaj plaws los tiv thaiv melanogenic thiab cov nyhuv antioxidant dua li qub polysaccharide. ECPS tau nthuav tawm kev ua haujlwm los tiv thaiv melanogenic los ntawm kev txo qis kev qhia ntawm tyrosinase, MITF, thiab TRP-1 yam tsis muaj teebmeem cytotoxic hauv B16F10 melanoma hlwb. Hauv kev xaus, ECPS muaj peev xwm los ua cov khoom siv tawv nqaij dawb. Raws li kev tshawb fawb cuam tshuam,cistancheyog ib hom tshuaj ntsuab uas hu ua "cov tshuaj ntsuab miracle uas ua rau lub neej ntev". Nws lub ntsiab tivthaiv yog cistanoside, uas muaj ntau yam teebmeem xws li antioxidant, tiv thaiv kev mob, thiab kev tiv thaiv kev ua haujlwm. Lub mechanism ntawmcistanchethiabtawv nqaij dawbnyob rau hauv cov nyhuv antioxidant ntawm cistanche glycosides.MelaninNyob rau hauv tib neeg daim tawv nqaij yog tsim los ntawm oxidation ntawm tyrosine catalyzed los ntawmtyrosinase, thiab cov tshuaj tiv thaiv oxidation yuav tsum muaj kev koom tes ntawm cov pa, yog li cov pa oxygen dawb radicals hauv lub cev dhau los ua ib qho tseem ceeb cuam tshuam rau kev tsim melanin. Cistanche muaj cistanoside, uas yog ib qho antioxidant thiab tuaj yeem txo qhov tsim ntawm cov dawb radicals hauv lub cev, yog liinhibiting melanin ntau lawm.

Ntsiab lus:Cuscuta chinensis polysaccharide; Kev ua haujlwm antimelanogenic; Enzymatic hydrolysis polysaccharide; Antioxidant kev ua haujlwm

Taw qhia 

Melanin yog L-tyrosine transformation kawg-cov khoom, uas yog tus txiav txim loj ntawm cov plaub hau thiab tawv nqaij xim thiab ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv kev raug mob ultraviolet (1). Txawm li cas los xij, kev sib sau ntawm melanin tuaj yeem koom nrog hauv cov pigmentation txawv txav thiab ua rau hyperpigmentation ntawm daim tawv nqaij, melasma, hnub ci melanosis, thiab ephelides (2). Lub biosynthesis ntawm melanin koom nrog ib ntu ntawm enzymatic thiab oxidative cov tshuaj tiv thaiv thiab tyrosinase plays lub luag haujlwm tseem ceeb hauv cov txheej txheem (3). Lub tyrosinase-related protein (TRP-1) txhawb kev tsim DHICA oxidase hauv melanin biosynthetic txoj kev (4). Intracellular microphthalmia-associated transcription factor (MITF) yog ib qho tseem ceeb ntawm kev sau ntawv

regulator ntawm melanin biosynthesis noob. MITF tseem koom nrog hauv kev tswj hwm melanocyte pigmentation, proliferation, thiab sib txawv (5). a-MSH-melanocortin 1 receptor signaling tshwm sim hauv melanogenic cov enzymes, suav nrog TRP-1; tyrosinase kuj tseem tswj hwm los ntawm MITF (5). Ntau cov tawv nqaij ua kom tawv nqaij ua rau muaj kev tiv thaiv melanogenic los ntawm kev tswj hwm tyrosinase qhia lossis inhibitory cuam tshuam ntawm kev ua haujlwm tyrosinase. Tsis tas li ntawd, cov tshuaj tiv thaiv kab mob hauv lub cev thiab cov dawb radical tsim kuj cuam tshuam rau cov ntsiab lus melanin (6). Yog li ntawd, tyrosinase inhibitors thiab antioxidant tebchaw feem ntau raug xaiv los ua cov tawv nqaij dawb. Cuscuta chinensis Lam., hu ua TuSiZi hauv Suav, yog ib hom tshuaj suav tshuaj feem ntau siv los ua zaub mov ua haujlwm thiab paub los txhim kho kev muaj peev xwm ntawm kev ua me nyuam (7). Nyob rau hauv xyoo tas los no, qee cov ntaub ntawv qhia tau hais tias nws siv los kho freckles thiab vitiligo (8). Lwm cov ntawv ceeb toom tau pom tias nws ua rau muaj txiaj ntsig zoo ntawm kev tiv thaiv tawv nqaij (9), thiab ua rau muaj kev cuam tshuam ntawm kev ua haujlwm tyrosinase (10).

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Polysaccharides yog cov khoom tseem ceeb ntawm cov dej extract ntawm C. chinensis Lam. noob, uas suav hais tias muaj kev tiv thaiv apoptosis (11) thiab kev tiv thaiv kab mob (12). Cov kev soj ntsuam yav dhau los tau qhia tias C. chinensis Lam. polysaccharide yog tsim los ntawm fructose, mannose, xylose, thiab arabinose; mannose yog cov ntsiab lus qab zib (13). Ntau tus kws tshawb fawb tau pom tias viscosity (14), molecular hnyav (Mw) faib (15), thiab monosaccharide feem (16) ntawm polysaccharides muaj txiaj ntsig zoo rau lawv cov bioactivity. Ntxiv mus, kev tshawb fawb tsis ntev los no tau pom tias cov polysaccharides uas tsis tshua muaj Mw nthuav tawm ntau dua antioxidant thiab tyrosinase-inhibiting kev ua ub no dua li thawj polysaccharide (17). Yog li, kev tsim cov Mw polysaccharide tsawg los ntawm C. chinensis Lam. noob yog tsim nyog los txhim kho nws bioactivity. Ntawm cov txheej txheem degradation sib txawv, qhov tseem ceeb ntawm kev degradation enzymatic yog substrate tshwj xeeb, kev xaiv siab, thiab mob me me, uas tsim cov hydrolysates nrog cov qauv zoo (18).

Raws li cov kev tshawb fawb tshuaj no, peb tau kwv yees tias C. chinensis polysaccharide (CPS) thiab enzymatically hydrolyzed C. chinensis polysaccharide (ECPS) yuav yog cov tshuaj botanical zoo rau kev txhim kho hyperpigmentation. Mannase tau siv kom tau txais Mw ECPS qis los ntawm cov noob. Tsis tas li ntawd, cov tshuaj tiv thaiv melanogenesis thiab antioxidant kev ua haujlwm ntawm polysaccharides nrog Mw sib txawv tau kwv yees, thiab kev sib raug zoo ntawm bioactivities thiab Mw ntawm polysaccharides tau tshawb xyuas.

Khoom siv thiab cov txheej txheem

Reagents

Cov tshuaj rau enzyme thiab antioxidant kev ua ub no tau yuav los ntawm Sigma Co. (USA). Tag nrho lwm cov tshuaj reagents thiab tshuaj tau yuav los ntawm Aladdin (Tuam Tshoj).

Kev npaj CPS thiab ECPS

Cov khoom siv tshuaj ntawm Cuscuta chinensis Lam noob tau muab los ntawm Guang Dong Feng Chun Pharmaceutical CO., LTD (Suav teb). Txog 500 g ntawm cov ntaub ntawv qhuav yog hmoov, thiab soaked nrog 1200 mL 80 feem pua ​​​​ethanol rau 24 teev nyob rau hauv chav tsev kub kom tshem tawm lipids, oligosaccharides, thiab cov ntaub ntawv xim. Cov qauv pretreated tau infiltrated nrog daim ntaub, thiab tom qab ntawd cov khoom seem qhuav tau muab rho tawm nrog 3000 mL dej ntawm 90 degree peb zaug. Cov aqueous extracts tau muab cais tawm ntawm qhov seem los ntawm centrifugation (4000 g rau 5 min ntawm 22 degree) thiab tom qab ntawd concentrated ntawm 70 degree hauv lub tshuab nqus tsev; cov condensate tau precipitated nrog 60 feem pua ​​​​ethanol ntawm 3 degree rau 24 teev. Thaum kawg, cov precipitate tau deproteinated los ntawm txoj kev Sevag, dialyzed nrog 3500 Da membrane, lyophilized, thiab ces sau npe C. chinensis polysaccharide (CPS).

Cov enzymatically hydrolyzed C. chinensis polysaccharide (ECPS) tau los ntawm hydrolysis nrog mannose (0.1 feem pua ​​​​hauv sodium acetate tsis) hauv mannose rau substrate piv ntawm 5: 1 (v/w) ntawm 60 degree , pH 4,5h 6. Tom qab ntawd, cov tshuaj tiv thaiv catalysis tau txiav tawm hauv dej npau npau rau 10 feeb. Cov tshuaj tiv thaiv yog centrifuged ntawm 10, 000 g rau 15 min (4 degree), thiab cov supernatant tau sau rau kev lim ntshav ntawm 3 degree rau 3 hnub nrog 3500 Da membrane kom tshem tawm cov tshuaj molecular me me thiab tau lyophilized.

Cov ntsiab lus carbohydrate tau sim los ntawm txoj kev phenol sulfuric acid nrog cov piam thaj raws li cov khoom siv ntawm kev ntsuas nkhaus.

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SEC-MALLS-RI ntsuas

Qhov loj me me tsis suav nrog chromatography (Waters, USA) ua ke nrog ntau lub kaum sab xis laser lub teeb tawg (Wyatt, USA) thiab lub ntsuas ntsuas qhov ntsuas tsis pom tseeb (Waters, TEB CHAWS USA) (SEC-MALLS-RI) tau siv los txheeb xyuas qhov hnyav nruab nrab molecular hnyav ntawm polysaccharides. SEC-MALLS-RI tau ua nrog Phenomenex Polysep-GFC-Linear kem (8 mm × 3{{10}}}0 mm); cov qauv (2 mg / mL) tau yaj nrog theem mobile, uas muaj 0.1 M sodium chloride. Cov koob txhaj tshuaj yog 100 mL thiab tus khub tau teem rau ntawm 0.7 mL / min.

Mushroom tyrosinase inhibition assay

Mushroom tyrosinase inhibition (19) tau ua raws li yav dhau los qhia nrog kev hloov kho. Luv luv, 25 mL ntawm Kojic acid (zoo tswj) los yog cov qauv kev daws teeb meem (25 mL ntawm 10 mM L-tyrosine, 25 mL ntawm 0.5 mM L-DOPA, thiab 875 mL ntawm 50 mM phosphate buffer (pH 6.5) tov) tau tov. Tom qab ntawd, 38 mL ntawm 2100 U / mL nceb tyrosinase tau ntxiv thiab vortexed. Tom qab 0. Qhov inhibition feem pua ​​​​ntawm tyrosinase kev ua haujlwm tau suav los ntawm cov qauv hauv qab no: feem pua ​​​​tyrosinase inhibition=[(A-control – A-sample) / A-control] × 100, qhov twg A-tswj sawv cev rau qhov nqus ntawm 475 nm yam tsis muaj qauv thiab A-piv txwv sawv cev rau qhov nqus ntawm 475 nm nrog cov qauv.

Cell kab lis kev cai thiab kev soj ntsuam kev muaj sia nyob

Murine B16F10 melanoma hlwb tau yuav los ntawm Biochemistry thiab Cell Biology (Tuam Tshoj). Cells tau khaws cia nyob rau hauv Dulbecco's Modified Eagle Medium (DMEM) ntxiv nrog 10 feem pua ​​​​fetal bovine serum (FBS), 100 mg/mL streptomycin, thiab 100 IU/mL penicillin ntawm 37 degree nyob rau hauv ib tug humidified tshwm sim uas muaj 5 feem pua ​​CO2. Cells tau cog rau ntawm kab lis kev cai daim phiaj thiab ntxiv nrog ntau qhov sib txawv ntawm cov qauv thiab a-melanocyte stimulating hormone (a-MSH) rau 72 h los ntsuas qhov kev ua haujlwm ntawm intracellular tyrosinase thiab quantitate melanin cov ntsiab lus.

Qhov kev ntsuam xyuas 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tau ua los ntsuas kev muaj peev xwm ntawm tes (20). Luv luv, 96-cov phiaj zoo tau cog nrog murine B16F10 melanoma hlwb. Ib qhov ntim ntawm 50 mL ntawm 2 mg / ml MTT tau pauv mus rau hauv txhua qhov dej tom qab kev kho mob nrog 100 mL ntawm cov qauv sib txawv rau 24 teev. Tom qab 4-h incubation, cov tshuaj tiv thaiv raug txiav thiab dimethyl sulfoxide tau ntxiv kom yaj cov insoluble resultant. Absorbance tau ntsuas ntawm 590 nm nrog tus nyeem ntawv microplate.

Kev ntsuas cov ntsiab lus melanin

Kev kuaj pom cov ntsiab lus melanin tau ua nrog ib qho kev hloov me ntsis (21). Tom qab ntxuav nrog iced PBS, melanoma hlwb (2 × 104 hlwb rau ib qhov dej) tau noob nyob rau hauv ib tug 96- zoo phaj thiab incubated ntawm 37 degree rau 48 h. Tom qab ntawd, 100 mL NaOH (1N) tau ntxiv rau txhua qhov dej kom yaj melanoma hlwb ntawm 80 degree rau 30 min. Lub lysate tau centrifuged ntawm 15, 000 g rau 15 min (4 degree ). Tom qab ntawd, qhov nqus tau raug ntsuas nrog tus nyeem ntawv microplate ntawm 405 nm. Txhua qhov kev sim tau ua tiav hauv triplicate.

Intracellular tyrosinase kev ua ub no

Intracellular tyrosinase kev ua ub no tau ua raws li cov ntaub ntawv yav dhau los nrog kev hloov pauv me me (22). Luv luv, melanoma hlwb raug lysed nrog lysis buffer (1 mM PMSF, 1 feem pua ​​Triton X-100, 20 mM sodium phosphate) los ntawm khov-thawing. Tom qab centrifugation ntawm lysate ntawm 15, 000 g rau 10 min (4 degree), cov protein cov ntsiab lus ntawm supernatant tau txiav txim los ntawm kev ntsuam xyuas bicinchoninic acid (BCA). Lub supernatant protein (10 mg) tau pauv mus rau 100 mL ntawm cov tshuaj tiv thaiv sib tov (0.1 feem pua ​​L-DOPA thiab 0.1 M phosphate tsis). Tom qab 60 min incubation ntawm 37 degree, tyrosinase kev ua haujlwm tau ntsuas nrog tus nyeem ntawv microplate ntawm 450 nm. Txhua qhov kev sim tau ua tiav hauv triplicate.

Ferric hlau txo hwj chim

Kev ntsuam xyuas ferric hlau-txo lub zog tau ua raws li txoj kev luam tawm yav dhau los nrog kev hloov kho me me (23). Qhov sib txawv ntawm cov qauv (2 mL) los yog Vc (ib qho kev tswj zoo) tau tov nrog 2 mL poov tshuaj ferricyanide (1 feem pua, W/V) thiab 2 mL phosphate buffer (0.2 M, pH 6.8). Tom qab incubation ntawm 5{19}} degree rau 30 min, 2 mL trichloroacetic acid (10 feem pua ​​, W/V) raug xa mus rau hauv cov tshuaj tiv thaiv sib tov thiab centrifuged ntawm 4000 g rau 15 min (22 degree ). Lub supernatant (2 mL) yog tov nrog cov sib tov uas muaj 2 mL distilled dej thiab 0.4 mL FeCl3 (0.1 feem pua, W / V). Tom qab 10 min incubation ntawm 37 degree, qhov nqus tau ntsuas nrog lub microplate nyeem ntawv ntawm 700 nm.

DPPH radical-scavenging kev soj ntsuam

Qhov kev ntsuam xyuas DPPH-scavenging tau ua raws li tau tshaj tawm yav dhau los nrog qee qhov kev hloov kho (24). Luv luv, 2 mL ntawm cov qauv tau ntxiv rau 2 mL 0.1 mM DPPH tov thiab vortexed. Tom qab 30 min incubation nyob rau hauv qhov tsaus ntuj, lub absorbance yog ntsuas nrog lub microplate nyeem ntawv ntawm 517 nm.

Protein expression analysis los ntawm western blot

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Tom qab kev kho mob nrog ntau qhov sib txawv ntawm ECPS rau 72 h, cov hlwb raug ntxuav nrog PBS thiab lysed hauv RIPA tsis (150 mM NaCl hauv 50 mM pH 8.0 Tris-HCl , 0.5 feem pua ​​sodium deoxycholate, 1.0 feem pua ​​nondiet P-40, thiab 0.1 feem pua ​​sodium dodecyl sulfate). Tom qab centrifugation ntawm 10, 000 g rau 25 min (4 degree), lub supernatant ntawm lysates tau sau. Cov proteins tau raug rau 12 feem pua ​​SDS-PAGE thiab tom qab ntawd hloov mus rau polyvinylidene difluoride membrane. Kev thaiv yog nqa tawm hauv Tris-buffered saline nrog Tween -20 thiab 2 feem pua ​​​​skim mis hmoov (TBST), thiab tom qab ntawd incubated rau 12 teev ntawm 4 degree . Thawj cov tshuaj tiv thaiv uas siv yog: anti-actin (1: 5000), anti-TRP-1 (1: 500), anti-tyrosinase (1: 500), thiab anti-MITF (1: 1000). Thawj cov tshuaj tiv thaiv raug tshem tawm, thiab cov ntaub so ntswg tau ntxuav ob zaug nrog TBST. Tom qab ntawd, daim nyias nyias nrog horseradish peroxidase-conjugated thib ob cov tshuaj tiv thaiv kab mob (Santa Cruz, USA) tau muab tso rau hauv chav sov li 60 feeb. Cov ntaub qhwv protein tau raug ntxuav nrog TBST dua thiab pom nrog cov khoom siv ECL (Amersham Pharmacia Biotech, USA) siv UVP imaging system (UVP, USA).

Kev txheeb cais

Tag nrho cov txiaj ntsig tau tshaj tawm raws li txhais tau tias ± SD thiab qhov kev sim tau rov ua dua peb zaug. Kev sib piv ntawm cov pab pawg tau kwv yees siv ANOVA ua raws li Dunnett qhov kev xeem. Ib qho kev sib piv ntawm ob pawg tau ua los ntawm Tub Ntxhais Kawm T-test. Txhua qhov kev txheeb xyuas txheeb cais tau tsim los siv SPSS software (version 16.0). Po0.05 feem ntau yog suav hais tias yog qhov tseem ceeb.

Cov txiaj ntsig

Mw thiab tag nrho polysaccharides ntawm ECPS thiab CPS

Tag nrho cov ntsiab lus polysaccharide ntawm ECPS thiab CPS ntsuas los ntawm phenol-sulfuric acid assay yog 89.17 thiab 90.26 feem pua. Lub caij no, Mw ntawm ECPS thiab CPS tau ntsuas los ntawm SEC-MALLS-RI. Mw ntawm ECPS yog 211.7 kDa, uas yog qis dua CPS (434.6 kDa). Daim duab 1A qhia txog qhov sib txheeb sib ze (RI) rau ECPS thiab CPS; Tom qab enzymatic hydrolysis los ntawm mannose, lub sijhawm khaws cia siab tshaj plaws ntawm ECPS tau ntev dua li ntawm CPS. Raws li pom nyob rau hauv daim duab 1B, qhov sib txawv qhov hnyav ntawm cov polysaccharides tau piav qhia raws li kev ua haujlwm ntawm molar loj rau cov qauv. Cov molar loj faib ntawm polysaccharides hloov pauv los ntawm enzymatic hydrolysis. Qhov sib txawv ntawm qhov hnyav ntawm ECPS hauv thaj av qis Mw tau nce, uas qhia tias CPS tau degraded enzymatically rau hauv qis Mw polysaccharide.

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Antioxidant kev ua haujlwm ntawm polysaccharides

DPPH dawb-radical scavenging peev xwm ntawm ECPS thiab CPS tau tshaj tawm hauv daim duab 2A. Cov kev ua si dawb-radical scavenging ntawm cov qauv polysaccharide thiab Vc tau nthuav tawm cov kev ua ub no ntawm koob tshuaj. Nyob rau hauv txoj kev tshawb no tam sim no, cov dawb-radical scavenging peev xwm ntawm CPS yog qis dua li ntawm ECPS. Txawm li cas los xij, ob qho tib si tau nthuav tawm qis dua dawb-radical scavenging nyhuv dua li cov qauv zoo. IC50 qhov tseem ceeb ntawm ECPS thiab CPS yog 0.39 thiab 0.51 mg/mL, raws li. Raws li pom nyob rau hauv daim duab 2B, tag nrho cov tshuaj tiv thaiv antioxidant tuaj yeem raug soj ntsuam los ntawm kev sim ferric hlau-txo lub zog. Cov concentrations txawv ntawm 0.1 txog 1 mg / mL; ob qho tib si cov qauv polysaccharide thiab Vc nthuav tawm cov kev ua haujlwm antioxidant nyob rau hauv koob tshuaj. Ntxiv mus, tus nqi absorbance ntawm ECPS yeej ib txwm siab dua CPS's ntawm tib qhov concentration.

Kev cuam tshuam ntawm ECPS thiab CPS ntawm nceb tyrosinase kev ua haujlwm thiab cell viability

Raws li pom hauv daim duab 2C, tyrosinase inhibitory kev ua ntawm polysaccharides (0.1B1 mg/mL) qhia txog kev sib raug zoo ntawm koob tshuaj. Ntxiv mus, cov nyhuv inhibitory ntawm ECPS yeej ib txwm siab dua CPS's ntawm tib qhov concentration. Kev soj ntsuam MTT tau ua los ntsuas cov teebmeem cytotoxic ntawm ECPS thiab CPS hauv B16F10 melanoma hlwb. Raws li pom hauv daim duab 2D, tsis muaj qhov hloov pauv tseem ceeb hauv B16F10 cell viability nrog ntau qhov sib txawv (0B320 mg / mL) ntawm ECPS thiab CPS. Raws li cov txiaj ntsig no, peb siv cov concentration ntau hauv

kev tshawb fawb ntxiv.

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Qhov cuam tshuam ntawm ECPS thiab CPS ntawm kev ua haujlwm ntawm tyrosinase intracellular thiab cov ntsiab lus melanin

Txhawm rau sib piv cov teebmeem ntawm ECPS thiab CPS ntawm kev ua haujlwm ntawm intracellular tyrosinase thiab melanogenesis hauv B16F10 melanoma cell qauv, inhibitory potency ntawm ECPS thiab CPS ntawm melanin cov ntsiab lus thiab tyrosinase kev ua hauv a-MSH-stimulated B16F1{{ 16}} hlwb raug kuaj. Raws li pom nyob rau hauv daim duab 3, melanin cov ntsiab lus thiab tyrosinase kev ua ntawm B16F1{18}} hlwb tau nce ntau dua thaum piv rau cov tsis muaj zog B16F10 hlwb (Po 0.01). Ntawm qhov siab ntawm 40 mg / mL (ECPS) thiab 160 mg / mL (CPS), qhov nce hauv cov ntsiab lus melanin tuaj yeem txo tau raws li kev siv tshuaj (Po0.01 thiab Po0.05). Ib yam li ntawd, kev kho mob nrog ECPS (40 mg / mL) thiab CPS (160 mg / mL) inhibited tyrosinase kev ua ntawm B16F10 hlwb (Po 0.01 thiab Po0.05). Ntxiv mus, ECPS tau nthuav tawm ntau dua tyrosinase inhibitory kev ua ntawm melanogenesis dua CPS. ECPS (160 thiab 320 mg / mL) tau siv cov tshuaj tiv thaiv melanogenesis piv rau kev tswj zoo (Kojic acid), uas tau siv dav los ua cov tawv nqaij-dawb bioactive compound.

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Cov nyhuv ntawm ECPS ntawm tyrosinase, MITF, thiab TRP-1 qib protein ntau hauv B16F10 hlwb

Raws li pom nyob rau hauv daim duab 4, ECPS txo qis tyrosinase, MITF, thiab TRP-1 cov protein qhia qib hauv B16F10 hlwb nyob rau hauv ib koob tshuaj (Po0.05 thiab Po0. 01). Cov txiaj ntsig no qhia tau tias ECPS inhibited qhov kev qhia ntawm tyrosinase los ntawm kev txo qis cov protein qhia ntawm TRP-1 thiab MITF.

Kev sib tham

Lub ntuj polysaccharides los ntawm C. chinensis tau txais kev saib xyuas los ntawm lawv cov txiaj ntsig zoo ntawm tyrosinase inhibition, dawb radical scavenging, thiab kev tiv thaiv ntawm daim tawv nqaij (25–27). Txawm li cas los xij, kev tshawb fawb me me tau tsom mus rau kev ua haujlwm antimelanogenesis ntawm kev hloov kho enzymatic ntawm polysaccharides. Cov kev tshawb fawb yav dhau los tau pom tias degraded polysaccharides los ntawm enzymatic hydrolysis txheej txheem nthuav tawm superior dawb radical scavenging nyhuv (28). Ntxiv mus, cov kev ua ub no lom ntawm polysaccharides yog ze ze rau lawv cov Mw distributions. Raws li txoj cai, Mw polysaccharides qis dua muaj zog dua li siab Mw polysaccharides vim lawv cov cuab yeej nkag siab ntawm cov cell membranes (29,30). Txawm li cas los xij, cov nyhuv antimelanogenesis ntawm ECPS ntawm B16F10 hlwb tseem tsis tau kawm. Qhov qis Mw polysaccharide tau npaj los ntawm enzymatic hydrolysis nrog mannose.

Oxidative stress tuaj yeem tsim cov dawb radicals ntau dhau thiab ua rau oxidative raug mob. Cov kev tshawb fawb yav dhau los tau ua pov thawj tias kab mob ntawm daim tawv nqaij muaj feem cuam tshuam nrog kev sib sau ntawm cov dawb radicals (31). Tsis tas li ntawd, cov dawb radicals ntau dhau ua lub luag haujlwm tseem ceeb hauv kev txo qis melanogenesis ntawm melanoma hlwb thiab kev loj hlob ntawm melanocytes (32). Tyrosinase yog ib tug multifunctional oxidant enzyme uas muaj bronze thiab tseem ceeb heev rau kev txhawb melanin biosynthesis (33). Txawm li cas los xij, daim tawv nqaij pigmentation thiab ntau yam kab mob ntawm daim tawv nqaij muaj feem cuam tshuam nrog kev sib txuam ntawm melanin thiab ua rau muaj teeb meem loj (34).

Cov khoom xyaw nquag nrog antioxidant thiab anti-tyrosinase peev xwm tuaj yeem tiv thaiv tawv nqaij thiab inhibit melanogenesis (35). Peb cov txiaj ntsig tau pom tias qis Mw ntawm enzymatically hloov kho polysaccharides nthuav tawm cov tshuaj tiv thaiv antioxidant thiab tiv thaiv tyrosinase ntau dua li qub polysaccharides hauv vitro. Qhov kev txhim kho yog los ntawm thaj chaw ntau dua thiab cov dej solubility zoo dua, uas tau ua raws li kev tshawb fawb yav dhau los (17) uas tau pom tias cov polysaccharide degraded los ntawm Sargassum fusiforme muaj cov kev ua haujlwm zoo tshaj plaws los tiv thaiv tyrosinase thiab kev ua haujlwm antioxidant dua li thawj polysaccharide.

cistanche reddit whitening

Ib txwm melanocytes pw ntawm qhov sib txuas ntawm epidermis thiab dermis ntawm daim tawv nqaij thiab tsim melanin, uas yog pauv mus rau keratinocytes (36). Nyob rau hauv txoj kev tshawb no, murine B16F10 melanoma hlwb tau siv vim lawv muaj melanogenic mechanisms, paub tias muaj intracellular tyrosinase, thiab muaj peev xwm tsim melanin, uas muaj feem xyuam rau a-MSH stimulation thiab melanogenesis (37). Tyrosinase kev ua ub no, cov ntsiab lus melanin, thiab kev muaj peev xwm ntawm tes yog cov kev ntsuam xyuas hauv vitro siv los tshuaj xyuas antimelanogenesis hauv txoj kev tshawb no. CPS thiab ECPS tau nthuav tawm cov tshuaj tiv thaiv kab mob ntawm cov tshuaj tyrosinase thiab melanin synthesis hauv B16F10 hlwb. ECPS pom muaj zog tiv thaiv melanin synthesis thiab anti-tyrosinase nyhuv.
Tyrosinase-related protein-1 (TRP-1) thiab tyrosinase ua lub luag haujlwm tseem ceeb hauv melanin biosynthesis thiab melanogenesis txoj hauv kev (38). MITF yog ib qho cellular transcription ntawm tyrosinase noob, uas koom nrog hauv melanogenesis. Feem ntau, kev ua kom TRP-1 thiab tyrosinase txhim kho MITF protein qhia thiab ua rau muaj kev nce hauv melanin synthesis (39). Yog li, cov tshuaj pleev xim rau ntawm daim tawv nqaij yuav muaj cov cuab yeej ntawm inhibiting txoj kev taw qhia kev koom tes hauv kev ua kom TYP-1 lossis tyrosinase. Yog li, peb tau tshawb xyuas qhov cuam tshuam ntawm ECPS ntawm TRP-1, cellular tyrosinase, thiab MITF protein qhia los kawm txog cov txheej txheem hauv qab ntawm inhibition ntawm tyrosinase kev ua si thiab melanogenesis. Cov txiaj ntsig ntawm qhov kev ntsuam xyuas sab hnub poob tau pom tias ECPS tau txwv tsis pub qhia txog TRP-1, tyrosinase, thiab MITF hauv B16F10 hlwb thiab txhais tau tias ECPS txo melanogenesis los ntawm kev tswj hwm tyrosinase, MITF, thiab TRP{12}} qhia nyob rau hauv B16F10 melanoma hlwb. Qhov tshwm sim yog los ntawm kev tshawb fawb yav dhau los uas qhia tau hais tias cov aqueous extract los ntawm Cuscuta japonica noob tau inhibited a-MSH-induced melanin synthesis thiab tyrosinase kev ua los ntawm suppressing p38 MAPK phosphorylation, inhibiting cAMP qib, thiab tom qab ntawd txo qhov kev qhia ntawm TRP thiab MITF (40) .
Nyob rau hauv cov ntsiab lus, cov enzymatically hloov polysaccharide muaj superior antioxidant thiab anti-melanogenic teebmeem dua li thawj polysaccharide. Tsis tas li ntawd, qhov tshuaj tiv thaiv melanogenic ntawm ECPS tau kho los ntawm kev tawm tsam ntawm TRP-1, tyrosinase, thiab MITF qhia hauv murine B16F10 hlwb. ECPS tuaj yeem siv tau rau kev siv hauv cov khoom siv tshuaj pleev ib ce thiab kho mob.

Kev lees paub

Txoj haujlwm no tau txais kev txhawb nqa los ntawm National Natural Science Foundation ntawm Tuam Tshoj (grant No. 81373640).

Cov ntaub ntawv

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