Isoastilbin Inhibits Neuronal Apoptosis Thiab Oxidative Stress nyob rau hauv Arat Qauv Ntawm Ischemia-reperfusion Injury nyob rau hauv lub hlwb: Kev koom tes ntawm SIRT1/3/6

Yog xav paub ntxiv Emailtina.xiang@wecistanche.com

Abstract

Keeb kwm.Isoastilbin (IAB) has been shown to haveantioxidativethiab anti-apoptotic muaj nuj nqi. Kev tshawb fawb tsis ntev los no pom tias IAB tuaj yeem txo tauoxidativekev ntxhov siab hauv Alzheimer's disease. Txawm li cas los xij, seb puas muaj kev ua haujlwm ntawm IAB los tiv thaiv lwm yam kab mob hauv hlwb tseem tsis paub.
Lub hom phiaj.Txhawm rau tshawb xyuas lub luag haujlwm thiab cov txheej txheem hauv qab ntawm IAB hauv nruab nrab cerebral artery occlusion reperfusion (MCAO/R) hauv cov nas.
Cov ntaub ntawv thiab cov txheej txheem.Txiv neej Wistar nas raug muab faib ua 5 pawg: pawg sham, MCAO/R pawg, thiab 3 MCAO/R pawg tswj hwm IAB (20 mg / kg, 40 mg / kg lossis 80 mg / kg) ib hnub ib zaug rau 3 hnub. Infarction loj, hloov kho Neurological Severity Score (mNSS),oxidativekev ntxhov siabmarkers, thiab neuronal apoptosis markers tau siv los ntsuas qhov ua haujlwm ntawm IAB.
Cov txiaj ntsig.Piv nrog rau pawg MCAO/R, kev tswj hwm ntawm IAB txo qhov loj me thiab mNSS cov qhab nia hauv MCAO/R nas. Isoastilbin kuj txo qis qib ntawm malondialdehyde (MDA) thiab txhim kho kev ua haujlwm ntawm catalase (CAT), superoxide dismutase (SOD), thiab glutathione peroxidase (GSH-PX). Kev kho Isoastilbin txo qis MCAO / R-induced neuronalapoptosispiv nrog pawg MCAO/R, raws li qhia los ntawm cov txiaj ntsig ntawm lub davhlau ya nyob twg deoxynucleotide transferase-mediated X-dUTP nick-end (TUNEL) thiab western blot assays. Isoastilbin kuj thim rov qab MCAO / R-induced downregulation ntawm SIRT1/3/6 protein qhia.
Cov lus xaus.Cov kev soj ntsuam no qhia tias IAB tiv thaivoxidativeKev ntxhov siab thiab neuronal apoptosis hauv cov nas tom qab cerebral ischemia-reperfusion (I / R) raug mob los ntawm kev txhim kho ntawm SIRT1/3/6, qhia tias IAB tuaj yeem yog tus kws kho mob cog lus rau kev raug mob hlwb I / R.
Ntsiab lus:apoptosis, oxidative kev nyuaj siab, isoastilbin, focal cerebral ischemia-reperfusion raug mob

Keeb kwm

Ischemic thiab hemorrhagic stroke yog ib qho ua rau muaj kev tsis taus thiab kev tuag thoob plaws ntiaj teb.1,2Ischemic stroke yog tshwm sim los ntawm kev cuam tshuam hauv cov hlab ntsha, yog li ua rau lub hom phiaj ntawm lub cev muaj kev pheej hmoo ntawm kev tuag ntawm tes.3Txawm hais tias txoj kev kho mob zoo tshaj plaws rau cerebral ischemia yog sai heev. rov muab cov ntshav rov qab, kev kho thrombolytic tuaj yeem ua rau qhov loj me me thiab ua rau muaj kev puas tsuaj rau ischemia-reperfusion (I / R) nyob rau hauv lub hlwb.1,4 Yog li ntawd, yuav tsum tau tsim cov tshuaj tshiab rau kev kho mob cerebral I / R raug mob.

Ischemia-reperfusion raug mob nyob rau hauv lub hlwb muaj txoj kev pathophysiology, nrog rau qhov tshwm sim ntawmoxidativekev ntxhov siab, apoptosis, thiab adenosine triphosphate (ATP) depletion, tag nrho cov no tuaj yeem ua rau muaj kev raug mob neuronal.5 Reactive oxygen hom (ROS) yog tsim thaum lub sij hawm ischemia, ua rau neuronal.apoptosisthiab neurological dysfunction.6 Tshwj xeeb, NADPH oxidases tsim ROS thaum I/R raug mob, uas ua rauoxidativekev ntxhov siab. Qhov kev ntxhov siab no yog cim los ntawm kev nce lipid peroxidation, txo qis catalase (CAT) thiab superoxide dismutase (SOD) kev ua haujlwm, thiab txo qis glutathione (GSH) qib.7-11 Siab ROS thiaboxidativekev ntxhov siabyuav, nyob rau hauv lem, ua rau cell tuag los ntawm mitochondrial apoptotic txoj kev tom qab cerebral I/R raug mob.12 Mitochondrial permeability hloov pores yuav qhib raws li qhov tshwm sim ntawm lub superfluous tsub zuj zuj ntawm ROS, uas ua rau qis mitochondrial transmembrane muaj peev xwm thiab nce zus tau tej cov caspase activators. raws li cytochrome c, thaum kawg pib lub caspase cascade thiab ua rau cell tuag.12 Geng li al. npaj siab tias cov apoptosis ntawm cov hlwb neuronal yog yam tsawg kawg ib nrab kho los ntawm txoj kev mitochondrial apoptosis hauv cerebral I/R raug mob.13 Yog li ntawd, qhov inhibition lossis thim rov qab ntawm oxidative kev nyuaj siab thiab neuronal apoptosis tau cog lus tshiab txoj hauv kev kom txo qis cerebral I/R raug mob.

Txhawm rau tshawb xyuas cov txheej txheem los ntawm qhov uas IAB attenuates I / R-induced kev puas tsuaj, peb tsom mus rau SIRT signaling cascade. Cov kev tshawb fawb tsis ntev los no tau qhia txog kev tiv thaiv lub luag haujlwm ntawm SIRT1/3/6 hauv ntau yam kab mob, suav nrog neural degeneration hauv lub hlwb, cov hlab ntsha o, thiab rog lub cev. Qhov tseeb, peb pom tias IAB kev kho mob nce qib SIRT qhia thaum lub sij hawm I / R raug mob, qhia tias IAB tuaj yeem tiv thaiv cellapoptosislos ntawm upregulating SIRT qhia. Ua ke, peb cov txiaj ntsig tau qhia tias IAB yog qhov muaj txiaj ntsig zoo rau kev kho mob cerebral I / R raug mob.

Lub hom phiaj

Peb lub hom phiaj tag nrho yog los tshuaj xyuas cov teebmeem thiab cov txheej txheem hauv qab ntawm IAB hauv I / R raug mob siv MCAO / R tus qauv tsiaj.

Cov ntaub ntawv thiab cov txheej txheem

Tsiaj txhu

Txiv neej Wistar nas tau txais los ntawm Shanghai Laboratory Animal Company (Shanghai, Suav). Cov nas muaj hnub nyoog 60-90 hnub nrog lub cev hnyav (200-250 g) tau noj cov zaub mov thiab dej thiab khaws cia nyob rau hauv ib puag ncig zoo ib yam (25 ± 2 degree, 40 ± 10 feem pua ​​​​ntawm cov av noo, thiab 12 h lub teeb / tsaus voj voog) . Tsiaj txhu tau ua raws li cov lus qhia rau kev saib xyuas thiab siv cov tsiaj kuaj ntawm Jiamusi College, Heilongjiang University of Traditional Chinese Medicine, Suav. Pawg Saib Xyuas Tsiaj Tsiaj thiab Siv Lub Tsev Kawm Ntawv ntawm Jiamusi College, Heilongjiang University of suav tshuaj pom zoo cov txheej txheem tsiaj (pom zoo No. 20190815).

Tsim MCAO/R raug mob thiab IAB kev kho mob

Lub toxicity ntawm IAB tau txiav txim raws li yav tas los piav.15 Isoastilbin (Chengdu Purechem-Standard Co., Ltd., Chengdu, Tuam Tshoj) yog diluted mus rau zaum kawg concentrations ntawm 20 mg / kg, 40 mg / kg, thiab 80 mg / kg hauv phosphate - buffered solution (PBS). Tsiaj txhu tau muab faib ua 5 pawg: 1. sham; 2. MCAO/R; 3. MCAO/R 20 mg/kg IAB; 4. MCAO/R 40 mg/kg IAB; and 5. MCAO/R 80 mg/kg IAB.

Cov nas tsuag tau tshuaj loog siv intraperitoneal (ip.) txhaj ntawm 50 mg/kg sodium pentobarbital (Sinopharm Chemical Reagent, Beijing, Suav). Txhawm rau qhia MCAO/R, peb nthuav tawm cov hlab ntsha carotid, uas yog sab nraud carotid artery (ECA), txoj cai carotid artery (CCA), thiab sab hauv carotid artery (ICA). Lub ECA thiab CCA tau ze ze ligated. Lub ICA tau ligated siv 0.285 hli monofilament suture, tso rau hauv lub lumen ntawm ICA txog li 18 hli los ntawm ECA stump los cuam tshuam rau nruab nrab cerebral artery (MCA). Ib LaserDoppler (USCN KIT INC., Wuhan, Tuam Tshoj) tau siv los xyuas kom meej tias cov ntshav ntws tau txo qis dua 20 feem pua ​​​​ntawm cov qib ib txwm muaj. Pawg sham tau ua haujlwm zoo ib yam; Txawm li cas los xij, cov hlab ntsha tsis tau ligated. Tom qab 2 h ntawm occlusion, cov hlab ntsha ligated tau rov ua dua thiab cov nas tau muab tshuaj nrog IAB (20 mg / kg, 40 mg / kg, lossis 80 mg / kg) lossis PBS txhua hnub rau 3 hnub sib law liag. Thaum kawg, 72 teev tom qab MCAO/R, qhov muaj peev xwm ntawm cov nas tau raug soj ntsuam.

Ntsuam xyuas nas hlwb infarct ntim

Tom qab IAB kev kho mob, lub hlwb tau nrawm nrawm los ntsuas lawv qhov hnyav ntub. Tom qab ntawd, lub paj hlwb tau qhuav dej ntawm 100 degree rau 24 teev los ntsuas lawv qhov hnyav qhuav. Qhov concentration ntawm cov dej hauv lub hlwb raug xam raws li: (( ntub dej - qhov hnyav qhuav) / ntub qhov hnyav) × 100 feem pua.

Modified Neurological Severity Score (mNSS)

Sensory, motor, balance, and reflex neurological functions raug soj ntsuam 72 h tom qab MCAO kev phais siv mNSS.17 Qhov hnyav ntawm cov hlab ntsha tsis txaus tau nyob rau ntawm qhov ntsuas ntawm 0 mus rau 10, nrog cov qhab nia siab dua qhia tias muaj kev puas tsuaj loj dua rau lub cev. paj hlwb.

Kev ntsuas ntawm superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), thiab glutathione peroxidase (GSH-PX)

Tom qab IAB kev kho mob, cov ntsiab lus protein tau txiav txim los ntawm cov ntaub so ntswg hlwb. Cov protein ntau ntawm cortical homogenates tau txiav txim siab siv cov khoom siv BCA (Beyotime, Shanghai, Suav). SOD, MDA, CAT, thiab GSH-PX tau kuaj pom siv cov khoom siv sib raug (catalog No. A003-1, A001-3, A007, thiab A005; Nanjing Jiancheng Bioengineering Institute, Nanjing, Suav). Luv luv, lub supernatant ntawm cortical homogenate tau sau. Rau SOD, cov qauv raug tsim nrog WST-8/enzyme ua hauj lwm tov ntawm 37 degree rau 30 min. Kev nqus tau ntsuas ntawm lub wavelength ntawm 450 nm. Ib qho SOD enzymatic kev ua ub no (U) tau txhais raws li tus nqi ntawm cov qauv xav tau kom ua tiav 50 feem pua ​​​​ inhibition tus nqi ntawm WST-8 formazan dye. Rau MDA, cov qauv tau muab sib xyaw nrog cov tshuaj ua haujlwm thiab tom qab ntawd rhaub ntawm 100 degree rau 15 min. Qhov nqus ntawm supernatant tau ntsuas ntawm lub wavelength ntawm 532 nm. Rau CAT, cov qauv sib xyaw nrog CAT nrhiav tsis pom thiab cov tshuaj hydrogen peroxide thiab tom qab ntawd incubated ntawm 25 degree rau 5 min. Kev nqus tau pom ntawm lub wavelength ntawm 520 nm. Rau GSH-PX, cov qauv sib xyaw nrog GSH ntawm 37 degree rau 5 min. Qhov nqus ntawm supernatant tau ntsuas ntawm lub wavelength ntawm 412 nm. Ib chav (U) ntawm enzyme kev ua haujlwm tau txhais tias yog tus nqi ntawm GSH-PX hauv 1 mg ntawm cov protein uas catalyzed noj 1 μmol / L GSH thaum txiav tawm cov nyhuv ntawm cov tshuaj tiv thaiv tsis-enzyme.

Terminal deoxynucleotidyl transferase dUTP nick kawg labeling (TUNEL) staining

Ipsilateral hemisphere hlwb cov ntaub so ntswg raug cais, tsau nyob rau hauv 4 feem pua ​​paraformaldehyde, thiab embedded nyob rau hauv paraffin. Tom qab ntawd cov ntaub so ntswg raug txiav rau hauv 5-μm slices thiab antigen raug tom qab 10 min ntawm microwave cua sov hauv citrate tsis. Neuronal apoptosis tau soj ntsuam siv TUNEL kev soj ntsuam (Hauv Situ Cell Detection Kit; Roche, Penzberg, Lub teb chaws Yelemees), raws li cov chaw tsim khoom cov lus qhia. Hauv luv luv, lub hlwb hlais tau muab tso rau hauv cov dej khov txias 4 feem pua ​​​​ntawm paraformaldehyde rau 30 min thiab tom qab ntawd, tau muab tso rau hauv qhov tsaus ntuj nrog TUNEL cov tshuaj tiv thaiv sib xyaw ntawm 37 degree rau 1 h. Tom qab ntawd, cov nuclei tau stained nrog 6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). Cov qauv tau ua duab thiab qhov ntsuas apoptosis raug xam raws li tus naj npawb ntawm TUNEL-zoo hlwb faib los ntawm tag nrho cov cell.

Western blot

Cov ntaub so ntswg nas tau lysed siv xov tooj cua-immunoprecipitation assay (RIPA) tsis (Beyotime, Dalian, Tuam Tshoj). Proteins tau raug rau 12-15 feem pua ​​​​sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) thiab tom qab ntawd xa mus rau polyvinylidene fluoride (PVDF) daim nyias nyias (Millipore, Billerica, USA). Membranes raug thaiv siv 5 feem pua ​​​​ntawm cov mis nyuj tsis muaj rog thiab incubated nrog thawj cov tshuaj tiv thaiv ntawm 4 degree thaum hmo ntuj. Horseradish peroxidase (HRP)-conjugated thib ob cov tshuaj tiv thaiv kab mob tau tom qab incubated ntawm chav tsev kub rau 2 h. Cov cim qhia tshwj xeeb ntawm cov ntawv sau cov protein tau kuaj pom siv lub tshuab chemiluminescence. Cov -actin tau siv los normalize cov protein qhia theem ntawm Bcl-2, Bax thiab Sirtuin (SIRT1/3/6). Cov duab tau txheeb xyuas siv ImageJ software.

Kev txheeb cais

Cov ntaub ntawv raug nthuav tawm raws li qhov nruab nrab ± tus qauv sib txawv (SD) los ntawm 3 qhov kev sim ywj pheej. Cov ntaub ntawv los ntawm txhua pab pawg tau lees paub ua raws li kev faib tawm ib txwm siv Shapiro-Wilk xeem. Qhov sib txawv ntawm cov pab pawg tau soj ntsuam siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA), ua raws li Tukey's post hoc test siv Prism v. 8 software (GraphPad Software, San Diego, USA). Tus nqi ntawm p< 0.05 indicated="" statistical="" significance.="" a post="" hoc="" power="" analysis="" was="" performed="" using="" g*power="" 3.1.9.7="" software="" (heinrich="" heine="" university,="" düsseldorf,="">

Cov txiaj ntsig

Isoastilbin tiv thaiv neurons los ntawm MCAO/R-induced raug mob

Txhawm rau tshawb xyuas yog tias IAB tiv thaiv cov neurons raug mob hlwb I / R, qhov concentration ntawm cov dej hauv lub hlwb thiab cov ntim infarct raug tshuaj xyuas. Nyob rau hauv cov tsiaj tswj, tsis muaj infarction thiab cov dej concentration kuj tsawg (Fig. 1B, C; sham pawg). Tom qab kev phais MCAO/R, peb tau pom qhov nce ntxiv ntawm qhov infarct loj thiab dej concentration (Fig. 1B, C). Infarction thiab ischemic edema qhia tias peb txoj hauv kev tau ua tiav cov qauv I / R hauv cov nas. Interestingly, thaum peb siv ntau qhov sib txawv ntawm IAB (20 mg / kg, 40 mg / kg, thiab 80 mg / kg) tam sim ntawd tom qab raug mob, peb tau pom cov koob tshuaj rov qab los ntawm infarction thiab ischemic cerebral edema (Fig. 1B, C). Qhov txiaj ntsig no qhia tias IAB tiv thaiv ischemia-induced hlwb puas. Tom ntej no, peb soj ntsuam cellapoptosissiv mNSS assay. Raws li pom nyob rau hauv daim duab 1D, cov nas uas ua haujlwm sham tsis pom muaj qhov tsis txaus ntseeg ntawm lub paj hlwb, qhov MCAO/R nas tau pom cov qhab nia mNSS nce ntxiv. Raws li kev hloov pauv morphological qhia hauv daim duab 1B thiab daim duab 1C, IAB kev kho mob tau txo qis mNSS qhab nia ntawm MCAO/R nas (Fig. 1D). Ua ke, cov txiaj ntsig no muab pov thawj tias IAB muaj cov haujlwm tiv thaiv kab mob hauv Vivo nas qauv ntawm cerebral I / R raug mob. Muab hais tias qhov siab dua IAB muaj kev tiv thaiv zoo dua yam tsis muaj kev phiv tshwm sim, peb siv 80 mg / kg rau cov kev sim hauv qab no.

Isoastilbin txo qis oxidative kev nyuaj siab hauv MCAO/R nas

To assess the influence of IAB onoxidative kev nyuaj siab, GHS-PX, MDA, SOD, thiab CAT qib raug soj ntsuam tom qab IAB kev kho mob. Vim qhov nce hauv oxidative kev nyuaj siab, nas raug rau MCAO/R muaj ntau dua MDA piv nrog cov nas tsuag. Cov nyhuv no tau txo qis los ntawm IAB kev kho mob (Fig. 2A). Tsis tas li ntawd, CAT, SOD, thiab GHS-PX qib tau txo qis hauv MCAO/R nas piv rau sham nas (Fig. 2B, D). Kev kho mob nrog IAB nce qib ntawm CAT, SOD, thiab GHS-PX hauv MCAO/R nas (Fig. 2B, D), qhia tias IAB tiv thaiv kev raug mob cerebral I/R los ntawm attenuating oxidative stress.

Isoastilbin inhibits neuronal apoptosis hauv MCAO/R nas

Tom ntej no, peb siv TUNEL kev ntsuam xyuas los ntsuas cov neuronalapoptosistom qab cerebral I/R raug mob. Raws li tau piav qhia hauv daim duab 3A, muaj ntau cov TUNEL-zoo hlwb hauv MCAO/R nas, uas tau txo los ntawm IAB kev kho mob (Fig. 3A). Txhawm rau tshawb xyuas cov txheej txheem hauv qab ntawm IAB kev tiv thaiv, peb tau tshuaj xyuas cov lus qhia ntawm apoptosis proteins Bax thiab Bcl-2 siv cov western blot. Hauv cov nas raug rau MCAO/R, qhov kev qhia ntawm Bax (pro-apoptotic marker) tau raug kho, thaum lub Bcl -2 (anti-apoptotic marker) raug downregulated, piv nrog cov sham pawg (Fig. 3B, C. ). Txawm li cas los xij, IAB kev kho mob tiv thaiv MCAO/R induced hloov (Fig. 3B, C). Cov txiaj ntsig no qhia tias IAB inhibited neuronal apoptosis hauv MCAO/R nas.

Isoastilbin upregulates SIRT qhia

Tom ntej no, peb nrhiav kev tshuaj xyuas seb IAB tiv thaiv neurons li casoxidative kev nyuaj siab. SIRT, NAD ntxiv rau -dependent deacetylase feem ntau nyob hauv mitochondria, tau pom tias muaj feem cuam tshuam nrog cell ciaj sia taus thiab apoptosis, cell metabolism, thiab teb rau kev ntxhov siab. SIRT tuaj yeem qhib cov cim mitochondrial thiab txoj hauv kev los txhawb mitochondria proliferation thiab ATP tiam. Nws kuj tseem koom nrog hauv kev mob, nyob rau hauv ib txoj hauv kev uas txo qis hauv SIRT ua rau muaj qhov nce hauv qhov mob ntev, xws li NFκB thiab RelA/p65 kev ua haujlwm.18-20 Txaus siab, ntau qhov kev tshawb fawb tau pom tias SIRT ua rau muaj kev tiv thaiv neural thiab attenuates oxidative. Kev ntxhov siab hauv pathophysiology ntawm cerebral I/R raug mob.21 Ntawm no, peb tshawb xyuas lub hlwb qhia ntawm SIRT1/3/6 hauv cov nas raug rau MCAO/R. Peb pom tias MCAO/R raug mob txo cov protein ntau ntawm SIRT1/3/6, thaum kho IAB txo cov teebmeem no (Fig. 4A–D). Cov ntaub ntawv no qhia tias IAB tuaj yeem tiv thaiv hauv MCAO/R nas los ntawm kev txhim kho SIRT1/3/6 qhia hauv kev tiv thaiv oxidative kev nyuaj siab.

Kev sib tham

ROS concentration nce mus rau lub ncov, uas tej zaum yuav induce apoptosis los yog cell necrosis.25–28 Muaj ob peb yam zoo-tsim cim ntawm oxidative kev nyuaj siab. Piv txwv li, MDA, cytotoxic compound uas ua los ntawm lipid peroxidation, 29 yog nce hauv nas cardiomyocytes tom qab I/R puas.30 Cov antioxidant enzymes SOD, CAT, thiab GHS-PX plays lub luag hauj lwm tseem ceeb hauv scavenging superoxides thiab tiv thaiv oxidative puas tsuaj.31, 32 Tsis tas li ntawd, kev hloov pauv hauv kev ua haujlwm ntawm cov enzymes no kuj muaj feem xyuam rau oxidative kev nyuaj siab. Attenuating oxidative stress yog ib txoj hauv kev tiv thaiv cov ntaub so ntswg los ntawm I / R raug mob.

Ib txoj kev tshawb fawb tsis ntev los no tau pom tias cov txiaj ntsig neuroprotective ntawm IAB tuaj yeem yog vim kev hloov pauv ntawm oxidative kev nyuaj siab. Tshwj xeeb, txoj kev tshawb fawb pom tias IAB inhibited ROS tiam thiab induced SOD thiab GSH-PX rau ameliorate oxidative puas nyob rau hauv tus nas AD model.15 Txawm li cas los xij, yog thiab yuav ua li cas IAB tiv thaiv I/R raug mob hauv lub hlwb, tseem tsis tau paub. Hauv txoj kev tshawb fawb tam sim no, peb tau pom tias IAB tau ua tiav kev raug mob cerebral I / R los ntawm kev txo qis qhov ntim thiab cov paj hlwb tsis txaus tom qab MCAO / R raug mob. Txhawm rau kuaj seb qhov cuam tshuam ntawm IAB yog vim qhov txo qis ntawm ROS hom, peb ntsuas ntau cov cim ntawm oxidative kev nyuaj siab. Isoastilbin kev kho mob tom qab MCAO/R tau pom tias txo qis MDA qib, tab sis nce CAT, SOD, thiab GSH-PX kev ua haujlwm. Cov txiaj ntsig no qhia tias lub hauv paus txheej txheem ntawm IAB-mediated neuroprotection tiv thaiv I / R raug mob yog los ntawm kev txo qis oxidative kev nyuaj siab.

Tseeb tiag, cerebral I/R muaj peev xwm induce apoptosis ntawm neurons.33,34 Kev ua kom cov pro-apoptotic proteins (Bax thiab Bak) thiab parallel inactivation ntawm anti-apoptotic proteins (xws li Bcl-2) tshwm sim thaum lub sij hawm cerebral. I/R raug mob. Ob leeg, Bcl-2 thiab Bax tau pom nyob rau sab nrauv ntawm mitochondrial membrane thiab koom nrog kev hloov pauv ntawm cell apoptosis.35–37 Cov kev tshawb fawb yav dhau los tau pom tias Bcl-2 overexpression blocks neuronal tuag hauv vitro thiab in vivo.38Erfani et al. pom tias Bax/Bcl{10}} piv tau nce ntxiv thaum lub paj hlwb I/R raug mob, ua rau cov neuronal apoptosis.39 Ntxiv mus, Bax upregulation thiab Bcl{12}} downregulation tau pom nyob rau hauv nas hlwb raug MCAO/R. 40 Tsis tas li ntawd, IAB tau pom los hloov kho cov protein qhia theem ntawm Bcl-2 thiab Bax los txhim kho cov kab ke redox hauv cov nas nrog AD, qhia txog lub luag haujlwm tiv thaiv apoptotic ntawm IAB hauv qhov xwm txheej no.15 Tam sim no txoj kev tshawb fawb, peb tau pom tias kev tswj hwm ntawm IAB txo I / R-induced neuronal apoptosis hauv vivo los ntawm downregulating Bax thiab upregulating Bcl-2. Cov kev tshawb pom no qhia tau hais tias lub peev xwm neuroprotective ntawm IAB yog nyob ntawm nws cov kev tiv thaiv apoptotic hauv nas. Txawm li cas los xij, I/R kuj tseem tuaj yeem ua rau necrosis, uas tsis yog kho los ntawm oxidative stress.25–28 Nws yog qhov txaus siab los tshuaj xyuas seb IAB puas tuaj yeem ua haujlwm los ntawm inhibiting necrosis kom txo qis I / R raug mob.

Txhawm rau tshuaj xyuas cov txheej txheem los ntawm IAB kom txo qis kev ntxhov siab oxidative, peb tsom mus rau SIRT cov proteins. Ib daim ntawv tshaj tawm yav dhau los tau pom tias overexpression ntawm SIRT3 inhibited mitochondrial fission los tiv thaiv cerebral I/R raug mob. Tsis tas li ntawd, SIRT6 tuaj yeem tiv thaiv lub hlwb tiv thaiv I / R kev puas tsuaj los ntawm kev tawm tsam ntawm oxidative stress.2 Ntawm no, peb pom tias SIRT1/3/ 6 tau txo qis hauv MCAO/R nas, qhov kev tswj hwm ntawm IAB tau nce qhov kev qhia ntawm SIRTl/3/6. Rau qhov zoo tshaj plaws ntawm peb txoj kev paub, txoj kev tshawb no yog thawj zaug qhia tias IAB tuaj yeem nce SIRT1/3/6protein qhia kom txo qis oxidative stress thiab neuronal apoptosis induced los ntawm cerebral I / R raug mob.

Kev txwv

Muaj ntau txoj hauv kev tiv thaiv oxidative kev nyuaj siab. Ntawm no, peb pom tau hais tias lub teeb liab mechanism ntawm IAB kev tiv thaiv yog los ntawm SIRT 1/3/6. Txawm li cas los xij, lwm cov txheej txheem tiv thaiv oxidative kev nyuaj siab lossis cerebral I / R raug mob yuav tsum tau piav qhia ntxiv. Thaum I / R raug mob, ob qho tib si apoptosis thiab necrosis koom nrog; Txawm li cas los xij, peb tsuas yog tsom rau apoptosis. Yog li, kev tshawb fawb ntxiv txog lub luag haujlwm ntawm IAB hauv necrosis yuav tsim nyog los piav qhia txog kev tiv thaiv kev ua haujlwm ntawm IAB thaum I / R raug mob.

Cov lus xaus

Ntawm no, peb tau pom tias IAB tuaj yeem txo cov kev ntxhov siab oxidative thiab apoptosis ntawm cov neurons hauv cov nas uas raug rau cerebral I / R. Ntxiv mus, cov tshuaj tiv thaiv oxidative kev nyuaj siab thiab kev tiv thaiv apoptotic ua haujlwm ntawm IAB tuaj yeem tshwm sim los ntawm kev tswj hwm ntawm SIRT1/3/6 qhia. Ua ke, peb cov ntaub ntawv qhia tias IAB yog tus neeg sib tw kho mob rau lub hlwb I / R raug mob.

ORCID iDs Lifeng An  https://orcid.org/0000-0002-4835-1817 Dandan Zhu  https://orcid.org/0000-0001-9119-9956 Xin Zhang  https://orcid.org/0000-0002-9496-1691 Jingwen Huang  https://orcid.org/0000-0001-6449-5362 Guangbao Lu https://orcid.org/0000-0002-2465-8996

Lifeng Ib1,B,D, Dandan Zhu2,B, Xin Zhang2,C, Jingwen Huang1,C, Guangbao Lu1,A,F

1 Jiamusi College, Heilongjiang University of Suav Tshuaj, Suav

2 Kawm Tiav Tsev Kawm Ntawv, Heilongjiang University of Suav Tshuaj, Jiamusi, Suav

A - kev tshawb fawb lub tswv yim thiab tsim qauv; B - sau thiab / lossis sib sau ua ke ntawm cov ntaub ntawv; C - kev txheeb xyuas cov ntaub ntawv thiab txhais lus;

D - sau tsab xov xwm; E - kev hloov kho tseem ceeb ntawm tsab xov xwm; F – kev pom zoo zaum kawg ntawm tsab xov xwm

Cov ntaub ntawv

1. Rossi DJ, Brady JD, Mohr C. Astrocyte metabolism thiab signaling thaum lub hlwb ischemia. Neeb Neurosci. 2007; 10(11): 1377–1386. doi: 10.1038/ nn2004
2. Ribeiro PW, Cola PC, Gatto AR, et al. Kev sib raug zoo ntawm dysphagia, National Institutes Of Health Stroke Scale qhab nia, thiab kev kwv yees ntawm mob ntsws tom qab ischemic stroke. J Stroke Cerebrovasc Dis. 2015; 24(9):2088–2094. doi: 10.1016/j.jstrokecerebrovasdis.2015.05.009
3. Hu X, De Silva TM, Chen J, Faraci FM. Kab mob cerebral vascular thiab kev raug mob neurovascular hauv ischemic stroke. Circ Res. 2017; 120(3): 449–471. doi: 10.1161/circresaha.116.308427
4. Tabassum R, Vaibhav K, Shrivastava P, et al. Perillyl cawv txhim kho kev ua haujlwm thiab cov txiaj ntsig histological tawm tsam ischemia-reperfusion raug mob los ntawm kev txo qis ntawm oxidative kev nyuaj siab thiab kev tsuj ntawm COX-2, NOS-2, thiab NF-κB hauv nruab nrab cerebral artery occlusion nas. Eur J Pharmacol. 2015; 747:190–199. doi: 10.1016/j.ejphar.2014.09.015.
5. Woodruff TM, Thundyil J, Tang SC, Sobey CG, Taylor SM, Arumugam TV. Pathphysiology, kev kho mob, thiab tsiaj txhu thiab cellular qauv ntawm tib neeg ischemic stroke. Mol Neurodegener Cov. 2011; 6(1):11. doi: 10.1186/1750- 1326-6-11
6. Gonzalez-Rodriguez PJ, Xiong F, Li Y, Zhou J, Zhang L. Fetal hypoxia ua rau muaj qhov tsis zoo ntawm hypoxic-ischemic hlwb raug mob hauv cov nas me me: Lub luag haujlwm ntawm glucocorticoid receptors. Neurobiol Dis. 2014; 65:172–179. doi: 10.1016/j.nbd.2014.01.020.
7. Rodrigo R, Fernández-Gajardo R, Gutiérrez R, et al. Oxidative stress thiab pathophysiology ntawm ischemic stroke: Novel therapeutic opportu nities. CNS Neurol Disord Drug Targets.2013;12(5):698–714. doi: 10.2174/1871527311312050015 ib.
8. Zhang C, Ling CL, Pang L, et al. Direct macromolecular tshuaj xa mus rau cerebral ischemia cheeb tsam siv neutrophil-mediated nanoparticles. Theranostics. 2017; 7(13): 3260–3275. doi: 10.7150/thno.19979
9. Kahles T, Luedike P, Endres M, et al. NADPH oxidase plays lub luag haujlwm tseem ceeb hauv ntshav-hlwb thaiv kev puas tsuaj hauv kev sim mob stroke. Mob stroke. 2007; 38(11): 3000–3006. doi: 10.1161/strokeaha.107.489765
10. Chen H, Song YS, Chan PH. Inhibition ntawm NADPH oxidase yog neuroprotective tom qab ischemia-reperfusion. J Cereb Ntshav Flow Metab. 2009; 29(7):1262–1272. doi: 10.1038/jcbfm.2009.47
11. Kapoor M, Sharma N, Sandhir R, Nehru B. Cov nyhuv ntawm NADPH oxidase inhibitor apocynin ntawm ischemia-reperfusion hippocampus raug mob hauv nas hlwb. Biomed Pharmacother. 2018; 97:458–472. doi: 10.1016/j. biopha.2017.10.123
12. Yu S, Wang C, Cheng Q, et al. Ib qho tseem ceeb ntawm Achyranthes bidentata polypeptides muab neuroprotection los ntawm inhibition ntawm mitochondrial-dependent apoptotic pathway nyob rau hauv kab lis kev cai neurons thiab nyob rau hauv cov tsiaj qauv ntawm cerebral ischemia. PLoS Ib. 2014; 9(10):e109923. doi: 10.1371/journal.pone.0109923
13. Geng HX, Li RP, Li YG, et al. 14, 15-EET suppresses neuronal apoptosis hauv ischemia-reperfusion los ntawm txoj kev mitochondrial. Neurochem Res.2017; 42(10): 2841–2849. doi: 10.1007/s11064-017-2297-6
14. Du Q, Li L, Jerz G. Purification ntawm astilbin thiab isoastilbin nyob rau hauv cov extract ntawm smilax glabra rhizome los ntawm high-speed counter-tam sim no chromatography. J Chromatogr A. 2005; 1077(1:98–101. doi: 10.1016/j.chroma. Xyoo 2005.04.072
15. Yu H, Yuan B, Chu Q, Wang C, Bi H. Kev tiv thaiv lub luag haujlwm ntawm isoastilbin tiv thaiv Alzheimer's kab mob los ntawm Nrf2-kev kho cov tshuaj tiv thaiv oxidation thiab anti-apoptosis. Int J Mol Med.2019;43(3):1406–1416. doi: 10.3892/ib. 2019.4058
16. Zhou X, Xu Q, Li JX, Chen T. Kev kho cov qauv ntawm ob flavanonol glycosides los ntawm Smilax glabra. Planta Med. 2009; 75(6): 654–655. ua: 10. 1055/s -0029-1185360
17. Morimoto J, Yasuhara T, Kameda M, et al. Hluav taws xob stimulation txhim khu lub peev xwm tsiv teb tsaws ntawm cov pob txha pob txha stromal hloov hauv cov qauv nas ischemic stroke. Cell Physiol Biochem. 2018; 46(1):57–68. doi: 10.1159/000488409
18. Torrens-Mas M, Pons DG, Sastre-Serra J, Oliver J, Roca P. SIRT3 silencing sensitizes mis mob qog noj ntshav mus rau cytotoxic kev kho mob los ntawm ib tug increment nyob rau hauv ROS ntau lawm. J Cell Biochem. 2017; 118(2): 397–406. doi: 10.1002/jcb.25653
19. Nassir F, Arndt JJ, Johnson SA, Ibdah JA. Kev tswj hwm ntawm mitochondrial trifunctional protein modulates nonalcoholic fatty siab kab mob hauv cov nas. J Lipid Res. 2018; 59(6): 967–973. doi: 10.1194/jlr.M080952
20. Torrens-Mas M, Hernández-López R, Oliver J, Roca P, Sastre-Serra J. Sirtuin 3 silencing txhim kho oxaliplatin kev ua tau zoo los ntawm acetyla tion ntawm MnSOD hauv kev mob qog noj ntshav. J Cell Physiol. 2018; 233(8): 6067–6076. doi: 10.1002/jcp.26443
21. Hernández-Jiménez M, Hurtado O, Cuartero MI, et al. Silent informationa tion regulator 1 tiv thaiv lub hlwb tiv thaiv cerebral ischemic puas. Mob stroke. 2013; 44(8): 2333–2337. doi: 10.1161/strokeaha.113.001715
22. Chen GY, Nuñez G. Sterile o: hnov ​​thiab hnov ​​​​mob rau kev puas tsuaj. Nat Rev Immunol. 2010; 10(12): 826–837. doi: 10.1038/nri2873
23. Eltzschig HK, Eckle T. Ischemia thiab reperfusion: Los ntawm cov txheej txheem rau kev txhais lus. Nat Med. 2011; 17(11): 1391–1401. doi: 10.1038/nm.2507
24. Dziedzic T. Kab mob o raws li kev kho mob lub hom phiaj nyob rau hauv mob ischemic stroke. Expert Rev Neurother.2015;15(5:523–531. doi: 10.1586/14737175.2015.1035712 dr hab.
25. Yemisci M, Gursoy-Ozdemir Y, Vural A, Can A, Topalkara K, Dalkara T. Pericyte contraction induced los ntawm oxidative-nitrative stress impairs capillary reflow txawm muaj kev vam meej qhib ntawm occluded cerebral hlab ntsha. Nat Med. 2009; 15(9): 1031–1037. doi: 10.1038/nm.2022
26. Chen H, Yoshioka H, ​​Kim GS, et al. Oxidative stress nyob rau hauv ischemic hlwb puas: Mechanisms ntawm cell tuag thiab muaj peev xwm molecular lub hom phiaj rau neuroprotection. Antioxidant Redox teeb liab. 2011; 14(8): 1505–1517. doi: 10.1089/ars.2010.3576
27. Olmez I, Ozyurt H. Reactive oxygen hom thiab kab mob ischemic cerebrovascular. Neurochem Int. 2012; 60(2): 208–212. doi: 10.1016/j. nco.2011.11.009
28. Guo J, Cheng C, Chen CS, et al. Overexpression of Fibulin-5 attenuates ischemia/reperfusion raug mob tom qab cerebral artery occlusion nyob rau hauv nas. Mol Neurobiol. 2016; 53(5):3154–3167. doi: 10.1007/s 12035- 015-9222-2
29. Qiang M, Xu Y, Lu Y, et al. Autofluorescence ntawm MDA-hloov cov proteins ua ib qho hauv vitro thiab hauv vivo soj ntsuam hauv oxidative kev nyuaj siab. Protein Cell. 2014; 5(6):484–487. doi: 10.1007/s 13238-014-0052-1
30. Hou S, Zhao MM, Shen PP, et al. Neuroprotective nyhuv ntawm salvianolic acids tiv thaiv cerebral ischemia / reperfusion raug mob. Int J Mol Sci. 2016; 17(7). doi: 10.3390/ijms17071190
31. Adibhatla RM, Hatcher JF. Lipid oxidation thiab peroxidation hauv CNS kev noj qab haus huv thiab kab mob: Los ntawm cov txheej txheem molecular mus rau kev kho mob. Antioxidant Redox teeb liab. 2010; 12(1): 125–169. doi: 10.1089/ ars.2009.2668
32. Staroń A, Mąkosa G, Koter-Michalak M. Oxidative stress in erythrocytes los ntawm cov neeg mob rheumatoid mob caj dab. Rheumatol Int. 2012; 32 (2): 331–334. doi: 10.1007/s 00296-010-1611-2
33. Aşcı S, Demirci S, Aşcı H, Doğuç DK, Onaran İ. Neuroprotective teebmeem ntawm pregabalin ntawm cerebral ischemia thiab reperfusion. Balkan Med J. 2016;33(2:221–227. doi: 10.5152/balkanmedj.2015.15742
34. Tao T, Li CL, Yang WC, et al. Kev tiv thaiv cov teebmeem ntawm propofol tiv thaiv tag nrho cerebral ischemia / reperfusion raug mob hauv cov nas los ntawm inhibition ntawm apoptosis-inducing factor pathway. Brain Res.2016; 1644:9–14. doi: 10.1016/j.brainres.2016.05.006
35. Martinou JC, Youle RJ. Mitochondria hauv apoptosis: Bcl-2 cov neeg hauv tsev neeg thiab mitochondrial dynamics. Dev Cell. 2011; 21(1): 92–101. doi: 10.1016/j.devcel.2011.06.017
36. Yug C, Andrews DW. Lub apoptotic pore ntawm mitochondria: Puas yog peb tawg lossis tseem daig? Cell tuag txawv. 2014; 21(2): 187–191. doi: 10.1038/cdd.2013.169
37. Siddiqui WA, Ahad A, Ahsan H. Qhov paub tsis meej ntawm BCL2 tsev neeg: Bcl-2 proteins thiab apoptosis. Kev hloov tshiab. Arch Toxicol. 2015; 89(3): 289–317. doi: 10.1007/s00204-014-1448-7
38. Maes ME, Schlamp CL, Nickells RW. BAX rau hauv paus: Yuav ua li cas BCL2 noob tsev neeg tswj kev tuag ntawm retinal ganglion hlwb. Prog Retin Qhov muag Res. 2017; 57:1–25. doi: 10.1016/j.preteyeres.2017.01.002
39. Erfani S, Khaksari M, Oryan S, Shamsaei N, Aboutaleb N, Nikbakht F. Nampt/PBEF/visfatin exerts neuroprotective teebmeem tiv thaiv ischemia/reperfusion raug mob los ntawm kev hloov pauv ntawm Bax/Bcl-2 piv thiab kev tiv thaiv ntawm caspase{ {2}} qhib. J Mol Neurosci. 2015; 56(1): 237–243. doi: 10.1007/s12031-014-0486-1

40. Chen L, Cao J, Cao D, thiab al.Cov nyhuv tiv thaiv ntawm dexmedetomidine

tiv thaiv ntshav qab zib hyperglycemia-exacerbated cerebral ischemia/reperfusion raug mob: Ib qho kev kawm hauv vivo thiab hauv vitro, Life Sdi, 2019: 235: 116553.doi; 10.1016/ib.2019116553

41. Zhao H, Luo Y, Chen L, et al. Sirt3 inhibits cerebral ischemia-reperfusion raug mob los ntawm normalizing Wnt / -catenin txoj kev thiab thaiv mitochondrial fission. Cell Stress Chaperones. 2018; 23(5): 1079–1092. doi:10.1007/s12192-018-0917-y

42. Zhang W, Wei R, Zhang L, Tan Y, Qian C. Sirtuin 6 tiv thaiv lub hlwb los ntawm cerebral ischemia/reperfusion raug mob los ntawm NRF2 ua kom. Neuroscience.2017; 366:95–104. doi: 10.1016/j.neuroscience.2017.09.035.