Echinacoside tiv thaiv 6-Hydroxydopamine-Induced Mitochondrial Dysfunction Thiab Inflammatory Responses hauv PC12 Cells Los Ntawm Kev Txo ROS Ntau Lawm

Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com

Yue-Hua Wang, 1,2 Zhao-Hong Xuan, 3 Shuo Tian, ​​1 thiab Guan-Hua Du 1,2

Parkinson's disease (PD) yog ib qho kab mob neurodegenerative uas tshwm sim los ntawm kev poob ntawm dopaminergic (DA) neurons ntawm substantia nigra. Mitochondrial dysfunction thiab inflammatory teb yog koom nrog hauv cov txheej txheem ntawm kev puas hlwb hauv PD. 6-Hydroxydopamine (6-OHDA), ib qho tshuaj dopamine analog, tshwj xeeb yog kev puas tsuaj rau dopaminergic neurons.Echinacoside(ECH) yog ib qho phenylethanoid glycoside cais tawm ntawm stemsCistanche salsa, qhia ntau yam ntawm cov teebmeem neuroprotective hauv kev tshawb fawb yav dhau los. Txoj kev tshawb fawb tam sim no yog los tshawb xyuas nws cov txiaj ntsig tawm tsam 6-OHDA-induced neurotoxicity thiab cov txheej txheem ua tau hauv PC12 hlwb. Cov txiaj ntsig tau pom tias 6-OHDA txo qis cell viability, txo qis oxidation-txo kev ua, txo cov mitochondrial membrane peev xwm, thiab induced mitochondria-mediated apoptosis piv nrog cov tsis kho PC12 hlwb. Txawm li cas los xij,echinacoside cov tshuajKev kho mob tau txo qis cov kev hloov pauv no tshwm sim los ntawm 6-OHDA. Ntxiv rau,echinacoside cov tshuajkuj tseem tuaj yeem txo cov kev mob tshwm sim los ntawm 6-OHDA. Kev tshawb fawb ntxiv tau pom tiasechinacoside cov tshuajtuaj yeem txo 6-OHDA-induced ROS ntau lawm hauv PC12 hlwb. Cov txiaj ntsig no qhia tias lub hauv paus mechanism ntawmechinacoside cov tshuajtawm tsam 6-OHDA-induced neurotoxicity tej zaum yuav koom nrog hauv attenuating mitochondrial dysfunction thiab inflammatory teb los ntawm kev txo ROS ntau lawm.

1. Taw qhia

Parkinson tus kab mob (PD)yog ib qho teeb meem neurodegenerative tshwm sim los ntawm kev maj mam zuj zus degeneration ntawm dopamine neurons nyob rau hauv substantia nigra pars compacta [1]. Txawm hais tias cov txiaj ntsig ntawm neuropathological ntawm PD tau piav qhia zoo, etiology tseem tsis tau txhais. Qhov tseem ceeb, ntau qhov kev soj ntsuam qhia tias mitochondrial dysfunction yog koom nrog hauv pathogenesis ntawm PD thiab degeneration ntawm dopaminergic neurons [2, 3]. Mitochondria muaj ntau cov enzymes redox thiab ib txwm tshwm sim tsis zoo ntawm oxidative phosphorylation tsim reactive oxygen hom (ROS) [4]. Tsis ntev los no, nce cov pov thawj los ntawm tib neeg thiab tsiaj cov kev tshawb fawb tau qhia tias neuroinflamation kuj yog ib qho tseem ceeb rau kev poob ntawm neuronal hauv PD [5].

6-Hydroxydopamine (6-OHDA) yog tshwj xeeb rau dopaminergic neurons hauv intrastriatal nas qauv thiab hauv qee lub hlwb [6–11]. PC12 cell tau dhau los ua qauv xovtooj nrov rau kev tshawb fawb PD vim tias cov kab ntawm tes no muaj ntau yam ntawm DAergic neurons. Nws tau raug siv los ua tus qauv hauv vitro rau txoj kev tshawb fawb ntawm PD thiab txiav txim siab cov nyhuv ntawm cov tshuaj tiv thaiv thiab kho. Echinacoside (Daim duab 1) yog phenylethanoid glycoside cais thiab purified los ntawm stemsofCistanche salsa[12], cov kab mob cab ib txwm nyob rau sab qaum teb sab hnub poob Tuam Tshoj, uas yog siv los ua cov txheej txheem hauv qab ntawm echinacoside tawm tsam 6-OHDA- induceddamageinPC12cells.

2. Cov ntaub ntawv thiab cov txheej txheem

2.1. Khoom siv. Echinacoside tau txais los ntawm National Institutes for Food and Drug Control (Beijing, Suav). RPMI-1640 thiab fetal bovine serum tau txais los ntawm Gibco

(Grand Island, NY, USA). 6-OHDA, MTT, 2,7-dichlorodihydrofluorescein diacetate, propidium iodide (PI), thiab resazurin tau yuav los ntawm Sigma-Aldrich (St. Louis, MO, USA). Cov khoom siv ATP bioluminescent assay tau txais los ntawm Promega (Madison, USA). 5,5,6,6 -Tetrachloro-1,1,3,3 -tet-raethyl benzimidazolyl carbocyanine iodide (JC-1) cov khoom siv tshuaj ntsuam xyuas tau txais los ntawm Beyotime Institute of Biotechnology (Haimen, Jiangsu, Suav teb). Cov khoom siv Cytochrome C ELISA thiab fluorogenic substrate Ac-DVD-AMC tau txais los ntawm Invitrogen (Carlsbad, CA, USA). IL-1 thiab IL-6 ELISA cov khoom siv tau txais los ntawm Boster Bio-engineer Limited tuam txhab (Wuhan, Suav). Spectramax M5 microplate nyeem ntawv tau yuav los ntawm Molecular Devices tuam txhab (Sunnyvale, CA, USA).

2.2. Cell Kab lis kev cai thiab kev kho mob. PC12 hlwb tau loj hlob hauv RPMI-1640 ntxiv nrog 10 feem pua ​​nees ntshav thiab 5 feem pua ​​​​fetal bovine serum. Cov xwm txheej tau khaws cia ntawm 37 ∘C nyob rau hauv ib qho chaw humidified uas muaj 5 feem pua ​​CO2 [19]. Cov hlwb raug plated rau hauv 96- cov phiaj zoo ntawm qhov ceev ntawm 2 × 105 hlwb / mL. Tom qab 80 feem pua ​​confluence, cov hlwb tau preincubated nrog txawv concentrations ntawm echinacoside nyob rau hauv ib tug serum-dawb RPMI -1640 nruab nrab rau 1 h. Tom qab ntawd, 6-OHDA tau ntxiv rau hauv qhov dej ntawm qhov kawg ntawm 100 M thiab incubated rau lwm Tom qab kev kho mob, cov kev hloov morphological tau pom nyob rau hauv lub tshuab tsom. Cell viability raug soj ntsuam los ntawm MTT assay [19]. Luv luv, tom qab kev kho mob, MTT ntawm qhov kawg siab ntawm 0.5 mg / mL tau ntxiv rau txhua qhov dej thiab incubated rau 4 h ntawm 37 ° C.

2.3.Morphological Changes and Viability Assay of PC12 Cells. Tom qab kev kho mob, kev hloov morphological tau pom nyob rau hauv lub tshuab tsom. Cell viability raug soj ntsuam los ntawm MTT assay [19]. Luv luv, tom qab kev kho mob, MTT ntawm qhov kawg siab ntawm 0.5 mg / mL tau ntxiv rau txhua qhov dej thiab incubated rau 4 teev ntawm 37∘C. Tom qab ntawd, cov supernatant raug tshem tawm thiab cov khoom formazan tau yaj hauv 100 L dimethyl sulfoxide nrog nplawm rau 15 min ntawm microtiter phaj shaker thiab cov absorbance tau nyeem ntawm 570 nm siv Spectramax M5 microplate nyeem. Cov ntaub ntawv pov thawj-based Complementary thiab Alternative Medicine

2.4. Intracellular ATP Level Assay. Cellular ATP theem tau ntsuas nrog ATP bioluminescent assay cov khoom raws li cov chaw tsim khoom cov lus qhia. Luminescence tau saib xyuas ntawm tus nyeem ntawv microplate hauv hom luminescence tom qab qhov sib ntxiv ntawm 100 L ntawm luciferin thiab luciferase [20].

2.5. Kev ntsuas ntawm Mitochondria Oxidation-Reduction Activity los ntawm Resazurin. Tom qab kev kho mob, resazurin ntawm qhov kawg concentration ntawm 5 M tau ntxiv rau hauv qhov dej thiab kev siv tshuaj fluorescence raug tshuaj xyuas ntawm qhov excitation ntawm 530 nm thiab emission ntawm 590 nm [21].

2.6. Kev ntsuas ntawm Mitochondrial Membrane Potential los ntawm JC-1. Lub peev xwm mitochondrial membrane tau ntsuas los ntawm JC-1 cov khoom siv ntsuas raws li cov chaw tsim khoom cov lus qhia. Tom qab kev kho mob, kab lis kev cai nruab nrab raug tshem tawm thiab thauj nrog JC-1 tov rau 15 feeb ntawm 37∘C hauv qhov tsaus ntuj. Kev siv fluorescence ntawm qhov sib piv liab / ntsuab tau txiav txim siab ntawm qhov excitation ntawm 490 nm thiab emission ntawm 530 nm (ntsuab fluorescent monomers) thiab 590 nm (liab fluorescent aggregates), feem [22].

2.7. Quantification ntawm Cytochrome C tso tawm. Cov qib cytosolic cytochrome C tau ntsuas siv cov khoom siv ELISA raws li cov chaw tsim khoom cov lus qhia [23]. Luv luv, tom qab kev kho mob, cov hlwb tau lysed thiab centrifugated ntawm 1000 g rau 15 min, thiab tom qab ntawd cov supernatant tau sau ua ib feem ntawm cytoplasmic thiab siv rau kev ntsuas cytochrome C tso tawm.

2.8. Kev txiav txim siab ntawm Caspase-3 Kev Ua Si. Caspase-3 kev ua haujlwm tau ua raws li cov lus qhia nrog kev hloov kho [24]. Luv luv, tom qab kev kho mob, lub lysis buffer wasaddedtoeach zoo thiab ua raws li centrifugation ntawm 12 000 g rau 10 min. Caspase-3 kev ua haujlwm tau raug sim hauv qhov ua rau supernatants los ntawm kev ntsuas cov proteolytic cleavage ntawm fluorogenic substrate Ac-DVD-AMC nrog ib qho excitation wavelength ntawm 380 nm thiab emission wavelength ntawm 460 nm.

2.9. Flow Cytometry Assay hauv PC12 Cells. Tom qab kev kho mob, cov hlwb raug trypsinized, ntxuav ob zaug nrog PBS, centrifuged ntawm 1000 r / min rau 5 min, thiab tom qab ntawd kho nrog 70 feem pua ​​​​ethanol hmo ntuj ntawm 4∘C. Cov hlwb raug centrifuged thiab ntxuav ob zaug nrog PBS, resuspended hauv PBS (muaj 50 g / mL RNaseA), stained nrog PI ntawm qhov kawg concentration ntawm 10 M rau 30 min ntawm chav tsev kub nyob rau hauv qhov tsaus ntuj nti, thiab nyeem nyob rau hauv ib tug flow cytometer ntawm 488. nm kev xav [10].

2.10. Kev ntsuas ntawm IL-1 thiab IL-6 Qib los ntawm ELISA Assay. Cov kab mob PC12 hlwb nruab nrab tau sau tom qab kev kho mob, thiab IL-1 thiab IL-6 concentrations raug ntsuas los ntawm cov khoom siv ELISA cov khoom lag luam raws li tus neeg tsim khoom raws tu qauv.

2.11. Kev txiav txim siab ntawm Intracellular Reactive Oxygen Hom Qib. Tom qab incubation nrog 6-OHDA, cov hlwb tau ntim nrog 10M H2DCFH-DA rau 30 min ntawm 37∘C hauv qhov tsaus ntuj. Qhov kev siv fluorescence ntawm DCF tau ntsuas ntawm qhov ua kom muaj zog

Cov ntaub ntawv pov thawj-raws li Kev Sib Txuas Ntxiv thiab Lwm Cov Tshuaj Kho Mob wavelength ntawm 495 nm thiab emission wavelength ntawm 530 nm ntawm tus nyeem ntawv microplate [25].

2.12. Kev txheeb cais. Tag nrho cov ntaub ntawv raug qhia raws li qhov nruab nrab ± SD. Kev txheeb xyuas ntawm cov txiaj ntsig tau ua los ntawm ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA), ua raws li -test. Kev lees paub qhov tseem ceeb ntawm <>

3. Cov txiaj ntsig

3.1. EffectofEchinacosideonMorphologyand Cell Viabilityin PC12 Cells puas los ntawm 6-OHDA. Raws li pom nyob rau hauv daim duab 2(A), qhov muaj peev xwm ntawm PC12 hlwb tsis muaj qhov sib txawv ntawm qhov concentration ntawm 0.1∼10 M ntawm echinacoside kev kho mob rau

24 h ua. Raws li pom nyob rau hauv daim duab 2(B), nyob rau hauv 24 teev ntawm kev kho mob nrog 6-OHDA ib leeg, feem ntau ntawm PC12 hlwb tau hloov morphological xws li membrane blebbing thiab cell shrinkage. Kev kho mob nrog echinacoside tiv thaiv cov hlwb los ntawm 6-OHDA kev puas tsuaj. MTT kev soj ntsuam tau pom tias kev kho mob ntawm PC12 hlwb nrog 6-OHDA ib leeg ua rau muaj kwv yees li 25 feem pua ​​​​ntawm cov cell ciaj sia nyob hauv 24 teev, thaum kho nrog {{10}}.1 M, 1 M, thiab 10 M echinacoside pom qhov txo qis ntawm 6-OHDA-mediated cytotoxicity (tag nrho <0.01, daim="" duab="">

3.2. Cov nyhuv ntawm Echinacoside ntawm Mitochondria Dysfunction Induced los ntawm 6-OHDA hauv PC12 Cells. Cellular ATP qib tau txo qis tom qab 24 teev incubation nrog 100M 6-OHDA, thaum kho nrog 1 Mand 10 M ntawm echinacoside alleviated 6-OHDA-mediated ATP txo qis anabolic hloov hauv PC12 hlwb . Cov txiaj ntsig tau pom tias kev kho mob nrog 1 Mand 10 M ntawm echinacoside tuaj yeem txo qis mitochondria oxidation-txo kev ua haujlwm tsis zoo cuam tshuam los ntawm 6-OHDA (ob leeg <0.05 thiab="" daim="" duab="">

Raws li pom hauv daim duab 3(c), kev kho mob nrog 100 M ntawm 6- OHDA rau 24 h ua rau muaj qhov txo qis ntawm cov mitochondrial membrane muaj peev xwm piv nrog cov pab pawg tsis kho ( < {="" {11}}.01).="" txawm="" li="" cas="" los="" xij,="" qhov="" txo="" qis="" ntawm="" mitochondrial="" membrane="" muaj="" peev="" xwm="" tshwm="" sim="" los="" ntawm="" 6-ohda="" tau="" txo="" qis="" los="" ntawm="" cotreatment="" nrog="" 0.1="" m,="" 1="" m,="" thiab="" 10="" m="" ntawm="" echinacoside="" (tag="" nrho=""><>

3.3. Anti-Mitochondria-Mediated Apoptosis ntawm Echinacoside hauv PC12 Cells tiv thaiv 6-OHDA Neurotoxicity. Cov txiaj ntsig tau pom tias 6-OHDA raug cuam tshuam ua rau muaj kev nce hauv cytosolic cytochrome C los ntawm 167 feem pua ​​​​ntawm cov pab pawg tsis kho ( < 0.{{10}}1).="" echinacoside="" ntawm="" koob="" tshuaj="" 0.1="" m,="" 1="" m,="" thiab="" 10m="" tuaj="" yeem="" txo="" cov="" cytochrome="" c="" tso="" rau="" hauv="" cytosol="" induced="" los="" ntawm="" 6-ohda="">< 0.05,="">< 0.01,="" thiab=""><0.01, resp.;="" daim="" duab="">

Caspase{{0}} kev ua si tau nce li ntawm 3.3-fold piv nrog cov pab pawg tsis kho tom qab 6-OHDA raug ( < 0="" 01="" ).="" hauv="" qhov="" sib="" piv,="" kev="" kho="" mob="" nrog="" 1="" m="" thiab="" 10="" m="" ntawm="" echinacoside="" pom="" qhov="" txo="" qis="" hauv="" caspase-3="" kev="" ua="" haujlwm="" piv="" nrog="" 6-ohda-kho="" hlwb,="" raws="" li="" (ob="" leeg=""><0.05 thiab="" daim="" duab="" 4(="" b)).pi="" yog="" daim="" nyias="" nyias="" permeability="" thiab="" feem="" ntau="" tsis="" suav="" nrog="" cov="" hlwb="" siv="" tau.="" yog="" li="" nws="" yog="" feem="" ntau="" siv="" los="" txheeb="" xyuas="" cov="" hlwb="" tuag="" hauv="" cov="" neeg.="" cov="" txiaj="" ntsig="" tau="" pom="" tias="" apoptosis="" tus="" nqi="" tau="" nce="" ntau="" ntxiv="" tom="" qab="" 24="" teev="" incubations="" nrog="" 100="" m="" 6-ohda,="" thaum="" kho="" nrog="" 1="" thiab="" 10="" m="" echinacoside="" txo="" qis="" 6-ohda-induced="" apoptosis="">< 0.05,="">< 0.01,="" resp.,="" daim="" duab="">

3.4. Anti-Inflammatory Effect of Echinacoside in PC12 Cells Induced by 6-OHDA. Cov txiaj ntsig tau pom tias 6-OHDA exposureresultedinanincreaseinIL-1 qib piv nrog cov pab pawg tsis kho ( < 0="" 05;="" daim="" duab="" 4(a)).="" txawm="" li="" cas="" los="" xij,="" 10="" m="" ntawm="" echinacoside="" tuaj="" yeem="" txo="" qis="" il="" -1="" tso="" tawm="" los="" ntawm="" 6-ohda="">< 0.05;="" daim="" duab="">

IL {{0}} qib kuj tseem nce ntxiv piv nrog pab pawg tsis kho ( < 0="" 01).="" txawm="" li="" cas="" los="" xij,="" kev="" kho="" mob="" nrog="" 0.1,="" 1,="" thiab="" 10="" m="" ntawm="" echinacoside="" txhua="" qhov="" txo="" qis="" il-6="" tso="" tawm="" piv="" nrog="" 6-ohda-kho="" hlwb="" (ob="" leeg=""><0.05 thiab="" daim="" duab="">

3.5. Qhov cuam tshuam ntawm ECH ntawm ROS Production Induced los ntawm 6-OHDA hauv PC12 Cells. Cells kho nrog 6-OHDA pom qhov nce ntxiv (kwv yees li 1.4-fold) ntawm intracellular ROS piv nrog cov hlwb tsis kho (< 0.01).="" qhov="" kev="" nce="" no="" tau="" txo="" qis="" los="" ntawm="" coincubation="" nrog="" 0.1,="" 1,="" thiab="" 10="" m="" echinacoside="" (="">< 0.05,="">< 0.01,="" thiab="">< 0.01,="" resp.;="" daim="" duab="" 6)="">

4. Kev sib tham

Echinacoside yog ib qho ntawm cov tseem ceeb ntawm phenylethanoid glycosides muab rho tawm los ntawm ib txwm muaj npe nrov tshaj tawm echinacoside muaj cov tshuaj tiv thaiv apoptotic thiab neuroprotective hauv cov kab mob neurodegenerative [18]. Hauv vivo txoj kev tshawb fawb tau tshaj tawm tias echinacoside muaj cov teebmeem neuroprotective thiab kev txhim kho tus cwj pwm hauv tus nas qauv ntawm MPTP- induced dopaminergic neuronal puas, thaum ntxiv kev tshawb fawb qhia tias echinacoside induced downregulation ntawm caspase -3 thiab caspase -8 ua rau MPP ntxiv -induced apoptosis ntawm cerebellar granule neurons [16]. Lwm qhov hauv vivo txoj kev tshawb fawb qhia txog cov teebmeem neuroprotective ntawm echinacoside ntawm striatal dopaminergic neurons raug mob los ntawm 6-OHDA [17]. Txawm li cas los xij, cov txheej txheem ntawm tes thiab molecular uas ua rau cov kev ua no tsis nkag siab tag nrho, tshwj xeeb tshaj yog rau lawv cov kev cuam tshuam ntawm mitochondrial kev ua haujlwm thiab cov lus teb rau qhov mob. Yog li peb txoj kev tshawb fawb tam sim no tau tshawb xyuas qhov cuam tshuam ntawm echinacoside ntawm mitochondrial kev ua haujlwm thiab cov lus teb rau qhov mob kom elucidate nws cov txheej txheem tiv thaiv PD. Cov txiaj ntsig tau pom thawj zaug pom tias echinacoside txo qis 6-OHDA-induced neurotoxicity hauv PC12 hlwb los ntawm kev tiv thaiv mitochondrial muaj nuj nqi thiab tiv thaiv kab mob.

Mitochondrial dysfunction tau ntev tau cuam tshuam txog kev tuag ntawm nigrostriatal dopaminergic neurons hauv Parkin-son tus kab mob thiab nws cov qauv sim [26]. Mitochondria muaj kev koom tes hauv kev tsim cov pa oxygen reactive (ROS) los ntawm cov khoom siv hluav taws xob ntawm cov kab ua pa hauv cov kab mob neurodegenerative [27]. Ntau yam ntawm ROS yog tsim nyob rau hauv cov chav kawm ntawm ib txwm metabolism hauv cov kab mob lom, tab sis lawv tsub zuj zuj tshaj lub

Cov nyiaj xav tau tuaj yeem ua rau cov lipids, cov protein, thiab nucleic acids puas tsuaj.

6-OHDA, ib qho catecholamine analog, ua rau qee qhov degeneration ntawm substantia nigra neurons. Neurotoxicity induced los ntawm 6-OHDA yog, ib feem, vim yog tsim los ntawm ROS [28–32].

OHDA ua rau kev tuag neuronal los ntawm kev cuam tshuam mitochondria muaj nuj nqi, suav nrog kev txo qis hauv ATP ntau lawm, cuam tshuam ntawm ROS metabolism, thiab induction ntawm apoptosis [10, 33, 34]. Hauv kev tshawb fawb tam sim no, peb cov txiaj ntsig kuj tau pom tias 6-OHDA puas mitochondrial oxidative-txo muaj nuj nqi, txo mitochondrial membrane peev xwm, induced mitochondria-mediated apoptosis, thiab nce ROS ntau lawm hauv PC12 hlwb.

Resazurin yog qhov taw qhia fluorescent uas tsuas yog nruab nrab ntawm qhov kawg txo qis ntawm molecular oxygen thiab cytochrome oxidase. Yog li ntawd, oxidative-txo lub peev xwm ntawm resazurin tso cai rau cov kab mob ua pa ua haujlwm kom ze rau qhov ua tiav, uas yuav muab qhov tseeb dua thiab muaj txiaj ntsig ntawm kev ua haujlwm mitochondrial. JC-1 yog ib daim nyias nyias muaj peev xwm-sensitive sojntsuam uas localizes mus rau lub puab mitochondrial membrane uas nws tsim yog monomers (ntsuab) los yog aggregates (liab) raws li mitochondrial membrane muaj peev xwm. Yog li, resazurin thiab JC-1 yog cov cuab yeej tshuaj ntsuam xyuas muaj txiaj ntsig rau kev tshuaj xyuas mitochondrial muaj nuj nqi [35]. Txoj kev tshawb no qhia tau hais tias kev txo qis hauv mitochondrial oxidative-txo kev ua haujlwm thiab mitochondrial membrane muaj peev xwm ua tau lub luag haujlwm tseem ceeb hauv 6-OHDA- induced dopaminergic neurotoxicity. Peb cov txiaj ntsig kuj tau pom mitochondrial oxidative-txo kev ua haujlwm thiab mitochondrial membrane muaj peev xwm. Cov txiaj ntsig no muab kev txhim kho ntawm mitochondrial dysfunction yuav yog txoj hauv kev zoo dua los ua kom qeeb dopaminergic neurodegeneration feem ntau cuam tshuam nrog PD.

Nws feem ntau lees txais tias cell apoptosis induced los ntawm 6- OHDA yog kho los ntawm mitochondrial dysfunction, tus cwj pwm los ntawm cytochrome C tso, thiab caspase -3 ua kom [36]. Ib qho apoptotic stimulus ua rau tso tawm ntawm cytochrome C los ntawm mitochondria mus rau hauv cytosol qhov chaw uas nws ces activates caspase -3 thiab lwm yam downstream caspases. Hauv kev tshawb fawb tam sim no, cov txiaj ntsig tau pom tias PC12 hlwb raug rau 100 M ntawm 6-OHDA txhim kho cytochrome C tso tawm thiab caspase nyob ntawm inhibited cytochrome C tso tawm thiab caspase- 3 ua kom muaj zog. Cov txiaj ntsig no qhia tias echinacoside tuaj yeem cuam tshuam mitochondria-mediated apoptosis los ntawm kev txo cov cytochrome C tso tawm ntawm mitochondria mus rau cytosol thiab caspase -3 ua kom.

Inflamation yog ib tug neuropathological feature ntawm Parkinsonian hlwb thiab kuj nyob rau hauv kev sim ua qauv ntawm tus kab mob. Cov tswv yim kho mob tsom mus rau txo qhov mob thiab inhibiting qhov tsim ntawm cov kab mob inflammatory no tej zaum yuav yog ib qho kev cog lus neuroprotective zoo rau kev kho mob Parkinson tus kab mob thiab lwm yam mob [37]. Ntau qhov kev tshawb fawb kuj tau ua nyob rau hauv cov kua cerebrospinal thiab thaum lub sij hawm postmortem tsom xam nyob rau hauv cov neeg mob Parkinsonian qhia tau hais tias nce qib ntawm cytokines, xws li interleukin -1 (IL-1) thiab interleukin -6 (IL{{4}). }), qhia txog kev ua kom aproinflammatory teb [38, 39]. Hauv txoj kev tshawb fawb tam sim no, cov txiaj ntsig tau qhia tias cov txiaj ntsig tau tshwm sim hauv PC12 lub hlwb (OHDAinduced IL-1) thiab IL Txawm li cas los xij, kev kho mob nrog echinacoside tuaj yeem txhaj tshuaj raws li inhibit IL-1 thiab IL{14}} tso tawm los ntawm 6-OHDA. Cov txiaj ntsig tau qhia tias txo cov lus teb inflammatory kuj koom nrog cov txheej txheem ntawm echinacoside tiv thaiv 6-OHDA-induced neurotoxicity.

Hauv kev xaus, cov txiaj ntsig ntawm kev tshawb fawb no tau pom tias echinacoside tuaj yeem thaiv 6-OHDA-induced neurotoxicity hauv PC12 hlwb, thiab cov txheej txheem yuav cuam tshuam txog kev tiv thaiv mitochondrial thiab tiv thaiv kab mob los ntawm kev txo qis ROS ntau lawm. Cov ntaub ntawv muab los ntawm kev ua haujlwm no yuav muaj txiaj ntsig los piav qhia txog cov txheej txheem ntawm echinacoside rau kev tiv thaiv PD. Nws tau txiav txim siab tias echinacoside yuav yog ib qho kev cog lus kho mob rau kev tiv thaiv thiab kho PD.

Tsis sib haum xeeb

Cov kws sau ntawv tau tshaj tawm tias tsis muaj kev cuam tshuam ntawm kev txaus siab

Kev lees paub

Txoj haujlwm no tau txais kev txhawb nqa los ntawm Cov Nyiaj Pab los ntawm National Natural Science Foundation ntawm Tuam Tshoj (81473383) thiab National Science thiab Technology Major Project (2012ZX09103101-078; 2013ZX09103-001-008; thiab 2013ZX09102106).