Los tiv thaiv kev laus ntawm Piper Cambodianum Cistanche Extract Ntawm Tib Neeg Dermal Fibroblast Cells Thiab Ib Tus Qauv Kho Mob hauv nas Ⅱ
Apr 27, 2023
Cistancheextract tiv thaiv NHDF hlwb los ntawm cov teebmeem ntawm UV irradiation
Cov hlwb ntawm daim tawv nqaij teb sai sai thaum raug UV lossis cytokines xws li interleukin (IL)-4 thiab IL-13.19Thaum raug UV, ntau yam tseem ceeb tau txhawb nqa, suav nrog EGFR,20,21 TNFR,22mitogen-activated protein (MAP) kinases, ERK, thiab c-Jun amino-terminal kinase (JNK).23Ob leeg JNK thiab p38 phosphorylate thiab qhib qhov kev hloov pauv hloov pauv c-Jun, uas, dhau los, nce c-Fos thiab nce qib ntawm cov txiaj ntsig transcription activator protein-1,24uas yuav tsum tau rau kev sau ntawv ntawm MMPs. Cov receptors no tau qhib los ntawm phosphorylation ntawm MAP kinase thiab lwm cov proteins hauv qab.25MAP kinases yog qhib los ntawm phosphorylation ntawm threonine thiab tyrosine residues.24,26UV raug ua rau muaj kev qhia siab ntawm c-Jun thiab c-Fos heterodimers, ntau cytokines, thiab kev loj hlob yam, uas yog phosphorylated los ntawm JNK thiab p38.27,28Ib yam li ntawd, p53 protein ntau tau nce siab, pib ntawm 2.5 teev mus rau 24 teev ua raws li kev raug UV.29Peb pom tau hais tias cov hlwb raug UV irradiation rau 24 teev qhia TNFR1 thiab EGFR protein ntau ntau, uas txo qis nyob rau hauv ib tug npaum li cas ntawm cov tshuaj thaum lub hlwb raug kho nrog.Cistanche. Peb tau tshuaj xyuas qhov cuam tshuam ntawm UV hluav taws xob ntawm ob qho tib si phosphorylation thiab kev ua haujlwm ntawm p38 (Thr180/Tyr182). Protein theem ntawm c-Jun thiab p53 tau downregulated heev nyob rau hauvCistanche-kho cov hlwb (Daim duab 4A thiab B).
Nyem qhov no kom tau txais cov ntaub ntawv ntau ntxiv txog Cistanche Rau Kev Tiv Thaiv Kev Laus
Cistanche extract tiv thaiv UV-irradiated NHDF hlwb los ntawm ROS
UVB teeb txhawb kev tsim cov ROS,30,31thiab UV raug rau ntawm daim tawv nqaij ua rau ROS xws li O2 ⋅−, H2 O2, thiab1 O2. 32Tsis kho thiabCistanche-kho cov hlwb raug UV irradiation rau 24 teev, thiab tiam ntawm ROS yog ntsuas los ntawm fluorescence-activated cell sorting (FACS).
Daim duab 3Kev tswj hwm ntawm ECM proteins hauvCistanche- kho cov hlwb.Nco tseg: (A) NHDF hlwb raug kho nrogCistancherau 24 teev. Cell lysates tau soj ntsuam los ntawm immunoblotting siv los tiv thaiv collagen, anti-elastin, anti-MMP-3, thiab anti-ERK-2 antibodies. Cov txiaj ntsig yog tus sawv cev ntawm peb qhov kev sim ywj pheej. (B) Actin, collagen, elastin, MMP-3, thiab ERK-2 hauv cov immunoprecipitates tau ntsuas los ntawm Western blotting. Bar qhov siab yog qhov sib piv± SDs ntawm peb qhov kev sim ywj pheej. *P,0.05; **P,0.01. Cov ntawv luv:Concentration; ECM, extracellular matrix; ERK, extracellular teeb liab tswj kinase; MMP, matrix metalloproteinase; NHDF, tib neeg dermal fibroblast;Cistanche, Piper cambodianumP. Plaub.; SDs, standard deviations.
Daim duab 4Cistanche extract inhibition ntawm ROS ntau lawm hauv UV-irradiated hlwb.Nco tseg: (A) NHDF hlwb raug UV irradiation ntawm 40 J rau 180 vib nas this thiab kho nrogCistancherau 24 teev. Cell lysates tau soj ntsuam los ntawm immunoblotting siv anti-TNFR6, anti-c-Jun, anti-c-Fos, anti-pp38, los yog anti-p-JNK tshuaj tiv thaiv. Cov txiaj ntsig yog tus sawv cev ntawm peb qhov kev sim ywj pheej. (B) Tag nrho thiab tyrosine-phosphorylated p38, JUK, actin, thiab TNFR6 hauv immunoprecipitates tau ntsuas los ntawm Western kev tshuaj ntsuam. Bar qhov siab yog qhov nruab nrab± SD ntawm peb qhov kev sim ywj pheej. *P,0.05; **P,0.01. Cov ntawv luv:Concentration; EGFR, epidermal growth factor receptor; JNK, c-Jun amino-terminal kinase; JNK, c-Jun N-terminal kinase; NHDF, tib neeg dermal fibroblast;Cistanche, Piper cambodianumP. Plaub.; ROS, reactive oxygen hom; SD, tus qauv sib txawv; TNFR, qog necrosis factor receptor; UV, ultraviolet.
UV irradiation nce ROS ntau lawm, thaumCistanche extract txo ROS ntau lawm hauv NHDF hlwb. ROS theem H2 O2 tau ntsuas siv cov cell permeants 2′,7′-dichlorodihydrofluorescein diacetate (CM-H2 DCFDA) (Daim duab 5A). UV irradiation nce ROS qib H2 O2 , whereas ROS theem raug txo los ntawmCistanche extract nyob rau hauv ib koob tshuaj raws li (Daim duab 5B). Cov txiaj ntsig no qhia tau tiasCistancheextract tiv thaiv dermal fibroblasts los ntawm UV irradiation los ntawm txo ROS tiam.
Macroscopic soj ntsuam ntawm qhov txhab kho hauv nas
Cov kab mob tau tsim nyob rau sab nraub qaum ntawm nas thiab tom qab ntawd kho nrogCistancherho tawm (0, 1, 10, thiab 50µg / mL); cov cheeb tsam qhov txhab raug ntsuas txhua hnub rau 4 hnub. Hnub 4, cov cheeb tsam ib puag ncig cov qhov txhab nyob hauvCistanche-kho nas pom qhov txo qis liab nyob rau hauv ib koob tshuaj raws li piv rau qhov chaw mob uas tsis kho (Daim duab 5). Cov cheeb tsam uas tau kho qhov txhab tau me dua qhov chaw tsis kho qhov txhab, uas qhia txog cov qauv ntawm koob tshuaj. Hnub 4, thaj chaw qhov txhab kho nrog 50µg / ml ntawmCistanchetau tag nrho los ntawm tshiab epithelium. Tsis muaj qhov txhab raug mob (Daim duab 6A thiab B). Qhov sib piv ntawm thaj chaw qhov txhab nyob rau hnub 0 thiab 4 raug xam (Daim duab 6C).
Kev sib tham
Kev siv cov khoom siv tshuaj ntsuab thiab cov nroj tsuag sib txawv tau nce ntxiv nrog kev nce qib hauv phytotherapy. Cov ntaub ntawv tshuaj ntsuab tau siv ntau yam xws li tshuaj mono- lossis polyherbal, cov khoom noj khoom haus, thiab cov khoom noj khoom haus, thiab ntau yam tau dhau los ua cov khoom lag luam zoo thiab muaj kev nyab xeeb.33Ntau yam nroj tsuag, suav nrogCistanche, tau raug tshawb xyuas rau lawv cov khoom muaj peev xwm pharmacological, tab sis tsis muaj kev tshawb fawb sib piv tau ua yav dhau los nrogCistanche rho tawm. Txoj kev tshawb no tau ua los txiav txim siabcov teebmeem ntawmCistanche extract ntawm qhov txhab-kho cov dej num siv incision qhov txhab qauv hauv nas. Kev kho qhov txhab cuam tshuam nrog kev rov tsim kho cov ntaub so ntswg ncaj ncees los ntawm cov txheej txheem cuam tshuam nrog kev mob, kev loj hlob, thiab kev hloov kho theem.34 Cov txheej txheem kho ntawm daim tawv nqaij yog tus cwj pwm los ntawm kev hloov kho thiab hloov kho ntawm collagen fiber ntau uas txhim kho tensile lub zog.35Cov txheej txheem kho yog tswj feem ntau los ntawm biosynthesis thiab deposition ntawm collagen tshiab thiab tom qab maturation.36Hauv peb qhov kev tshawb nrhiav qhov txhab,Cistanche extract tau pom muaj qhov txhab-kho qhov txhab zoo dua piv nrog kev tswj hwm (Daim duab 1C thiab 6). Cov kev kho zoo li no yuav yog vim qhov nce collagen concentration thiab stabilization ntawm fibers. Nyob rau hauv cov txheej txheem kho cov ntaub so ntswg, inflammatory hlwb txhim khu kev tsiv teb tsaws thiab proliferation ntawm endothelial hlwb, ua rau lub neovascularization ntawm connective cov ntaub so ntswg cell uas synthes extracellular matrices, xws li collagen thiab keratinocytes, ua rau reepithelialization ntawm cov ntaub so ntswg.37Qhov nrawm nrawm nrawm tuaj yeem ua rau muaj qhov sib txawv ntawm kev sib txuas lus hauv lub cev hauv fibroblasts thiab txhim kho qhov kev loj hlob sai ntawm cov ntaub so ntswg granulation.38Nyob rau hauv tus qauv incision qhov txhab, nas kho nrogCistancheextract tau pom qhov txhab kho sai dua piv nrog kev tswj hwm (Daim duab 6). Ntau cov kev tshawb fawb tau qhia tias kev txo qis ntawm p38 kinase txoj kev tiv thaiv ntau yam tsiaj qus p53- qhia cov kab ntawm tes los ntawm kev ntxhov siab genotoxic.39,40Peb cov ntaub ntawv qhia tau hais tias p38 qhia nyob rau hauv UV-irradiated hlwb raug downregulated los ntawmCistanche kev kho mob. Yog li ntawd,Cistanche extract tau los ntawm nplooj thiab stems qhia tau zoo kev kho mob nyob rau hauv lub qhov txhab kho.Cistancheextract txo ROS ntau lawm los ntawm UV irradiation los ntawm scavenging ROS nyob rau hauv NHDF hlwb (Figures 5A thiab B). Ntxiv mus,Cistancheextract ceev qhov txhab kho tom qab daim tawv nqaij irradiation ((Daim duab 6A-C).Cistanche extract tiv thaiv daim tawv nqaij los ntawm UV irradiation puas, thiab tej zaum yuav pab tau raws li ib tug tshiab raw khoom nyob rau hauv daim ntawv thov kom hniav zoo nkauj.
Daim duab 5Cistanche-mediated inhibition ntawm ROS ntau lawm hauv UV-irradiated hlwb.Nco tseg: (A) Cells raug kho nrog UVB irradiation (40 J) rau lub sij hawm qhia thiab ces incubated nrog 2′,7′-dichlorodihydrofluorescein diacetate rau 30 feeb. Lub cim ntawm ROS tau ntsuas los ntawm FACS. (B) Cells raug UVB (40 J) rau 180 vib nas this thiab kho nrog qhov qhia pom ntawmCistancherau 24 teev. ROS ntau lawm tau kwv yees nrog staining ntawm 2′,7′-dichlorodihydrofluorescein diacetate thiab tshuaj xyuas los ntawm FACS.Cov ntawv luv:FACS, fluorescence-activated cell sorting; FITC, fluorescein isothiocyanate; NHDF, tib neeg dermal fibroblast;Cistanche, Piper cambodianumP. Plaub.; ROS, reactive oxygen hom; UV, ultraviolet; UVB, ultraviolet B.
Daim duab 6Kev kho mob rau ntawm daim tawv nqaij tsis zoo nas.Nco tseg: (A) Cov kab mob ntawm 5- hli txoj kab uas hla tau tsim rau ntawm daim tawv nqaij dorsal ntawm 7-cov txiv neej hnub nyoog C57Bl/6 nas (n=3). Kev kho mob ntawm qhov txhab raug tshuaj xyuas nrogCistanchekev kho mob hauv qhov sib txawv ntawm qhov sib txawv (kev tswj tsis zoo thiab tom qab kev kho mob nrogCistanchentawm 1, 10, thiab 50µg / mL). (B) Qhov chaw qhov txhab tau txiav txim siab los ntawm kev ntsuas lub cheeb ntawm cov qhov txhab qhib. (C) Qhov chaw qhov txhab raug suav nrog qhov dav ntawm qhov qhib qhov txhab; *P,0.05; **P,0.01. Cov ntawv luv:Cistanche, Piper cambodianumP. Plaub; Concentration, ua.
Xaus
Qhov kev tshawb nrhiav no tau tshawb nrhiavmuaj peev xwm tiv thaiv kev laus thiab qhov txhab-kho los ntawm Cistanche qia thiab nplooj extracthauv NHDF hlwbthiab tus nas qauv ntawm qhov txhab kho. CovKev kho ntawm NHDF hlwb nrog Cistanche rho tawmtxhawb kev rov qab los ntawm qhov chaw khawbnyob rau hauv ib koob tshuaj raws li kev txhim kho ntawm ECM gene qhia. Tsis tas li ntawd, Cistancherho tawmmuaj peev xwmtiv thaiv lub cell los ntawm UV raug los ntawm kev txo ROS ntau lawm. Thaum kawg, cov kev soj ntsuam no tau soj ntsuam ntxiv thiab lees paub hauv cov nas qauv rau kev kho mob hauv vivo. Yog li ntawd,Cistanche extract sawv cev rau ib qho kev cog lus tshiab kho tus neeg sawv cev rau kev tiv thaiv kev laus thiab kho qhov txhab.
Kev lees paub:Txoj kev tshawb no tau txais kev txhawb nqa los ntawm Kev Tshawb Fawb Kev Tshawb Fawb Txog Kev Tshawb Fawb los ntawm National Research Foundation of Kauslim (NRF), tau nyiaj los ntawm Ministry of Education (NRF-2014R1A1A3050752). Cov kws sau ntawv xav ua tsaug rau Kauslim Kev Tshawb Fawb Lub Tsev Haujlwm ntawm Bioscience & Biotechnology (KRIBB) rau kev muab rauCistancherho tawm.
Qhia tawm Cov neeg sau ntawv tshaj tawm tsis muaj qhov tsis sib haum xeeb ntawm kev txaus siab hauv txoj haujlwm no.
Cov ntaub ntawv
1. Slominski AT, Zmijewski MA, Semak I, et al. Cytochromes p450 thiab mob qog noj ntshav ntawm daim tawv nqaij: lub luag haujlwm ntawm txoj hauv kev endocrine hauv zos.Anticancer Agents Med Chem. 2014;14(1):77–96.
2. Gilchrest IB. Daim tawv nqaij laus thiab photoaging.Dermatol Nurs. 1990; 2(2):79–82.
3. Ha TY. Kev tsim cov khoom noj khoom haus ua haujlwm rau lub neej noj qab haus huv.Korean J Crop Sci. 2006;51:26–39.
4. Makrantonaki E, Zouboulis CC. Molecular mechanisms ntawm daim tawv nqaij laus: lub xeev ntawm daim duab.Ann NY Acad Sci. 2007;1119:40–50.
5. Seo MY, Chung SY, Choi WK, et al. Anti-aging nyhuv ntawm nplej wine nyob rau hauv cultured human fibroblasts thiab keratinocytes.J Biosci Bioeng. 2009; 107(3):266–271.
6. Kligman AM. Kev puas tsuaj thaum ntxov ntawm lub hnub ci ntawm tib neeg daim tawv nqaij.JAMA. 1969;210(13):2377–2380.
7. Oikarinen A, Karvonen J, Uitto J, Hannuksela M. Connective cov ntaub so ntswg alterations nyob rau hauv daim tawv nqaij raug rau tej yam ntuj tso thiab kho UV-kua hluav taws xob.Photodermatol. 1985;2(1):15–26.
8. Bech-Thomsen N, Wulf HC. Carcinogenic thiab melanogenic cuam tshuam ntawm cov lim hlau halide UVA qhov chaw thiab tubular fluorescent UVA tanning qhov chaw nrog lossis tsis muaj hnub ci-simulated UV hluav taws xob ntxiv hauv cov nas tsis muaj plaub hau.Fotochem Cov. 1995;62(4):773–779.
9. De MS, Lahiri SC. Kev tshawb fawb ntawm nplooj ntawm Piper betle.Bull Calcutta Sch Trop Med. 1971;19(3):69–72.
10. Ferreres F, Oliveira AP, Gil-Izquierdo A, Valentão P, Andrade PB. Piper betle nplooj: profiling phenolic tebchaw los ntawm HPLC/DAD-ESI/ MS(n) thiab kev tiv thaiv cholinesterase.Phytochem Qhov Ncauj. 2014;25(5): 453–460.
