Lub Cim Acylated Flavonol Glycoside Los Ntawm Prunus Persica (L.) Var. Florida Tub Vaj Ntxwv: Tus Tshiab Khoom Lipid Nanoparticle Cosmetic Formulation Rau Skincare Part 1

Abstract: Polyphenols paub txog kev noj haus antioxidants. Lawv tsis ntev los no tau nyiam kev txaus siab rau kev siv los tiv thaiv kev laus ntawm daim tawv nqaij thiab hyperpigmentation uas tshwm sim los ntawm hnub ci UV irradiation. Prunus persica (L.) nplooj yog suav tias yog cov khoom lag luam thiab tau tshaj tawm tias muaj cov tshuaj tiv thaiv antioxidant zoo tshaj plaws vim lawv cov ntsiab lus siab ntawm polyphenols. Txoj kev tshawb no tsom rau kev txhim kho cov tshuaj pleev ib ce los tiv thaiv kev laus thiab cov tawv nqaij ntxuav hniav dawb siv cov ethanol nplooj extract ntawm Prunus persica (L.) (PPEE) loaded nyob rau hauv cov khoom lipid nanoparticles (SLNs) los txhim kho daim tawv nqaij. Kev tshawb nrhiav tshuaj lom neeg ntawm PPEE tau pom tias muaj cov ntsiab lus phenolic thiab flavonoid ntau heev nrog cov haujlwm tseem ceeb ntawm cov tshuaj antioxidant (DPPH, ABTS, thiab -carotene assays). Ib qho tshwj xeeb acylated kaempferol glycoside nrog cov qauv tsis tshua muaj, kaempferol 3-O- -4C1-(600 -O-3,4-dihydroxyphenylacetyl glucopyranoside) ( KDPAG) raug cais tawm thawj zaug thiab nws cov qauv tau qhia meej meej. Nws sawv cev rau thawj piv txwv ntawm acylation nrog 3, {{10}}dihydroxy phenylacetic acid hauv flavonoid chemistry. Cov kev tshawb fawb hauv vitro cytotoxicity tawm tsam tib neeg keratinocytes cell kab qhia qhov tsis muaj tshuaj lom ntawm PPEE thiab PPEE-SLNs. Ntxiv mus, PPEE, PPEE-SLNs, thiab KDPAG tau pom zoo los tiv thaiv elastase kev ua si, piv rau cov N (Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone. Tsis tas li ntawd, PPEE-SLNs thiab KDPAG tau pom zoo (p <0.001) ntau dua los tiv thaiv collagenase thiab kev tiv thaiv tyrosinase hauv kev sib piv rau EDTA thiab kojic acid, raws li. Cov qauv PPEE-SLNs sib txawv (2 feem pua thiab 5 feem pua ) tau raug tshuaj xyuas rau qhov ua tau los tiv thaiv kev ua xua tiv thaiv UV-induced photoaging nyob rau hauv tus qauv nas siv txoj kev wrinkle scoring thiab tau pom tias muaj kev tiv thaiv tseem ceeb heev tiv thaiv UV, raws li pov thawj. los ntawm cov ntaub so ntswg biomarkers (SOD) thiab cov kev tshawb fawb histopathological. Yog li, txoj kev tshawb fawb tam sim no qhia tau hais tias Prunus persica nplooj los ntawm cov khoom lag luam muab kev nthuav dav, muaj txiaj ntsig zoo rau cov khoom xyaw ntuj kom zoo nkauj. Qhov no muab cov ntaub ntawv muaj feem xyuam rau kev kawm ntxiv txog kev siv PPEE-SLNs nyob rau hauv cov tshuaj pleev ib ce los tiv thaiv kev laus nrog kev txhim kho ntawm daim tawv nqaij permeation zog.

Raws li kev tshawb fawb cuam tshuam,cistancheyog ib hom tshuaj ntsuab uas hu ua "cov tshuaj ntsuab miracle uas ua rau lub neej ntev". Nws lub ntsiab tivthaiv yogcistanoside, uas muaj ntau yam teebmeem xws liantioxidant, anti-inflammatory, thiabkev tiv thaiv kev ua haujlwm txhawb nqa. Lub mechanism ntawm cistanche thiabtawv nqaij dawbnyob rau hauv cov nyhuv antioxidant ntawmcistanche glycosides. Melanin nyob rau hauv tib neeg daim tawv nqaij yog tsim los ntawm oxidation ntawm tyrosine catalyzed los ntawmtyrosinase, thiab cov tshuaj tiv thaiv oxidation yuav tsum muaj kev koom tes ntawm cov pa, yog li cov pa oxygen dawb radicals hauv lub cev dhau los ua ib qho tseem ceeb cuam tshuam rau kev tsim melanin. Cistanche muaj cistanoside, uas yog ib qho antioxidant thiab tuaj yeem txo qhov tsim ntawm cov dawb radicals hauv lub cev, yog liinhibiting melanin ntau lawm.

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Nyem rau ntawm Cistanche Supplement For Whitening

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Ntsiab lus:kaempferol 3-O- -4C1-(600 -O-3,4-dihydroxyphenylacetyl glucopyranoside) los ntawm cov khoom; khoom lipid nanoparticles; antioxidant; in-vitro daim tawv nqaij ntsig txog enzymes; nyob rau hauv vivo anti-wrinkle

1. Taw qhia

Kev laus ntawm daim tawv nqaij yog cov txheej txheem ntau yam kev ua haujlwm uas ua rau lub cev hloov pauv ntawm daim tawv nqaij thiab cov nqaij sib txuas [1]. Nws tau muab cais rau hauv kev laus thiab kev laus.

Kev laus ntawm daim tawv nqaij feem ntau tshwm sim los ntawm ib puag ncig, xws li cov pa phem, haus luam yeeb, thiab kev nyuaj siab hauv lub neej, lossis feem ntau los ntawm kev rov raug UV hluav taws xob (duab duab) [2].

Overexposure rau UV hluav taws xob nkoos lub overproduction ntawm reactive oxygen hom (ROS) uas ua rau endogenous oxidative kev nyuaj siab nyob rau hauv daim tawv nqaij cov ntaub so ntswg ua rau degradation ntawm extracellular matrix (ECM) Cheebtsam. ECM degradation yog txuas ncaj qha rau ntawm daim tawv nqaij laus thiab yog lub luag haujlwm rau kev nce hauv kev ua haujlwm ntawm qee cov enzymes xws li collagenase, elastase, thiab tyrosinase uas koom nrog hauv kev laus ntawm daim tawv nqaij. Cov dermal enzyme activations ua rau txo qis hauv qib ntawm elastin, thiab collagen uas ua rau poob ntawm elasticity thiab lub zog ntawm daim tawv nqaij thiab cov tsos ntawm wrinkles [3]. Tsis tas li ntawd, induction ntawm ntau melanin ntau lawm thiab daim tawv nqaij tanning ua rau hyperpigmentation ntawm daim tawv nqaij.

Yog li ntawd, kev laus ntawm daim tawv nqaij tuaj yeem tiv thaiv tau los ntawm cov tshuaj tua kab mob antioxidant nrog cov dawb radical scavenging kev ua haujlwm uas yuav muaj qhov tseem ceeb hauv kev tiv thaiv thiab kho cov kab mob hnub nyoog uas cuam tshuam nrog ROS [4]. Lwm txoj hauv kev los tiv thaiv kev laus yog siv elastase, collagenase, thiab tyrosinase inhibitors uas yog cov neeg sib tw muaj zog heev rau kev tiv thaiv wrinkle thiab tawv nqaij ua kom tawv nqaij ua kom pom zoo rau kev khaws cia ntawm daim tawv nqaij elasticity thiab tuaj yeem yog ib txoj hauv kev los daws cov teeb meem pigmentation [5].

Nws tau raug tshaj tawm tias cov tshuaj tua kab mob ntuj tsim tawm los ntawm cov nroj tsuag ua rau muaj kev pheej hmoo ntawm kev yees duab los ntawm UV irradiation ob qho tib si hauv-vitro thiab hauv-vivo [6] thiab lawv zoo dua li cov khoom xyaw cosmeceutical dua li hluavtaws antioxidants ntawm cov nqi thiab cov kev mob tshwm sim. Tsis tas li ntawd, ntau qhov kev tshawb fawb hauv vitro thiab hauv vivo tau pom tias cov tshuaj phenolic xws li phenolic acids, flavonoids, thiab tannins tuaj yeem tshem tawm cov dawb radicals thiab inhibit elastase, collagenase, thiab tyrosinase enzymes [7].

Kev siv cov tshuaj antioxidants ntuj nyob rau hauv cov tshuaj pleev ib ce los tiv thaiv daim tawv nqaij tiv thaiv oxidative kev nyuaj siab los ntawm extrinsic yam tau raug tshaj tawm [8]. Txawm li cas los xij, feem ntau ntawm cov tshuaj antioxidant molecules yog innately tsis ruaj khov thiab tuaj yeem yooj yim oxidize los tsim cov tshuaj tsis muaj zog ua ntej mus txog qhov chaw ua haujlwm uas ua rau lawv nyuaj rau kev tsim cov khoom tsim nyog thiab ruaj khov. Cov kev tshawb pom no ua pov thawj qhov tsim nyog ntawm kev xa khoom tshwj xeeb los txhim kho cov qauv tshuaj antioxidant [9].

Ntau yam cuam tshuam rau permeability ntawm topically thov polyphenols; phenolic subclass, nws cov qauv, nws cov molecular loj, thiab daim ntawv glycoside los yog aglycone, nrog rau lwm yam formulation Cheebtsam. Txawm li cas los xij, kev sib cuam tshuam nrog cov tawv nqaij los ntawm kev sib xyaw ntawm lub cev thiab tshuaj txoj kev tuaj yeem txhim kho permeability los ntawm kev cuam tshuam rov qab ntawm stratum corneum txheej txheej txheej txheej thaiv. Encapsulation mus kom ze yog ib qho ntawm cov permeability-txhim kho txoj kev uas feem ntau siv los yooj yim stabilize polyphenolics thaum lub sij hawm cia, ua rau lawv cov teebmeem antioxidant, dermal absorption, thiab nkag mus rau hauv tshuaj pleev ib ce thiab tshuaj pleev ib ce. Tshiab tsim encapsulation technologies muaj xws li nanoemulsions, transferases, khoom lipid nanoparticles, nanocrystals, thiab ribosomes [10].

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Khoom lipid nanoparticles (SLNs) yog cov khoom xa tuaj ruaj khov rau cov khoom siv tawv nqaij. SLNs raug suav hais tias yog cov khoom lag luam pheej hmoo rau cov tshuaj pleev ib ce vim lawv cov duab tsis muaj zog, tswj cov khoom tso tawm, occlusivity, tsw npog qhov ncauj qhov ntswg, thiab kev nkag mus ntawm cov khoom siv hauv cov tawv nqaij los ntawm kev ua kom nws cov dej tsis muaj cov tsos mob ntawm daim tawv nqaij, ntxiv rau qhov yooj yim. ntawm ntau lawm, tsis tshua muaj toxicity, thiab lub cev stability [9,11].

Yog li ntawd, nws yog ib qho tseem ceeb heev uas yuav tau nrhiav ib qho tseem ceeb ntawm cov nroj tsuag polyphenols raws li kev kho mob los tiv thaiv wrinkle thiab cov tawv nqaij whitening agents siv ib tug nanocarrier xa system, nrog rau nws muaj peev xwm los txhim kho lub cev antioxidant system.

Nyob rau hauv lub ntiaj teb no, muaj txog 2000 peach cultivars [12]. Prunus (Rosaceae) yog cov nroj tsuag nto moo uas suav nrog phenolic-nplua nuj hom [13-15]. Qee hom ntawm cov genus no tau cog qoob loo hauv tebchaws Iziv rau lawv cov txiv hmab txiv ntoo noj tau lossis ua khoom siv [16].

Prunus persica (L.) Batch (PP) yog ib tsob ntoo me me nrog cov ceg tawv thiab lub cev luv, nthuav dav, puag ncig puag ncig [12] thiab feem ntau cog rau hauv West Asia, Europe, India, thiab North Africa [17]. Kev lig kev cai, nws cov nplooj tau siv los ua diuretics, laxatives, vermifuges, tshuaj tua kab, sedatives, rau hnoos hawb pob, rau kev kho mob ntawm leukoderma, thiab raws li febrifuges [17]. Tsis tas li ntawd, cov kev tshawb fawb pharmacological ntawm nplooj pom lawv cov in-vivo los tiv thaiv ntshav qab zib, cov nyhuv spasmogenic, tiv thaiv inflammatory, anticoagulant, hepatoprotective, antimalarial, anti-asthmatic, thiab in-vitro cytotoxic, antimicrobial, thiab nitric oxide inhibitory nrog cov haujlwm tseem ceeb los tiv thaiv oxidant. [18–22].

Ntau cov kev tshawb fawb tau ua tiav los tshawb xyuas cov phenolic profile ntawm PP cov thwjtim, cov txiv hmab txiv ntoo, thiab cov noob, lawv cov kev ua haujlwm antioxidant, thiab cov tshuaj pharmacological thiab cov khoom noj muaj txiaj ntsig [13,14,23,24]. Txawm li cas los xij, cov ntaub ntawv tsawg heev tau muaj nyob rau ntawm nplooj. Txog rau hnub tim, ib daim ntawv tshaj tawm muaj qhia txog kev cais ntawm tsib flavonol glycosides los ntawm ethanol nplooj extract [25]. Tsis tas li ntawd, phenolic tebchaw tau ua tus cwj pwm, suav nrog phenolic acids thiab flavonoids siv HPLC-MS tsom xam nrog flavonols ua cov tshuaj tseem ceeb ntawm txhua qhov kev txiav txim siab [20,26].

Ntxiv mus, txawm hais tias PP noob, txiv hmab txiv ntoo, cov thwjtim, thiab lwm yam tsiaj tau tshaj tawm tias muaj kev tiv thaiv hauv vivo tiv thaiv UV-induced photoaging thiab in-vitro anti-wrinkle thiab tawv nqaij ua haujlwm dawb [15,23,27–32], qhov no tsis tau tshaj tawm rau PP nplooj extract.

Cov nplooj loj ntawm PP yog cov khoom lag luam tau los ntawm kev cog ntoo txiv duaj thiab cov txiv hmab txiv ntoo hauv kev lag luam [33]. Cov khoom los ntawm PP cov noob, txiv hmab txiv ntoo, thiab lwm hom nplooj tau raug tshuaj xyuas hauv cov tshuaj pleev ib ce thiab ua zaub mov noj rau qhov muaj peev xwm valorization [32,34,35]. Txawm li cas los xij, rau qhov zoo tshaj plaws ntawm peb txoj kev paub, cov nplooj los ntawm cov khoom ntawm PP var. Florida Prince yeej tsis tau tshawb xyuas thiab tuaj yeem yog qhov nthuav dav ntawm phytochemicals. Piv nrog rau lwm cov kev lag luam, kev lag luam tshuaj pleev ib ce yog qhov nkag tau yooj yim dua thiab nthuav dav thiab tuaj yeem yog qhov chaw rau valorization ntawm cov khoom lag luam [15]. Kev tu tawv nqaij thiab cov khoom siv tawv nqaij laus yog qee qhov tseem ceeb tshaj plaws ntawm kev ua kom zoo nkauj.

Nyob rau hauv kev txuas ntxiv ntawm peb cov kev tshawb fawb ntawm polyphenolics ntawm Egyptian edible nroj tsuag [36] thiab txij li thaum PP nplooj yog suav hais tias yog los ntawm cov khoom thiab tau tshaj tawm tias muaj cov tshuaj antioxidant tseem ceeb vim lawv cov ntsiab lus flavonols siab, txoj kev tshawb no tsom mus rau PPEE rau ib qho phytochemical uas nws kim heev. tsom xam ntawm nws cov phenolic profile thiab muab PPEE rau hauv loaded SLNs raws li ib tug tshwj xeeb daim tawv nqaij xa system. Tsis tas li ntawd, qhov kev tiv thaiv kev laus thiab cov tawv nqaij ua kom zoo nkauj zoo nkauj tau raug soj ntsuam los ntawm kev tshawb xyuas cov tshuaj tua kab mob hauv vitro ntawm PPEE, PPEE-SLNs, thiab cov khoom sib cais, nrog rau kev soj ntsuam ntawm lawv cov teebmeem inhibitory tiv thaiv cov enzymes ntawm daim tawv nqaij. Ntxiv nrog rau kev koom ua ke ntawm PPEE-SLNs rau hauv cov tshuaj pleev ib ce los tiv thaiv kev laus thiab tshuaj xyuas cov khoom lag luam kev nyab xeeb nrog rau kev soj ntsuam ntawm qhov ua tau hauv vivo los tiv thaiv wrinkle kev ua ntawm PPEE-SLNs qab zib formulations tiv thaiv UV-induced photoaging hauv ib nas qauv. Qhov no tuaj yeem muab cov ntaub ntawv rau kev kawm ntxiv txog kev siv PPEE-SLN nplooj hauv cov tshuaj pleev ib ce nrog kev txhim kho ntawm daim tawv nqaij permeation.

2. Cov khoom siv thiab cov txheej txheem

2.1. General

NMR spectra were acquired in DMSO-d6 on an Avance 400 NMR spectrometer (Bruker bio spin Gmbh, Rheinstetten, Germany) at 400 MHz. Standard pulse sequence and parameters were used to obtain one-dimensional 1H and 13C-APT, and two-dimensional HSQC and HMBC spectra. 1H chemical shifts (δ) were measured in ppm, relative to TMS, and 13C-NMR chemical shifts to DMSO-d6 and were converted to TMS scale by adding 39.49. ORAC measurements were performed on a FLUOstar Omega Microplate Reader (BMG LABTECH Gmbh, Ortenberg, Germany). A Shimadzu UV–Visible-1601 spectrophotometer (Shimadzu, Kyoto, Japan) was used for recording the UV spectra. Chromatographic analysis (PC) was carried out on Whatman No. 1 paper, using solvent systems: (1) H2O; (2) 6% HOAc; (3) BAW (n-BuOH–HOAc–H2O, 4:1:5, upper layer). For 2-DPC, solvent 2 was used for the first way and solvent 3 for the second way. Materials: chemicals, solvents, elastase, collagenase, tyrosinase enzymes, and reference drugs (EDTA, kojic acid, and N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone with purities >95 feem pua) tau txais los ntawm Sigma-Aldrich (Sigma-Aldrich, MO, USA).

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2.2. Khoom siv cog

Nplooj ntawm Prunus persica (L.) Batsch var. Florida Tub Vaj Ntxwv tau sau thaum Lub Rau Hli 2020 los ntawm EL-Hashash Farm, El-Bastan nkag, Cairo-Alexandria suab puam txoj kev K. 103, 18 k. sab hauv, Egypt. Cov voucher specimen (4643) tau muab tso rau hauv Herbarium ntawm Flora thiab Phytotaxonomy Researches Department (CAIM), Horticultural Research Institute, Agricultural Research Center, Giza, Egypt. Prof. Dr. Abdel-Haleem Abdel-Mogaly, Prof. ntawm Botany ntawm Agricultural Research Center, tau lees paub nws tus kheej.

2.3. Kev npaj cog Extract

Extraction ntawm PP nplooj qauv (2 kg) tau ua tiav siv kub EtOH / H2O (3: 1, peb zaug, 3 L txhua, rau 8 h, nyob rau hauv reflux) ua raws li hnyav evaporation nyob rau hauv txo siab ntawm 50 ◦C yielding ib amorphous extract. , PPEE, nrog xim av tsaus nti (101 g).

2.4. Kev txiav txim siab ntawm Tag Nrho Polyphenols Cov ntsiab lus (TPC)

Folin-Ciocalteu reagent raws li tau piav qhia los ntawm Li et al., [37] tau siv los kwv yees tus nqi ntawm tag nrho cov ntsiab lus phenolic hauv PPEE, nthuav tawm li mg sib npaug gallic acid / g qhuav extract.

2.5. Kev txiav txim siab ntawm tag nrho cov ntsiab lus Flavonoid (TFC)

Cov txheej txheem aluminium chloride colorimetric piav qhia los ntawm Bahromun li al., [38] tau siv los kwv yees tag nrho cov ntsiab lus flavonoid. Cov txiaj ntsig tau nthuav tawm raws li mg sib npaug quercetin / g qhuav extract.

2.6. Fractionation of PPEE thiab Isolation of Compound 1

Cov khoom qhuav qhuav (101 g), yaj hauv 150 mL MeOH, tau siv rau hauv MCI gel kem (CHP20P, 75-150 mm; Mitsubishi Chemical Co., Düsseldorf, Lub teb chaws Yelemees) thiab eluted los ntawm MeOH / H2O sib tov ntawm txo polarities. Qhov no ua rau plaub feem loj (I–IV). Elution ntawm feem kuv tau ua tiav siv 30 feem pua MeOH/H2O, 22 g; feem II yog eluted nrog 50 feem pua aqueous MeOH, 3.1 g; feem III, nrog 70 feem pua aqueous MeOH (4.6 g); feem IV, tau eluted nrog 90 feem pua aqueous MeOH (1.9 g). Ib qho kev daws teeb meem ntawm feem III tau tso tseg thaum hmo ntuj ntawm chav tsev kub, muab cov qauv ntawm cov khoom sib xyaw 1 (765 mg). Qhov no yog ua raws li kev pom thiab rov ua kom rov ua kom sov hauv EtOH, tom qab ntawd sab laug los sawv ntawm chav tsev kub kom them taus cov qauv xim daj-daj ntshiab ntawm 1 (530 mg).

2.7. Kev txheeb xyuas ntawm kaempferol 3-O- -4C1-(6"-O-3,4-dihydroxyphenylacetyl glucopyranoside) (1)

R f qhov tseem ceeb: 34 (H2O), 39 (15% HOAc), 44 (BAW). UV hauv MeOH λmax nm: 268, 316, 368; NaOAc: 267, 317, 369; NaOAc plus H3BO3: 268, 318, 340; NaOMe: 277, 370. Cov kua qaub hydrolysis ib txwm muab cov piam thaj (Co-PC), kaempferol (Co-PC), thiab 3, 4-dihydroxy phenylacetic acid (UV thiab 1H NMR spectra), UV λmax (MeOH): 224, 282 nm; λmax (MeOH ntxiv rau A1C13): 220, 250 (sh), 291 nm; (MeOH plus NaOAc plus H3BO3) 211, 234 (sh), 288 nm. 1H NMR: δ ppm: 3.40 (s, 2H, C-H2), 6.60 (m, 3H aromatic, H-2, H-5, H-6), -glucosidase kev kho mob ntawm 1 [lyophilized, chromatographically ntshiab, ntsev-dawb enzyme los ntawm almond, BDH Merck, Poole Dorset, UK (EC 3.2.1.21)] rau 24 h, ntawm 37 ◦C hauv acetate tsis, pH 5.1: cov compound tau zoo li qub tsis hloov. 1H-NMR ntawm 1: δppm: kaempferol moiety: 8.0.3 (d, J=8 Hz, H-20 thiab H-60 ), 6.89 (d, J=8) Hz, H-30 thiab H-50 ), 6.45 (d, J=2 Hz, H-6), 6.21 (d, J=2 Hz, H -8); Glucose moiety: 5.42 (d, J=8 Hz, anomeric proton H-100), 4.04 (dd, J=12 Hz thiab J=4.5 Hz, H{{ 88}} a), 4.2 (d, J=12 Hz, H-600 b); 3,4-dihydroxyphenyl acetic acid moiety: 3.46 (broad singlet, ∆ν1/2=4 Hz, CH2-7), 6.67 (m, 3H, H-2000 , H{ {104}} , H-6000 ). 13C-NMR ntawm 1: δ ppm: kaempferol moiety: 156.65 (C-2), 133.5 (C-3), 178.01 (C-4), 161.69 (C-5) ), 99.13 (C-6), 164.58 (C-7), 93.90 (C-8), 156.71 (C-9), 104.39 (C-10) ), 121.34 (C-10), 133.66 (C-20 thiab C-60), 161.63 (C-40), 115.20 (C-30 thiab C{ {149}}); qabzib moiety: 99.13 (C{152}}), 73.85 (C-200), 76.88 (C-300), 70.35 (C{161}}), 76.96 (C{164}) } ), 62.25 (C-600 ); 3, 4-dihydroxy phenyl acetic acid moiety: 127.87 (C-1000), 116.35 (C-2000), 145.33 (C-3000 ), 148.92 (C{{181}) } ), 121.57 (C{184}} ), 117.0 (C-6000 ), 176.26 (C=O), 40.02 (C-H2).

2.8. In-Vitro Studies

2.8.1 ib. Kev ntsuam xyuas ntawm Cytotoxicity hauv tib neeg Keratinocytes

Tib neeg keratinocytes cell kab tau txais los ntawm VACSERA CO. (Lub Tuam Txhab Egyptian rau Kev Tsim Tshuaj Tiv Thaiv, Sera, thiab Tshuaj, Giza, Egypt). Qhov kev ntsuam xyuas MTT tau ua tiav rau PPEE thiab PPEE-SLNs raws li txoj kev Mostafa li al. [36].

2.8.2 ib. In-Vitro Antioxidant Assay

Qhov kev ntsuam xyuas DPPH tau ua los ntawm kev siv Yardpiroon li al. [39], thiab Vitamin C tau siv los ua kev tswj xyuas zoo. Qhov kev ntsuam xyuas ABTS tau ua raws li txoj kev ntawm Re et al., [40], Vitamin C tau siv los ua tus qauv. Kev ntsuam xyuas -carotene bleaching siv cov qauv piav qhia los ntawm Soulef li al., [14] nyob rau hauv lub xub ntiag ntawm kev tswj zoo BHT. Tus nqi rau txhua tus qauv kuaj tau nthuav tawm raws li qhov cuam tshuam qhov nkhaus ntawm 50 feem pua lossis IC50.

2.8.3 ib. Kev kwv yees ntawm Anti-Elastase, Anti-Collagenase, thiab Anti-Tyrosinase Activities

Kev tiv thaiv kev laus thiab kev ua kom tawv nqaij dawb tau soj ntsuam raws li cov txheej txheem qhia los ntawm Mostafa li al. [41]. Rau cov tshuaj tiv thaiv elastase, 1 µg / mL ntawm tib neeg leukocyte elastase tau tsim nrog HEPES tsis pH 7.5 thiab txhua qhov kev sim tshuaj los yog 1.4 mg / mL N-Methoxysuccinyl-Ala-Ala-Pro-chloromethyl ketone (tus qauv inhibitor) hauv a 96-zoo phaj rau 20 min ntawm chav tsev kub ua ntej qhov sib ntxiv ntawm 100 µL ntawm 1 mM N-Methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide ua substrate. Tom qab 40 min incubation, lub absorbance yog ntsuas ntawm 405 nm nrog ib tug zoo devoid ntawm extract/inhibitor pab raws li ib tug dawb paug. Cov qauv kuaj tau siv nyob rau hauv qhov ntau ntawm 25-300 µg / mL.

Rau anti-collagenase, 1 mg / mL ntawm collagenase hom 1 los ntawm Clostridium histolyticum, nws qhov tsis zoo (pH 7.4), thiab txhua qhov kev sim tshuaj los yog 1.4 mg / mL EDTA (tus qauv inhibitor) tau ua rau 20 min ntawm 37 ◦C ua ntej. ntxiv 100 µL ntawm FALGPA ua cov substrate. Qhov sib tov yog incubated rau lwm 1 h ntawm 37 ◦C ua ntej ntxiv 200 µL 2 feem pua ninhydrin nyob rau hauv 200 mM citrate tsis (pH 5) thiab muab tso rau hauv ib lub boiling dej da dej rau 5 feeb. Cov tshuaj tau tso cai rau txias. 200 µL ntawm 50 feem pua isopropanol tau ntxiv thiab qhov nqus tau raug ntsuas ntawm 540 nm. Cov qauv kuaj tau siv nyob rau hauv qhov ntau ntawm 25-300 µg / mL.

Rau cov tshuaj tiv thaiv tyrosinase, 5600 units / mL ntawm nceb tyrosinase yog incubated nrog txhua yam ntawm cov tshuaj rho los yog 1.4 mg / mL kojic acid (tus qauv inhibitor) rau 15 min ntawm 37 ◦C ua ntej ntxiv 1 mM L-DOPA ua substrate. . Qhov nqus ntawm cov dopachrome tsim tau ntsuas ntawm 475 nm. Cov qauv kuaj tau siv nyob rau hauv qhov ntau ntawm 75-500 μg / mL.

2.9. Formulation and Characterization of PPEE-SLNs

Lub formulation tau ua raws li txoj kev ntawm Choubey li al. [11]. Ua ntej, glyceryl monostearate tau yaj hauv chloroform-methanol sib tov (1: 1) thiab tom qab ntawd cov ethanolic extract tau tawg rau hauv cov tshuaj ntawd. Tom qab ntawd cov organic solvents raug tshem tawm siv rotovap thiab cov tshuaj tau rhuab. Tween 80 ces ntxiv thiab cov sib tov yog stirred ntawm 3000 rpm rau 30 min thiab ces homogenized rau 4 h ua ntej lim thiab ziab. Tus cwj pwm ntawm PPEE-SLNs tau tshuaj xyuas los ntawm kev kwv yees cov duab thiab qhov chaw morphology, particle loj, polydispersity index (PDI), zeta muaj peev xwm tsom xam, feem pua entrapment efficiency (PEE), thiab Fourier transform infrared (FT-IR) spectroscopy kev tshawb fawb.

2.10. Kev npaj ntawm PPEE Topical Cream Formula

2.10.1. Formulation of PPEE-SLNs Cream

Ob hom tshuaj nplaum (2 feem pua & 5 feem pua ntawm PPEE-SLNs) tau npaj raws li qhia hauv Table 1 raws li txoj kev ntawm Mahawar li al. [42].

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2.10.2. Kev soj ntsuam Parameters ntawm PPEE-SLNs Cream

Organoleptic zog, pH ntawm cov qab zib, kev tshawb fawb txog kev sib kis, kev ntsuas viscosity, kuaj homogeneity, patch test (Burchard test), in-vitro skin permeation exam, and stability studies (ICH guide) tau soj ntsuam raws li Matangi li al. [43]. Kev kuaj microbial txwv tau ua raws li Sekar et al. [44].

2.11. In-Vivo Anti-Wrinkle Study of PPEE-SLNs Cream

2.11.1. Kev sim tsiaj

Tsib caug txiv neej plaub hau tsis muaj nas (HR{{0}}}) (4-lub lim tiam), 17–24 g tau los ntawm VACSERA. Qhov kev sim tau ua tiav tom qab kev pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Lub Kaum Hli University rau Kev Tshawb Fawb Txog Niaj Hnub Nyoog thiab Kev Ua Yeeb Yam (MSA), Tus Qauv Zauv (PG1/EC1/2020PD) tus qauv loj tau txiav txim siab raws li (G. Power) software. Ib lub lis piam ua ntej qhov kev tshawb fawb hauv vivo los tiv thaiv wrinkle, nas tau nkag mus rau zaub mov, thiab dej thiab ua kom haum rau hauv chav cua txias (23 ± 2 ◦C). Cov nas (50) tau muab faib ua tsib pawg, random (n=10) raws li qhia hauv Table 2. Kev kho mob tau ua tiav 40 hnub, peb zaug ib lub lim tiam, nrog 0.5 g ntawm PPEE-SLNs qab zib los yog lwm yam kev kho mob thiab siv. mus rau dorsal nas daim tawv nqaij thiab tom qab raug UV (365 nm).

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2.11.2. Wrinkle Score Measurement

Ua ntej thiab tom qab kho cov tshuaj pleev ib ce, ob zaug / lub lis piam, cov duab ntawm daim tawv nqaij dorsal nas raug coj. Kev ntsuas ntawm daim tawv nqaij wrinkle tau soj ntsuam siv Bissett qhov muag pom qhov muag [45]. Lub hauv paus rau qhov ntsuas qhov ntsuas yog qhov kev ntxhib los mos ntawm wrinkle (zoo los yog ntxhib) thiab keratosis, muab faib ua tsib qib: zuj zus (−2), me ntsis zuj zus (−1), tsis hloov (0), me ntsis txhim kho ( ntxiv 1), thiab txhim kho (ntxiv rau 2).

2.11.3. Kev ntsuam xyuas Histological

Kev ntsuam xyuas histological tau ua raws li txoj kev ntawm Txwj Laug li al. [46].

2.11.4 ib. Superoxide Dismutase (SOD) Kev Ua Haujlwm

SOD kev ua haujlwm tau kwv yees raws li tus qauv ntawm Ukeda li al. [47].

2.12. Kev txheeb cais

Tag nrho cov qauv tau txheeb xyuas hauv triplicates. Cov ntaub ntawv tau sawv cev raws li txhais tau tias ± tus qauv sib txawv (SD). IC50 qhov tseem ceeb (kev xav tau rau 50 feem pua inhibition) raug xam los ntawm kev ua cov kab rov tav rov qab nkhaus uas qhia cov qauv concentrations thiab feem pua inhibition. Kev sib piv kev txheeb cais tau ua tiav siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua tiav los ntawm Tukey qhov kev sim rau ntau qhov sib piv. Cov txiaj ntsig sib txawv ntawm kev ua haujlwm antioxidant thiab kev soj ntsuam enzyme inhibition tau muab piv nrog Pearson correlation test thiab qhov sib txawv tau suav tias yog qhov tseem ceeb ntawm p <0.05. GraphPad Prism 5.03 rau Windows (GraphPad Software, San Diego, CA, USA) tau siv los ua cov kev tshuaj ntsuam no.

3. Cov txiaj ntsig

3.1. Tag nrho cov ntsiab lus Phenolic thiab Flavonoid

TPC tau txiav txim siab spectrophotometrically hauv PPEE li 387.5 ± 4.28 mg GAE / g extract. Tsis tas li, TFC tau kwv yees li 241.7 ± 3.25 mg QE / g extract. PPEE tau pom tias muaj ntau cov phenolic thiab flavonoid compounds. Ob qhov txiaj ntsig tau ua raws li cov txiaj ntsig yav dhau los tau tshaj tawm rau PP nplooj extracts lees paub tias flavonoids yog cov tshuaj tseem ceeb hauv PP nplooj [20].

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3.2. Isolation thiab Structure Elucidation ntawm kaempferol

Tom qab txheeb xyuas tag nrho cov txheej txheem paub hauv cov extract, kev tshawb fawb tshwj xeeb, muaj peev xwm ua kom muaj roj ntsha muaj txiaj ntsig zoo dua. Raws li tau txais 2-DPC cov ntaub ntawv txheeb xyuas cov phenolics ntawm PPEE tau pom tias yog qhov zoo tshaj plaws fractionated tshaj MCI gel kem eluted los ntawm MeOH / H2O cov khoom sib xyaw ntawm cov polarities txo qis, ib txheej txheem uas tau them plaub feem loj ntawm kab. Cov khoom siv amorphous raug cais tawm ntawm qhov kub siab ntawm feem III (desorbed los ntawm kem los ntawm 70 feem pua aqueous MeOH) ntawm kev sawv hmo ntuj ntawm chav tsev kub. Crystallization ntawm cov khoom no ob zaug los ntawm boiling EtOH yielded ib tug chromatographically ntshiab xim av-daj amorphous hmoov 1. Nws pom chromatographic zog (tsaus liab doog-ntawm PC nyob rau hauv UV teeb, tig txiv qaub daj thaum fumed nrog ammonia vapor nrog nruab nrab migration nyob rau hauv aqueous thiab organic solvents. ) thiab cov tshuaj tiv thaiv xim (ib xim txiv qaub-daj nrog Naturstoff reagent), uas tau pom tias muaj kaempferol derivative bearing ib pawg dawb 40 hydroxyl thiab O-substituted 3-O-txoj hauj lwm. Lub UV spectra ntawm 1 hauv MeOH (268 nm, 316 nm, 360 nm) thiab tom qab sib ntxiv ntawm cov tshuaj ntsuam xyuas hloov reagents [48] yog ib yam ntawm cov ntawm 3-O-glycosylated kaempferol (zoo ua haujlwm nrog NaOAc, ua haujlwm ruaj khov nrog NaOMe). Ib txwm acid hydrolysis ntawm 1 (2 N aqueous HCl, 3 h, 100 ◦C) yielded qabzib (piv cov ntawv chromatography, Co-PC), kaempferol, thiab 3,4-dihydroxy phenylacetic acid (UV absorption thiab 1H-NMR spectra, rau ib tus neeg kuaj, cais los ntawm kev npaj daim ntawv chromatograms ntawm ethyl acetate extract ntawm hydrolysate). Yog li ntawd, 1 yog kaempferol 3-O-(3,4-dihydroxy phenyl acetyl glucoside). Qhov sib xyaw ua ke tau zoo tsis hloov pauv tom qab tau tsim nrog -glucosidase enzyme rau 24 teev, yog li ua pov thawj tias glucosyl moiety yog acylated. Cov mis mos molecular ntawm 1 tau txiav txim siab yog C29H26O14 los ntawm nws qhov tsis zoo HRESI huab hwm coj spectrum uas pom ib [MH]− ion ntawm m/z=597.5302 (calc. rau C29H25O14, 597.5015). Qhov kev sim ESI-MS (qhov tsis zoo-ion hom) tau muab quasi-molecular ion ncov [MH] - ntawm m / z: 597 qhia qhov hnyav molecular ntawm 598 rau 1. Ntxiv fragment ion peaks hauv ESI-MS-MS spectrum yog pom ntawm m / z: [MH] −: 447, 167, thiab 285 sib raug rau qhov poob ntawm dihydroxy phenyl acetate los ntawm niam txiv compound 1 thaum cov fragment ions ntawm m / z 285 thiab m / z 167 raug ntaus nqi rau kaempferol thiab dihydroxyphenylacetic acid moieties, feem. Txhawm rau txiav txim siab qhov chaw ntawm qhov txuas ntawm txhua qhov moieties hauv molecule ntawm 1 thiab tso cai rau tag nrho cov haujlwm ntawm cov pa roj carbon thiab proton resonances, NMR spectroscopic tsom xam ntawm 1, suav nrog 1D- 1H thiab 13C, thiab 2D- HSQC thiab HMBC , tau ua tiav. Los ntawm tsib lub cim hauv thaj av qab zib ntawm δ ppm 62.25 thiab 76.9 thiab los ntawm cov teeb liab anomeric carbon nyob ntawm 99.13 ppm, nws tau ua pov thawj tias cov piam thaj moiety yuav tsum tau txuas rau txoj hauj lwm 3 ntawm kaempferol vim tias qhov C-3 carbon teeb liab tau hloov chaw upfield thiab qhov sib thooj ortho thiab para-carbon signals tau hloov downfield (saib kev sim). Cov kev hloov pauv zoo sib xws yog paub zoo los ntawm kev ua haujlwm ntawm Nawwar li al. [48]. Qhov no tau lees paub ntxiv los ntawm 3 J qhov kev sib raug ntev ntev tau lees paub hauv HMBC spectrum, uas yog, ib qho teeb meem hla tau pom muaj kev cuam tshuam ntawm cov piam thaj hauv ntshav proton H-100 teeb liab ntawm δ 5.42 rau flavonol C-3 carbon. teeb liab ntawm δ 133.5. Cov-configuration ntawm cov piam thaj moiety tau muab los ntawm C-l00 tshuaj hloov ntawm 99.13 ppm. Cov tshuaj hloov pauv qhov tseem ceeb ntawm cov piam thaj carbons tau lees paub daim ntawv pyranose ntawm no moiety [48]. 1H-NMR spectrum ntawm 1 kuj yog los ntawm cov qauv tsim. Kev hloov tshuaj lom neeg ntawm cov teeb liab anomeric proton ntawm δ 5.42 ppm (d, J=8 Hz) qhia tias cov pa roj carbon monoxide txuas mus rau kaempferol moiety ntawm C -3 (δppm 133.5) thiab txiav txim siab sib txuas tas li. 8 Hz, ua pov thawj -configuration ntawm cov piam thaj moiety. Lub conformation ntawm cov piam thaj moiety yog 4C1 raws li ua raws li cov-configurations tham saum toj no.

Tsis tas li ntawd, txuas ntawm 3, 4-dihydroxy phenylacetic acid moiety rau C-600 methylene glucopyranose moiety ua raws li los ntawm downfield hloov ntawm lub teeb liab ntawm cov pa roj carbon no rau δ ppm 62.25 hauv 13C-NMR spectrum. Qhov no tau lees paub ntxiv los ntawm tus ntoo khaub lig ncov nyob rau hauv HMBC spectrum, correlating methylenic qabzib protons signals ntawm δ 4.04 (H-600 a) thiab δ 4.2 ppm (H-600 b) rau carbonyl carbon ntawm 3, 4-dihroxyphenylacetic acid moiety ntawm δ 176.26 ppm. Cov no thiab cov ntaub ntawv sau saum toj no thaum kawg tau lees paub cov qauv ntawm cov khoom sib xyaw 1 los ua cov kaempferol tshiab 3-O- - 4C1-(600 -O-3,{ {24}}dihydroxyphenylacetyl glucopyranoside) (KDPAG), tshaj tawm thawj zaug hauv qhov xwm txheej, raws li nws sawv cev rau thawj acylation nrog 3,4-hydroxyphenyl acetic acid nyob rau hauv koom nrog flavonoid chemistry (Daim duab 1).

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