Neuroprotective Effects Ntawm Plaub Phenylethanoid Glycosides Ntawm H2O2-Induced Apoptosis Ntawm PC12 Cells Ntawm Nrf2/ARE Pathway
Mar 04, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
Maiquan Li 1, Tao Xu 1, Fei Zhou 1, Mengmeng Wang 1, Huaxin Song 1, Xing Xiao thiab Baiyi Lu 1,*
1. Taw qhia
Oxidative kev nyuaj siab, uas yog qhov tsis txaus ntawm cov tshuaj tiv thaiv antioxidant homeostasis, induces lipid peroxidation, raug mob rau cov protein thiab DNA, cell aging, thiab cell tuag. Cov txheej txheem no yuav ua rau muaj ntau yam kab mob neurodegenerative, xws li Alzheimer's disease (AD), Parkinson's disease (PD), thiab ischemia / reperfusion [1]. Hydrogen peroxide (H2O2), uas yog ib qho tseem ceeb ntawm cov pa oxygen reactive (ROS), paub tias ua rau lipid peroxidation thiab DNA puas [2]. Ntxiv mus, H2O2 yog ib qho chaw endogenous ntawm hydroxyl dawb radicals uas ua rau cov theem tom ntej ntawm cellular oxidative stress [3,4]. Yog li, cov tswv yim kho mob rau kev tiv thaiv oxidative kev ntxhov siab vim apoptosis tuaj yeem muaj peev xwm hauv kev kho mob neurodegenerative.
Nuclear factor erythroid 2-related factor 2 (Nrf2), yog ib qho kev hloov pauv uas cuam tshuam nrog kev ntxhov siab oxidative. Kev ua kom Nrf2 induces transcription ntawm ntau cov tshuaj antioxidant thiab detoxification noob, suav nrog heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1, anti-inflammatory [13], thiab immunomodulatory [14] ] bioactivities. Osmanthusfragrans yog ib qho khoom muaj nyob hauv ntau cov zaub mov Esxias thiab tau siv ntev. Peb yav dhau los tau pom tias O.fragrans paj rho tawm txhim kho kev kawm spatial thiab nco, inhibited oxidative puas tsuaj, thiab nthuav tawm cov kev tiv thaiv neuroprotective hauv d-galactose-induced aging hauv ICR nas qauv [15]. Salidroside, acteoside, thiab isoacteoside yog cov lus teb PhGs loj rau cov haujlwm antioxidant ntawm O.fragrans paj rho tawm [16].
Kev tshawb fawb txog cov nyhuv neuroprotective ntawm PhGs tau txais txiaj ntsig zoo. Salidroside txo qis cell apoptosis ntawm PC12 hlwb uas tau nthuav tawm hauv MPP ntxiv [17,18]. Acteoside kuj txo MPP ntxiv rau -induced apoptosis thiab oxidative stress nyob rau hauv PC12 hlwb [19] thiab A p25-35-induced SH-SY5Y cell raug mob [20]. Echinacoside tau tshawb xyuas ntawm qog necrosis factor-a (TNFa)-induced apoptosis hauv SH-SY5Y hlwb [21], MPTP-induced dopaminergic toxicity nyob rau hauv nas [22], glutamate-injured thawj kab lis kev cai ntawm nas cortical hlwb [23], thiab { {19}}OHDA-vim kev puas tsuaj hauv PC12 hlwb [24]. Cov txiaj ntsig tau qhia tias PhGs tau nthuav tawm cov txiaj ntsig cytoprotective thiab yog cov neeg ua haujlwm muaj peev xwm los kho cov kab mob neurodegenerative. Cov kev tshawb fawb tau pom cov khoom antioxidant ntawm PhGs hauv qab ntau lwm yam bioactivities rau cov tebchaw no [25]. Txawm li cas los xij, qee qhov kev tshawb fawb tau tshawb xyuas cov txheej txheem molecular ntawm PhGs tiv thaiv oxidative toxicity.
Hauv peb txoj kev tshawb fawb, peb xaiv plaub yam PhGs raws li hauv qab no: salidroside (phenylethanoid monosaccharides), acteoside (phenylethanoid disaccharides), isoacteoside (phenylethanoid disaccharides), thiab echinacoside (phenylethanoid trisaccharides). Peb siv tus qauv ntawm kev tuag neuronal siv cov PC12 sib txawv [26] los tshawb xyuas cov txiaj ntsig tiv thaiv thiab cov txheej txheem molecular ntawm PhGs ntawm H2O2-induced PC12 cell model. Peb tau pom tias PhGs tau qhib txoj hauv kev Nrf2/ARE los ntawm kev khi rau Kelch-zoo li ECH-koom nrog protein 1 (Keap1). Cov txheej txheem no txhawb nqa cov enzymes antioxidant thiab ua rau muaj zog ntawm PC12 hlwb rau oxidative kev nyuaj siab.
Kev kho cov kab mob neurodegenerative:PhGs los ntawm cistanche
2. Cov txiaj ntsig
2.1. PhGs Txhaum H2O2-Induced Cytotoxicity hauv PC12 Cells
Cytotoxic teebmeem ntawm H2O2 thiab PhGs (0.1,1,5, thiab 10 卩g/mL) ntawm PC12 hlwb raug kuaj. Cov txiaj ntsig tau pom tias H2O2 ua rau poob ntawm PC12 cell viability nyob rau hauv concentration-dependent thiab lub sij hawm-dependent yam ntxwv (Daim duab 1A). Kev nthuav tawm ntawm PC12 hlwb rau 200 H2O2 rau 2 h ua rau lub cev muaj zog ntawm 57.4 feem pua. Kev kho cov hlwb nrog PhGs ntawm 0.1,1, 5, thiab 10 昭/mL tsis muaj kev cuam tshuam rau cell viability (Daim duab 1B) thiab muaj kev tiv thaiv PC12 hlwb los ntawm H2O2-induced kev puas tsuaj los ntawm kev txhim kho cov cell viability as 9.549-22.141 feem pua , 12.092-25.289 feem pua , 1.470-9.289 feem pua , thiab 3.411-11.441 feem pua , raws li ( Daim duab 1C). Txawm li cas los xij, salidroside (0.1 ^ g / mL), isoacteoside (0.1, 1, 5, thiab 10 ^ g / mL), echinacoside (0.1,1, thiab 5 ^ g / mL) pretreatment tsis pom qhov txawv ntawm HzOz-induced cell. raug mob. Pretreatment ntawm hlwb nrog PhGs kuj ameliorated morphological yam ntxwv induced los ntawm H2O2 (Daim duab 1D).
Daim duab 1.PhGs suppressed H2O2-induced cytotoxicity hauv PC12 hlwb. Cell viability tau kuaj pom los ntawm MTT kev soj ntsuam. Cytotoxic nyhuv ntawm H2O2 (A) thiab PhGs (B) ntawm qhov sib txawv ntawm PC12 hlwb. (C) PhGs attenuated H2O2-vim txo qis hauv cell viability. PC12 hlwb raug incubated nrog PhGs (0.1, thiab 10 mg/mL) rau 24 teev, thiab ces incubated nrog 200 pM H2O2 rau lwm 2 h tom qab PhGs raug tshem tawm. (D) Kev soj ntsuam morphological. Cells tom qab kev kho mob tau pom los ntawm lub sij hawm-contrast microscope (x100), CK: ib txwm pab pawg neeg, H2O2: H2O2 kho pab pawg, HL: salidroside tsawg npaum li cas kho pab pawg, HH: salidroside siab ntau npaum kho pab pawg, ML: acteoside tsawg npaum li cas kho pab pawg, MH: acteoside high dosage treated group, IL: isoacteoside low dosage treated group, IH: isoacteoside high dosage treated group, SL: echinacoside low dosage treated group, SH: echinacoside siab npaum li cas kho pab pawg. ** p <0.01 piv="" rau="" pab="" pawg="" tsis="" kho;="" #="" p="">< 0.05,="" piv="" rau="" h2o2="" kho="" pab="" pawg;="" ##="" p=""><0.01, piv="" rau="" h2o2="" pab="" pawg="">
2.2. PhGs Ncua tseg H2O2-Induced Intracellular Accumulation ntawm ROS, Lipid Peroxidation (MDA), thiab nce Superoxide Dismutase (SOD) Kev Ua Haujlwm hauv PC12 Cells
Kev tawm ntawm PC12 hlwb rau 200 pM H2O2 rau 2 h nce ROS qib, MDA cov ntsiab lus, thiab txo qis kev ua SOD (Daim duab 2). PhGs pretreatment attenuated ROS theem, salidroside, thiab acteoside, thiab siab npaum li cas ntawm isoacteoside thiab echinacoside pretreatment ho attenuated ROS theem (p <0.01). salidroside="" pretreatment="" tsis="" cuam="" tshuam="" rau="" mda="" cov="" ntsiab="" lus,="" tab="" sis="" acteoside="" pretreatment="" txo="" qis="" mda="" cov="" ntsiab="" lus="" (p=""><0.05). isoacteoside="" thiab="" echinacoside="" pretreatment="" txo="" qis="" mda="" cov="" ntsiab="" lus="" rau="" ntau="">
Daim duab 2.PhGs thaiv ROS thiab MDA tsub zuj zuj thiab nce cov dej num ntawm SOD hauv PC12 hlwb. PC12 hlwb raug incubated nrog PhGs (0.1, thiab 10 4g/mL) rau 24 teev, thiab ces incubated nrog 200 |^M H2O2 rau lwm 2 teev tom qab PhGs raug tshem tawm. (A) PhGs thaiv ROS thiab MDA tsub zuj zuj. (B) PhGs thaiv MDA tsub zuj zuj. (C) PhGs tau nce cov haujlwm ntawm SOD. CK: pawg ib txwm, Qauv: H2O2 kho pab pawg, Salidroside: salidroside kho pab pawg, Acteoside: acteoside kho pawg, Isoacteoside: isoacteoside kho pawg, Echinacoside: echinacoside kho pawg. ** p < 0.01="" tiv="" thaiv="" pab="" pawg="" tsis="" kho;="" #="" p="">< 0.05,="" piv="" rau="" h2o2="" kho="" pab="" pawg,="" ##="" p="">< 0.01,="" piv="" rau="" h2o2="" kho="">
2.3.PhGs Reversed H2O2-Induced Apoptosis hauv PC12 Cells
Kev kho H2O2 (200 |1M) rau 2 h nce apoptosis hauv PC12 hlwb, nrog rau tag nrho apoptotic tus nqi txog li 16.02 feem pua (Daim duab 3). Txawm li cas los xij, pretreatment nrog PhGs (0.1 thiab 10 卩g / mL) rau 24 h txo tus nqi apoptosis nyob rau hauv ib tug concentration-dependent yam (p <0.01). salidroside,="" acteoside,="" isoacteoside,="" thiab="" echinacoside="" tau="" txo="" qis="" feem="" pua="" ntawm="" cov="" cell="" apoptosis="" los="" ntawm="" 4.="" 1.530-7.510="" feem="">
Daim duab 3. PhGs thim rov qab H2O2-induced apoptosis hauv PC12 hlwb. PC12 hlwb raug incubated nrog PhGs (0.1, thiab 10 卩g/mL) rau 24 teev, thiab ces incubated nrog 200 pM H2O2 rau lwm 2 teev tom qab lub PhGs raug tshem tawm. Tom qab ntawd, apoptosis tau ntsuas los ntawm kev ntws cytometry siv PI / FITC fluorescent sojntsuam. CK: pawg ib txwm, Qauv: H2O2 kho pab pawg, Salidroside: salidroside kho pab pawg, Acteoside: acteoside kho pawg, Isoacteoside: isoacteoside kho pawg, Echinacoside: echinacoside kho pawg. ** p <0.01 piv="" rau="" pab="" pawg="" tsis="" kho;="" ##="" p=""><0.01, piv="" rau="" h2o2="" pab="" pawg="">
2.4. PhGs Reversed H2O2-Induced Downregulation of Protein Expression of HO-1, NQO1, GCLC, and GCLM
HO{{0}}, NQO1, thiab glutamate-cysteine ligase (GCL) yog ib qho tseem ceeb cellular antioxidant enzymes, thiab HO-1, NQO1, thiab catalytic los yog hloov subunits ntawm GCL (GCLC los yog GCLM) yog Nrf2- tswj cov noob caj noob ces [27]. Protein expression of HO-1, NQO1, GCLC, thiab GCLM tau soj ntsuam tom qab kho. Qhov sib txawv pom tseeb tau pom ntawm cov protein qhia ntawm HO-1 thiab NQO1 nrog lossis tsis muaj H2O2 (Daim duab 6A-C) (p < 0.01).="" phgs="" (0.1="" thiab="" 10="" p^g/ml)="" thim="" rov="" qab="" h2o2-induced="" downregulation="" ntawm="" protein="" qhia="" ntawm="" ho-1="" (tshwj="" tsis="" yog="" salidroside="" ntawm="" 0="" }.1="" pg/ml),="" nqo1="" (tshwj="" tsis="" yog="" acteoside="" ntawm="" 0.1="" pg/ml)="" (p="">< {{40}}="" 01).="" h2o2="" kuj="" txo="" qis="" gclc="" thiab="" gclm="" protein="" qhia="" (p="">< 0.05)="" (daim="" duab="" 6a,d,e).="" phgs="" (0.1="" thiab="" 10="" pg/ml)="" thim="" rov="" qab="" h2o2-induced="" downregulation="" ntawm="" protein="" qhia="" ntawm="" gclc="" (tshwj="" tsis="" yog="" echinacoside="" ntawm="" 0.1="" pg/ml)="" (p=""><0.01) thiab="" gclm="" (tshwj="" tsis="" yog="" salidroside="" ntawm="" 0.1="" pg="" ml)="" (p=""><0.01). tom="" qab="" ntawd,="" cov="" tshuaj="" inhibitors="" rau="" ho-1="" tau="" siv="" los="" ntsuas="" ntxiv="" lub="" luag="" haujlwm="" ntawm="" cov="" enzymes="" antioxidant="" hauv="" kev="" tswj="" hwm="" kev="" tiv="" thaiv="" phgs="" tiv="" thaiv="" h2o2-induced="" cytotoxicity.="" phgs="" (0.1="" thiab="" 10="" pg/ml)="" tiv="" thaiv="" h2o2-induced="" cytotoxicity,="" tab="" sis="" cov="" kev="" tiv="" thaiv="" zoo="" li="" no="" tau="" thim="" rov="" qab="" los="" ntawm="" ho-1="" inhibitor="" znpp="" (p=""><0.01) ntawm="" 20="" pm="" (daim="" duab="" 6f,="" p="">< 0.01)="">
2.5. Keap1 Kev Qhia thiab Molecular Docking Analysis
Raws li lub cev muaj zog, Keap1 ua raws li cov protein repressor ntawm Nrf2 los ntawm kev khi rau Neh2 lub npe ntawm Nrf2 thiab tsom Nrf2 rau Cul3-raws li E3 ubiquitin ligase rau ubiquitination thiab tom qab degradation los ntawm 26S proteasome [28]. Kev khi lub peev xwm rau Keap1 ntawm PhGs tau soj ntsuam los ntawm kev soj ntsuam molecular docking los tshawb xyuas cov txheej txheem raws li lawv cov nyhuv antioxidant.
Antioxidant nyhuv: CistanchePhGs
3. Kev sib tham
Peb tau tshawb xyuas cov neuroprotection ntawm PhGs ntawm H2O2 induced-cytotoxicity hauv PC12 hlwb. Cov txiaj ntsig tau pom tias PhGs pretreatment cuam tshuam HzOz-induced cytotoxicity, attenuated intracellular ROS theem, txhim kho qib ntawm intracellular antioxidant enzymes, thiab thaum kawg thim rov qab H2O2-induced cytotoxicity hauv PC12 hlwb. Ntxiv mus, PhGs tau nce qhov kev hloov pauv ntawm Nrf2, thim rov qab HzOz-induced downregulation ntawm cov protein qhia ntawm HO-1, NQO1, GCLC, thiab GCLM. Tsis tas li ntawd, PhGs tau pom tias muaj peev xwm cuam tshuam nrog Nrf2 khi qhov chaw hauv Keap1 protein.
Hauv H2O2-induced PC12 cell raug mob, lipid peroxidation, uas yog hais txog oxidative degradation ntawm lipid, nce permeability ntawm daim nyias nyias, ua rau cell puas [29]. MDA tsim yog dav siv raws li qhov ntsuas ntawm lipid peroxidation [30]. H2O2 txhim kho ROS ntau lawm thiab tso tawm cov tshuaj tiv thaiv antioxidant, xws li SOD, catalase, thiab GPx. Cov txheej txheem no ua rau muaj kev ntxhov siab oxidative [31], uas ua lub luag haujlwm tseem ceeb hauv kev ua rau thiab kev loj hlob ntawm feem ntau ntawm cov kab mob neurodegenerative. Raws li cov kev tshawb fawb yav dhau los, peb tau pom qhov nce qib ntawm ROS, txo cov enzymes intracellular antioxidant, thiab txhim kho apoptosis ntawm PC12 hlwb tom qab kho H2O2. PhGs pretreatment ho txo qis HzOz-induced nce hauv intracellular ROS, txhim kho intracellular antioxidant enzymes, thiab thaum kawg thim rov qab HzOz-induced cytotoxicity hauv PC12 hlwb.
Kuang et al. [32] tau tshaj tawm tias echinacoside tau pom tias muaj txiaj ntsig zoo rau cov neuroprotective ntawm H2O2-induced cytotoxicity hauv PC12 hlwb los ntawm txoj kev mitochondrial apoptotic. Hauv txoj kev tshawb no, peb pom tias echinacoside, salidroside, acteoside, thiab isoacteoside tau pom cov teebmeem neuroprotective los ntawm kev txhim kho cov kev ua haujlwm antioxidant ntawm PC12 hlwb vim tias lawv tau nce cov kev ua haujlwm ntawm Nrf2 thiab txhawb nqa cov ntsiab lus qis qis ntawm HO-1, NQO1, GCLC, thiab GCLM. Ntau cov kev tshawb fawb tau pom tseeb tias kev ua kom Nrf2 lub hom phiaj, tshwj xeeb yog HO-1, hauv astrocytes thiab neurons tiv thaiv kev mob, oxidative puas tsuaj, thiab kev tuag ntawm tes. HO-1 system tau raug tshaj tawm tias muaj zog heev nyob rau hauv lub hauv paus paj hlwb, thiab nws qhov kev hloov kho pom tau tias ua lub luag haujlwm tseem ceeb hauv cov kab mob ntawm cov kab mob neurodegenerative [33]. Cov kev tshawb fawb tsis ntev los no kuj tau qhia meej txog lub luag haujlwm ntawm Nrf2 hauv kev nce qib thiab kev pheej hmoo ntawm PD [9] thiab Keap1 raws li lub hom phiaj zoo rau kev rov ua haujlwm ntawm Nrf2 hauv AD [34]. Cov txiaj ntsig txhawb nqa cov pov thawj tshiab rau Nrf2 raws li lub hom phiaj kho mob hauv cov kab mob neurodegenerative.
Molecular docking tsom xam pom tias PhGs tuaj yeem khi rau Keap1, nrog cov peev txheej hauv qab no: echinacoside> isoacteoside> acteoside> salidroside. Raws li cov txiaj ntsig no, PhGs pretreatment coj mus rau Nrf2 nuclear translocation, nrog cov Nrf2 qhia hauv qab no: echinacoside> isoacteoside * acteoside> salidroside. Peb xav tias tus naj npawb ntawm glycosides cuam tshuam rau txoj kev sib khi ua tau ntawm PhGs thiab Keap1, thiab hom kev khi txuas ntxiv ua rau kev tso tawm Nrf2 los ntawm Keap1. Cov txheej txheem no ua rau muaj kev ua kom Nrf2 thiab cov noob caj noob ces thiab thaum kawg tiv thaiv PC12 hlwb los ntawm H2 O2 -induced oxidative stress.
Neuroprotective teebmeem ntawm cistanchePhGs
4. Cov ntaub ntawv thiab cov txheej txheem
4.1. Chemical Compounds thiab Reagents
Salidroside (CAS No. {{{0}}}), acteoside (CAS No. 61276-17-3), isoacteoside (CAS No. 61303-13-7), thiab echinacoside (CAS No. {{3}). }) tau yuav los ntawm YYuanye Biotechnology Company (Shanghai, Suav). Cov PhGs tau yaj hauv PBS los tsim cov tshuaj 10 mg / mL, uas tau khaws cia ntawm -20 degree . H2O2 tau yuav los ntawm Aladdin® (Shanghai, Suav). RPMI-1640 nruab nrab thiab fetal bovine serum tau yuav los ntawm Hyclone (Logan, UT, USA), thiab 0.5 feem pua trypsin EDTA, penicillin, thiab streptomycin tau yuav los ntawm Keyi (Hangzhou, Suav). MDA, SOD cov khoom siv kuaj mob, MTT, thiab DCFH-DA tau yuav los ntawm Beyotime Institute of Biotechnology (Nanjing, Jiangsu, Suav). Annexin V-FITC / PI ob chav staining khoom tau yuav los ntawm Solarbio Life Sciences (Beijing, Suav). Antibodies rau Nrf2, Histone H3, Keap1, HO-1, NQO1, GCLC, GCLM, thiab p-actin, anti-nas-horseradish peroxide (HRP) IgG, thiab tshuaj tiv thaiv luav-HRP-IgG tau muas los ntawm Abcam (London, UK). Inhibitors ntawm HO-1 thiab ZnPP tau yuav los ntawm Sigma Chemical Co. (St. Louis, MO, USA). RNAiso Plus, PrimeScript™ RT reagent Kit nrog gDNA Eraser, thiab SYBR® Premix Ex Taq™ II tau yuav los ntawm Takara (Shiga, Nyiv). Lipofectamine® RNAiMAX Transfection Reagent tau yuav los ntawm Thermo Fisher Scientific (Waltham, UK). Nrf2 siRNA sequences tau raws li nram no: pem hauv ntej, CCGAAUUAACUGUCUUAA; and reverse, UAAGACACUGUAAUUGGG. Meanwhile, tswj siRNA sequences raws li nram no: rau pem hauv ntej, UUCUCCGAACGUGUCACGU; thiab thim rov qab, ACGUGACACGUUCGGAGAA.
4.2. Cell Culture
Nas adrenal pheochromocytoma kab (PC12 hlwb) tau txais los ntawm Lub Tsev Haujlwm ntawm Biochemistry thiab Cell Biology, SIBS, (CAS, Shanghai, Suav). Cov hlwb tau khaws cia hauv RPMI-1640 (Hyclone) uas muaj 10 feem pua fetal bovine serum (Hyclone), 100 U/mL penicillin, thiab 0.1 mg/mL streptomycin ntawm 37 degree nrog 5% CO2. Qhov nruab nrab tau hloov txhua hnub.
4.3. Cell ViabilityAssay
PC12 hlwb tau cog rau hauv 96-cov phiaj zoo ntawm 2 x 104 cell/well. Tom qab txuas, cov hlwb raug preincubated nrog los yog tsis muaj inhibitor rau 20 min, incubated nrog los yog tsis muaj PhGs rau 24 teev, thiab ces incubated nrog H2O2 rau lwm 2 teev tom qab lub PhGs raug tshem tawm. Tom qab incubation, cov hlwb raug kho nrog 5 mg / mL MTT rau 4 h ntawm 37 degree, thiab cov xov xwm tau ua tib zoo tshem tawm. Cov pob zeb formazan uas tau tsim los ntawm cov hlwb tseem muaj sia nyob tau yaj hauv 150 rL ntawm DMSO los tsim cov xim xiav [35], thiab qhov nqus tau ntsuas ntawm 570 nm ntawm lub phaj nyeem ntawv. Cov tswj tau siv tib qhov concentration ntawm nruab nrab nrog DMSO ib leeg. Cell viability tau normalized raws li feem pua ntawm kev tswj.
Cov concentrations ntawm PhGs (0.1,1,5, thiab 10 Rg/mL) raug xaiv nyob ntawm seb qhov kev tshuaj xyuas cytotoxicity ntawm PhGs thiab cov txiaj ntsig cytoprotective ntawm echinacoside [32]. Raws li tsab ntawv ceeb toom, tsis muaj cytotoxicity tshwm sim hauv qab 10 Rg / mL, thiab echinacoside tau tshaj tawm los qhia cov nyhuv cytoprotective hauv HzOz-mob mob cell qauv.
4.4. Apoptosis Assay
Apoptosis tau kuaj pom nrog Annexin V-FITC / PI ob chav staining Kit (Solarbio). PC12 hlwb tau cog rau hauv 6-zoo daim hlau ntawm 2 x 105 cells/well. Tom qab txuas, hlwb raug kho nrog PhGs (0.1 thiab 10 Rg / mL) rau 24 teev thiab incubated nrog H2O2 rau lwm 2 teev tom qab PhGs raug tshem tawm. Tom qab incubation, lub hlwb raug ntxuav nyob rau hauv txias PBS, centrifuged ob zaug ntawm 1500 rpm rau 10 min, thiab resuspended nyob rau hauv 500 rL ntawm binding tsis. FITC-labeled Annexin V (5 rL) thiab propidium iodide.
Cov ntawv luv
GCLC glutamate-cysteine ligase-catalytic subunit
GCLM glutamate-cysteine ligase-catalytic modifier subunit
H2O2 hydrogen peroxide
HO-1 heme oxygenase 1
Keap1 Kelch ECH koom haum protein 1
NQO1 NAD(P)H quinone oxidoreductase 1
Nrf2 nuclear factor erythroid 2-txog yam 2
PhGs salidroside, acteoside, isoacteoside, thiab echinacoside
ROS reactive oxygen hom
ZnPP zinc protoporphyrin
Cistanche deserticola extract: PHG ntawm cistanche
Cov ntaub ntawv
1. Sun, AY; Chen, YM Oxidative stress thiab neurodegenerative disorders. J. Biomed. Sci. Xyoo 1998, 5, 401-414. [CrossRef] [PubMed]
2. Maheshwari, A.; Micro, MM; Aggarwal, UA; Sharma, RK; Nandan, D. Pathways koom nrog hauv cov kab mob qog nqaij hlav ntawm tes apoptosis tshwm sim los ntawm H2O2 hauv vitro. FEBS J. 2009,276, 870-881. [CrossRef] [PubMed]
3. Halliwell, B. Reactive oxygen hom thiab lub hauv paus paj hlwb. J. Neeb. Xyoo 1992, 59, 1609-1623. [CrossRef] [PubMed]
4. Richardson, JS; Subbarao, KV; Ang, LC Ntawm lub luag haujlwm ua tau ntawm hlau-vim dawb radical peroxidation hauv neural degeneration hauv Alzheimer's kab mob. Ann. NY Acad. Sci. Xyoo 1992, 648, 326-327. [CrossRef] [PubMed]
5. Zhang, DD Mechanistic kev tshawb fawb ntawm Nrf2-Keap1 txoj kev taw qhia. Drug Metab. Rev. 2006, 38, 769-789. [CrossRef] [PubMed]
6. Ib., K.; Tong, KI; Yamamoto, M. Molecular mechanism activating Nrf2-Keap1 txoj kev nyob rau hauv cov kev cai ntawm adaptive teb rau electrophiles. Dawb Radic. Biol. Med. 2004, 36, 1208-1213. [CrossRef] [PubMed]
7. Kong, AN; Aws, E.; Yus, R.; Hebbar, V.; Chen, C.; Hu, R.; Mandlekar, S. Induction ntawm xenobiotic enzymes los ntawm daim ntawv qhia kinase txoj kev thiab cov antioxidant los yog electrophile teb lub caij (ARE/EpRE). Drug Metab. Rev. 2001, 33, 255-271. [CrossRef] [PubMed]
8. Buendia, ib.; Michalska, P.; Navarro, E.; Gameiro, ib.; Eeg, J.; Leon, R. Nrf2-YOG txoj kev: Ib lub hom phiaj tawm tsam oxidative stress thiab neuroinflamation hauv cov kab mob neurodegenerative. Pharmacol. Ther. Xyoo 2016, 157, 84-104. [CrossRef] [PubMed]
9. Skibinski, G.; Hwang, V; Ando, DM; Daub, A.; Li, AK; Ravisankar, UA; Modan, S.; Finucane, MM; Shabby, BA; Finkbeiner, S. Nrf2 mitigates LRRK2- thiab a-synuclein-induced neurodegeneration los ntawm modulating proteostasis. Proc. Natl. Acad. Sci. USA 2016, 114, 1165-1170. [CrossRef] [PubMed]
10. Jimenez, C.; Riguera, R. Phenylethanoid glycosides hauv cov nroj tsuag: Cov qauv thiab kev ua haujlwm lom neeg. Nat. Prod. Rep. 1994, 11, 591-606. [CrossRef] [PubMed]
11. Georgiev, M.; Alipieva, K.; Orhan, ib.; Abrashev, R.; Denev, P.; Angelova, M. Antioxidant thiab cholinesterases inhibitory kev ua ub no ntawm Verbascum xanthophoeniceum Griseb. thiab nws cov phenylethanoid glycosides. Khoom noj khoom haus Chem. Xyoo 2011, 128, 100-105. [CrossRef] [PubMed]
12. Li, N.; Wang, J.; Ma, J.; Gu, Z.; Jiang, C.; Yus, L.; Fu, X. Neuroprotective Effects ofCistanches HerbaKev kho mob rau cov neeg mob uas muaj tus kab mob Moderate Alzheimer/s. Evid.-Based Complement. Hloov. Med. Xyoo 2015, 2015, 103985. [CrossRef] [PubMed]
13. Liu, YL; He, WJ; Mo, L.; Shi, MF; Zhu, YY; Pan, S.; Li, XR; Xu, QM; Yang, SL Antimicrobial, tiv thaiv kev ua ub no thiab toxicology ntawm phenylethanoid glycosides los ntawm Monochasma savatieri Franch. ua Maxim. J. Ethnopharmacol. Xyoo 2013, 149, 431-437. [CrossRef] [PubMed]
14. Dong, Q.; Yaj, J.; Fang, JN; Ding, K. Cov yam ntxwv ntawm tus yam ntxwv thiab kev tiv thaiv kab mob ntawm ob lub dej txias-dej extractable polysaccharides los ntawmCistanche deserticola YC Ma. Carbohydrate. Res. Xyoo 2007, 342, 1343-1349. [CrossRef] [PubMed]
15. Xiong, L.; Mao, S.; Lu, B.; Yaj, J.; Zhou, F.; Hu, Y.; Jiang, Y.; Shen, C.; Zhao, Y. Osmanthusfragrans Paj Extract thiab Acteoside tiv thaiv d-Galactose-Induced Aging hauv ICR Mouse Model. J. Med. Zaub mov 2016, 19, 54-61. [CrossRef] [PubMed]
16. Jiang, Y.; Mao, S.; Huang, W. Lu, B.; Cai, Z.; Zhou, F.; Li, M.; Lou, T.; Zhao, Y. Phenylethanoid Glycoside Profiles thiab Antioxidant Activities ntawm Osmanthusfragrans Lour. Paj los ntawm UPLC/PDA/MS thiab Simulated Digestion Model. J. Agric. Khoom noj khoom haus Chem. Xyoo 2016, 64, 2459-2466. [CrossRef] [PubMed]
17. Li, X.; Yog, X.; Li, X.; Sun, X.; Liang, Q.; Tau, L.; Kang, X.; Chen, J. Salidroside tiv thaiv MPP ntxiv -induced apoptosis hauv PC12 hlwb los ntawm inhibiting NO txoj kev. Lub hlwb Res. 2011, 1382, 9-18. [CrossRef] [PubMed]
18. Zhou, L.; Ding, W.; Sun, H.; Zhou, Q. Huang, J.; Li, X.; Xie, Y.; Chen, J. Salidroside tiv thaiv PC12 hlwb los ntawm MPP ntxiv rau -induced apoptosis los ntawm kev ua kom txoj hauv kev PI3K/Akt. Khoom noj khoom haus Chem. Toxicol. Xyoo 2012, 50, 2591-2597. [CrossRef] [PubMed]
19. Sheng, GQ; Zhang, JR; pua, XP; Ma, J.; Li, CL Tiv thaiv cov nyhuv ntawm verbascoside ntawm 1-methyl-4- phenylpyridinium ion-induced neurotoxicity hauv PC12 hlwb. Eur. J. Pharmacol. 2002, 451, 119-124. [CrossRef]
20. Wang, H.; Xu, Y.; Yog, J.; Zhao, X.; Sun, X.; Zhang, YJ; Guo, J.; Zhu, C. Acteoside tiv thaiv tib neeg neuroblastoma SH-SY5Y hlwb tiv thaiv beta-amyloid-induced cell raug mob. Lub hlwb Res. Xyoo 2009, 1283, 139-147. [CrossRef] [PubMed]
21. Min, D.; Jin, YZ; Yog, J.; Zheng, BL; Yao, HW Echinacoside cawm SHSY5Y neuronal hlwb los ntawm TNFa-induced apoptosis. Chin. Pharmacol. Bull. 2005, 505, 11-18.
22. Zhao, Q.; Gao, JP; Li, WW; Cai, DF Neurotrophic thiab neurorescue teebmeem ntawm Echinacoside hauv subacute MPTP nas qauv ntawm Parkinson tus kab mob. Lub hlwb Res. 2010, 1346, 224-236. [CrossRef] [PubMed]
23. Koo, KA; Sung, SH; Park, JH; Kim, SH; Li, KY; Kim, YC In vitro neuroprotective kev ua ub no ntawm phenylethanoid glycosides los ntawm Callicarpa dichotoma. Planta Med. Xyoo 2005, 71, 778-780. [CrossRef] [PubMed]
24. Liu, YG; Li, X.; Xiong, C.; Yus, B.; pua, X.; Yog lawm, XS Synthetic phenylethanoid glycoside derivatives ua cov tshuaj tiv thaiv neuroprotective. Eur. J. Med. Chem. Xyoo 2015, 95, 313-323. [CrossRef] [PubMed]
25. Fu, G.; Pang, H.; Wong, YH Ib txwm tshwm sim phenylethanoid glycosides: Muaj peev xwm ua rau cov kev kho tshiab. Curr. Med. Chem. Xyoo 2008, 15, 2592-2613. [CrossRef] [PubMed]
26. Greene, LA; Tischler, AS Tsim los ntawm noradrenergic clonal kab ntawm nas adrenal pheochromocytoma hlwb uas teb rau cov paj hlwb loj hlob. Proc. Natl. Acad. Sci. USA 1976, 73, 2424-2428. [CrossRef] [PubMed]
27. Liang, QN; Sheng, YC; Jiang, P. Ji, LL; Xia, YY; Min, Y.; Wang, ZT Qhov sib txawv ntawm glutathione antioxidant system nyob rau hauv tshiab weaned thiab hluas nas daim siab thiab nws kev koom tes nyob rau hauv isoline-induced hepatotoxicity. Arch. Toxicol. Xyoo 2011, 85, 1267-1279. [CrossRef] [PubMed]
28. Kobayashi, A.; Kang, M.; Okawa, H.; Tsoo, M.; Zenke, Y; Chib, T.; Igarashi, K.; Yamamoto, M. Oxidative Stress Sensor Keap1 Muaj nuj nqi ua ib qho Adapter rau Cul3-Raws li E3 Ligase los tswj cov Proteasomal Degradation ntawm Nrf2. Mol. Cell. Biol. Xyoo 2004, 24, 7130-7139. [CrossRef] [PubMed]
29. Cornelius, C.; Crupi, R.; Calabrese, V; Graziano, UA; Milon, P. Pennisi, G.; Radak, Z.; Calabrese, EJ; Cuzzocrea, S. Mob hlwb raug mob: Oxidative stress thiab neuroprotection. Antioxidant. Redox teeb liab. Xyoo 2013, 19, 836-853. [CrossRef] [PubMed]
30. Hu, Z.; Luo, W.; Sun, X.; Hao, S.; Zhang, YJ; Xu, F.; Wang, Z.; Liu, B. Hydrogen-nplua nuj saline tiv thaiv oxidative kev puas tsuaj thiab kev txawj ntse tsis txaus tom qab mob me ntsis mob hlwb. Lub hlwb Res. Bull. Xyoo 2012, 88, 560-565. [CrossRef] [PubMed]
31. Ansari, MA; Roberts, KN; Scheff, SW Ib lub sij hawm kawm ntawm contusion-induced oxidative kev nyuaj siab thiab synaptic proteins nyob rau hauv cortex nyob rau hauv ib tug nas qauv ntawm TBI. J. Neurotrauma 2008, 25, 513-526. [CrossRef] [PubMed]
32. Kuang, R.; Sun, Y.; Yuan, W.; Li, L.; Zheng, X. Tiv thaiv cov teebmeem ntawm echinacoside, ib qho ntawm phenylethanoid glycosides, ntawm H2O2-induced cytotoxicity hauv PC12 hlwb. Planta Med. Xyoo 2009, 75, 1499-1504. [CrossRef] [PubMed]
33. Scapagnini, G.; Vasto, S.; Abraham, NG; Caruso, C.; Zella, D.; Fabio, G. Modulation ntawm Nrf2/ARE txoj hauv kev los ntawm cov zaub mov polyphenols: Lub tswv yim noj zaub mov zoo rau kev paub txog kev paub thiab kev puas hlwb neurodegenerative. Mol. Neurobiol. Xyoo 2011, 44, 192-201. [CrossRef] [PubMed]
34. Kerr, F.; Sofolaadesakin, O.; Ivanov, DK; Gatliff, J.; Gomez, PB; Bertrand, HC; Martinez, P. Callard, R.; ib. Snoeren, ib.; Cochem, HM Direct Keap1-Nrf2 cuam tshuam raws li lub hom phiaj kho mob Alzheimer/s. PLoS Genet. Xyoo 2017, 13, e1006593. [CrossRef] [PubMed]
35. Hansen, MB; Nielsen, SE; Berg, K. Kev tshuaj xyuas dua thiab kev txhim kho ntxiv ntawm txoj kev zas xim kom meej thiab nrawm rau kev ntsuas kev loj hlob ntawm tes / cell tua. J. Immunol. Methods 1989, 119, 203-210. [CrossRef]
36. Xovxwm, C. Kev tshuaj ntsuam xyuas Virtual hauv Kev Tshawb Nrhiav Tshuaj; Crc Xovxwm: Boca Raton, FL, USA, 2005.
37. Muegge, I.; Martin, YC; Hajduk, PJ; Fesik, SW Kev Ntsuam Xyuas ntawm PMF qhov qhab nia hauv docking tsis muaj zog ligands rau FK506 binding protein. J. Med. Chem. 1999, 42, 2498–2503. [CrossRef] [PubMed]
38. Kuntz, ID; Blaney, JM; Oatley, SJ; Langridge, R.; Ferrin, TE Ib txoj hauv kev geometric rau macromolecule-ligand kev sib cuam tshuam. J. Mol. Biol. 1982, 161, 269-288. [CrossRef]
39. MD, E.; Murray, CW; Auton, TR; Paolini, GV; Mee, RP Empirical scoring functions: I. Txoj kev loj hlob ntawm qhov ceev ceev empirical scoring muaj nuj nqi los kwv yees cov affinity ntawm ligands nyob rau hauv receptor complexes. J. Comput.-Aided Mol. Des. 1997, 11, 425-445.
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