Mitochondrial Proteins Unveil The Mechanism Los Ntawm Lub Cev Lub Cev Lub Cev Ameliorates Memory, Kev Kawm Thiab Lub Cev Muaj Zog hauv Hypoxic Ischemic Encephalopathy Rat Model Part 2

2.3. Mitochondrial Proteins nyob rau hauv Nuclei ntawm Hippocampus

Cov ntsuas mitochondrial apoptosis nyob rau hauv qhov chaw nuclear, cytochrome c, apoptosis-inducing factor (AIF), thiab cleaved caspase-3, nrog rau Smac/Diablo thiab OPA1, nce qhov tseem ceeb hauv HIE, raws li pom los ntawm Western blot semi-quantification. (Daim duab 3).

Cytochrome yog ib qho tseem ceeb neurochemical uas pab txhawb kev sib txuas lus ntawm cov neurons thiab pab tib neeg tswj kev nco zoo. Ntau cov kev tshawb fawb tau pom tias cytochromes muaj kev cuam tshuam loj heev rau kev ua haujlwm ntawm tib neeg lub hlwb thiab yog qhov tseem ceeb hauv kev kawm, kev nco, kev paub, thiab lwm yam.

Ua ntej tshaj plaws, cytochromes tuaj yeem txhim kho tib neeg kev mloog thiab kev xav, uas yog ib qho tseem ceeb hauv kev txhim kho kev nco. Lub luag haujlwm ntawm cytochromes tau pom meej dua thaum peb yuav tsum nco ntsoov qee cov ntaub ntawv thiab peb yuav tsum tau mloog zoo rau nws nrog tag nrho cov concentration. Nws tuaj yeem koom tes ua haujlwm ntawm cov neurons thiab txhim kho kev sib kis ntawm cov ntaub ntawv ntawm ntau qhov chaw, yog li pab peb kawm thiab nco qab zoo dua.

Thib ob, cytochromes tuaj yeem txhawb kev sib txuas ntawm lub hlwb neurons thiab ntxiv dag zog rau kev ua haujlwm ntawm cov ntaub ntawv xa mus. Lub xub ntiag ntawm cytochromes modulates synaptic kis tau tus mob thiab ntxiv dag zog rau kev sib txuas synaptic. Cov txheej txheem no txhawb kev sib cuam tshuam thiab cov ntaub ntawv hloov pauv ntawm cov neurons, ua rau lub hlwb hloov pauv thiab ua haujlwm tau zoo.

Thaum kawg, cytochromes tseem tuaj yeem pab tib neeg ua haujlwm zoo dua thiab khaws cov ntaub ntawv nco. Kev tshawb fawb qhia tau hais tias cytochromes ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm kev tsim thiab khaws cia. Nws tuaj yeem ntxiv dag zog rau kev sib txuas thiab kev sib cuam tshuam ntawm cov neurons, yog li pab peb khaws thiab nco qab cov ntaub ntawv zoo dua thiab txhim kho kev ua tau zoo thiab ua haujlwm ntev ntawm kev nco.

Piv txwv li, cytochromes muaj kev cuam tshuam tseem ceeb heev rau lub hlwb kev ua haujlwm thiab kev nco. Peb yuav tsum tsom ntsoov rau kev tswj hwm thiab txhawb kev tsim cov cytochrome, tswj lub hlwb kev noj qab haus huv, thiab txhim kho kev nco los ntawm kev noj zaub mov kom zoo, qoj ib ce, thiab pw tsaug zog zoo. Nws tuaj yeem pom tias peb yuav tsum txhim kho kev nco, thiab Cistanche deserticola tuaj yeem txhim kho kev nco, vim Cistanche deserticola muaj antioxidant, tiv thaiv kev mob, thiab tiv thaiv kev laus, uas tuaj yeem pab txo qis oxidation thiab inflammatory tshwm sim hauv lub hlwb, yog li tiv thaiv cov kab mob. kev noj qab haus huv ntawm lub paj hlwb. Tsis tas li ntawd, Cistanche deserticola kuj tseem tuaj yeem txhawb kev loj hlob thiab kho cov paj hlwb, yog li txhim kho kev sib txuas thiab kev ua haujlwm ntawm neural networks. Cov teebmeem no tuaj yeem pab txhim kho kev nco, kev kawm, thiab kev xav nrawm, thiab tseem tuaj yeem tiv thaiv kev loj hlob ntawm kev paub tsis meej thiab kab mob neurodegenerative.

Nyem paub cov tshuaj ntxiv los txhawb kev nco

Cytochrome c (Daim duab 3B), cleaved caspase-3 (Daim duab 3C), thiab Smac/Diablo (Daim duab 3D) tau hais txawv txawv hauv HIE nrog thiab tsis muaj kev tawm dag zog tab sis tsis muaj kev txheeb cais tseem ceeb sib txawv thaum piv rau SHAM, NT . Txawm li cas los xij, nuclear AIF thiab OPA1 (Daim duab 3A, E) tau pom qhov tseem ceeb.

2.4. Mitochondrial Proteins hauv Cytoplasm ntawm Cerebral Cortex

Cov ntsuas mitochondrial apoptosis hauv cytoplasmic feem, apoptosis-inducingfactor (AIF), cytochrome c, cleaved caspase-3, nrog rau Smac/Diablo thiab OPA1, nce qhov tseem ceeb hauv HIE, raws li pom los ntawm Western blot semi-quantification (Fig 4).

AIF (Daim duab 4A), Cytochrome c (Daim duab 4B), thiab Smac/Diablo (Daim duab 4D) tau qhia txhua yam txawv hauv HIE nrog thiab tsis muaj kev tawm dag zog tab sis tsis muaj qhov sib txawv ntawm qhov sib txawv thaum piv rau SHAM, NT. Txawm li cas los xij, cleaved caspase-3 (Daim duab 4C) thiab OPA1 (Daim duab 4E) tau pom cov kev hloov pauv tseem ceeb.

2.5. Mitochondrial Proteins hauv Nuclei ntawm Cerebral Cortex

Cov ntsuas mitochondrial apoptosis nyob rau hauv qhov chaw nuclear, cytochrome c, apoptosis-inducing factor (AIF), thiab cleaved caspase-3, nrog rau Smac/Diablo thiab OPA1, nce qhov tseem ceeb hauv HIE, raws li pom los ntawm Western blot semi-quantification. (Daim duab 5), AIF (Daim duab 5A), Cytochrome c (Daim duab 5B), cleaved caspase-3 (Daim duab 5C), thiab OPA1 (Daim duab 5E) tau tag nrho cov qhia txawv nyob rau hauv HIE thiab kev tawm dag zog thiab tau statistically tseem ceeb piv rau SHAM, NT.

Kev hloov pauv hauv nuclear Smac/Diablo (Daim duab 5D), ntawm qhov tod tes, tsis yog qhov tseem ceeb.

Ua ke, cov kev tshawb pom no txhawb cov lus pom zoo tias kev tawm dag zog txhim kho lub cev muaj zog, kev kawm, thiab kev nco rov qab los ntawm kev tawm tsam hippocampal thiab corticalmitochondrial apoptotic protein qhia hauv lub hlwb ntawm HIE nas.

2.6. Immunofluorescence Analysis ntawm Proteins nyob rau hauv lub cev muaj zog Cortex

Daim duab 6 qhia cov duab sawv cev immunofluorescence los ntawm lub cev muaj zog cortex ntawm cheeb tsam cerebral cortex stained rau AlF, Cytochrome C, Cleaved caspase-3, Smac, thiab OPAl Cov kev soj ntsuam ntau ntawm qhov nruab nrab fluorescent siv (MFI) ntawm cov proteins kuj tau sau tseg. nyob rau hauv daim duab 6 thiab 7. $wimming ce ua rau ib tug tseem ceeb txo nyob rau hauv cov kev qhia ntawm cov proteins. cov duab thiab qhov nruab nrab fluorescent siv (MFI) graphs.

Daim duab 6. Immunofluorescence cov duab thiab txhais tau tias fluorescent intensities ntawm cov proteins nyob rau hauv lub motorcortex. (A) Cov duab sawv cev immunofluorescence thiab MFI ntawm AIF nyob rau hauv lub cev muaj zog cortex ntawm txhua pab pawg. AIF molecules yog ntsuab. AIF molecules tau txheeb xyuas hauv lub cev muaj zog cortex.

Cov txiaj ntsig tau nthuav tawm raws li Qhov Ntsuas ± SEM ntawm 3 nas los ntawm txhua pab pawg. (** p < 0.001 thiab *** p < 0.0001) piv nrog SHAM, NT pawg; (## p < 0.001 piv nrog pawg HIE, NT; txhua pab pawg, n=3). Scale bar, 100 µm. (B) Cov duab sawv cev immunofluorescence thiab MFI ntawm Cytochrome C hauv motorcortex ntawm txhua pab pawg. Cytochrome C molecules (ntsuab) tau txheeb xyuas hauv lub cev lub cev.

Cov ntaub ntawv raug nthuav tawm raws li Qhov Ntsuas ± SEM ntawm 3 nas los ntawm txhua pab pawg. (** p < 0.001 thiab *** p < 0.0001) piv nrog SHAM, NT pawg; (## p < 0.001 piv nrog HIE, NT pawg); txhua pab pawg, n=3. Scalebar, 100 µm. (C) Cov duab sawv cev immunofluorescence thiab MFI ntawm Cleaved caspase-3 nyob rau hauv themotor cortex ntawm txhua pab pawg. Cleaved caspase{10}} cov molecules (ntsuab) tau txheeb xyuas hauv lub cev lub cev.

Cov txiaj ntsig tau nthuav tawm raws li Qhov Ntsuas ± SEM ntawm 3 nas los ntawm txhua pab pawg. (* p < 0.05 thiab *** p < 0.0001) piv nrog SHAM, NT pawg); txhua pab pawg, n=3. Scale bar, 100 µm. (D) Cov duab sawv cev immunofluorescence thiab MFI ntawm SMAC nyob rau hauv lub cev muaj zog cortex ntawm txhua pab pawg.

SMAC molecules (ntsuab) tau soj ntsuam nyob rau hauv lub cev muaj zog cortex. Cov txiaj ntsig tau nthuav tawm raws li Qhov Ntsuas ± SEM ntawm 3 nas ntawm txhua pawg. (** p < 0.001 thiab *** p < 0.0001) piv nrog SHAM, NT pawg); txhua pab pawg, n=3.Scale bar, 100 µm. (E) Cov duab sawv cev immunofluorescence thiab MFI ntawm OPA1 hauv motorcortex ntawm txhua pab pawg. OPA1 molecules (ntsuab) tau soj ntsuam nyob rau hauv lub cev muaj zog cortex.

Cov ntaub ntawv tus kheej tau nthuav tawm raws li Qhov Ntsuas ± SEM los ntawm 3 nas hauv txhua pab pawg. (*** p < 0 0001) piv nrog pawg SHAM-NT; (## p < 0.001 piv nrog HIE, NT pawg); txhua pab pawg, n=3. Scale bar, 100 µm.

3. Kev sib tham

Vim tias mitochondria yog qhov tseem ceeb rau kev tsim hluav taws xob hauv hlwb [23,24], peb tau tshawb xyuas cov txiaj ntsig ntawm kev ua luam dej ntawm mitochondrial apoptotic thiab dynamic signals hauv HIE.

Txhawm rau ua kom tiav qhov no, peb tau siv cov kev tawm dag zog niaj hnub thiab soj ntsuam tsib lub mitochondrial muaj nuj nqi regulators (AIF, Cytochrome c, Cleaved caspase-3, Smac/Diablo, thiab OPA1). Peb cov txiaj ntsig tau pom tias kev cob qhia kev tawm dag zog cuam tshuam rau cov protein no tsis hais txog kev noj qab haus huv ntawm lub xeev (uas yog, hauv HIE nas qauv lossis pawg sham).

Kev ua luam dej ua rau txo qis kev qhia ntawm mitochondrial apoptosis-txog cov proteins; AIF, cytochrome c, thiab cleaved caspase-3 nyob rau hauv cytosol thiab nuclei ntawm lub hippocampus, thiab cortex, uas yog raws li cov nqe lus uas tau ua los ntawm Moore li al. [25] uas lub cev ua haujlwm cuam tshuam txhua yam ntawm mitochondrion.

Tsis tas li ntawd, plaub lub lis piam ntawm kev tawm dag zog ua rau txo qis cov lus qhia ntawm cov cim fission Smac / Diablo thiab cristae remodel-ing protein OPA1, uas tau ua ke nrog kev txhim kho lub cev muaj zog, kev kawm, thiab kev nco qab.

Nyob rau hauv lub hippocampus thiab cerebral cortex ntawm HIE thiab sedentary nas, cov theem ntawm mitochondrial apoptotic ntsuas; cytochrome c, AIF, thiab cleaved caspase-3 nce qhov tseem ceeb. Hauv kev sib piv rau SHAM, NT nas, thaum HIE thiab cov nas ib txwm raug ua luam dej tawm dag zog, qib ntawm AIF, cytochrome c, thiab cleaved caspase-3 poob qis, qhov kev soj ntsuam hauv tandem nrog [24,26] kev tshawb pom tias kev tawm dag zog rov qab los. mitochondria ua haujlwm hauv cov kab mob neurodegenerative.

Nrog AIF tau taw qhia tias yog ib qho tseem ceeb contributorto neuron poob hauv lub hlwb tsis paub qab hau tom qab hypoxia-ischemia thiab hypomorphicmutation ua rau txo qis AIF qhia qhia los tiv thaiv neonatal hypoxicischemia [27], peb pom ntawm no tias kev ua luam dej muaj txiaj ntsig zoo ib yam hauv HIErat lub hlwb los ntawm inhibiting cov protein's cytosolic thiab nuclear translocation.

Qhov cuam tshuam antiapoptotic ntawm kev tawm dag zog sib raug rau kev txhim kho hauv kev ua haujlwm ntawm lub cev muaj zog, kev kawm, thiab kev nco, qhia tias kev cob qhia kev tawm dag zog cuam tshuam mitochondrial malfunction andapoptosis.

Qhov no piav qhia txog cov txheej txheem tiv thaiv neuronal tau tshaj tawm tom qab kev tawm dag zog uas txhawb nqa neurogenesis thiab kho myelin hauv penumbra tom qab mob stroke [28], vim tias cov no yog cov txheej txheem xav tau lub zog siab uas yuav tsum tau ua haujlwm ruaj khov mitochondria.

Muab kev tawm dag zog lub peev xwm los hloov kho mitochondrial protein stabilization, thiab txhim kho lub cev muaj zog, kev kawm, thiab kev nco, nws yuav yog qhov zoo tshaj plaws los txiav txim siab seb qhov kev tawm dag zog ntsig txog metabolite, lactate, uas hla cov ntshav-hlwb barrier [29] contributessignificantly rau kev tawm dag zog cov txiaj ntsig zoo ntawm suppressing apoptosis. thiab txhawb nqa motorfunction, kev kawm, thiab kev nco hauv HIE.

Kev ua luam dej ua tau zoo txhim kho lub cev muaj zog, kev nco, thiab kev kawm inrats nrog HIE los ntawm kev txo qis mitochondrial apoptosis los ntawm Cyto.C/Cleaved Caspase-3 thiab AIF cov kev taw qhia.

Ib cag ntawm qhov ntawd, kev cuam tshuam kev ua luam dej tau raug qhia kom ruaj khov mitochondrial cristae thiab daim nyias nyias muaj peev xwm hauv HIE nas, raws li pov thawj los ntawm Smac / Diablo thiab OPA1 thim rov qab hauv cov tsiaj no, uas zoo ib yam nrog [30,31] nrhiav pom qhov tseem ceeb ntawm OPA1 hauv mitochondrial cristae. ruaj khov.

Nrog rau cov txheej txheem neurobiological hauv qab ntawm kev tawm dag zog-induced neuroplasticity tseem feem ntau tsis txaus ntseeg [8], peb cov kev tshawb pom tau muab lub platform zoo nkauj thiab plausible rau kev tshawb fawb yav tom ntej uas txheeb xyuas cov txheej txheem molecular ntxiv uas tuaj yeem hloov kho kom zoo tswj HIE thiab qhov tshwm sim tsis zoo.

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