Phoenix Dactylifera L. Noob Extract Exhibits Antioxidant Effects Thiab Attenuates Melanogenesis

Huey-Chun Huang1 , Shr-Shiuan Wang2, Tsang-Chi Tsai3, Wang-Ping Koj3 and Tsong-Min Chang2,*

Abstract:Keeb kwm: Hom kev ua ntawm Phoenix dactylifera noob extract hauv kev tu tawv nqaij tsis tau tshawb pom. Txoj kev: P. dactylifera L. noob tau muab rho tawm los ntawm ultrasonic extraction. Cov yam ntxwv antioxidant ntawm cov extract tau txiav txim siab los ntawm 2,2-diphenyl-1-picrylhydrazyl (DPPH) thiab 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS ntxiv) kev soj ntsuam thiab cov txheej txheem scavenging. Tag nrho cov ntsiab lus phenolic, txo lub peev xwm, hlau (II) ion-chelation, thiab intracellular reactive oxygen hom (ROS)-scavenging capacities kuj tau tshawb xyuas. Cov teebmeem ntawm P. dactylifera L. noob extract ntawm melanogenesis tau soj ntsuam spectrophotometrically los ntawm kev soj ntsuam kev ua si ntawm nceb tyrosinase, kev txiav txim siab ntawm intracellular tyrosinase kev ua, thiab cov ntsiab lus melanin. Kev qhia theem ntawm melanogenesis-txog cov proteins tau txheeb xyuas los ntawm Western blotting. Cov txiaj ntsig: Cov txiaj ntsig tau pom tias P.dactylifera L.noob extract exerted pom meej antioxidant muaj peev xwm thiab txo qis intracellular ROS cov ntsiab lus ntawm cov concentrations ntawm {{0}}.245 thiab 0.49 (mg/mL). Tsis tas li ntawd, cov extract txo qis kev qhia ntawm melanocortin 1 receptor (MC1R), microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP1), thiab tyrosinase-related protein-2 ( TRP2), thiab inhibited melanogenesis hauv B16F10 hlwb. Cov ntsiab lus: Peb cov txiaj ntsig tau qhia tias P. dactylifera L. noob extract attenuated melanogenesis hauv B16F10 hlwb los ntawm kev txo qis protein kinase A (PKA) cov kev taw qhia. Li no, cov extract yuav siv tau raws li ib hom ntawm daim tawv nqaij-dawb tus neeg sawv cev nyob rau hauv cov khoom tu tawv nqaij.

Lo lus tseem ceeb: Phoenix dactylifera;tyrosinase; melanin; ROS; PKA

cistanche pharma tshwj xeebyog ib hom ntawm daim tawv nqaij-dawb tus neeg sawv cev.

1. Taw qhia

Antioxidants tau dav siv los tiv thaiv lossis kho oxidative kev ntxhov siab cuam tshuam txog kev ua kom zoo nkauj thiab dermatological. Ob peb xyoos dhau los, cov tshuaj antioxidants kuj tau siv hauv kev lag luam tshuaj pleev ib ce los tiv thaiv lossis ncua kev laus ntawm daim tawv nqaij. Nws tau tshaj tawm tias cov dawb radicals thiab reactive oxygen hom (ROS) cuam tshuam nrog ntau yam kab mob, xws li kev laus thiab cov kab mob uas muaj hnub nyoog [1]. Interestingly, dawb radical puas ntawm daim tawv nqaij tshwm sim los ntawm UV-irradiation kev nyuaj siab thiab ROS ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm daim tawv nqaij photoaging [2,3]. Antioxidants tau tshaj tawm tias cuam tshuam nrog cov txheej txheem oxidation los ntawm kev tshem tawm cov dawb radicals thiab ROS los yog los ntawm chelating oxidation-catalytic hlau [4]. Li no, tau muaj ntau qhov kev siv ntawm cov tshuaj antioxidants los yog cov khoom noj khoom haus antioxidant kom txo tau oxidative kev nyuaj siab los yog oxidative kev nyuaj siab vim kev puas tsuaj hauv lub cev [5,6]. Txawm li cas los xij, qee cov tshuaj antioxidants, suav nrog sodium ascorbate, tert-butyl hydroxyanisole (BHA), sodium erythorbate, thiab tert-butyl hydroxytoluene (BHT), tau pom los txhawb cov teebmeem carcinogenic rau tib neeg noj qab haus huv [7]. Yog li ntawd, kev loj hlob sai ntawm cov kev tshawb fawb ntawm cov nroj tsuag-derived natural antioxidants tau tshwm sim ntau xyoo dhau los. Qhov tseem ceeb, nws tau pom tias nce qib ntawm ROS tuaj yeem ua kom tawv nqaij pigmentation. Ntawm cov ROS muab tau los ntawm melanocytes thiab keratinocytes, nitric oxide, NO, txhawb kev tsim cov melanin los ntawm kev txhim kho cov protein qhia qib ntawm tyrosinase thiab tyrosinase-related protein 1 (TRP1) [8,9]. Cov nyhuv ntawm ROS ntawm melanin ntau lawm tau kawm siv ntau yam antioxidants, xws li N-acetyl cysteine, tshem tawm qhov kev txiav txim ntawm UVB-induced -melanocyte-stimulating hormone (-MSH) [10].

Melanin yog tsim thiab secreted los ntawm melanocytes uas yog faib nyob rau hauv lub basal txheej ntawm daim tawv nqaij epidermis [11]. Thawj ob kauj ruam ntawm txoj kev melanogenesis hauv tib neeg yog hydroxylation ntawm L-tyrosine rau 3-4-dihydroxy phenylalanine (L-DOPA) thiab oxidation ntawm L-DOPA rau o-dopaquinone. Tyrosinase (EC 1.14.18.1) yog tus nqi-txheej enzyme uas catalyzes ob qho tib si ntawm thawj ob qhov kev tshwm sim thaum lub sij hawm melanogenesis [12]. Melanin plays lub luag haujlwm tseem ceeb hauv kev tiv thaiv daim tawv nqaij tiv thaiv tshav ntuj ultraviolet (UV) kev puas tsuaj thiab tseem muaj lub luag haujlwm rau xim plaub hau, tawv nqaij, thiab ob lub qhov muag. Nws tau raug tshaj tawm tias ntau yam kab mob dermatological, xws li lub hnub nyoog melasma, post-inflammatory melanoderma, freckles, thiab qhov chaw ntawm actinic kev puas tsuaj, uas tshwm sim los ntawm tsub zuj zuj ntawm ntau dhau ntawm epidermal melanin [13]. Inhibitors ntawm melanin synthesis tau siv ntau dua hauv cov khoom tu tawv nqaij rau kev kho lossis tiv thaiv cov tawv nqaij hyperpigmented mob [14,15]. Tsis tas li ntawd, cov tshuaj antioxidants, xws li arbutin lossis kojic acid [16], tau siv rau hauv kev kho mob ntawm hyperpigmentation [17]. Tsis ntev los no, daim tawv nqaij-dawb tshuaj pleev ib ce tau accounted rau ib tug loj feem ntawm cov tshuaj pleev ib ce. Tsis tsuas yog cov poj niam uas muaj tawv nqaij tsaus, tab sis kuj yog cov neeg uas tab tom sim daws qhov tsaus me ntsis ntawm lawv cov tawv nqaij uas tshwm sim los ntawm UV teeb, cev xeeb tub, los yog muaj hnub nyoog nce zuj zus siv cov khoom siv tawv nqaij dawb.

Melanogenesis yog tswj los ntawm ntau yam. Piv txwv li, tyrosinase-related protein-1 (TRP1), microphthalmia-associated transcription factor (MITF), thiab tyrosinase-related protein-2 (TRP2) tau tshaj tawm los tswj kev tsim cov melanin [18–20. ]. Lub melanocortin 1 receptor (MC1R) kuj tseem muaj qhov tseem ceeb hauv -MSH-induced melanin synthesis [21]. Muaj ntau txoj hauv kev qhia txog kev koom tes hauv kev tsim melanin. Ntawm cov kev taw qhia sib txawv uas tswj hwm melanin ntau lawm, cyclic AMP (cAMP)-mediated protein kinase A (PKA) ua kom muaj lub luag haujlwm tseem ceeb hauv kev tswj hwm ntawm melanogenesis [22]. Hauv cAM-PKA kev taw qhia txoj hauv kev, cAMP induces lub activation ntawm PKA [23], uas yog ua raws li phosphorylation ntawm cAMP teb cov ntsiab lus-binding protein (CREB). CREB yog ib qho intracellular transcription yam uas txhawb kev qhia ntawm microphthalmia factor (MITF) noob, uas yog ib qho tseem ceeb hauv melanogenesis. MITF yog ib qho kev hloov pauv uas khi rau thaj chaw txhawb nqa ntawm cov noob melanogenic tyrosinase, TRP1, thiab TRP2, txhawb nqa lawv cov lus qhia [24,25]. Tom qab ntawd, intracellular melanin cov ntsiab lus nce [26]. Nws tau raug tshaj tawm tias -MSH nce qib cAMP thiab feem ntau yog siv los qhib cov phosphorylation ntawm CREB thiab tom qab ntawd txhim kho MITF protein ntau [27].

MITF qhia tau tswj los ntawm ntau txoj hauv kev qhia. c-Jun N-terminal kinase (JNK) thiab p38 mitogen-activated protein kinase (MAPK) tau koom nrog hauv kev ua kom MITF qhia thiab qhov ua rau muaj kev nthuav qhia tyrosinase. Extracellular responsive kinase (ERK) activation signals kuj nce CREB phosphorylation thiab tom qab MITF qhia, uas modulates melanin synthesis [28,29]. Yog li ntawd, ntau tus neeg ua kom tawv nqaij dawb tau tshaj tawm tias inhibit MITF transcriptional kev ua haujlwm los ntawm kev txo qis cov protein qhia qib ntawm tyrosinase, TRP1, thiab TRP2 los ntawm kev txo qis ntawm p38MAPK-mediated MITF phosphorylation. Tsis tas li ntawd, txoj kev ERK tau piav qhia tias yuav koom nrog MITF txoj cai thaum lub sij hawm melanogenesis [30,31]. Ua kom ERK phosphorylates MITF, ua rau degradation ntawm MITF, ua rau txo cov ntsiab lus melanin [32,33].

Kev ntxhov siab oxidative tuaj yeem yog qhov tshwm sim ntawm ROS overproduction lossis txo cov tshuaj tiv thaiv antioxidant [1]. Kev tshawb nrhiav thiab kev siv cov tshuaj antioxidants ntuj tseem nyob hauv kev tsom mus rau ntau pab pawg tshawb fawb thoob plaws ntiaj teb. Tsis ntev los no, tau muaj kev txaus siab rau kev tshawb fawb thiab kev loj hlob ntawm phytochemicals, xws li ntuj antioxidants thiab phenolic tebchaw los ntawm ntau yam ntuj tsim, uas yuav siv tau raws li nutraceuticals, anti-aging tshuaj pleev ib ce, thiab cov khoom siv tshuaj [34]. Phoenix dactylifera L. yog ib qho ntawm cov txiv hmab txiv ntoo loj hlob nyob rau hauv Middle East thiab North Africa [35,36]. Cov kev tshawb fawb yav dhau los tau tshaj tawm tias P. dactylifera L. txiv hmab txiv ntoo muaj cov tshuaj tua kab mob antioxidant, dawb radical-scavenging, tshuaj tua kab mob, thiab kev ua haujlwm tiv thaiv kab mob [37,38]. Cov teebmeem kev ua haujlwm ntawm P. dactylifera L. tsis yog txwv rau cov txiv hmab txiv ntoo lawv tus kheej nkaus xwb tab sis kuj rau lawv cov noob, uas yog ib yam khoom ntawm cov txiv hmab txiv ntoo [39,40]. Ua tsaug rau nws cov khoom antioxidant, P. dactylifera L. noob rho tawm tuaj yeem ua lub hauv paus ntawm cov tshuaj antioxidant uas ua rau oxidative kev nyuaj siab [41–43]. Txawm li cas los xij, ob peb txoj kev tshawb fawb tau ua tiav ntawm daim ntawv thov ntawm P. dactylifera L. noob rau kev txhim kho cov khoom siv tshuaj pleev ib ce. Cov noob no feem ntau yog muab pov tseg raws li pov tseg, tab sis lawv qhov tseem ceeb raws li cov khoom siv tshuaj pleev ib ce yog qhov tseem ceeb ntawm no. Txog rau tam sim no, tsis muaj ntawv ceeb toom hais txog cov txheej txheem ntawm kev txiav txim ntawm P. dactylifera L. noob extract nyob rau hauv lub tshav pob ntawm cov tshuaj pleev ib ce los yog cov teebmeem ntawm cov noob extract ntawm melanin ntau lawm. Lub hom phiaj ntawm txoj kev tshawb fawb tam sim no yog los txiav txim siab txog lub peev xwm ntawm P. dactylifera L. noob extract raws li ib qho melanogenesis inhibitor rau kev lag luam tshuaj pleev ib ce.

Cov txheej txheem hauv qab ntawm cov teebmeem inhibitory ntawm P. dactylifera L. noob extract ntawm melanogenesis tseem tsis tau tshawb xyuas. Lub hom phiaj ntawm txoj kev tshawb fawb tam sim no yog txhawm rau tshawb xyuas cov yam ntxwv antioxidant thiab muaj peev xwm ntawm kev ua ntawm P. dactylifera L. noob extract ntawm melanogenesis los ntawm kev tshuaj xyuas MITF transcription regulators thiab phosphorylation ntawm regulators ntawm MAPK thiab PKA signaling pathways.

tshiab cistanche

2. Cov ntaub ntawv thiab cov txheej txheem

2.1. Tshuaj thiab Reagents

Cov tshuaj reagents siv nyob rau hauv txoj kev tshawb no tau yuav los ntawm Sigma-Aldrich Chemical Co. (St. Louis, MS, USA). Cov tshuaj tiv thaiv yog los ntawm Santa Cruz Biotech (Santa Cruz, CA, USA) thiab ECL reagent yog los ntawm Millipore (Billerica, MA, USA).

2.2. Kev npaj thiab Extraction ntawm P. dactylifera Seed Extract

Cov txiv hmab txiv ntoo qhuav P. dactylifera L. tau sau nyob rau hauv 2019 los ntawm cov khw muag khoom ib txwm nyob hauv Medina City, Saudi Arabia. P. dactylifera L. cov noob raug ntxuav kom huv si, raug tshav ntuj, cua qhuav rau ib hnub, thiab tom qab ntawd qhuav ntawm 80 ◦C rau 2 teev hauv qhov cub. Cov noob lub cev qhuav dej tau pulverized rau cov hmoov zoo (#50 mesh) nrog lub tshuab cog lus (Retsch Ultra Centrifugal Zeb thiab Sieving Tshuab, Hom ZM1, Haan, Lub Tebchaws Yelemees). Cov hmoov tau muab sau rau hauv lub khob ntim ntim thiab khaws cia ntawm 25 ◦C kom txog thaum siv. Pulverized desiccated P. dactylifera L. noob hmoov (2 g) tau muab tso rau hauv ib lub raj mis extraction uas muaj 10 mL dej distilled. Tom ntej no, ultrasonic extraction tau nqa tawm nrog ULTRAsonikTM 57H qauv (250 W, C, thiab A Muag Khoom Muag Khoom, Yucaipa, CA, USA). Kev ua haujlwm zaus yog 45 kHz rau 30 min. Tom qab rho tawm, cov qauv tau centrifuged ntawm 4500 rpm rau 50 min ntawm 25 ◦C nrog Eppendorf Centrifuge 5810 R (Hamburg, Lub teb chaws Yelemees). Cov supernatants tau sau thiab lim nrog cov nylon lim (pore loj: 0.45 μm) thiab cov filtrate yog concentrated rau 9.8 mg / mL.

2.3. Ultra-Performance Liquid Chromatography (UPLC) Kev Ntsuam Xyuas ntawm P. dactylifera Seed Extract thiab Ferulic Acid

P. dactylifera noob extract tau txheeb xyuas siv ACQUITY UPLC H chav kawm nrog lub photodiode array detector (Waters, Milford, MA, USA). Kem yog ACQUITY UPLC BEH C18 kem, 130 Å, 1.7 μm, 2.1 mm × 100 mm. Txawb theem A yog 0.5 feem pua ​​​​acetic acid, thiab theem B yog acetonitrile. Rau lub sijhawm cov ntsiab lus 0 thiab 5 min, qhov feem pua ​​​​ntawm A / B yog 95/5; rau 20 min, qhov feem pua ​​​​ntawm A / B yog 5/95; rau lub sijhawm cov ntsiab lus 21 thiab 25 min, qhov feem pua ​​​​ntawm A / B yog 95/5. Ferulic acid (1 ppm) tau siv los ua tus qauv zoo.

2.4. DPPH Kev Tshawb Fawb Kev Ua Si

Hauv qhov kev ntsuam xyuas no, vitamin C ({{0}}.5 mg/mL) thiab BHA ({4}}.1 mg/mL) tau siv los ua cov qauv tshuaj antioxidant. P. dactylifera L. noob extract ntawm ntau yam concentrations (0.0049, 0.0245, thiab 0.049 mg/mL) tau ntxiv rau 2.9 mL ntawm DPPH (60 μM) tov. Antioxidant Cheebtsam hauv cov noob extract tuaj yeem pub hydrogen rau DPPH thiab hloov DPPH rau hauv daim ntawv txo. Qhov ua rau txo qis hauv absorbance ntawm 517 nm tau sau tseg siv UV-Vis spectrophotometer [44].

2.5. ABTS ntxiv rau Kev Tshawb Fawb Txog Kev Tshawb Fawb

Lub ABTS ntxiv rau kev tshem tawm ntawm P. dactylifera L. noob extract tau muab piv nrog cov vitamin C ({0}}.9 mg/mL) thiab BHA ({{10}}}. 9 mg/ml). Rau qhov kev xeem no, ABTS ntxiv tau siv. Cation yog thawj zaug tsim los ntawm reacting cov tshuaj ABTS (7 mM) nrog poov tshuaj persulfate (2.45 mM), thiab cov sib tov tau tso cai sawv ntsug nyob rau hauv qhov tsaus ntuj nti rau tsawg kawg yog 6 teev ua ntej siv. Qhov nqus ntawm 734 nm tau ntsuas 10 min tom qab sib xyaw ntawm qhov sib txawv ntawm cov noob extract (0.0098, 0.049, thiab 0.098 mg / mL) nrog 1 mL ntawm ABTS ntxiv rau cov tshuaj [45].

2.6. Kev txiav txim siab ntawm tag nrho cov ntsiab lus Phenolic

Tag nrho cov ntsiab lus phenolic tau txiav txim siab siv Folin-Ciocalteu reagent [46], thiab gallic acid (2 ug / mL) tau ua haujlwm raws li tus qauv zoo. Kev sib txawv ntawm P. dactylifera L. noob extract (0.0196, 0.098, thiab 0.196 mg/mL) tau npaj hauv 80 feem pua ​​​​methanol; 100 μL ntawm extract tau pauv mus rau hauv lub raj kuaj thiab 0.5 mL ntawm Folin-Ciocalteu reagent (yav dhau los diluted 10- quav nrog dej deionized) tau ntxiv thiab sib tov. Cov sib tov tau tso cai sawv ntsug ntawm chav tsev kub rau 5 min; 1.5 mL ntawm 20 feem pua ​​​​sodium carbonate tau ntxiv. Tom qab sawv ntawm chav sov li 2 h, qhov nqus tau nyeem ntawm 760 nm siv lub UV-Vis spectrophotometer.

2.7. Kev txiav txim siab txo qhov peev xwm

Vitamin C ({{0}}}008 mg/mL) los yog BHA (4.8 mg/mL) tau siv los ua cov qauv zoo hauv qhov kev ntsuam xyuas no. Lub peev xwm txo qis ntawm cov noob extract tau txiav txim siab raws li txoj kev piav qhia los ntawm Oyaizu [47]. Ntau qhov ntau ntawm P. dactylifera L. noob extract (0.0073, 0.036, 0.073 mg/mL) tau tov nrog phosphate buffer (2.5 mL, 0.2 M, pH 6.6). ) thiab potassium ferricyanide [K3Fe (CN)6] (2.5 mL, 1 feem pua ​​w/v). Qhov sib tov yog incubated ntawm 50 ◦C rau 20 min. Trichloroacetic acid (2.5 mL, 10 feem pua ​​w / v) tau ntxiv rau qhov sib tov, uas yog tom qab ntawd centrifuged ntawm 1000 × g rau 10 min. Cov txheej txheej sab saud (2.5 mL) yog tov nrog dej distilled (2.5 mL) thiab FeCl3 (0.5 mL, 0.1 feem pua ​​w / v), thiab qhov nqus ntawm 700 nm tau ntsuas.

2.8. Kev ntsuas ntawm Hlau (II) Ion Chelating Capacity

Rau kev ntsuas ntawm Fe2 ntxiv -ion chelating muaj peev xwm ntawm P. dactylifera L. noob extract, EDTA (0.02, 0.04, thiab { {10}} 08 mg/mL) tau siv los ua tus qauv zoo. Qhov sib txawv ntawm cov noob extract (0.0196, 0.098, thiab 0.196 mg/mL) tau ntxiv rau cov tshuaj FeCl2 (0.05 mL, 1 mM). Cov tshuaj tiv thaiv sib xyaw yog reacted nrog ferrozine (0.1 mL, 1 mM), ces cov sib tov yog quantified rau 1 mL nrog methanol thiab incubated ntawm 25 ◦C rau 10 min. Lub absorbance ntawm cov tshuaj tiv thaiv sib tov yog ntsuas ntawm 562 nm [48].

2.9. Cell Viability Assay

Kev ntsuam xyuas cell viability tau ua los ntawm MTT txoj kev [49]. Cov hlwb raug nthuav tawm rau ntau qhov ntau ntawm P. dactylifera L. noob extract (0.049, 0.245, thiab 0.49 mg/mL) rau 24 teev ntawm 37 ◦C thiab 5 feem pua ​​​​CO2 nyob rau hauv lub tshuab nqus tsev vacuum thiab MTT tov tau ntxiv rau hauv qhov dej. Lub insoluble derivative ntawm MTT yog solubilized nrog ethanol-DMSO (1: 1 tov tov). Qhov nqus dej ntawm qhov dej ntawm 570 nm tau ntsuas los ntawm kev nyeem ntawv microplate. B16F10 hlwb (BCRC60031) tau txais los ntawm Bioresource Collection thiab Research Center (BCRC), Hsinchu nroog, Taiwan. Cov hlwb tau khaws cia hauv DMEM (Hyclone, Logan, UT, USA) ntxiv nrog 10 feem pua ​​​​fetal bovine serum thiab 1 feem pua ​​​​cov tshuaj tua kab mob. Tsis tas li ntawd, cov nyhuv ntawm P. dactylifera L. noob ntawm B16F10 cell viability kuj tau soj ntsuam los ntawm trypan xiav cais tawm txoj kev ntsuam xyuas. Tom qab kev kho mob nrog cov noob extract, hlwb tau trypsinized thiab centrifuged los sau cov pellet. Lub cell pellet tau rov raug ncua hauv 300 μL ntawm DMEM cell kab lis kev cai xov xwm. Ib qho me me aliquot ntawm kev ncua ntawm tes (20 μL) tau sib tov maj mam nrog 10 μL trypan xiav (4 feem pua ​​​​hauv PBS). Cov xov tooj ntawm tes raug suav (cov hlwb uas tsis tuaj yeem yog xiav thiab cov hlwb siv tau tsis muaj xim) los ntawm kev siv hemocytometer nyob rau hauv lub tshuab tsom.

2.10. Kev ntsuas ntawm Mushroom Tyrosinase Kev Ua Haujlwm

Cov tshuaj nceb tyrosinase (10 μL, 200 units) tau ntxiv rau 96-zoo microplate. Cov tshuaj tiv thaiv sib xyaw muaj 5 mM ntawm L-DOPA yaj hauv phosphate-buffered saline (PBS) (50 mM, pH 6.8), thiab P. dactylifera L. noob extract (0.049, 0.245 , thiab 0.49 mg / mL), kojic acid (200 mM; 0.03 mg / mL), los yog arbutin (2 mM; 0.54 mg / mL). Qhov sib tov sib xyaw tau muab tso rau ntawm 37 ◦C rau 30 min thiab qhov nqus ntawm dopachrome tsim tau ntsuas ntawm 490 nm. Kev ntsuas ntawm nceb tyrosinase kev ua haujlwm tau ua raws li tau piav qhia yav dhau los [50].

2.11. Kev txiav txim siab ntawm Melanin Cov ntsiab lus

Cov hlwb B16F10 tau kho nrog -MSH (100} nM) rau 24 teev thiab tom qab ntawd kho nrog P. dactylifera L. noob extract (0.0245–0.147 mg/mL) los yog arbutin (0.54 mg / mL) ntxiv rau 24 teev. Tom qab kev kho mob, cov cell pellets tau solubilized hauv NaOH tov (1 N) ntawm 60 ◦C rau 60 min. Cov ntsiab lus melanin tau kuaj pom ntawm 405 nm, raws li yav dhau los tau piav qhia los ntawm Tsuboi [51].

Cistanche inhibits melanin ntau lawm.

2.12. Kev ntsuas ntawm Intracellular Tyrosinase Kev Ua Haujlwm

Intracellular tyrosinase kev ua haujlwm tau txiav txim siab raws li tau piav qhia yav dhau los [52]. Cov hlwb raug kho nrog -MSH (100} nM) rau 24 teev thiab tom qab ntawd nrog P. dactylifera L. noob extract (0.0245–0.147 mg / mL) lossis arbutin (0.54 mg / mL) rau 24 teev. Tom qab kev kho mob, cov cell rho tawm (100 μL) tau tov nrog freshly npaj L-DOPA tov (0.1 feem pua ​​​​hauv PBS) thiab incubated ntawm 37 ◦C, thiab cov absorbance ntawm 490 nm tau ntsuas.

2.13. Western Blotting Experiment

Cov hlwb raug kho nrog P. dactylifera L. noob extract ({{0}}}0245, 0.0368, 0. {14}}49, thiab 0.147 mg/mL) los yog arbutin (0.54 mg/mL) thiab ces lysed nyob rau hauv PBS uas muaj nondiet P—40 (1 feem pua), sodium deoxycholate (0.5 feem pua), sodium dodecyl sulfate (SDS, 0.1 feem pua), aprotinin (5 ug/mL), phenylmethylsulfonyl fluoride (100 ug/mL), pepstatin A (1 ug/mL), thiab EDTA (1 mM) ntawm 4 ◦C rau 20 feeb. Tag nrho cov lysates tau suav nrog siv cov khoom siv micro BCA (Thermo Fisher Scientific, Waltham, MA, USA). Proteins (30 ug) tau daws los ntawm SDS-polyacrylamide gel electrophoresis thiab electrophoretically pauv mus rau polyvinylidene fluoride membrane. Lub membrane raug thaiv hauv 5 feem pua ​​​​cov mis nyuj tsis muaj roj hauv PBST (PBS nrog 0.05 feem pua ​​Tween-20) thiab incubated ntawm 4 ◦C thaum hmo ntuj nrog cov tshuaj tiv thaiv hauv qab no diluted hauv PBST: MITF Ab (1: 1000), TRP1 Ab (1:6000), TRP2 Ab (1:1000), MC1R Ab (1:500), GAPDH Ab (1:1500), tyrosinase Ab (1:2000), p-p38 Ab (1:500), p38 Ab (1:500), p-JNK Ab (1:500), JNK Ab (1:500), p-ERK Ab (1:500), ERK Ab (1:500), p-CERB Ab (1: 500), and CERB Ab (1:200), all from Santa Cruz Biotech (Dallas, TX, USA). Cov tshuaj tiv thaiv kab mob tau raug kuaj pom los ntawm horseradish peroxidase-conjugated Secondary antibody (Amersham Corp.) ua raws li ECL kev kuaj pom qhov system (Amersham) raws li cov chaw tsim khoom cov lus qhia.

2.14. PKA Inhibitor Assay

Cov hlwb raug kho nrog -MSH (100 nM) rau 24 h tom qab 1 h ntxiv ntawm 10 μM ntawm PKA regulator H89. Tom qab kev kho mob, P. dactylifera L. noob extract (0.147 mg/mL) thiab H89 tau ntxiv rau cov hlwb thiab incubated rau 23 teev ntxiv. Cov ntsiab lus melanin tau txiav txim siab raws li tau piav qhia saum toj no.

2.15. Kev txheeb cais

Kev txheeb xyuas kev txheeb xyuas tau ua tiav siv Tukey's post hoc test, uas yog siv rau kev sib piv ntawm cov ntaub ntawv ntsuas, siv Cov Txheej Txheem Txheej Txheem rau Social Sciences (SPSS) version 22.0 statistical software (SPSS Inc., Chicago, IL, USA). * p < 0="" 05="" tau="" suav="" tias="" yog="" qhov="" tseem="" ceeb,="" **="" p="">< 0.01,="" ***="" p=""><>

3. Cov txiaj ntsig

3.1. UPLC Kev Ntsuam Xyuas ntawm P. dactylifera L. Seed Extract

Cov txiaj ntsig ntawm UPLC tsom xam tau pom hauv daim duab 1A, B. Daim duab 1A qhia txog qhov sib tshooj ntawm P. dactylifera L. noob extract thiab ferulic acid ntawm 330 nm. Daim duab 1B qhia zoo sib xws scan spectra ntawm 220 thiab 500 nm rau P. dactylifera L. noob extract thiab ferulic acid.

3.2. Antioxidant Yam ntxwv ntawm P. dactylifera L. Seed Extract

Kev ua haujlwm antioxidant ntawm P. dactylifera L. noob extract hauv vitro qhia pom muaj peev xwm antioxidant. Hauv kev soj ntsuam DPPH, vitamin C ({{0}}}05 mM; 0.53 mg/mL) thiab tert-butyl hydroxyanisole (BHA) (0 .1 mg/mL) tau siv los ua cov qauv zoo antioxidant. Lub peev xwm DPPH scavenging ntawm cov extract yog 49.97 ± 2.9 feem pua ​​​​, 81.36 ± 0.56 feem pua ​​, thiab 78.53 ± 3.83 feem pua ​​​​ntawm kev tswj rau cov extract concentrations ntawm 0.0{{ 31}}49, 0.0245, thiab 0.049 (mg / mL), feem. Hauv kev sib piv, cov peev txheej ntawm cov vitamin C thiab BHA yog 90.12 ± 0.31 feem pua ​​thiab 90.44 ± 0.49 feem pua, raws li (Daim duab 2A).

Qhov no ABTS radical cation yog xiav xim thiab yog reactive rau feem ntau antioxidants. Thaum lub sij hawm no cov tshuaj tiv thaiv, lub xiav ABTS radical cation yog hloov dua siab tshiab rau nws cov xim tsis muaj xim. Cov tshuaj tiv thaiv tau saib xyuas spectrophotometrically. ABTS ntxiv rau kev tshem tawm lub peev xwm ntawm cov extract yog 5.69 ± 1.36 feem pua, 18.81 ± 0.68 feem pua, thiab 66.82 ± 8.51 feem pua ​​​​ntawm kev tswj ntawm qhov ntau ntawm 0.0{{16 }}98, 0.{20}}49, thiab 0.098 mg/mL, raws Hauv qhov sib piv, ABTS ntxiv rau qhov muaj peev xwm ntawm cov vitamin C (0.9 mg / mL) thiab BHA (0.9 mg / mL) yog 95.52 ± 0.12 feem pua ​​thiab 40.3 ± 0.83 feem pua. Cov txiaj ntsig hauv daim duab 2B qhia tau hais tias cov noob rho tawm ua rau ib qho tseem ceeb ntawm ABTS ntxiv rau radical.

Txhawm rau txiav txim siab tag nrho cov ntsiab lus phenolic ntawm P. dactylifera L. noob extract, gallic acid (2 ug / mL) tau siv los ua tus qauv zoo. Cov txiaj ntsig hauv daim duab 2C qhia tias tag nrho cov ntsiab lus phenolic hauv 0.0196, 0.{{20}}98, thiab 0. 196 (mg / mL) ntawm cov extract yog 33.43 ± 2.33 feem pua, 117.22 ± 7.81 feem pua, thiab 195.4 ± 10.91 feem pua, feem. Lub gallic acid sib npaug (GAE) ntawm cov lus sau tseg ntawm cov noob rho tawm yog 0.068 ± 0.01, 0.365 ± 0.01, thiab 0.642 ± 0.03, raws li (Daim duab 2C).

3.3. Inhibitory Effects ntawm P. dactylifera L. Noob Extract ntawm Melanogenesis

Cov txiaj ntsig tau pom hauv daim duab 4A qhia tau tias 0.049 mg/mL ntawm P. dactylifera L. noob extracttsis muaj kev cuam tshuam inhibitory ntawm nceb tyrosinase kev ua haujlwm. Ntawm qhov tod tes, cov enzyme inhibition feem pua ​​​​yog 8.34 ± 2.67 feem pua ​​​​thiab 33 ± 2.36 feem pua ​​​​ntawm kev tswj hwm rau 0.245 thiab 0.49 mg / mLP. dactylifera L. noob extract kho, feem. Tsis tas li ntawd, tyrosinase inhibition feem pua ​​ntawm kojic acid (200 μM; 0.0}3 mg/mL) thiab arbutin (2 mM; 0.54 mg/mL) ) yog 56.26 ± 5.06 feem pua ​​​​thiab 64.56 ± 2.88 feem pua ​​​​ntawm kev tswj hwm, raws li (Daim duab 4A). Yog li, qhov siab dua ntawm P. dactylifera L. noob extract (0.245 thiab 0.49 mg / mL) tuaj yeem sawv cev rau qib inhibitory ntawm nceb tyrosinase.

Hauv daim duab 4B, cov txiaj ntsig tau qhia tias tsuas yog 0.147 mg/mL ntawm P. dactylifera L. noob extract tuaj yeem txo cov ntsiab lus melanin hauv B16F10 melanoma hlwb. Tom qab kev kho mob, cov ntsiab lus melanin hauv B16F10 hlwb yog 80.66 ± 5.43 feem pua ​​​​ntawm cov tswj. Rau cov qauv zoo, arbutin ({29}}.54 mg / mL), cov ntsiab lus ntawm cov tshuaj melanin tseem tshuav yog 61.19 ± 8.17 feem pua ​​​​ntawm cov tswj. Cov txiaj ntsig tau qhia tias 0.147 mg / mL ntawm P. dactylifera L. noob extract pom cov teebmeem me dua arbutin. Qhov tseem ceeb ntawm intracellular tyrosinase kev ua ub no yog 85.1 ± 8.1 feem pua ​​​​thiab 73.7 ± 11.9 feem pua ​​​​rau 0.098 thiab 0.147 mg / mL ntawm P. dactylifera L. noob extract, feem. Cov kev ua haujlwm ntawm intracellular tyrosinase ntxiv yog 64.3 ± 14.9 feem pua ​​​​ntawm kev tswj xyuas tom qab cov hlwb tau kho nrog arbutin (Daim duab 4C). Cov txiaj ntsig tau qhia tias ntau dua ntawm P. dactylifera L. noob extract tseem pom qhov cuam tshuam me me ntawm -MSH-induced tyrosinase kev ua hauv B16F10 hlwb dua li arbutin.

3.4. P. dactylifera L. noob Extract Inhibits qhov nthuav qhia theem ntawm cov protein koom nrog hauv melaninogenesis

Western blots tau siv los tshuaj xyuas cov theem qhia ntawm melanogenesis ntsig txog cov proteins hauv B16F10 hlwb (Daim duab 5A). Cov txiaj ntsig tau qhia tias kev kho mob nrog ntau qhov ntau ntawm P. dactylifera L. noob extract ua rau txo qis ntawm MC1R, MITF, tyrosinase, TRP1, thiab TRP2. Cov teebmeem inhibitory ntawm extract ntawm cov protein qhia tau pom meej ntawm qhov concentration ntawm {{10}}.147 mg/mL. Qhov kev hloov pauv hauv qib protein qhia rau MC1R yog 0.74 ± 0.02; rau MITF nws yog {{20}}.51 ± 0.03; rau tyrosinase nws yog 0.54 ± 0.26; rau TRP-1 nws yog 0.57 ± 0.15; thiab rau TRP-2 nws yog 0.49 ± 0.1 (Daim duab 5B).

Cov theem qhia ntawm melanogenesis ntsig txog cov cim qhia cov proteins tau tshuaj xyuas siv Western blots (Daim duab 5C). Cov txiaj ntsig tau qhia tias 0.147 mg/mL ntawm P. dactylifera L. noob extract kev kho mob pom tau tias ua rau txo qis ntawm p-p38, p-JNK, p-ERK, thiab p-CREB. Kev hloov pauv ntawm cov protein qhia qib rau p-p38 yog 0.88 ± 0.15; rau p-JNK nws yog 0.68 ± 0.39; rau p-ERK nws yog 0.65 ± 0.23; thiab rau p-CREB nws yog 0.44 ± 0.07 (Daim duab 5D).

3.5. P. dactylifera L. Seed Extract Tsis cuam tshuam rau Tyrosinase, TRP1, TRP2, lossis MITF Gene Expression

Cov noob qhia theem ntawm melanogenesis ntsig txog cov proteins, xws li tyrosinase, TRP1, TRP2, thiab MITF tau tshuaj xyuas los ntawm ntau npaum li cas tiag-lub sij hawm polymerase saw cov tshuaj tiv thaiv (RT-PCR). Qhov txheeb ze ntawm cov lus qhia ntawm tyrosinase/GAPDH rau 0.0367, 0.049, thiab 0.147 mg/mL ntawm P. dactylifera L. noob extract kev kho mob yog 3.51 ± 0.52, 2.38 ± 0.24, thiab 1.64 ± 0.55, feem. Rau TRP-1/GAPDH, tus txheeb ze cov lus qhia theem rau 0.0367, 0.049, thiab {{4{{43} }}}.147 mg/mL ntawm P. dactylifera L. noob extract kev kho mob yog 1.64 ± 0.2, 1.15 ± 0.02, thiab 0. 89 ± 0 07, raws. Tus txheeb ze ntawm kev qhia theem ntawm TRP-2/GAPDH rau 0.0367, 0.049, thiab 0.147 mg /mL ntawm P. dactylifera L. noob extract yog 1.08 ± 0.13, 0.85 ± 0.01, thiab 0.7 ± 0.05, raws li. Rau MITF/GAPDH, cov txheeb ze qhia tau zoo rau 0.0367, 0.049, thiab 0.147 mg/mL ntawm P. dactylifera L. noob extract kho yog 1.48 ± 0.19, 1.03 ± 0.02, thiab 0.75 ± 6.0.

4. Kev sib tham

Nws tau raug tshaj tawm tias melanogenesis nce cellular oxidative kev nyuaj siab. Tsis tas li ntawd, ntau cov antioxidants, ROS scavengers, inhibitor scavengers, thiab inhibitors yuav inhibit UV-mediated melanin synthesis [54]. Yog li ntawd, antioxidants, ROS scavengers, thiab inhibitors ntawm melanogenesis tau nce ntxiv rau cov tawv nqaij-dawb tshuaj pleev ib ce rau kev tiv thaiv ntawm undesirable daim tawv nqaij hyperpigmentation [14]. Nws kuj tau pom tias stimulation ntawm metallothionein, ib qho antioxidant endogenous yuav inhibit melanogenesis hauv melanocytes [55]. Tib neeg cov tawv nqaij feem ntau raug rau ib puag ncig oxidizing kuab paug los yog UV teeb thiab puas los ntawm extrinsic ib puag ncig yam. Tshwj xeeb, UV irradiation tau raug tshaj tawm los ua rau cov tiam ntawm ROS hauv daim tawv nqaij thiab txhawb nqa cell membrane lipid peroxidation [56]. Txhawm rau tiv thaiv oxidative kev nyuaj siab, daim tawv nqaij yog nruab nrog tshwj xeeb antioxidant systems [57].

Txhawm rau elucidate cov kev ua antioxidant ntawm P. dactylifera L. noob extract, DPPH thiab ABTS ntxiv rau cov kev ua si radical scavenging, tag nrho cov ntsiab lus phenolic, txo lub peev xwm, thiab hlau ion-chelating muaj peev xwm ntawm cov extract tau txiav txim siab raws li yav tas los piav [47,48]. P. dactylifera L. noob extract exerted ntau yam antioxidant kev ua ub no nyob rau hauv tag nrho cov aforementioned analytical kev tshawb fawb. Cov txiaj ntsig tau pom tias muaj peev xwm antioxidant ntawm P. dactylifera L. noob extract hla ntau qhov sib txawv nrog cov txiaj ntsig sib txawv. DPPH-scavenging kev ua ntawm cov noob extract tau pom nyob rau hauv daim duab 2A yog txawv los ntawm ABTS ntxiv rau hauv daim duab 2B, uas tej zaum yuav tau tshwm sim los ntawm txawv mechanisms ntawm lub antioxidant-radical kev sib tshuam. Tsis tas li ntawd, stoichiometry ntawm cov tshuaj tiv thaiv ntawm cov tshuaj tiv thaiv antioxidant hauv cov noob extract yuav txawv, uas ua rau muaj qhov sib txawv ntawm cov dawb radical-scavenging peev [58]. Lub peev xwm antioxidants hauv P. dactylifera L. noob extract tuaj yeem hloov pauv Fe3 ntxiv / ferricyanide complex rau hauv daim ntawv ferrous, thiab txo lub peev xwm ua haujlwm raws li qhov qhia txog kev ua haujlwm antioxidant ntawm cov noob extract qhia hauv daim duab 2D. Nws tau pom tias kojic acid [59] ua raws li tyrosinase inhibitor vim kojic acid ua cov hlau chelator rau chelate cov tooj liab ions thiab inhibit cov ions nkag mus rau qhov chaw nquag ntawm tyrosinase. Yog li ntawd, antioxidants nyob rau hauv cov noob extract yuav tsim insoluble hlau complexes nrog ferrous ions thiab ces inhibit kev sib cuam tshuam ntawm hlau ions thiab lipids. Qhov siab dua hlau ion-chelating muaj peev xwm ntawm cov noob extract tau qhia tias nws muaj peev xwm ua haujlwm antioxidant thiab muaj peev xwm cuam tshuam rau kev ua haujlwm tyrosinase (Daim duab 2E).

Lub hauv paus ntsiab lus ntawm kev soj ntsuam ntawm intracellular ROS yog tias DCFH-DA diffuses los ntawm cov cell membrane thiab yog enzymatically hydrolyzed rau DCFH los ntawm esterase; ces DCFH reacts nrog ROS (xws li H2O2) yield DCF. Kev nce nrawm hauv DCF qhia tau hais tias oxidation ntawm DCFH los ntawm intracellular radicals [60]. Lub tswv yim tom qab tshawb nrhiav cov tshuaj antioxidants tshiab rau kev ua kom tawv nqaij dawb nyob hauv qhov kev xav tias oxidative kev nyuaj siab uas tshwm sim los ntawm UV irradiation yuav ua rau muaj kev txhawb nqa ntawm melanin synthesis. UV irradiation tau tshaj tawm los tsim ROS hauv cov ntaub so ntswg uas tuaj yeem ua rau melanin tsim los ntawm kev ua kom tyrosinase [61]. Tsis tas li ntawd, nws tau tshaj tawm tias ntau cov tawv nqaij dawb tuaj yeem tiv thaiv melanogenesis los ntawm kev cuam tshuam nrog tooj liab ions ntawm qhov chaw nquag ntawm tyrosinase lossis nrog o-quinones los thaiv cov polymerization ntawm intermediates hauv melanin synthetic txoj kev [62]. Tsis tas li ntawd, vitamin C los yog vitamin E tuaj yeem txo cov oxidation ntawm cov khoom melanin uas twb muaj lawm. Yog li, cov vitamins no tau siv dav hauv cov khoom siv tawv nqaij dawb [63]. Interestingly, peb cov txiaj ntsig tau pom cov yam ntxwv antioxidant ntawm P. dactylifera L. noob extract, qhia tias cov noob extract ntawm P. dactylifera L. tuaj yeem ua cov khoom siv tawv nqaij dawb hauv cov tshuaj pleev ib ce.

Qhov kev ntsuam xyuas MTT yog ib qho kev ntsuam xyuas colorimetric paub zoo uas ntsuas kev ua haujlwm ntawm cellular NADH / NADPH-dependent oxidoreductase enzymes uas txo MTT mus rau ntshav-xim xim formazan dyes. Qhov kev ntsuam xyuas no tuaj yeem siv los tshuaj xyuas qhov muaj peev xwm cytotoxicity ntawm cov tshuaj siv tshuaj thiab cov khoom siv tshuaj lom, vim tias cov tshuaj no txhim kho lossis inhibit cell viability. Cov txiaj ntsig tau pom hauv daim duab 3A tau ua raws li qhov ntsuas ntawm Trypan xiav cais tawm hauv daim duab 3B, uas qhia tias P. dactylifera L. noob extract tsis muaj cytotoxic cuam tshuam rau B16F10 melanoma cell viability.

Mushroom tyrosinase feem ntau yog siv los ua lub hom phiaj enzyme rau kev tshuaj xyuas cov peev xwm inhibitors ntawm melanogenesis. Nws tau pom thawj zaug tias qhov ntau npaum li cas ({{0}}}.049–0.49 mg/mL) ntawm P. dactylifera L. noob extract tuaj yeem inhibit qhov kev ua ntawm nceb tyrosinase. Cov txiaj ntsig tau pom nyob rau hauv daim duab 4A qhia tias cov noob extract tau nthuav tawm qis inhibitory cuam tshuam rau nceb tyrosinase kev ua haujlwm dua kojic acid. Tyrosinase plays lub luag haujlwm tseem ceeb hauv thawj ob kauj ruam ntawm txoj kev melanogenesis. Txhawm rau elucidate qhov tseeb inhibitory nyhuv ntawm P. dactylifera L. noob extract ntawm melanin ntau lawm, B16F10 melanin cov ntsiab lus thiab intracellular tyrosinase kev ua tau raug txiav txim. Cov txiaj ntsig tau nthuav tawm hauv daim duab 4B qhia tau hais tias ntau dua ntawm P. dactylifera L. noob extract exerted pom meej inhibitory nyhuv ntawm melanin tsim. Cov ntaub ntawv qhia tau hais tias P. dactylifera L. noob extract tiag tiag blocks melanogenesis hauv melanoma hlwb. Nrog rau qhov no, cov txiaj ntsig tau tshwm sim hauv daim duab 4C tau ua raws li cov txiaj ntsig tau pom hauv daim duab 4B. Hauv cov lus hais saum toj no, -MSH tau siv los ua ib qho inducer los txhawb melanin synthesis. -MSH tau tshaj tawm los khi melanocortin 1 receptor (MC1R) thiab ua kom cov kab mob hauv lub cev adenylate cyclase, uas nyob rau hauv lem catalyzes ATP rau cAMP thiab activates cAMP-dependent PKA. Cov txiaj ntsig hauv daim duab 5A tau qhia tias P. dactylifera L. noob extract inhibited kev qhia ntawm MC1R, yog li thaiv melanin ntau lawm induced los ntawm -MSH mediated intracellular cAMP upregulation. Tsis tas li ntawd, cAMP-mediated PKA signaling txoj hauv kev tsis ua haujlwm thiab melanogenesis raug txwv. Txhawm rau kom paub meej qhov kev xav, peb tau ua qhov kev sim PKA inhibitor, thiab cov ntaub ntawv qhia hauv daim duab 4D qhia tias inhibitory nyhuv ntawm P. dactylifera L. noob extract (0.147 mg / mL) ntawm melanogenesis hauv B16F10 hlwb raug txwv los ntawm H{{ 24}}kev kho mob. H89 (N-[2-p-bromocinnamylamino-ethyl]-5-isoquinolinesulphonamide) yog ib qho kev xaiv thiab muaj zog inhibitor ntawm PKA. Cov txiaj ntsig tau qhia tias cAMP-mediated PKA signaling tau cuam tshuam los ntawm P. dactylifera L. noob extract.

Tyrosinase, TRP1, thiab TRP2 yog peb lub enzymes loj uas tswj melanin synthesis hauv mammalian hlwb [64]. Tsis tas li ntawd, MITF yog tus tswj hwm loj ntawm tyrosinase, TRP1, thiab TRP2, thiab yog tus tswj hwm tseem ceeb tshaj plaws ntawm melanocyte pigmentation [65]. Cov txiaj ntsig hauv daim duab 5A qhia tias P. dactylifera L. noob extract txo qis cov protein qhia cov qib ntawm cov proteins uas cuam tshuam nrog melanogenesis, inhibited tyrosinase kev ua, thiab txo melanin cov ntsiab lus hauv B16F10 hlwb.

Hauv txoj kev tshawb no, P. dactylifera L. noob extract suppressed CREB phosphorylation thiab PKA ua kom. Cov ntaub ntawv no qhia tias txoj hauv kev cAMP-PKA tuaj yeem ua lub luag haujlwm rau P. dactylifera L. noob extract-induced anti-melanogenesis (Daim duab 5C).

MAPKs tau tshaj tawm los hloov kho melanogenesis [66]. Tsev neeg MAPK muaj peb hom protein kinases, suav nrog cov xov tooj ntawm tes teb kinase (ERK), c-Jun N-terminal kinase (JNK), thiab p38 MAPK. Lub p38 MAPK tuaj yeem qhib cAMP cov lus teb cov ntsiab lus-binding protein (CREB thiab CREB activates MITF qhia, uas muaj txiaj ntsig zoo rau melanin synthesis [67]. Cov txiaj ntsig hauv daim duab 5C muab pov thawj tias P. dactylifera L. noob extract tuaj yeem ua rau CREB, JNK, thiab p38, nyob rau hauv lem inhibiting MITF qhia (Daim duab 5A). Cov kev tshwm sim no qhia tias cov. feature prominently pib ntawm ob peb daim tawv nqaij mob, nrog rau scaling, wrinkling, thiab hyperpigmentation [68].

Txhawm rau ntsuas cov txiaj ntsig ntawm P. dactylifera L. noob extract ntawm cellular melanin ntau lawm, peb txiav txim siab mRNA qib hauv B16F10 hlwb kho nrog -MSH thiab P. dactylifera L. noob extract. Cov hlwb -MSH-kho tau pom tau nce qib ntawm MITF, tyrosinase, TRP-1, thiab TRP-2, raws li xav tau. Interestingly, P. dactylifera L. noob extract-kho hlwb pom ib tug txo nyob rau hauv lub mRNA qhia ntawm cov noob caj noob ces melanogenesis. Txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb ntawm cov kab ke, uas tau qhia tias P. dactylifera L. noob extract tsis cuam tshuam rau cov noob qhia theem ntawm MITF, tyrosinase, TRP1, thiab TRP2.

Qhov no yog peb cov khoom.

Yog xav paub ntxiv, thov nyem qhov no.

Nws tau raug tshaj tawm tias qhov loj phenolic acid pom hauv P. dactylifera L. noob yog ferulic acid [69]. Ferulic acid tau pom tias muaj txiaj ntsig zoo inhibit melanin synthesis hauv B16 melanoma hlwb los ntawm inhibiting casein kinase 2-induced phosphorylation ntawm tyrosinase nyob rau hauv ib koob tshuaj [70]. Cov ntawv ceeb toom no txhawb nqa peb cov txiaj ntsig tau pom hauv daim duab 1-ferulic acid hauv cov dej extract ntawm P. dactylifera L. noob tau pab txhawb rau inhibition ntawm melanogenesis hauv B16F10 hlwb. Muaj tseeb tiag, lwm yam khoom siv hauv cov noob extract yuav tsum tau tshawb xyuas yav tom ntej.

Peb cov txiaj ntsig tau qhia tias P. dactylifera L. noob extract inhibited melanogenesis nyob rau hauv B16F10 hlwb los ntawm downregulating PKA signaling pathways. Li no, P. dactylifera L. noob extract yuav siv tau raws li ib tug zoo ntawm daim tawv nqaij-whitening neeg sawv cev.

5. Cov lus xaus

Qhov no yog thawj daim ntawv tshaj tawm txog cov txheej txheem ntawm kev ua ntawm P. dactylifera L. noob dej extract hauv melanin synthesis. Txoj kev tshawb fawb tam sim no tau qhia txog kev koom tes ntawm cAMP/PKA/CREB txoj hauv kev hauv cov nyhuv depigmenting ntawm P. dactylifera L. noob extract. Peb cov txiaj ntsig tau qhia tias P. dactylifera L. noob extract inhibited melanogenesis hauv B16F10 hlwb los ntawm kev txo qis PKA kev taw qhia txoj hauv kev. Li no, P. dactylifera L. noob extract yuav siv tau raws li ib tug tshiab dermatological antimelanogenesis tus neeg saib xyuas thiab ib tug zoo ntawm daim tawv nqaij-dawb tus neeg sawv cev.