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Inhibitory Effect Ntawm Gastrodia Elata Blume Extract Ntawm Alpha-melanocyte Stimulating Hormone-induced Melanogenesis hauv Murine B16F10 Melanoma

May 17, 2023

COV NTAUB NTAWV / Hom phiaj:Gastrodia elata Blume (GEB), tshuaj ntsuab tshuaj ntsuab, tau siv los kho ntau yam kab mob hauv lub paj hlwb (xws li, tuag tes tuag taw thiab mob stroke) thiab teeb meem ntawm daim tawv nqaij (xws li, atopic dermatitis thiab eczema) hauv cov tshuaj oriental. Txoj kev tshawb no tau tsim los tshawb xyuas seb GEB extract inhibits melanogenesis kev ua hauv murine B16F10 melanoma.

Raws li cov kev tshawb fawb cuam tshuam, cistanche yog ib hom tshuaj ntsuab uas hu ua "cov tshuaj ntsuab muaj txuj ci 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. Cov txheej txheem ntawm cistanche thiab tawv nqaij dawb yog nyob rau hauv cov nyhuv antioxidant ntawm cistanche glycosides. Melanin nyob rau hauv tib neeg daim tawv nqaij yog tsim los ntawm oxidation ntawm tyrosine catalyzed los ntawm tyrosinase, thiab cov tshuaj tiv thaiv oxidation yuav tsum muaj kev koom tes ntawm oxygen, yog li cov pa-dawb radicals nyob rau hauv lub cev ua ib qho tseem ceeb cuam tshuam rau melanin ntau lawm. Cistanche muaj cistanoside, uas yog ib qho antioxidant thiab tuaj yeem txo qhov tsim ntawm cov dawb radicals hauv lub cev, yog li inhibiting melanin ntau lawm.

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MATERIALS/MTHOD:Murine B16F10 hlwb tau kho nrog 0-5 mg / mL ntawm GEB extract lossis 400 ug / mL arbutin (kev tswj zoo) rau 72 teev tom qab kho nrog / tsis muaj 200 nM alpha-melanocyte stimulating hormone (-MSH) rau 24 teev . Melanin concentration, tyrosinase kev ua, mRNA qib, thiab cov protein qhia ntawm microphthalmia-txuas sib txuas lus transcription factor (MITF), tyrosinase, tyrosinase-related protein (Trp)1, thiab Trp2 tau txheeb xyuas hauv -MSH-tsis kho thiab -MSH-kho B16F10 hlwb.
TSEEM CEEBS:Kev kho mob nrog 200 nM -MSH induced yuav luag 2-fold melanin synthesis thiab tyrosinase kev ua si nrog rau nce mRNA qib thiab protein qhia ntawm MITF, tyrosinase, Trp1, thiab Trp2. Tsis hais txog -MSH stimulation, GEB extract ntawm koob tshuaj 0. Thaum cov koob tshuaj qis dua (0. } mg / mL) cuam tshuam rau tag nrho cov cim no hauv -MSH-kho B16F10 hlwb nyob rau hauv ib koob tshuaj. Cov teebmeem inhibitory ntawm GEB extract ntawm ntau dua cov ntsiab lus zoo ib yam li cov ntawm 400 ug / mL arbutin, ib tug paub zoo depigmenting tus neeg saib xyuas.
Cov lus xaus:Cov txiaj ntsig no qhia tias GEB qhia txog tshuaj tiv thaiv melanin synthesis los ntawm kev tawm tsam ntawm tyrosinase kev ua haujlwm nrog rau qib molecular ntawm MITF, tyrosinase, Trp1, thiab Trp2 hauv murine B16F10 melanoma. Yog li ntawd, GEB tej zaum yuav yog ib tug zoo thiab tej yam ntuj tso daim tawv nqaij-dawb tus neeg sawv cev rau daim ntawv thov nyob rau hauv kev lag luam tshuaj pleev ib ce.
Ntsiab lus: Gastrodia elata, melanogenesis, tyrosinase

Taw qhia

Tib neeg cov tawv nqaij xim yog cuam tshuam los ntawm noob caj noob ces thiab ntau yam, feem ntau yog los ntawm cov nqi thiab hom melanin. Melanin yog tsim nyob rau hauv melanosomes ntawm melanocytes thiab pauv los ntawm melanocytes mus rau keratinocytes nyob rau hauv lub basal txheej ntawm lub epidermis nyob rau hauv teb rau cov raug ntawm daim tawv nqaij rau tshav ntuj, kev nyuaj siab, thiab melanin-stimulating yam tseem ceeb [piv txwv li, alpha-melanocyte stimulating hormone (-MSH). , nitric oxide, thiab endothelin -1] [1,2]. Piv txwv li, -MSH txhawb cov melanogenesis los ntawm microphthalmia-associated transcription factor (MITF) induction, uas yog ib tug muaj zog inducer ntawm melanocyte-specific enzymes xws li tyrosinase, tyrosinase-related protein (Trp)1, thiab Trp2 [1]. Melanin ntau lawm yog tswj hwm los ntawm tyrosinase, tus nqi-liming enzyme rau melanogenesis, uas catalyzes hydroxylation ntawm tyrosine mus rau 3, 4-dihydroxyphenylalanine (L-DOPA) los ntawm mycophenolate thiab oxidation ntawm L-DOPA rau dopaquinone los ntawm diphenolase, uas yog ib kauj ruam rau ob qho tib si eumelanin thiab pheomelanin biosynthesis. Cov kauj ruam tom ntej yog koom nrog ob qho ntxiv melanocyte-specific enzymes, Trp1 thiab Trp2 (tseem hu ua dopachrome tautomerase), los tsim eumelanin (xim av / dub pigment) thiab cysteine ​​/ glutathione-koom nrog uas tsis yog-enzymatic kauj ruam los tsim pheomelanin (liab / daj. pigment), raws. Tyrosinase inhibitors xws li hydroquinone [3,4], kojic acid [5], ascorbic acid [6], thiab arbutin [7] tau siv los kho cov kab mob hyperpigmentation tab sis lawv tsis ntev los no tau tshaj tawm txog ntau yam kev mob tshwm sim hauv tib neeg [1, 2]. Yog li ntawd, kev saib xyuas ntau yog them rau kev nrhiav kev nyab xeeb thiab zoo dua melanogenic inhibitors los ntawm cov khoom ntuj tsim uas tuaj yeem ua rau depigmentation yam tsis muaj kev phiv.

Gastrodia elata Blume (GEB), ib tsob nroj orchid uas tsis muaj chlorophyll, xav tau cov symbiotic fungi, Mycena osmundicolaand Armillaria mellea rau nws cov germination thiab loj hlob / maturation, feem [8,9]. GEB tau siv los ua cov tshuaj kho mob oriental los kho cov kab mob neurodegenerative (xws li, tuag tes tuag taw, vertigo, mob stroke, qaug dab peg, thiab dementia) [9,10]. Ntau yam bioactive compounds tau raug cais los ntawm GEB, qhov tseem ceeb tshaj plaws yog cov phenolic compounds xws li gastrodin, 4-hydroxybenzyl cawv, 4-hydroxybenzaldehyde, vanillin, vanillyl cawv, thiab vanillic acid [{{6} }}]. Nws tau raug tshaj tawm tsis ntev los no tias bioactive compounds ntawm GEB muaj qhov inhibitory potency rau tyrosinase kev ua haujlwm [13-16]. Ntxiv mus, cov kev tshawb fawb tsis ntev los no tau txheeb xyuas ergothioneine (Ergo) [8,11] thiab diosgenin [17] hauv GEB. Ergo feem ntau pom nyob rau hauv nceb [18,19], thaum diosgenin nyob rau hauv qus yam [20]. Ob leeg ntawm lawv tau teev nyob rau hauv 10 botanical cov khoom xyaw uas muaj nyob rau hauv anti-aging cream nyob rau hauv 2010 vim lawv muaj zog antioxidant kev ua si [21], hais tias GEB yuav siv tau raws li ib tug functional cosmetic neeg sawv cev thiab nyob rau hauv cov zaub mov tshuaj.

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Txawm hais tias muaj cov lus ceeb toom ntawm cov khoom xyaw ntawm khov-qhuav thiab steam-dried GEB [22], nws cov khoom siv tshuaj [11,12,23], thiab antioxidant [12], anti-platelet thiab anti-thrombotic [24] cuam tshuam, me ntsis paub txog cov teebmeem inhibitory ntawm GEB extract ntawm melanin synthesis thiab nws cov enzymes cuam tshuam hauv B16F10 melanoma. Txij li thaum GEB tau siv los ua ib qho kev kho mob rau cov kab mob hauv lub paj hlwb thiab cov teeb meem ntawm daim tawv nqaij vim nws cov bioactive compounds thiab antioxidant zog, peb tau soj ntsuam cov ntsiab lus melanin thiab tyrosinase kev ua si nrog rau cov qib molecular ntawm transcription yam thiab kev tswj enzymes hauv murine B16F10 melanoma.

Cov khoom siv thiab cov txheej txheem

Kev npaj ntawm Gastrodia elata Blume (GEB) extract

GEB tau txais los ntawm Hyewonchunma Farm (Chuncheon, Kauslim). GEB qhuav (42 degree rau 16 h) yog hauv av thiab ces GEB hmoov (10 g) tau muab rho tawm hauv 200 mL ntawm de-ionized H2O ntawm 90 degree rau 3 h, ua raws li centrifugation ntawm 4,500 rpm. Lub supernatant tau lim siv cov ntawv lim nrog 6 μm pore loj (No. 3; Whatman, Little Chalfont, England), thiab concentrated nyob rau hauv lub tshuab nqus tsev evaporator (Rotavapor R-100; Buchi, Flawil, Switzerland). Cov dej khov khov tau evaporated rau cov hmoov qhuav qhuav rau 72 teev siv lub tshuab ziab khaub ncaws (Freezone plus 6; Labconco, Kansas City, MO, USA). GEB extract hmoov tau khaws cia ntawm -20 degree ua ntej siv.

Cell kab lis kev cai

Murine B16F10 melanoma kab (Korean Cell Line Bank, Seoul, Kauslim Teb) tau coj mus kuaj hauv Dulbecco's Modified Eagle Medium (Gibco, Grand Island, NY, USA) muaj 10 feem pua ​​​​fetal bovine serum (Gibco) thiab 1 feem pua ​​penicillin-streptomycin (Gibco) hauv 5% CO2 huab cua. B16F10 hlwb tau coj mus kuaj kom txog 100 feem pua ​​​​ntawm txhua qhov kev sim.

Cell viability assay

B16F10 hlwb tau plated ntawm qhov ceev ntawm 4 × 104 hlwb/zoo nyob rau hauv ib tug 24-zoo phaj. Cov 100 feem pua ​​​​ntawm cov hlwb raug pretreated nrog 0-25 mg/mL ntawm GEB extract rau 24 teev tom qab kho nrog / tsis muaj 200 nM -MSH (Sigma-Aldrich Co., St. Louis, MO, USA). Qhov nruab nrab tau hloov nrog ib qho 3-(4,5-dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide (MTT; Amresco, Solon, OH, USA ) dye tov [5 mg / mL hauv phosphate-buffered saline (PBS)] ntawm 37 degree rau 3 h. Lub supernatant tau decanted thiab cov ntsev formazan tau yaj hauv 500 μL isopropanol. Absorbance tau ntsuas ntawm 570 nm siv lub spectrophotometer (Gen5.2; Biotek, Winooski, VT, USA).

Kev tsom xam cov ntsiab lus melanin

B16F10 hlwb tau kho nrog 0-5 mg / mL ntawm GEB extract lossis 400 ug / mL arbutin (ib qho kev tswj zoo; Sigma-Aldrich) rau 72 teev tom qab kho nrog / tsis muaj 200 nM -MSH rau 24 teev. Cov hlwb raug ntxuav ob zaug nrog 1X PBS, ua raws li lysis hauv radioimmunoprecipitation assay (RIPA) lysis buffer (Thermo Scientific, Rockford, IL, USA) thiab centrifugation ntawm 14, 000 rpm rau 10 min ntawm 25 degree . Cov pellet (kwv yees li 105 hlwb) tau solubilized hauv 1 mL ntawm 1N NaOH nrog 10 feem pua ​​dimethyl sulfoxide rau 2 h ntawm 80 degree. Lub absorbance ntawm lub supernatant yog ntsuas ntawm 470 nm. Cov melanin concentration nyob rau hauv cov qauv yog muab los ntawm ib tug hluavtaws melanin (Sigma-Aldrich) tus qauv nkhaus. Txhua tus qauv tau ua kom zoo rau tag nrho cov cellular protein cov ntsiab lus siv Pierce® BCA txoj kev (Thermo Scientific).

Kev ntsuas kev ua haujlwm tyrosinase

B16F10 hlwb tau kho nrog 0-5 mg/mL ntawm GEB extract los yog 400 ug/mL arbutin (ib qho kev tswj zoo) rau 72 teev tom qab kho nrog / tsis muaj 200 nM -MSH rau 24 teev. Cov hlwb raug ntxuav ob zaug nrog 1X PBS, ua raws li lysis hauv RIPA lysis buffer (Thermo Scientific) thiab centrifugation ntawm 15, 000 rpm rau 20 min ntawm 4 degree . Lub supernatant (100 μL) tau muab tso rau hauv 96- lub phaj zoo thiab sib xyaw nrog 100 μL ntawm 5 mM L-DOPA thiab incubated ntawm 37 degree rau 2 h. Lub absorbance yog ntsuas ntawm 420 nm. Txhua tus qauv tau ua kom zoo rau tag nrho cov cellular protein cov ntsiab lus siv Pierce® BCA txoj kev (Thermo Scientific).

Kev soj ntsuam ntawm lub sijhawm tiag tiag-polymerase saw cov tshuaj tiv thaiv (lub sijhawm tiag tiag-PCR)

B16F10 hlwb tau kho nrog 0-5 mg/mL ntawm GEB extract los yog 400 ug/mL arbutin (ib qho kev tswj zoo) rau 72 teev tom qab kho nrog / tsis muaj 200 nM -MSH rau 24 teev. Tag nrho RNA ntawm cov hlwb raug cais tawm siv Trizol reagent (Invitrogen, Carlsbad, CA, USA) thiab cov khoom siv rho tawm RNA (RNase Mini cov khoom siv; Qiagen, Hilden, Lub teb chaws Yelemees). Tag Nrho RNA (2 ug) tau thim rov qab siv cov ntawv sau rov qab tus tswv premix (ELPIS, Daejeon, Kauslim). Lub sijhawm tiag tiag-PCR (LightCycler 96; Roche, Indianapolis, IN, USA) tau ua tiav siv cov primers: murine MITF rau pem hauv ntej 5'-GGCCAAAGGCAGAGCAACTT-3', rov qab 5'-GCCCATGGTGGCAAGCT-3'; tyrosinase rau pem hauv ntej 5′-ATAGGTGCATTGGCTTTGG-3′, rov qab 5′-CCAACGATCCCATTTTTTTT-3′; Trp1 forward 5′-GAGTGACATCCTGTGGCTCA-3′, rov qab 5′-CGATACCCTGGGAACACTTT-3′; Trp2 forward 5′-GCATTCTGTGGAAGGGTTGTT-3′, rov qab 5′-ACTCCTTCCTGAATGGGACC-3′; glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rau pem hauv ntej 5′-TCAATGAAGGGGTCGTTGAT-3′, rov qab 5′-CGTCCCGTAGACAAAATGGT-3′. Cov theem qhia tau normalized rau ntawm GAPDH.

Western blot tsom xam

B16F10 hlwb tau kho nrog 0-5 mg/mL ntawm GEB extract los yog 400 ug/mL arbutin (ib qho kev tswj zoo) rau 72 teev tom qab kho nrog 200 nM -MSH rau 24 teev. Tag nrho cov cell lysates (20 ug protein txhua) tau daws los ntawm 10 feem pua ​​SDS-PAGE, pauv mus rau polyvinylidene difluoride membrane (Roche), thiab soj ntsuam nrog thawj cov tshuaj tiv thaiv tshwj xeeb rau MITF, tyrosinase, Trp1, thiab Trp2 lossis -actin (Santa Cruz). Biotechnology, Dallas, TX, USA). Tom qab ntxuav, daim nyias nyias tau incubated nrog horseradish peroxidase-conjugated secondary antibody (Santa Cruz). Immunodetection tau ua tiav siv txoj kev chemiluminescence (SuperSignal; Pierce Biotechnology, Rockford, IL, USA) thiab tom qab ntawd normalized nrog -actin.

Kev txheeb cais

Kev txheeb xyuas txheeb cais tau ua tiav siv IBM SPSS software (Ver. 23; IBM-SPSS, Armonk, NY, USA). Ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li kev soj ntsuam tom qab hoc nrog Tukey qhov kev sim tau siv los txheeb xyuas qhov sib txawv ntawm pawg sim. Tus nqi ntawm P < 0.05 tau suav tias yog qhov tseem ceeb.

TSEEM CEEB

Cov teebmeem ntawm Gastrodia elata Blume (GEB) ntawm cell viability hauv B16F10 hlwb

Txhawm rau tshuaj xyuas qhov muaj peev xwm toxicity ntawm GEB extract, peb thawj zaug ntsuas GEB cytotoxicity los ntawm incubating B16F10 hlwb nrog ntau yam concentrations (0-25 mg/mL) ntawm GEB extract tom qab kho nrog / tsis muaj 200 nM -MSH rau 24 h. Tsis hais txog ntawm -MSH stimulation, cell viability tsis inhibited los ntawm GEB concentrations qis (0.5-5 mg/mL), whereas ntau dua concentrations (10-25 mg/mL) ho induced toxicity (Fig. 1). Cell viability ntawm 10 thiab 25 mg / mL ntawm GEB yog 85.9 feem pua ​​​​thiab 68.7 feem pua ​​​​ntawm cov kev tswj tsis tau kho, feem. Cell viability ntawm 10 thiab 25 mg / mL ntawm GEB tom qab -MSH stimulation yog 89.9 feem pua ​​​​thiab 65.3 feem pua ​​​​ntawm cov kev tswj tsis tau kho, feem. Yog li ntawd, qhov concentration ntawm 0-5 mg/mL tau siv rau hauv kev sim ntxiv.

cistanche norge

Cov nyhuv ntawm Gastrodia elata Blume (GEB) ntawm melanin synthesis thiab tyrosinase kev ua hauv B16F10 Cells

Txhawm rau ntsuas qhov kev ua haujlwm melanogenesis ntawm GEB extract, peb ntsuas melanin cov ntsiab lus hauv ob leeg -MSH-tsis kho thiab -MSH-kho B16F10 hlwb. Cov hlwb raug pretreated nrog / tsis muaj -MSH rau 24 teev, ua raws li kev kho mob nrog GEB extract ntawm koob tshuaj 0-5 mg/mL lossis 400 ug/mL arbutin rau 72 teev. GEB kev kho mob tau txo qis cov cellular melanin cov ntsiab lus nyob rau hauv koob tshuaj raws li nyob rau hauv ntau dua koob tshuaj (2-5 mg/mL) nyob rau hauv -MSH-kho pab pawg thiab tag nrho cov -MSH-tsis kho cov pab pawg. -MSH ib leeg nce melanin cov ntsiab lus los ntawm 180 feem pua ​​​​piv rau kev tswj hwm, thiab GEB kev kho mob pom tau tias inhibited melanin synthesis nyob rau hauv -MSH-induced hyperpigmentation xeev nyob rau hauv ib koob tshuaj raws li piv rau -MSH-tsis kho cov pab pawg (Fig. 2).

Txij li tyrosinase yog tus nqi-txheej enzyme ntawm melanin synthesis, peb txiav txim siab inhibitory nyhuv ntawm GEB extract ntawm cellular tyrosinase kev ua nyob rau hauv B16F10 hlwb. -MSH induced tyrosinase kev ua si los ntawm 171.2 feem pua ​​​​hauv B16F10 hlwb. Tsis hais txog ntawm -MSH kev kho mob, GEB extract cuam tshuam kev ua haujlwm tyrosinase. Arbutin paub zoo tias yog ib qho inhibitor ntawm melanin synthesis ntawm tyrosinase txoj hauv kev [7], uas tau cuam tshuam loj heev los ntawm 35-40 feem pua ​​​​piv rau kev tswj tsis tau hauv ob pawg -MSH-tsis kho thiab -MSH-kho pab pawg (Fig. 3) . Cov ntsiab lus melanin thiab tyrosinase kev ua ub no ntawm GEB extract ntawm 2 thiab 5 mg / mL tau zoo ib yam li cov ntawm 400 ug / mL arbutin, qhia tias GEB qhia txog cov tshuaj tiv thaiv melanogenesis kev ua haujlwm rau qib ntawm kev tswj tau zoo thiab qhov no melanogenesis inhibition. los ntawm GEB extract muaj feem xyuam rau txo qis cellular tyrosinase kev ua haujlwm (Fig. 3).

cistanche nedir

Cov teebmeem ntawm Gastrodia elata Blume (GEB) ntawm mRNA qib thiab cov protein qhia ntawm MITF, tyrosinase, Trp1, thiab Trp2 hauv B16F10 hlwb

Txhawm rau txiav txim siab seb qhov inhibition ntawm melanin synthesis los ntawm GEB extract yog txuam nrog melanogenesis hais txog cov noob qhia, peb tau ua lub sijhawm tiag-PCR los tshuaj xyuas cov qib mRNA ntawm MITF, tyrosinase, Trp1, thiab Trp2. mRNA qib ntawm tag nrho cov noob no tau txo qis los ntawm GEB extract nyob rau hauv koob tshuaj nyob rau hauv ob qho tib si -MSH-tsis kho thiab -MSH-kho pab pawg, tab sis lawv tsis saib xyuas hauv -MSH-tsis kho cov hlwb. Cov qib molecular ntawm MITF, tyrosinase, Trp1, thiab Trp2 tau inhibited ntawm qis GEB concentrations (0.5-1 mg/mL) thiab tau pom zoo tab sis tsis cuam tshuam ntau dua ({15}} mg /mL) hauv -MSH-tsis kho cov hlwb. -MSH markedly induced cov noob 'mRNA qib tab sis GEB extract txo nws. Txawm tias nyob rau hauv -MSH-kho cov hlwb, qis dua ntawm GEB extract tended txo MITF, tyrosinase, Trp1, thiab Trp2 mRNA qib, qhov ntau dua ntawm GEB extract ho inhibited mRNA qib ntawm tag nrho cov noob no. Cov qib inhibition ntawm GEB extract ntawm cov koob tshuaj ntau dua yog zoo ib yam li arbutin, uas yog ib qho paub zoo ntawm cov tawv nqaij dawb hauv cov tshuaj pleev ib ce (Fig. 4).

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Tsis tas li ntawd, peb tau soj ntsuam cov protein qhia ntawm qhov kev hloov pauv no thiab cov enzymes melanogenic los ntawm kev txheeb xyuas sab hnub poob blot. -MSH kev kho mob ua rau muaj kev cuam tshuam cov protein ntau ntxiv nrog rau mRNA qib ntawm tag nrho cov noob no, uas tau zoo los ntawm kev siv tshuaj ntau npaum li cas. Thaum cov koob tshuaj qis dua ntawm GEB extract tended txo qis -MSH-vim cov protein qhia ntawm MITF, tyrosinase, Trp1, thiab Trp2, cov koob tshuaj ntau dua inhibited tag nrho cov cim no hauv -MSH-kho B16F10 hlwb nyob rau hauv ib koob tshuaj. Cov qib inhibition ntawm GEB extract ntawm koob tshuaj ntau dua tau zoo li arbutin (Fig. 5).

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Kev sib tham

Hauv txoj kev tshawb no, peb tau pom tias GEB extract muaj kev tiv thaiv kev tiv thaiv melanogenesis thiab kev ua haujlwm thiab molecular qhia ntawm nws cov enzymes cuam tshuam xws li MITF, tyrosinase, Trp1, thiab Trp2 hauv murine B16F10 melanoma hlwb. GEB rho tawm ntawm koob tshuaj 0 5-5 mg/mL inhibited melanogenesis-txog cov cim nyob rau hauv ib koob tshuaj. Thaum cov koob tshuaj qis dua (0. } mg/mL) ntawm GEB extract tau inhibited tag nrho cov cim cim hlwb nyob rau hauv ib koob tshuaj nyob rau hauv ob qho tib si -MSH-tsis kho thiab -MSH-kho B16F10. Cov teebmeem inhibitory ntawm GEB extract ntawm qhov siab dua tau zoo ib yam li cov 400 ug / mL arbutin.

Melanin yog tsim los ntawm L-tyrosine nyob rau hauv melanosomes thiab nws txiav txim cov yam ntxwv xim ntawm daim tawv nqaij, plaub hau, thiab ob lub qhov muag [1]. Txawm hais tias melanin ua lub luag haujlwm tiv thaiv kev tiv thaiv ultraviolet (UV) lub teeb thiab kev ntxhov siab, kev tsim tawm ntau dhau thiab sib sau ntawm cov xim melanin ua rau hyperpigmentation thiab ntau yam teeb meem ntawm daim tawv nqaij xws li freckles, hnub nyoog me ntsis, thiab melasma hauv cov tsiaj txhu [25]. Yog li ntawd, melanogenesis inhibition yog ib qho teeb meem ntawm kev txhawj xeeb rau kev kho mob thiab kho kom zoo nkauj ntawm daim tawv nqaij depigmentation thiab lightening. Kev khi ntawm -MSH rau melanocortin -1 receptor (MC1R) nce cyclic adenosine monophosphate (cAMP) thiab tom qab ntawd ua rau MITF. MIFT muaj txiaj ntsig zoo- tswj cov kev qhia ntawm melanogenic enzymes, suav nrog tyrosinase, Trp1, thiab Trp2, uas sib koom ua ke ntawm 40-45 feem pua ​​​​zoo li tus kheej [1,2]. Feem ntau ntawm tag nrho cov, tyrosinase inhibition yog txoj hauv kev zoo tshaj plaws kom ua tiav daim tawv nqaij hypopigmentation vim tias qhov enzyme no ua rau tus nqi txwv cov kauj ruam ntawm pigmentation. Ob qho tib si Trp1 thiab Trp2 tau koom nrog hauv qhov tshwm sim biosynthesis ntawm melanin [1,2].

cistanche supplement review

Tyrosinase, tseem hu ua monophenol monooxygenase los yog polyphenol oxidase yog ib tug multifunctional tooj liab-muaj enzyme [1]. Tyrosinase muaj ob yam kev ua haujlwm catalytic: monophenolase thiab diphenolase. Tyrosinase muaj ob lub tooj liab ions, uas koom tes nrog histidine residues nyob rau hauv lub active site thiab lawv tseem ceeb heev rau cov catalytic kev ua si [1]. Nws tau raug tshaj tawm tias tyrosinase yog inhibited los ntawm hydroquinone [3,4], kojic acid [5], ascorbic acid [6], thiab arbutin [7]. Txawm li cas los xij, cov neeg ua haujlwm no tau raug tshaj tawm tias ua rau tawv nqaij khaus, tawv nqaij tsis zoo, tsis muaj kev ruaj ntseg, thiab txawm tias muaj peev xwm carcinogenicity. Ntawm qhov teeb meem no, nws yog ib qho tsim nyog los tsim cov khoom ntuj tsim hauv kev lag luam tshuaj pleev ib ce.

Lee et al. [26] soj ntsuam 100 cov nroj tsuag rho tawm rau kev siv tshuaj pleev ib ce los ntawm kev ntsuas qhov inhibitory kev ua ub no ntawm tyrosinase thiab DOPA auto-oxidation thiab qhia tias muaj ntau cov nroj tsuag rho tawm xws li Gastrodia elata, Chaenomeles speciosa, Dryopteris crassirrhizoma, Glycyrrhizaglabra, Morusalba, Myristica andframegra. japonica pom inhibition ntawm nceb tyrosinase kev ua haujlwm. Chen et al. [2] kuj tau tshuaj xyuas 78 cov tshuaj ntsuab tshuaj ntsuab thiab pom tias rhizome ntawm GEB muaj peev xwm inhibit tyrosinase kev ua haujlwm. Txawm hais tias GEB tau siv los ua cov tshuaj oriental los kho cov kab mob neurodegenerative, nws muaj peev xwm siv GEB ua tyrosinase inhibitor los ntawm cov kev tshawb fawb soj ntsuam no.

Ib txoj kev tshawb fawb tsis ntev los no [9] tau tshaj tawm 64 GEB cov khoom siv siv ntau yam chromatography cov txheej txheem. Ntawm lawv, cov tshuaj phenolic tseem ceeb tshaj plaws yog gastrodin [4-(-d-glucopyranosyloxy) benzyl cawv], 4-hydroxy benzyl cawv (4-HBA), 4-hydroxy- benzaldehyde, vanillin (4-hydroxy-3-methoxybenzaldehyde), vanillyl cawv (4-(hydroxymethyl)-2-methoxyphenyl), thiab vanillic acid (4-}hydroxy{{14} }}methoxybenzoic acid) [8-12]. Cov tebchaw tshiab tshiab tau pom tsis ntev los no hauv GEB; bis ({16}} hydroxyphenyl) sulfide [2], Ergo [8], thiab diosgenin [17]. Cov qauv ntawm hydroquinone thiab cov phenolic tebchaw loj ntawm GEB zoo sib xws [27]. Cov pab pawg Hydroxyl (-OH) ntawm hydroquinone yog hloov nrog ib pawg hydroxymethyl (-CH2OH) hauv 4-HBA [27] thiab pawg carbonyl (-CHO) hauv 4-hydroxybenzaldehyde. Ib pawg methoxy (-OCH3) tau ntxiv rau hydroquinone los ua vanillyl cawv thiab rau 4-hydroxybenzaldehyde los ua vanillin, feem. Vanillic acid yog oxidized daim ntawv ntawm vanillin.

Cov kev tshawb fawb tsis ntev los no tau tshaj tawm tias feem ntau ntawm cov tshuaj no tawm dag zog tiv thaiv cov nceb tyrosinase thiab melanogenesis hauv B16 [14,28] thiab HM3KO melanoma hlwb [13], tib neeg lub cev nqaij daim tawv nqaij tawv nqaij [29], thiab hauv vivo zebrafish qauv [2,29]. Piv txwv li, cais phenolic feem los ntawm butanol-extracted GEB [14] exerted inhibitory teebmeem ntawm melanogenesis los ntawm kev txo qis kev ua ntawm tyrosinase, Trp1, thiab Trp2 raws li zoo raws li mRNA thiab protein qhia ntawm cov enzymes. Gastrodin [15] thiab 4-HBA [16] suppressed melanogenesis los ntawm inhibiting tyrosinase kev ua hauv B16 melanoma hlwb. Cov pab pawg hydroxyl ntawm gastrodin thiab 4-HBA cuam tshuam feem ntau nrog histidine residues hauv qhov chaw nquag ntawm tyrosinase, uas ua rau inhibition ntawm tyrosinase kev ua haujlwm [30]. Cov pa oxygen los ntawm phenolic OH ntawm 4-HBA cuam tshuam rau kev ua haujlwm catalytic ntawm tyrosinase los ntawm kev tawm tsam nucleophilic [31]. Hydroxy thiab methoxy pawg ntawm vanillic acid thiab vanillyl cawv ua raws li inhibitors ntawm diphenenolase kev ua ntawm tyrosinase [32] thiab tshwj xeeb tshaj yog hloov dua siab tshiab ntawm CH2OH nyob rau hauv vanillyl cawv rau -COOH (vanillic acid), uas ua rau ib tug cim nce nyob rau hauv tyrosinase inhibition [32] ]. Vanillic acid cais tawm los ntawm Origanum vulgare txo qis cellular tyrosinase kev ua, DOPA oxidase, thiab melanin cov ntsiab lus, nrog rau kev txo qis kev qhia ntawm MC1R, MITF, Trp1, thiab Trp2 hauv B16F10 hlwb txhawb nrog -MSH. Vanillin tsis qhia inhibition ntawm tyrosinase kev ua haujlwm [29]. Tsis ntev los no, ib yam khoom ntuj cais los ntawm GEB, bis (4-hydroxy benzyl) sulfide, ua raws li kev sib tw tyrosinase inhibitor los thaiv melanin synthesis nyob rau hauv tib neeg epidermal melanocytes thiab zebrafish qauv los ntawm kev qhia kev sib koom tes sulfur atom ntawm bis ({{ 33}}hydroxy benzyl) sulfide nrog tooj liab ions nyob rau hauv qhov chaw nquag ntawm tyrosinase los ntawm kev ua qauv molecular [2]. Cov txiaj ntsig no qhia tias ntau yam phenolic tebchaw ntawm GEB tuaj yeem ua raws li tyrosinase inhibitors los ntawm kev cuam tshuam nrog tooj liab ions thiab / lossis histidine residues hauv qhov chaw nquag ntawm tyrosinase.

Ergo thiab diosgenin raug xaiv los ua ib qho ntawm 10 cov khoom xyaw botanical nyob rau hauv 2010 anti-aging cream [21]. Ergo yog ib qho dej-soluble thiol amino acid [18,19], tsis ntev los no pom hauv GEB [8]. Ntawm physiological pH, Ergo muaj nyob hauv thione (= S) es tsis yog thiol (-SH) daim ntawv [19]. Nws qeeb degradation, tsis kam mus rau disulfide tsim, thiab nws pib-oxidization ua rau nws ruaj khov dua glutathione [18,19]. Lub xub ntiag ntawm cov sulfur-hloov imidazole nplhaib hauv Ergo inhibited nceb tyrosinase kev ua haujlwm nyob rau hauv koob tshuaj [33]. Lwm qhov sib xyaw, diosgenin los ntawm cov tsiaj qus (Dioscorea villosa) extract muaj cov nyhuv depigmenting thiab yog li siv tau rau hauv melasma, melanodermatitis, thiab sun lentigo [34]. Ib txoj kev tshawb fawb tau ua los ntawm cov hlwb melanoma tau pom tias cov nyhuv depigmenting cuam tshuam nrog kev ua kom cov cellular phosphatidylinositol- 4, 5-bisphosphate 3-kinase txoj hauv kev, qhia tias diosgenin yuav yog ib qho zoo inhibitor. hyperpigmentation ntawm lub ntsej muag [34] Tsis tas li ntawd, nws muaj anti-inflammatory thiab anti-aging zog los ntawm inducing anti-collagenase kev ua si [35].

Muaj tsawg kawg peb txoj kev teeb liab tau koom nrog hauv kev tswj hwm ntawm melanogenesis; cAMP-dependent, Wnt, thiab extracellular signal-regulated kinase signaling pathways [1]. Tag nrho cov teeb liab txoj hauv kev tswj hwm melanogenesis ntsig txog enzymes, piv txwv li, tyrosinase, Trp1, thiab Trp2 ntawm MITF qhia thiab / lossis kev ua haujlwm [1]. Lub peev xwm kev koom tes ntawm cov teeb liab txoj hauv kev hauv kev tawm tsam ntawm melanogenesis los ntawm GEB tseem yuav raug tshuaj xyuas yav tom ntej.

Hauv peb txoj kev tshawb fawb dhau los, peb tau pom tias GEB extract ameliorated daim tawv nqaij-photoaging cov cim los ntawm kev txhawb nqa kev ua haujlwm antioxidant nrog cov nyiaj txaus ntawm polyphenols, flavonoid, thiab ergo thiab los ntawm kev hloov pauv kev qhia / kev ua ntawm procollagen hom I, matrix metalloproteinase-1, thiab elastase -1 nyob rau hauv UVA- irradiated human dermal fibroblasts [36]. Hauv txoj kev tshawb no, GEB inhibited melanin synthesis los ntawm kev tawm tsam ntawm tyrosinase kev ua si thiab mRNA qib thiab cov protein qhia ntawm MITF, tyrosinase, Trp1, thiab Trp2 nyob rau hauv ib koob tshuaj nyob rau hauv murine B16F10 melanoma. Yog li ntawd, cov kev tshawb fawb no qhia tias GEB extracts yuav siv tau raws li zoo thiab tej yam ntuj tso daim tawv nqaij-dawb huv si, raws li zoo raws li anti-aging agents rau cosmeceutical daim ntaub ntawv.

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Kev tsis sib haum xeeb ntawm kev txaus siab

Cov kws sau ntawv tshaj tawm tias tsis muaj kev tsis sib haum xeeb ntawm kev txaus siab.

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