Kev soj ntsuam ntxiv ntawm Salvia Haenkei Raws li Lub Tswv Yim Tsim Kho Tshiab los tiv thaiv daim tawv nqaij daim duab-laus thiab rov ua kom muaj kev ncaj ncees

Thov hu rauoscar.xiao@wecistanche.comyog xav paub ntxiv

abstract:Daim tawv nqaij yog qhov teeb meem tseem ceeb ntawm tib neeg lub cev uas ua haujlwm ntau yam. Endogenous yam, nyob rau hauv kev hais kwv txhiaj nrog rau sab nraud assaults, tsis tu ncua cuam tshuam rau ntawm daim tawv nqaij kev ncaj ncees, ua rau cov kev hloov ntawm cov qauv uas cuam tshuam tsis tau tsuas yog cov tawv nqaij tsos, tab sis kuj nws ntau yam physiological functions. Kev hloov pauv ntawm cov kev cuam tshuam kev ua haujlwm ua rau muaj kev pheej hmoo ntau ntxiv ntawm kev tsim kab mob thiab cov kev cuam tshuam sab nraud, yog li qhov tseem ceeb ntawm kev tswj xyuas kev ncaj ncees ntawm cov kab mob epidermal thiab kev ua kom qeeb ntawm daim tawv nqaij laus yog qhov tseeb.cistanche noov loj hlobSalvia hankie (SH) tsis ntev los no tau txheeb xyuas tias yog tus neeg sawv cev tiv thaiv kev laus; nws cov extract muaj peev xwm txo qis qib ntawm senescent hlwb los ntawm kev cuam tshuam rau ll.1a tso tawm thiab txo cov reactive oxygen hom (ROS) tiam. Hauv txoj kev tshawb no, SH extract tau sim ntawm tib neeg keratinocyte cell kab (HaCaT) raug rau cov kev ntxhov siab cuam tshuam txog kev laus ntxov ntawm cov hlwb xws li dawb radicals thiab ultraviolet B hluav taws xob. Peb tau lees paub tias SH ua raws li cov khoom pov tseg ntawm ROS thiab pom nws lub peev xwm los kho cov tawv nqaij thaiv kev ncaj ncees los ntawm kev txhawb nqa cov qauv cytoskeleton, sealing cov junctions nruj, thiab nce cov kev tsiv teb tsaws ntawm cov hlwb. Muab cov txiaj ntsig no, txoj haujlwm no ua rau muaj feem cuam tshuam, txheeb xyuas Salvia hankie ua ib qho khoom siv rau kev tiv thaiv kev laus ntawm daim tawv nqaij hauv kev kho mob.

Thov nias ntawm no kom paub ntxiv

Taw qhia

Kev noj qab haus huv thiab kev ua haujlwm ntawm daim tawv nqaij yog qhov tseem ceeb los tiv thaiv lub cev qhuav dej thiab nkag mus ntawm ntau yam tiv thaiv 100 feem pua, kab mob, tshuaj tsis haum, tshuaj lom neeg, thiab hluav taws xob. Txhua hnub raug rau extrinsic agents ua ke nrog ib co intrinsic yam, xws li noob caj noob ces, cellular metabolism, hormone, thiab metabolic dab, ua rau lub ntuj aging ntawm daim tawv nqaij [1]. Tshwj xeeb, qhov tshwm sim ntawm daim tawv nqaij laus muaj kev cuam tshuam loj heev rau cov hluav taws xob ultraviolet, muaj kuab paug, siv cov leeg rov ua dua, thiab lub ntiajteb txawj nqus [2, 3]. Kev laus ntawm daim tawv nqaij, yog li ntawd, tshwm sim los ntawm kev hloov pauv ntawm cov qauv thiab lub cev thiab kev hloov pauv ntawm cov txheej txheem ntawm daim tawv nqaij [2, 4, 5]. Txawm hais tias kev laus tsis suav hais tias yog ib qho mob pathological, tab sis yog txheej txheem lom neeg thiab tsis tuaj yeem, nws txoj kev sib raug zoo nrog ntau yam kab mob ntawm daim tawv nqaij yog tam sim no tsim tau zoo. Xav txog tias cov kab mob uas muaj feem cuam tshuam rau hnub nyoog tshaj plaws yog cov kab mob degenerative, benign thiab malignant neoplasms, thiab lwm yam, qhov tseem ceeb ntawm kev tu tawv nqaij los kho lossis tiv thaiv qee yam kab mob ntawm daim tawv nqaij yuav tshwm sim tau tshwm sim.[6-8]. Niaj hnub no, cov tswv yim tiv thaiv kev laus ntawm daim tawv nqaij ua rau thim rov qab dermal thiab epidermal cov cim qhia ntawm daim duab / kev laus. Raws li lawv txoj hauv kev, lawv tuaj yeem raug cais raws li hauv qab no: kev saib xyuas cosmetological, tshuaj pleev ib ce, cov txheej txheem cuam tshuam, cov kab mob hauv lub cev, zam kev laus exogenous yam, kho txoj kev ua neej thiab tus cwj pwm, thiab tshuaj tiv thaiv [9]. Muaj ob pawg tseem ceeb ntawm cov neeg ua haujlwm uas tuaj yeem siv los ua cov khoom siv los tiv thaiv kev laus: 1. Cell regulators, xws li retinol, peptides, thiab kev loj hlob yam, ncaj qha ua haujlwm ntawm collagen metabolism thiab ntau lawm; 2. Antioxidants, zoo li cov vitamins, polyphenols, thiab flavonoids, muaj peev xwm txo tau collagen degradation [9].

Cov pov thawj tseem ceeb txhawb lub tswv yim tias cov tawv nqaij laus feem ntau yog tsav los ntawm ROS, tsim thaum lub sij hawm endogenous oxidative cell metabolism [10,11]. Nws paub tias ROS tsub zuj zuj yog lub luag haujlwm rau kev puas tsuaj ntawm tes nrog rau lipid peroxidation, membrane protein puas, thiab DNA hloov pauv uas ua rau muaj kev hloov pauv ntawm daim tawv nqaij thiab kev ua haujlwm. Nyob rau lub sijhawm, tib neeg lub hlwb ua rau muaj kev nce qib ntawm ROS, cuam tshuam nrog ib feem poob ntawm cov tshuaj tiv thaiv antioxidant. Qhov kev tsis pom zoo no tau txhawb nqa los ntawm txhua hnub ultraviolet irradiation (UVR) uas ua rau matrix metalloproteinases (MMPs) synthesis thiab, yog li, MMP-mediated collagen puas [12, 13]. Hauv vivo cov kev tshawb fawb pom tau tias muaj kev sib raug zoo ntawm mitochondrial oxidative puas, cellular senescence, thiab aging phenotypes [10, 14].

Cellular senescence txhais tau tias yog qhov ruaj khov ntawm kev loj hlob ntawm tes uas tshwm sim hauv txhua tus tib neeg lub hlwb thaum laus [15-17]. Raws li qhov tshwm sim ntawm cov xwm txheej ntxhov siab, xws li oncogene over-expression, ROS tiam, thiab DNA puas (piv txwv li induced los ntawm UVR, tshwj xeeb tshaj yog los ntawm UVB), cov hlwb yuav ntxov ntxov ua senescent [18].cistanche salsa cov txiaj ntsigHauv qhov xwm txheej no, nws pom tseeb tias qhov tseem ceeb ntawm kev tsim cov tswv yim tshiab tuaj yeem tiv thaiv kev sib sau ntawm cov hlwb senescent los yog xaiv tua lawv, yog li tiv thaiv kev laus thiab kev laus-txoj kev mob. Tsis ntev los no, peb tau txheeb xyuas Salvia hankie (SH), ib tsob nroj Bolivian nplua nuj nyob rau hauv vitamin B thiab antioxidant zog, raws li ib tug muaj peev xwm los tiv thaiv kev laus [18].

Kev sib sau ntawm cov hlwb senescent nyob rau lub sijhawm tau paub zoo tias yog ib qho ntawm cov txheej txheem tseem ceeb tshaj plaws los ntawm cov ntaub so ntswg mus rau cov txheej txheem kev laus, uas zoo nkaus li tau tshwm sim los ntawm kev pheej hmoo ntawm kev mob tshwm sim. Qhov tseeb, txawm tias senescent hlwb tsis tuaj yeem loj hlob tuaj, lawv muaj cov haujlwm metabolic siab thiab lawv ua tau zoo tsim ntau yam pro-inflammatory molecules [19]. Los ntawm kev siv tus qauv ntawm daim tawv nqaij tib neeg epidermis (EpiSkin), nws tau pom tseeb tias SH extract txo cov qib ntawm senescent hlwb los ntawm kev cuam tshuam rau ILla tso tawm thiab txo ROS tiam [18].

Nyob rau hauv homeostatic tej yam kev mob, daim tawv nqaij yog ib tug npaum barrier uas tiv thaiv los ntawm sab nraud assaults thiab endogenous yam thiab tswj tsis tau cov kua, electrolytes, thiab cov proteins. Lub luag haujlwm tseem ceeb no feem ntau yog sau rau ntawm lub epidermis, ib qho dynamic, heev stratified epithelium. Ob lub epidermis tseem ceeb yog stratum corneum, txheej txheej sab nraud, thiab nruj junctions (TJs), intercellular junctions uas kaw keratinocytes nyob ib sab hauv stratum granulosum [20]. Cell-cell adhesions yog tsim nyog los tswj cov epidermal-barrier kev ncaj ncees.cistanche tubulosa noj redditQhov kev ua haujlwm tseem ceeb no tau ua tiav los ntawm kev sib txuas ntawm tes tshwj xeeb: adherens junctions (AJs), desmosomes, thiab nruj junctions. Cov qauv no tsis tsuas yog tso cai rau kev sib txuas ntawm lub cev ntawm cov hlwb, tab sis tag nrho cov txheej txheem cytoskeletal, thiab hloov cov kev taw qhia kev koom tes hauv cov ntaub so ntswg, qauv, thiab lub cev. Kev tshawb nrhiav caj ces tsis ntev los no tau npaj txhij txog qhia txog lub luag haujlwm tsis yog canonical ntawm junctional proteins, coj tawm qhov tseem ceeb ntawm junctional crosstalk, interdependencies, thiab them nyiaj rau cov ntaub so ntswg robustness [21].

Ntawm no peb tau kawm txog cov haujlwm ntawm Salvia haenkei extract kom nkag siab txog cov txheej txheem molecular lub luag haujlwm rau nws cov teebmeem tiv thaiv ntawm daim duab / sijhawm ntawm daim tawv nqaij laus. Nrog rau qhov kev xav no, tib neeg spontaneously immortalized keratinocytes (HaCaT) tau siv raws li nyob rau hauv vitro qauv, thiab SH mechanism ntawm kev txiav txim tau soj ntsuam nyob rau hauv ib tug basal regimen raws li nyob rau hauv cov kev ntxhov siab vim los ntawm H202 thiab / los yog UVB irradiation.

TSEEM CEEB

Kev nyab xeeb profile ntawm SH extract thiab antiaging zog Oxidative kev nyuaj siab nyob rau hauv daim tawv nqaij plays lub luag hauj lwm tseem ceeb nyob rau hauv lub aging txheej txheem thiab nyob rau hauv lub pathogenesis ntawm ob peb daim tawv nqaij mob [22]. Matic et al. twb tau qhia txog kev tiv thaiv kev ua haujlwm ntawm Salvia haenkei kuj cuam tshuam rau nws cov khoom pov tseg [18]. Cov teebmeem kev tiv thaiv no yuav raug siv los tsim kho cov tawv nqaij tshiab tuaj yeem khaws cov kev ncaj ncees ntawm kev thaiv. Nrog rau lub hom phiaj ntawm kev tshawb xyuas SH cov khoom, cov kev ua haujlwm ntawm cov cell viability thiab ROS tiam hauv HaCaT cell qauv tau kuaj pom thawj zaug.

Cistanche tuaj yeem tiv thaiv kev laus

Kev ntsuam xyuas ntawm tes tau ua tiav los daws cov teebmeem cytotoxic ntawm Salvia haenkei extract ntawm keratinocytes. HaCaT hlwb raug kho nrog SH extract (001-10 ug/ml) rau 24 thiab 48 teev. Cell viability raug soj ntsuam los ntawm Trypan blue Exclusion Assay. Daim duab 1A thiab 1B qhia tias kev kho mob nrog SH extract tsis cuam tshuam rau cell viability lees paub qhov kev nyab xeeb ntawm cov extract ntawm HaCaT hlwb nyob rau hauv cov kev sim siv.

Cov dej num ntawm Salvia haenkei extract ntawm ROS tiam tau soj ntsuam hauv HaCaT hlwb tom qab 3-24 teev ntawm kev kho mob thiab ua ntej thiab tom qab H2O2 raug.

Cov txiaj ntsig hauv daim duab 2A qhia tau tias SH tuaj yeem txo qis ROS theem hauv cov hlwb, tshwj xeeb tshaj yog thaum lub sijhawm 3- teev-kev kho mob. Dhau li ntawm qhov tseem ceeb ntawm ROS theem tom qab H-Oz stimulation (Daim duab 2C), SH muaj peev xwm tiv thaiv kev tsim ntawm ROS. SH extract (0.01 ug/ml) txo qis (-25 feem pua ​​) ROS tiam induced los ntawm oxidative stimulus (Daim duab 2B), uas qhia cov nyhuv piv nrog NAC tom qab 3 teev ntawm kev kho mob. 24 teev ntawm SH kev kho mob tshwm sim ntau dua ROS muaj txiaj ntsig tiv thaiv kev nce ntxiv, hauv kev txo qis ROS qib ntawm 1 thiab 10 ug / ml (Daim duab 2B). Cov txiaj ntsig no lees paub Salvia haenkei

Cov teebmeem antioxidant, raws li yav dhau los pom [18], qhia txog cov nyhuv antioxidant basal twb pib los ntawm kev kho luv luv; cov nyhuv hauv kev tiv thaiv oxidative stimulus-induced nce yog feem ntau cuam tshuam rau lub sij hawm kho mob ntev.

Nws tau tshaj tawm tias cov hluav taws xob ultraviolet ua rau muaj qhov nce hauv cov qib ntawm cov cell ntawm ROS hauv tib neeg cov tawv nqaij ua rau kev puas tsuaj thiab kev laus ntxov ntxov thiab lwm yam pathologies [23-25]. Yog li, cov nyhuv ntawm SH extract ntawm UVB-mediated ROS ntau lawm tau raug soj ntsuam. Raws li qhov xav tau, cov txiaj ntsig hauv daim duab 3C qhia tias muaj kev nce ntxiv hauv cov cellular ROS qib tom qab UVB stimulation. Kev kho mob nrog SH, uas ua rau muaj kev nyiam los txo qis ROS theem hauv cov hlwb (Daim duab 3A), muaj peev xwm tiv thaiv qhov nce ntxiv los ntawm UVB stimulation (Daim duab 3B). 3 teev ntawm SH kev kho mob ua rau muaj kev nyiam los txo ROS qib; Kev kho mob SH ntev dua yog qhov ua tau zoo dua hauv kev tawm tsam oxidative kev nyuaj siab, qhia qhov txo qis raws li qhia hauv daim duab 3A. Cov ntaub ntawv no qhia txog kev ua haujlwm antioxidant ntawm Salvia haenkei kuj nyob rau hauv UVB-mediated oxidative stress.

Raws li tau hais, cov pov thawj nce ntxiv qhia tias cov hlwb senescent sau nyob rau hauv lub sijhawm thiab cov duab-hnub nyoog ntawm daim tawv nqaij, ua rau muaj hnub nyoog cuam tshuam ntawm daim tawv nqaij hloov pauv thiab pathologies [26]. Cov nyhuv ntawm Salvia hankie yog li tau soj ntsuam ntawm p21 thiab p27 senescence markers (uas tiv thaiv phosphorylation ntawm retinoblastoma protein (Rb) thiab nrog inhibitory kev ua si ntawm CDK2-cyclin E, feem [27]) tom qab yees duab-stress induced los ntawm ultraviolet hluav taws xob. Cov txiaj ntsig hauv daim duab 4 qhia tias UVB irradiation induces ib qho tseem ceeb nce nyob rau hauv p21 thiab p27 mRNA qib raws li piv rau unstimulated tswj.cistanche แอมเวIb qho piv txwv zoo sib xws hauv kev tiv thaiv kev nce ntxiv yog pom hauv cov hlwb kho nrog SH. SH 1 ug / ml induces ib tug tseem ceeb txo ntawm p21 qib piv rau UVB stimulated tswj, thaum lwm yam concentrations induce ib tug nyiam txo (Daim duab 4A). P27 qib tau txo qis los ntawm kev kho ua ntej nrog SH 0.01,0.1 thiab 10 ug/ml (Daim duab 4B).

Nws kuj tseem tsim tau tias cov hlwb tsis zoo cuam tshuam rau microenvironment los ntawm kev zais cov tshuaj tiv thaiv kab mob sib xyaw ntawm chemokines, cytokines, cov tshuaj hormones loj hlob thiab proteases, hu ua senescence-associated secretory phenotype (SASP) [28]. Cov txiaj ntsig hauv daim duab 5 qhia tias UVB stimulation induces ib tug tseem ceeb nce nyob rau hauv ILla (A), IL6 (B), thiab IL18 (C) mRNA qib nrog rau kev hwm rau cov unstimulated tswj. Pretreatment nrog SH nthuav tawm cov haujlwm muaj zog hauv kev tawm tsam qhov nce hauv qib cytokine. Raws li pom hauv daim duab 5A, SH 10 ug/ml txo qis ILla ntau ntau; qhov nce hauv IL6 yog qhov cuam tshuam loj heev los ntawm SH 0.01,0.1 thiab 10 ug / ml (Daim duab 5B) thaum tag nrho cov kev soj ntsuam muaj peev xwm txo qis IL18 qib (Daim duab 5C). Kev saib xyuas ntawm daim tawv nqaij Muab cov txiaj ntsig tau tshaj tawm saum toj no ntawm cov kab mob epithelial, cov txiaj ntsig ntawm SH extract ntawm ntau cov proteins thiab cov txheej txheem koom nrog hauv kev saib xyuas ntawm daim tawv nqaij thiab cov ntaub so ntswg puas tsuaj raug soj ntsuam.

Sirtuin-1 qhia

Ntau qhov kev tshawb fawb hauv vitro tau tshaj tawm tias UVB hluav taws xob ua rau txo qis hauv Sirtuinl (SIRT1) protein ntau ntawm tib neeg fibroblasts, qhia txog kev koom tes ntawm SIRT1 hauv UVB-mediated puas [29]. Yog li, cov nyhuv ntawm Salvia haenkei extract ntawm Sirtuinl protein qhia tau soj ntsuam los ntawm western blot assay (Daim duab 6). Raws li kev cia siab, UVB stimulation induced ib tug tseem ceeb txo nyob rau hauv SIRTI protein qhia nyob rau hauv HaCaT hlwb. Qhov kev ua ntawm SH extract ntawm SIRT1 qib tau tshawb xyuas. Interestingly, cov txiaj ntsig tau pom tias 1 ug / ml SH extract, piv rau kev tswj hwm, ua rau muaj kev nce ntxiv ntawm SIRTl qhia (ntxiv rau 40 feem pua) nyob rau hauv cov xwm txheej basal, qhia txog qhov muaj peev xwm txhawb nqa ntawm SIRT1 qib. Raws li kev cia siab, UVB stimulation induces ib tug tseem ceeb txo ntawm SIRT1 qib. Cov txiaj ntsig tau pom tias qhov kev kho ua ntej nrog 0.01 ug / ml SH extract nthuav tawm cov nyhuv ameliorative hauv Sirtuin 1 qib, piv rau kev tswj hwm UVB.

Kev kuaj mob kho mob

Keratinocytes yog cov khoom tseem ceeb ntawm cov epidermis, muab kev cuam tshuam ntawm lub cev thiab ib puag ncig. Yog li ntawd, lawv txoj kev loj hlob thiab kev tsiv teb tsaws chaw ua lub luag haujlwm tseem ceeb hauv kev saib xyuas qhov cuam tshuam zoo thiab kho qhov txhab tom qab raug mob [30, 31]. Yog li, qhov cuam tshuam ntawm SH ntawm HaCaT cell migration raug soj ntsuam. Cov duab hauv daim duab 7A qhia tau hais tias kev kho mob nrog SH ua rau txo qis hauv qhov tuab ntawm khawb qhia tias muaj kev cuam tshuam rau HaCaT cell migration. Daim duab 7B qhia txog kev tsim kho kymogram ntawm kev tsiv teb tsaws chaw ntawm HaCaT hlwb; dluab qhia ib tug concentration-dependant nce nyob rau hauv cell migration. Daim duab hauv daim duab 7C qhia tias muaj kev nce ntxiv hauv cell migration ceev tom qab 4-8 teev ntawm kev kho mob nrog 0.01 ug/ml SH extract. Qhov siab tshaj plaws nce mus txog rau lub sijhawm 8-12 teev, qhov twg tus nqi ntawm cov cell tsiv teb tsaws ceev yog li 165 feem pua ​​​​ntawm kev tswj hwm. Cov ntaub ntawv no, raws li cov txiaj ntsig tau txais los ntawm kev loj hlob ntawm tes, qhia tias qhov qis tshaj plaws ntawm SH extract tuaj yeem ua rau muaj kev nce hauv cell tsiv teb tsaws ceev tsis cuam tshuam nrog hyperproliferative phenotype.

Tight junctions'maintenance

Raws li zoo li stratum corneum, nruj junctions (TJs) ua lub luag haujlwm tseem ceeb hauv kev ncaj ncees ntawm daim tawv nqaij, ua qhov cuam tshuam rau cov dej thiab cov kuab tshuaj, thiab koom nrog hauv kev sib txawv, kev loj hlob, cell polarity, thiab cov teeb liab hloov cov txheej txheem ntawm cov hlwb. Cov kev tshawb fawb tsis ntev los no tau pom tias UV-lub teeb hloov pauv qhov kev qhia thiab hauv cheeb tsam ntawm tib cov proteins ntawm TJ qauv, uas ua rau cuam tshuam kev ua haujlwm ntawm daim tawv nqaij [32].

Txhawm rau ntsuas Salvia haenkei extract cov nyhuv los ua tus neeg sawv cev tiv thaiv kev cuam tshuam TJs, kev qhia ntawm occludin tau ntsuas los ntawm kev tiv thaiv kab mob tom qab raug UVB hluav taws xob. Cov duab hauv daim duab 8A, thiab 8B qhia tau hais tias kev kho mob nrog ob qho tib si ntawm SH induces ib qho kev nce hauv occludin protein qhia, txhawb kev sib txuas tsim muaj txiaj ntsig zoo rau kev saib xyuas ntawm epidermal barrier kev ncaj ncees. Raws li qhov xav tau, tom qab raug UVB hluav taws xob, cov hlwb pom qhov tsis xwm yeem thiab tsis tau txhais lub nplhaib (Daim duab 8A). SH kev kho mob, tshwj xeeb tshaj yog nyob rau hauv ntau dua concentration, khaws cia lub morphology ntawm daim nyias nyias tswj cov qauv piv rau unstimulated tswj. SH extract txhawb kev sib txuas ntxiv thiab tiv thaiv kev puas tsuaj los ntawm UVB raug, tswj kev ua haujlwm ntawm daim tawv nqaij barrier.

Filaggrin qhia

Cov nyhuv ntawm Salvia haenkei extract ntawm filaggrin (FLG) protein qhia tau soj ntsuam los ntawm immunocytochemistry. Filaggrin yog cov txheej txheem protein tsim nyog hauv kev txhim kho thiab kev saib xyuas ntawm daim tawv nqaij barrier. Cov txiaj ntsig hauv daim duab 9A, thiab 9B qhia txog kev nce siab ntawm filaggrin protein qhia tom qab kev kho mob rau 24 teev nrog SH extract. Tsis tas li ntawd, qhov kev qhia ntawm F-actin protein qhia txog kev faib tawm ntawm cov hlwb tsis tu ncua, tom qab kev kho mob nrog SH, uas ua rau muaj kev txhim kho zoo txuas ntxiv ntawm cov kab mob epithelial barrier. Cov ntaub ntawv no qhia tias SH muaj txiaj ntsig zoo rau ntawm daim tawv nqaij barrier, txhim kho filaggrin qhia, uas yog qhov tseem ceeb rau kev tsim kom raug thiab ua haujlwm ntawm daim tawv nqaij.

Metalloproteinase qhia

Cov tawv nqaij raug rau ultraviolet hluav taws xob induces impairments nyob rau hauv lub extracellular matrix (ECM) ua rau daim tawv nqaij wrinkling, sagging, thiab laxity, cov yam ntxwv tseem ceeb ntawm daim tawv nqaij laus. Matrix metalloproteinases (MMPs) yog lub luag haujlwm tseem ceeb rau kev rhuav tshem ECM cov khoom xws li collagen, elastin, fibronectin, thiab proteoglycans [33].npaum li cas cistanche cojNtau cov kev tshawb fawb qhia tias cov tawv nqaij raug UV irradiation induces upregulation ntawm txawv MMPs, cuam tshuam rau daim tawv nqaij tsos thiab noj qab haus huv [34-37].

Cov txiaj ntsig hauv daim duab 10 qhia tias UVB hluav taws xob nce mRNA qhia ntawm gelatinase MMP-2 uas degrades hauv qab daus daim nyias nyias collagens thiab denatured structural collagens [38]. Kev kho mob nrog SH tuaj yeem txo qis MMP-2 mRNA ntau ntau (SH 0.01-0.1-10 ug/ml) qhia txog kev ua haujlwm ntawm SH extract hauv kev saib xyuas ntawm cov qauv kev ntseeg siab. daim tawv nqaij, puas los ntawm kev yees duab-kev nyuaj siab.

Kev sib tham

Kev laus ntawm daim tawv nqaij yog cov txiaj ntsig ntawm cov khoom siv sab hauv thiab sab nrauv uas ua rau cov tawv nqaij poob qis, ua rau nyias nyias, tawv nqaij, thiab poob ntawm qhov yooj yim. Daim tawv nqaij yog thawj qhov cuam tshuam ntawm lub cev thiab ib puag ncig, yog li kev saib xyuas ntawm nws txoj kev ua haujlwm thiab kev ncaj ncees anatomical yog qhov tseem ceeb. Dhau li ntawm cov txheej txheem kev laus thiab kev laus ntawm cov kab mob xws li ntshav qab zib, menopause, thiab lwm yam, extrinsic yam tuaj yeem cuam tshuam rau cov qauv ntawm daim tawv nqaij; tshwj xeeb photodamage, kuab paug, lossis kev ua neej nyob (kev haus luam yeeb, noj zaub mov, kev ntxhov siab, thiab lwm yam) tuaj yeem cuam tshuam cov qauv ntawm daim tawv nqaij thiab kev ua haujlwm [39,40]. Keratinocytes yog cov ntsiab lus tseem ceeb ntawm cov epidermis, muab lub hauv paus ntawm cellular rau sab nraud; Hauv kev ua haujlwm no, HaCaT tib neeg keratinocyte cell kab tau raug cuam tshuam rau cov teeb meem kev ntxhov siab, xws li dawb radicals thiab ultraviolet B hluav taws xob, txhawm rau ua kom cov tawv nqaij laus zuj zus, yog li hloov qhov kev cuam tshuam ntawm kev ua haujlwm. Salvia hankie tsis ntev los no tau raug txheeb xyuas tias yog tus neeg sawv cev tiv thaiv kab mob uas muaj peev xwm tawm tswv yim tias nws muaj peev xwm siv tau rau hauv daim tawv nqaij laus thiab kev laus ntawm daim tawv nqaij; Yog li ntawd, cov nyhuv ntawm ib qho extract ntawm cov nroj tsuag no tau tshawb xyuas tob txog qhov tsis muaj feem cuam tshuam rau ntawm daim tawv nqaij laus, kev saib xyuas ntawm daim tawv nqaij ncaj ncees, thiab lub cev nqaij daim tawv. Kev tsim cov pa oxygen reactive lossis dawb radicals los ntawm cov txheej txheem endogenous metabolic lossis UVR thiab cov pa phem tau xav tias yuav pab txhawb rau cov txheej txheem kev laus ntawm daim tawv nqaij. Cov tawv nqaij muaj kev cuam tshuam rau ntau yam kev cuam tshuam redox, thiab redox tshuav nyiaj li cas yog qhov tseem ceeb hauv kev saib xyuas ntawm daim tawv nqaij homeostasis. Tsawg ROS cov ntsiab lus tawm dag zog lub cev, tab sis nce qib ntawm cov tsiaj no tau koom nrog hauv kev ua kom txoj hauv kev ua rau muaj kev nce ntxiv ntawm collagen, tsub zuj zuj ntawm elastin, ua kom cov matrix-degrading metalloproteases, thiab lwm yam. [41]. Ntxiv mus, ROS plays lub luag haujlwm tseem ceeb hauv cov txheej txheem pathological suav nrog kev laus, mob, raug mob, thiab kab mob ntawm ntau yam kab mob ntawm daim tawv nqaij xws li psoriasis thiab vitiligo [42]. Interestingly, Salvia haenkei extract tau pom tias yuav txo tau ROS theem nyob rau hauv basal tej yam kev mob, oxidative kev nyuaj siab tej yam kev mob, thiab kev ntxhov siab vim los ntawm UVB irradiation, qhia tau hais tias muaj peev xwm tiv thaiv cov nyhuv tiv thaiv ROS-mediated alterations [22].

Nyob rau hauv kev txhawb nqa ntawm yav tas los soj ntsuam cov nyhuv antiaging ntawm Salvia hankie, nyob rau hauv no ua hauj lwm, peb proved lub peev xwm ntawm SH extract los txo senescence nyob rau hauv keratinocytes stimulated los ntawm UVB hluav taws xob. Cov txiaj ntsig tau pom tias SH tuaj yeem txo cov noob qhia ntawm p21 thiab p27 senescence cov cim, uas tau txhawb nqa los ntawm UVB hluav taws xob. Interestingly, SH extract qhia tau hais tias kev ua haujlwm zoo hauv kev txo cov inflammatory teb fostered los ntawm photodamage hauv hlwb, los ntawm kev txo qis interleukins transcription. Tsis tas li ntawd, peb tau ua kom pom tseeb tias Salvia haenkei extract induced Sirtuinl protein qhia tsis tau tsuas yog tom qab kev ntxhov siab tab sis kuj nyob rau hauv basal tej yam kev mob, uas qhia tau hais tias muaj kev ua ub no nyob rau hauv slowing txoj kev laus. Sirtuin tsev neeg, thiab tshwj xeeb tshaj yog SIRT1, tau pom tias ua lub luag haujlwm hauv kev laus-photoaging, tshwj xeeb tshaj yog los ntawm inhibition ntawm MMPs thiab tom qab collagen degradation; Ntau qhov kev tshawb fawb qhia txog nws txoj haujlwm tiv thaiv hauv UVB-mediated photoaging. Cov khoom xyaw ntuj xws li juglone (5-hydroxy-1,4-napthalenedione) tau pom tias yuav rov qab SIRT1 mus rau qib qub tom qab kev kho UVB, qhia tias SIRTI tuaj yeem ua lub luag haujlwm tiv thaiv UVB-induced carcinogenesis [29]. Peb cov txiaj ntsig tau qhia zoo ib yam, muaj peev xwm koom tes ntawm sirtuins hauv cov txheej txheem kho tom qab UVB kev ntxhov siab thiab lawv lub luag haujlwm hauv kev saib xyuas ntawm epidermis homeostasis.

Cov kab mob inflammatory ntawm daim tawv nqaij, xws li atopic dermatitis thiab psoriasis, nrog rau cov txheej txheem kev laus, ua rau muaj kev cuam tshuam tsis zoo, thiab cov lus teb ntawm cov kab mob epidermal rau kev cuam tshuam kev cuam tshuam tuaj yeem ua rau hnyav dua, tswj xyuas, lossis txawm tias pib cov xwm txheej zoo li no. Cov txiaj ntsig ntawm txoj haujlwm no tau nthuav tawm lub peev xwm muaj zog ntawm SH los kho cov tawv nqaij thaiv kev ncaj ncees los ntawm kev hloov pauv ntawm ntau cov proteins koom nrog hauv cov txheej txheem no. Hauv particular, nws tau pom stimulation ntawm occludin thiab filaggrin transcription qhia ib qho kev txhawb zog ntawm adhesions ntawm hlwb. Inherited thiab tau txais filaggrin deficiency cuam tshuam rau epidermal barrier, hloov lub koom haum ntawm

cytoskeleton keratin filaments thiab cov qauv ntawm lub hnab ntawv cornified ntawm daim tawv nqaij [43]. Tsis tas li ntawd, qhov tsis xws luag hauv FLG qhia ua rau muaj qhov txo qis ntawm cov keratohyalin granules, ib qho kev poob qis hauv ntuj moisturizing factor (NMF) concentration, thiab alkalinization ntawm daim tawv nqaij pH [44]. Tsis tas li ntawd, kev hloov pauv hauv claudins thiab occludin proteins, uas yog cov Tight junctions (TJs) proteins, tau piav qhia zoo txog cov txheej txheem kev laus. TJs yog lub luag haujlwm rau kev sib khi qhov chaw sib txuas ntawm cov hlwb epithelial, tsim kom muaj kev cuam tshuam kev ua haujlwm, thiab koom nrog hauv kev loj hlob ntawm tes thiab kev sib txawv [45]. Salvia haenkei extract tau raug pom los txhim kho filaggrin qhia thiab khaws cov occludin localization ntawm cell membranes, impaired los ntawm UVB hluav taws xob. Ib qho kev ua ntxiv ntawm SH extract hauv kev saib xyuas ntawm daim tawv nqaij noj qab haus huv tau ua pov thawj los ntawm nws lub peev xwm los txo cov matrix metalloproteinases 2 (MMP-2) kev hloov pauv, txhawb nqa los ntawm UVB hluav taws xob. Lub upregulation ntawm MMPs pab txhawb kev laus thiab daim tawv nqaij pathologies, los ntawm degrading collagen thiab elastin ua rau impairments nyob rau hauv lub extracellular matrix (ECM) [33, 38], yog li txo cov proteinases yog tsim nyog nyob rau hauv lub ntsiab lus ntawm daim tawv nqaij laus.

Cov txiaj ntsig zoo ntawm SH kev kho mob tau pom nyob rau hauv txoj kev tshawb no tshaj qhov feem ntau ntawm cov tawv nqaij molecular tsis tau lees paub los ntawm kev soj ntsuam tsis ntev los no, raug tua nyob rau hauv ib pawg ntawm tsib caug Caucasian poj niam hnub nyoog ntawm 45 thiab 60 [46]. Salvia hankie tau raug tswj hwm nyob rau hauv cov tshuaj pleev ib ce thiab ntau yam tsis zoo, xws li daim tawv nqaij liab thiab elasticity, wrinkle qhov tob, thiab antioxidant muaj peev xwm, tau raug soj ntsuam los ntawm kev soj ntsuam thiab ntsuas ntsuas nrog rau kev ntsuas tus kheej los ntawm cov kev kawm. Cov txiaj ntsig ntawm txoj kev tshawb no qhia SH kev ua tau zoo hauv ameliorating daim tawv nqaij poob thaum siv txhua hnub hauv cov ntsiab lus ntawm cov tawv nqaij laus. Qhov tseeb, tom qab 84 hnub ntawm daim ntawv thov, cov poj niam ntawm daim tawv nqaij ua rau kom elastic dua thiab nthuav tawm tsawg dua ntawm thaj chaw liab thiab ntiav cov wrinkles. Tsis tas li ntawd, txoj kev tshawb fawb pom tau tias muaj kev nce qib hauv cov tawv nqaij antioxidant muaj peev xwm ua raws li kev thov txhua hnub ntawm cov tshuaj nplaum [46].

Raws li kev tiv thaiv vaj tse rau lub cev los ntawm ib puag ncig sab nraud, daim tawv nqaij raug mob tas li, thiab yog li kev kho qhov txhab yog qhov tseem ceeb rau kev ciaj sia ntawm txhua yam kab mob. Xav txog qhov muaj peev xwm ua pov thawj ntawm SH hauv kev saib xyuas cov tawv nqaij noj qab haus huv thiab tsis muaj teeb meem, peb xav tias cov extract no tuaj yeem pab txhim kho daim tawv nqaij rov qab los ntawm kev ua haujlwm zoo hauv kev kho cov tawv nqaij. Yog li, SH kev ua haujlwm ntawm keratinocytes tsiv teb tsaws, yog ib kauj ruam tseem ceeb hauv kev kho cov tawv nqaij barrier tom qab raug mob, thaum kawg txiav txim siab. SH extract kev kho mob tau nthuav tawm ib qho kev nthuav dav uas muaj peev xwm ua kom HaCaT ceev ntawm kev tsiv teb tsaws, ua kom pom kev muaj zog ntawm kev rov tsim kho ntawm epithelial raug mob tom qab qhov txhab. Raws li kev tsiv teb tsaws yog ib qho tseem ceeb hauv cov txheej txheem kho qhov txhab, peb cov ntaub ntawv, kuj tseem xav txog cov kev ua haujlwm tsis yog hyperproliferative ntawm cov extract, qhia txog qhov muaj peev xwm ntawm SH extracts hauv kev tawm tsam sai dua qhov cuam tshuam ntawm kev raug mob ntawm daim tawv nqaij barrier, txo qhov kev pheej hmoo ntawm kab mob.

Peb cov ntaub ntawv qhia txog Salvia haenkei extract raws li ib qho tseem ceeb compound rau ob qho tib si kev pab cuam thaiv kev rov tsim los yog dampening lub epidermal kev nyuaj siab teb. Concomitantly nrog rau kev sim tsis ntev los no nyob rau hauv tib neeg, qhov kev ua hauj lwm no muab kev nkag siab ntxiv rau cov txheej txheem koom nrog cov txiaj ntsig zoo ntawm cov extract ntawm daim tawv nqaij laus.

Txoj haujlwm no, txhawb kev ua haujlwm yav dhau los [18, 46], txheeb xyuas txoj hauv kev tshwj xeeb tshaj plaws ntawm cov txheej txheem molecular txuas rau kev ua tau zoo ntawm Salvia hankie hauv kev hloov pauv ntawm daim tawv nqaij tsis zoo, qhia txog qhov cuam tshuam ntawm Salvia hankie ua tus neeg sawv cev txaus siab rau kev tiv thaiv kev laus. thiab kev laus ntawm daim tawv nqaij mob.

MATERIALS THIAB METHODS Cell kab lis kev cai thiab UVB irradiation

HaCaT hlwb (tib neeg spontaneously immortalized keratinocytes) tau kab lis kev cai nyob rau hauv siab qabzib Dulbecco's modified Eagle's nruab nrab (DMEM) ntxiv nrog 10 feem pua ​​​​fetal bovine serum (Lub neej-Technologies, Waltham, MA, USA), 2 mM glutamine, 100in penicillin thiab ug/ml streptomycin (Lonza, Basilea, Switzerland). Cells tau khaws cia nyob rau hauv ib tug humidified huab cua ntawm 5 feem pua ​​CO2 nyob rau hauv cov huab cua thiab incubated ntawm 37 degree.

Thaum thov los ntawm cov txheej txheem kev sim, cov hlwb raug rau 30 KJ / m² UVB hluav taws xob, muab los ntawm Philips Medical Narrowband UVB PL-S teeb. Txhawm rau tiv thaiv lub teeb nqus los ntawm cov ntaub so ntswg-kev coj noj coj ua, qhov no tau raug tshem tawm ua ntej irradiation, thiab hloov nrog DMEM FBS thiab liab phenol-dawb, ua raws li irradiation raws tu qauv [47]. Tom qab UVB irradiation, hlwb tau noj nrog ib tug tshiab loj hlob nruab nrab los yog siv rau kev sim ua raws li cov txheej txheem.

Cell viability-Trypan blue exclusion assay 35 x10³ cov hlwb raug plated rau 12 lub phaj zoo thiab, tom qab kev tsim tawm thaum hmo ntuj, tau nthuav tawm ntau qhov sib txawv ntawm Salvia haenkei hydroethanolic extract raws li kev sim cov txheej txheem. Cov extract tau ua siab zoo muab los ntawm IBSA Farmaceutici Italia (Lodi, Ltalis) thiab nthuav tawm cov ntsiab lus ntawm cov cim rosmarinic acid ntawm 0.6 feem pua ​​w/w los ntawm HPLC. Tom qab kev kho mob, cov hlwb raug ntxuav, tshem tawm nrog 0.25 feem pua ​​trypsin -0.2 feem pua ​​​​EDTA, thiab raug tshem tawm hauv trypan xiav (Sigma-Aldrich, St Louis, MO, USA) ntawm ib qho l: Kuv piv rau hauv cov tshuaj nruab nrab. Cells raug suav siv lub chamber Burker hemocytometer.

ROS fluorescence xeem

ROS tau suav hais tias siv 2',7'-dichlorofluorescein-diacetate (Hz-DCF-DA, Sigma-Aldrich, St Louis, MO, USA), raws li tau piav qhia yav dhau los [48]. Luv luv, cov hlwb (5x10') tau cog rau hauv 96-cov phiaj zoo, tso cai rau ua raws li hmo ntuj, thiab tom qab ntawd raug rau SH extract rau 3 lossis 24 teev. N-acetylcysteine ​​5mM (Sigma-Aldrich, St Louis, MO, USA) tau siv los ua kev tswj xyuas zoo.

Rau oxidative kev nyuaj siab, ROS qib tau ntsuas tom qab qhov sib ntxiv ntawm 50 μM Hz-DCF-DA nyob rau hauv qhov tsis muaj lossis muaj H2Oz [48]. DCF fluorescence siv tau ntsuas ntawm excitation 485 nm-emission 535 nm, siv Multilabel Plate Reader VICTOR X3 (PerkinElmer, Waltham, MA, USA).

Rau kev ntsuam xyuas ntawm ROS qib hauv UVB kev ntxhov siab, cov txheej txheem tau hloov kho me ntsis, los ntawm kev tsim cov hlwb hauv SH kev kho mob - 50 μM Hz-DCF-DA tshuaj hauv PBS. Tom qab 25 feeb ntawm incubation ntawm 37 degree, lub phaj tau raug UVB hluav taws xob thiab DCF fluorescence siv tau tam sim ntawd ntsuas ntawm excitation 485 nm-emission 535 nm, siv Multilabel Plate Reader VICTOR X3 (PerkinElmer, Waltham, MA, USA).

Immunofluorescence microscopy

10x104 hlwb tau cog rau ntawm cov iav npog ua ntej-coated nrog collagen hauv 24- cov phiaj zoo, tso cai rau xa mus rau hmo ntuj, thiab kho nrog SH extract raws li kev cai. Rau kev kuaj pom ntawm -catenin thiab occludin proteins, hlwb raug pom los ntawm UVB hluav taws xob, raws li tau piav qhia hauv cov txheej txheem saum toj no. Tom qab irradiation, cov hlwb tau noj nrog qhov nruab nrab loj hlob tshiab los yog siv rau kev sim ua raws li txoj cai. Rau immunofluorescence assay, 4 teev tom qab UVB irradiation, hlwb raug ntxuav, kho nrog 4 feem pua ​​formaldehyde, permeabilized nrog 0.1 feem pua ​​Triton X-100 hauv PBS, thiab stained rau 1 teev ntawm 37 degree nrog txawv protein-specific antibodies: nas monoclonal anti-occludin (Invitrogen Life Technologies, Waltham, MA, USA), nas anti-filaggrin (Sigma-Aldrich, St Louis, MO, USA). Tom qab PBS ntxuav, cov hlwb raug incubated nrog lwm cov tshuaj tiv thaiv kab mob / fluorescein isothiocyanate (Alexa Fluor 488 anti-nas los yog Alexa Fluor 536 anti-Lavbit immunoglobulin G, Molecular Probes, Invitrogen Life Technologies, Waltham, MA, USA) thiab nrog phalloidin rau 1 teev. chav kub. Tom qab 10 feeb ntawm kev kho mob nrog RNase, cov ntawv npog tau muab tso rau ntawm cov iav slides los ntawm kev siv Mowiol 40-88 (Sigma-Aldrich, St Louis, MO, USA) thiab ntxiv propidium iodide. Cov duab tau txais los ntawm confocal microscope LSM 800, magnification 60X, software ZN 2.1 blue Edition (Carl Zeiss, Jena, Germany) thiab suav nrog siv ImageJ software.

Kev txheeb cais

Kev txheeb xyuas txheeb cais tau ua tiav siv GraphPad Prism version 7.02 rau Windows (GraphPad Software, San Diego, CA, USA). Tshwj tsis yog hais tias lwm yam, cov txiaj ntsig tau nthuav tawm raws li txhais tau tias ntxiv rau SEM. Cov tub ntxhais kawm qhov t-test tau siv thiab p tus nqi<0.05 were="" considered="" statistically="">

Cov ntawv luv

AJ:adherens kev sib tshuam; ECM: extracellular matrix; FLG: filaggrin; Hz-DCF-DA: 2',7'-dichlorofluorescein-diacetate; H2Oz: Hydrogen Peroxide; MMP: matrix metalloproteinase; NAC: N-acetylcysteine; NMF: yam; ROS: reactive oxygen natural moisturizing

hom; SIRTl: Sirtuinl; SH: Salvia hankie; TJ: nruj kev sib tshuam; UVB: Ultraviolet hluav taws xob B.

Kab lus no yog muab rho tawm los ntawm www.aging-us.com