Epigallocatechin-3-Gallate Ameliorated Iron Accumululation Thiab Apoptosis Thiab Txhawb Neuronal Regeneration Thiab Memory / Cognitive Functions nyob rau hauv Hippocampus Induced Los Ntawm Kev Raug Mob Rau Ib puag ncig High-Altitude Hypoxia Ib puag ncig

Abstract

Peb tsom los tshawb txog cov teebmeem kev tiv thaiv thiab muaj peev xwm kho cov txheej txheem ntawm Epigallocatechin-3-gallate (EGCG) nyob rau hauv cov tsiaj qauv ntawm kev raug mob ntev nyob rau hauv ib puag ncig high-altitude hypoxia (HAH) ib puag ncig. Kev hloov pauv tus cwj pwm tau raug soj ntsuam nrog Morris dej tshawb nrhiav. Cov hlau txuam hauv hippocampus tau kuaj pom los ntawm kev siv DAB-enhanced Perl's staining, MRI, qPCR, thiab colorimetry, feem. Oxidative stress (malondialdehyde, MDA), apoptosis (Caspase-3), thiab neural regeneration (brain-derived neurotrophic factor, BDNF) tau kuaj pom los ntawm kev siv ELISA thiab western blotting. Neural ultrastructural hloov pauv tau soj ntsuam los ntawm kev xa hluav taws xob microscopy (TEM).

Kev sib raug zoo ntawm apoptosis thiab nco tau ib txwm yog ib qho ntawm cov ntsiab lus kub ntawm cov kws tshawb fawb neuroscientists. Apoptosis yog hais txog cov txheej txheem ntawm kev tuag ntawm tes los ntawm cov txheej txheem kev tswj hwm sab hauv. Raws li ib txwm muaj, cell apoptosis tsis ua rau muaj kev phom sij rau tib neeg lub cev, tab sis pab tswj kev noj qab haus huv. Txawm li cas los xij, nyob rau hauv qee yam mob pathological, kev tswj tsis tau apoptosis tuaj yeem ua rau muaj ntau yam kab mob.

Tib lub sijhawm, kev nco kuj tau pom tias muaj feem cuam tshuam nrog cell apoptosis. Qee cov kev tshawb fawb qhia tias apoptosis tsim nyog ua lub luag haujlwm tseem ceeb hauv kev tsim thiab kev sib koom ua ke ntawm kev nco. Qhov kev sib raug zoo no ua raws li txoj cai yooj yim: qee qhov tsis tsim nyog lossis tas sij hawm cov neurons raug tshem tawm los ntawm apoptosis, tsim ntau qhov chaw thiab cov peev txheej rau kev tsim cov neurons tshiab thiab nco cia. Tsis tas li ntawd, apoptosis tsim nyog kuj tuaj yeem tiv thaiv cov neurons los ntawm kev puas tsuaj los ntawm kev tshem tawm cov tshuaj lom neeg hauv cov neurons.

Txawm li cas los xij, yog tias muaj ntau dhau lossis tsis txaus apoptosis, nws yuav cuam tshuam rau kev ua haujlwm ntawm kev nco. Ntau dhau apoptosis tuaj yeem ua rau ntau dhau ntawm cov neurons, ua rau nco tsis tau. Ntawm qhov tod tes, yog tias tsis muaj apoptosis txaus los tshem tawm cov neurons tsis tsim nyog thiab cov hlwb, cov hlwb no yuav siv ntau qhov chaw txwv thiab cov peev txheej, uas tuaj yeem ua rau kev nco poob. Yog li ntawd, nws yog ib qho tsim nyog kom tsis txhob muaj ntau dhau los yog tsis txaus apoptosis thaum tswj qhov qib ntawm apoptosis.

Apoptosis yog ib qho txheej txheem lom neeg uas muaj ntau yam cuam tshuam. Txawm hais tias kev sib raug zoo rau kev nco tsis tau nkag siab tag nrho, qhov tshwm sim no tso cai rau peb kom nkag siab tob txog lub paj hlwb thiab qhib txoj hauv kev tshiab rau kev txhim kho kev nco. Hauv cov ntsiab lus, kev paub qhov tsim nyog apoptosis yog qhov tseem ceeb rau kev noj qab haus huv ntawm kev nco. Nws pom tau tias peb yuav tsum txhim kho peb lub cim xeeb. Cistanche deserticola tuaj yeem txhim kho kev nco zoo vim Cistanche deserticola yog cov khoom siv tshuaj suav tshuaj suav nrog ntau yam tshwj xeeb, ib qho ntawm kev txhim kho kev nco. Kev ua tau zoo ntawm cov nqaij minced los ntawm ntau yam khoom xyaw uas nws muaj, nrog rau cov kua qaub, polysaccharides, flavonoids, thiab lwm yam. Cov khoom xyaw no tuaj yeem txhawb lub hlwb kev noj qab haus huv ntau txoj hauv kev.

Nyem paub txoj hauv kev los txhim kho lub hlwb

Cov txiaj ntsig tau pom tias kev kawm thiab kev nco ntawm nas tau txo qis thaum raug HAH ib puag ncig. Nws tau ua raws li cov hlau tsub zuj zuj, cov metabolism hauv cov hlau tsis zoo, txo BDNF, thiab kev txhim kho ntawm MDA thiab Caspase-3. TEM tau lees paub qhov hloov pauv ntawm cov txheej txheem hauv cov neurons thiab mitochondria. EGCG txo HAH-induced kev paub tsis meej, hlau deposition, oxidative kev nyuaj siab, thiab apoptosis thiab txhawb neuronal regeneration tiv thaiv mob HAH-mediated neural raug mob.

Ntsiab lus
High altitude · Hypoxia · Quantitative susceptibility mapping · Iron accumululation · Drug intervention.

Taw qhia
Tib neeg kev ua ub no hauv thaj chaw siab siab (ntau tshaj 3000 m) tsis ntev los no tau nce ntau [1]. Ntawm 140 lab tus tib neeg thoob ntiaj teb nyob mus tas li ntawm qhov chaw siab [2, 3] thiab lwm yam rau kev ncig xyuas lossis tiv thaiv ciam teb, 5-10% muaj kev pheej hmoo ntawm kev mob siab rau roob roob uas yog tus cwj pwm los ntawm ntau dhau ntawm erythrocytosis thiab hypoxemia hnyav [4, 5] . Lub hlwb, raws li ib qho ntawm cov khoom siv oxygen ntau tshaj plaws, yog rhiab rau hypoxia [6]. Tsis tas li ntawd, high-altitude hypoxia (HAH) ua rau muaj kev cuam tshuam loj rau kev ncaj ncees ntawm tus thawj xibfwb neurons thiab mitochondrial morphology hauv hippocampus [7]. Cov tsos mob tshwm sim los ntawm kev raug mob mus rau HAH ib puag ncig muaj xws li mob taub hau, kiv taub hau, pw tsaug zog, qaug zog, thiab tsis muaj lub siab xav [5, 8, 9]. Tsis tas li ntawd, HAH tseem tuaj yeem ua rau muaj kev tsis txaus ntseeg xws li kev kawm spatial, nco, thiab mus ob peb vas [10]. Kev kho mob ntawm HAH-induced neural raug mob li no tau los ua ib qho tseem ceeb ntawm kev mloog nyob rau hauv lub teb ntawm high-altitude tshuaj [11, 12].

Nws yog ib qho tseem ceeb heev kom nrhiav tau cov qauv tsim nyog rau kev tiv thaiv HAH-induced hlwb raug mob. Ntsuab tshuaj yej nplooj muaj (−)-epigallocatechin-3-gallate (EGCG) (50–60%), (−)-epigallocatechin (EGC) (15–20%), (−)-epicatechin-3- gallate (ECG) (10–15%) thiab (−)-epicatechin (EC) (5–10%) [13, 14]. EGCG, uas tau tshaj tawm tias muaj ntau dua hauv cov tshuaj yej nplooj (7.1 g ib 100 g) dua li hauv tshuaj yej oolong (3.4 g ib 100 g) thiab cov tshuaj yej dub (1.1 g ib 100 g) [15], muaj cov khoom muaj zog antioxidant vim rau yim pawg hydroxyl thiab ob pawg triphenolic hauv nws cov qauv yooj yim [16, 17]. Tsis tas li ntawd, nws tau pom tias muaj peev xwm hla cov ntshav-hlwb thaiv thiab ncav cuag lub hlwb parenchyma hauv kev tshawb fawb tsiaj [18, 19]. Muaj qee cov lus ceeb toom txog cov txheej txheem tiv thaiv neuroprotective ntawm EGCG xws li hlau chelation zog, oxidative kev nyuaj siab, o, apoptosis, thiab acceleration ntawm paj hlwb regeneration [20–22]. Zhang et al. tshuaj xyuas cov nyhuv ntawm EGCG rau ntau yam kab mob thiab taw qhia tias EGCG tiv thaiv cov hlwb neuronal los ntawm inducing autophagy. Lawv kuj tau sau tseg tias cov tshuaj tiv thaiv kab mob thiab antioxidant ntawm EGCG yog qhov tseem ceeb rau nws txoj haujlwm tiv thaiv kab mob hauv nruab nrab paj hlwb [23].

Txawm li cas los xij, qee qhov kev tshawb fawb tau tshaj tawm cov txiaj ntsig neuroprotective ntawm EGCG tiv thaiv HAH-mediated neural raug mob. Txhawm rau ua kom muaj qhov sib txawv no, hauv txoj kev tshawb no, peb tau tsim cov qauv nas ntawm kev raug mob mus rau HAH ib puag ncig hauv kev sim los txheeb xyuas cov txheej txheem kev kho mob ntawm EGCG. Tsis tas li ntawd, peb tau siv ntau qhov kev tiv thaiv kab mob (QSM), los ntawm gradient-echo MRI ntawm 7 T, uas tuaj yeem kov yeej cov txiaj ntsig tsis nyob hauv cheeb tsam ntawm cov hlau nplaum thiab muab qhov sib piv rau cov ntaub so ntswg hauv vivo, kom muaj nuj nqis ntawm lub hlwb hlau cov ntsiab lus [24].
Cov ntaub ntawv thiab cov txheej txheem

Tsiaj txhu

Tag nrho ntawm 120 tus txiv neej Sprague-Dawley nas hnyav 130-150 g tau txais los ntawm Chengdu Dashuo Laboratory Animal Co., Ltd. Lawv tau khaws cia hauv tsev tsiaj ntawm 18-22 degree hauv 12 teev lub teeb / tsaus ntuj nrog zaub mov thiab dej. muab ad libitum. Tag nrho cov txheej txheem ua rau tsiaj tau pom zoo los ntawm Pawg Saib Xyuas Tsiaj thiab Siv Khoom ntawm West China Tsev Kho Mob.

Kawm Tsim

Cov nas tsuag tau randomized thiab muab faib ua plaub pawg. Cov nas tsuag hauv pawg hypoxia thiab pawg h-EGCG tau noj thiab nyob hauv Yushu, Tuam Tshoj ntawm qhov siab ntawm 4500 m. Cov nas nyob rau hauv ib txwm altitude pawg (n pab pawg neeg) thiab cov n-EGCG pawg tau noj nyob rau hauv Chengdu, Tuam Tshoj ntawm qhov siab ntawm 500 m. Cov nas no tau noj ib txwm noj rau ib hlis, ua raws li kev kho mob sib txawv. (1) Hypoxia pawg: nas tau txhaj tshuaj intraperitoneally txhua hnub nrog physiological saline (0.9%) rau ib hlis. (2) h-EGCG pawg: nas tau txhaj tshuaj intraperitoneally txhua hnub nrog 50 mg / kg EGCG (purity, 98%; Cas, 989-51-5; Sigma-Aldrich; khaws cia ntawm 4 degree) rau ib hlis. (3) n pawg: nas tau txhaj tshuaj intraperitoneally txhua hnub nrog physiological saline (0.9%) rau ib hlis. (4) n-EGCG pawg: nas tau txhaj tshuaj intraperitoneally txhua hnub nrog 50 mg / kg EGCG rau ib hlis.

EGCG (5 mg / mL) tau yaj hauv dej ntawm qhov sib piv ntawm 1: 1. Qhov ntim uas tau txhaj rau hauv nas yog txiav txim siab los ntawm qhov hnyav ntawm cov nas, uas tau mus txog qhov kawg ntawm kev txhaj tshuaj ntawm 50 mg / kg. Tom qab kev kho mob ntawd, qee cov nas tau siv rau Morris dej tshawb nrhiav (n=10 rau txhua pab pawg), thiab kev tsom xam MRI ntawm lub hlwb (n=10 rau txhua pab pawg), feem. Qee lwm cov nas raug txi thiab cov ntaub so ntswg hlwb tau sau rau DAB-enhanced Perls 'staining (n=3 rau txhua pab pawg), rau Western blotting, biochemical ntsuam xyuas, thiab qPCR kev soj ntsuam (n=6 rau txhua pab pawg) , thiab kis electron microscope assay (n=1 rau txhua pab pawg).

Kev sim coj cwj pwm

Morris dej maze (MWM) tau ua raws li tau piav qhia yav dhau los los tshuaj xyuas kev kawm thiab kev nco ntawm nas [25], nws tau ua nyob rau tib qhov chaw uas cov nas tau nyob. MWM muaj ib lub pas dej ua ke (160 cm inch, 60 cm nyob rau hauv qhov siab, thiab 31 cm nyob rau hauv qhov tob) ntim nrog dej mus rau ib theem ntawm 1 cm saum toj no lub platform (10 cm inch thiab 30 cm nyob rau hauv qhov tob). ). Cov dej kub tau khaws cia ntawm 22 ± 2 degree thiab opacified nrog cov xim dub ci ntsa iab. Lub platform tau kho nyob rau hauv ib qho ntawm plaub lub quadrants teem thaum lub sij hawm kev cob qhia rau plaub hnub sib law liag. Qhov kev sim tau xaus thaum cov nas mus txog lub platform. Kev khiav tawm latency tau kaw. Yog tias cov nas tsis tuaj yeem ncav cuag lub platform hauv 60s, lawv tau raug coj mus rau lub platform thiab tso cai rau nyob twj ywm rau 15 vib nas this. Hnub thib tsib, tom qab lub platform raug tshem tawm, cov nas nkag mus rau hauv quadrant rov qab mus rau qhov qub platform. Tom ntej no, tus naj npawb ntawm kev hla platform thiab kev txav mus los tau raug kaw hauv 60 vib nas this.

MRI raws tu qauv

QSM tau soj ntsuam los ntawm MRI, uas tau tshaj tawm hauv kev tshawb fawb yav dhau los [26].MRI tau ua ntawm 7 Tesla scanner (BioSpec 70/30, Bruker, Germany). Ib qho peb-dimensional (3D) ntau ncha gradient-recalled ncha (GRE) raws li tau siv rau QSM. Cov kev sim ntsuas tau teeb tsa raws li hauv qab no: lub sijhawm rov ua dua (TR)=60 ms, fip kaum =15 degree, hlais thickness{10}} hli, tau txais matrix loj =256 × 256, teb of view (FOV)=32 mm × 32 mm, ncha lub sij hawm ntawm thawj ncha (TE1)=5 ms, ncha spacing (ΔTE)=5.77 ms, tus naj npawb ntawm ncha{19 }}, thiab bandwidth =50 kHz. Ob qhov loj thiab theem duab tau txais kev cawmdim rau QSM reconstruction. Muaj peb kauj ruam hauv QSM mapping algorithms ua nrog MATLAB R2014a (The Math Works, Natick, MA), unwrapping the wrapped phase, tshem tawm cov keeb kwm yav dhau los, thiab tsim cov ntawv qhia kev raug mob los ntawm cov ntaub so ntswg.

Kev npaj cov ntaub so ntswg

Cov nas nyob rau hauv txhua pab pawg tau muab tshuaj 10% chloral hydrate rau cov tshuaj loog sib sib zog nqus (1.5 mg / kg) thiab tom qab ntawd transcardially perfused nrog dej khov-txias saline (kwv yees li 30 min) los ntawm lub twj tso kua mis peristaltic (BT100-2 J, LongerPump, Shanghai, Suav teb). Rau DAB-enhanced Perls 'staining, lub paj hlwb raug txiav thiab kho tom qab 2.5% paraformaldehyde hmo ntuj ntawm 4 degree. Hnub tom qab, ntu coronal raug coj los ntawm hippocampus rau staining. Rau lwm qhov kev txiav txim siab sib nrug ntawm immunohistochemistry, hippocampus tau muab tshem tawm sai sai rau hauv dej khov thiab khaws cia ntawm 80 degree.

DAB Enhanced Perls 'Staining

DAB-enhanced Perls staining tau siv los kuaj xyuas cov hlau txuas ntawm tes [27]. Ntu ntawm cov ntaub so ntswg ntawm lub paj hlwb tau muab tso rau hauv dej distilled rau 3 min thiab tom qab ntawd incubated nrog freshly npaj Perls 'solution (2% potassium ferrocyanide / 2% hydrochloric acid) rau 30 min, ua raws li phosphate-buffered saline (PBS) ntxuav. Endogenous peroxidase kev ua haujlwm raug thaiv nrog 0.3% hydrogen peroxide tov hauv methanol rau 15 min, ua raws li 3 ntxuav hauv PBS. Cov teeb liab tau tsim los ntawm incubation rau 3 min hauv 3, 3-diaminobenzidine (DAB) thiab hematoxylin (Sigma-Aldrich) tau siv rau kev tiv thaiv.

Kev soj ntsuam biochemical

Lub hippocampus raug cais tawm thiab sib npaug sib npaug kom tau txais 10% homogenate uas yog tom qab ntawd centrifuged ntawm 30, 000-40,000 RPM rau 10 min kom tau txais ib qho supernatant los kwv yees lub hippocampal hlau los ntawm cov khoom siv xim xim (E-BCK139S, Elabscience, Wuhan, Tuam Tshoj), thiab malondialdehyde (MDA) tau kuaj pom nyob rau hauv supernatant los ntawm kev siv cov khoom siv ELISA (Elabscience, Wuhan, Suav).

Western Blotting

Cov protein concentration tau txiav txim siab siv cov khoom siv tshuaj bicinchoninic acid (BCA, Biosharp, Beijing, Suav). Cov proteins raug cais los ntawm 12% SDS-PAGE thiab tom qab ntawd hloov mus rau polyvinylidene fluoride (PVDF) daim nyias nyias. Cov tom kawg raug thaiv rau 2 h ntawm chav tsev kub hauv 5% skimmed mis nyuj hmoov diluted nrog tsis thiab ces incubated nrog thawj cov tshuaj tiv thaiv thaum hmo ntuj ntawm 4 degree, nrog rau luav anti-cleaved caspase -3 (1: 1000 , Affinity Biosciences, Jiangsu, Suav teb), luav anti-brain-derived neurotrophic factor (BDNF) (1: 1000, Affinity Biosciences, Jiangsu, Tuam Tshoj) thiab luav anti-Actin (1: 5000, Affinity Biosciences, Jiangsu, Suav). Hnub tom qab, cov ntaub so ntswg tau ntxuav peb zaug nrog TBST rau 5 feeb txhua zaus, thiab tom qab ntawd muab tshuaj txhuam nrog HRP-labeled tshis los tiv thaiv tus luav theem nrab cov tshuaj tiv thaiv kab mob (1: 10,000, Servicebio, Wuhan, Tuam Tshoj) rau 1 h, thiab ntxuav. peb zaug rau 5 feeb.

Quantitative Real-Time PCR

Tsiaj Tag Nrho RNA Isolation Kit (Foregene), 5 × All-In-One MasterMix (nrog AccuRT Genomic DNA Tshem Tawm Cov Khoom Siv) (abm), thiab EvaGreen Express2 × qPCR MasterMix-No Dye (abm) tau siv los ntawm cov lus qhia ntawm cov tuam ntxhab. Cov khub tshwj xeeb ntawm primers tau raws li hauv qab no: Fpn, forward primer, 5ʹ-CACCACAGGATATGCTTAAC TCAGG-3ʹ; rov qab primer, 5ʹ-GAGAACAGACAGTC CGAACAAGG-3ʹ; b-actin, pem hauv ntej primer, 5ʹ-TGTCAC CAACTGGGACGATA-3ʹ; rov qab primer: 5ʹ-GGGGTG TTGAAGGTCTCAAA-3ʹ. Fpn mRNA qib ntawm txhua tus qauv tau normalized rau ntawm b-actin mRNA.

Transmission Electron Microscopy (TEM)

Cov hippocampi tau tom qab kho hauv 2.5% glutaraldehyde electron microscope nyob twj ywm kua thiab tom qab ntawd lub cev qhuav dej hauv cov tshuaj acetone ntawm kev nce ntxiv thiab kos nrog Epox 812. Tom qab ntawd, cov seem tau stained nrog uranyl acetate thiab lead citrate. Ultrastructural dluab nyob rau hauv CA3 teb ntawm lub hippocampus ces ntes nrog ib tug kis tau tus mob electron microscope (TEM) nrog ib tug JEM-1400-FLASH (JEOL, Tokyo, Nyiv).

Cov txheej txheem txheeb cais

Cov ntaub ntawv raug nthuav tawm raws li tus qauv sib txawv (SD). Cov kev hloov pauv uas ua tau raws li qhov ntsuas ntsuas parametric tau ntsuas los ntawm kev siv Welch's t-test, ib-txoj kev lossis ob-txoj kev rov ntsuas ntsuas qhov sib txawv (ANOVA), tom qab los ntawm Tukey ntau qhov kev sib piv. Cov kev hloov pauv uas tsis tau raws li qhov kev ntsuas parametric tau ntsuas los ntawm kev siv Mann-Whitney U test, Kruskal-Wallis test, lossis Welch thiab Brown-Forsythe ANOVA. Tus nqi ntawm P<0.05 was considered statistically significant. Statistical analysis and figures were obtained by GraphPad Prism Version 9.0 (GraphPad Software, CA, USA).

Cov txiaj ntsig

Cov txiaj ntsig ntawm EGCG ntawm Kev Kawm thiab Kev Nco Hauv Cov Tsiaj Ua Rau Cov Hniav Hnyav Hnyav

Txhawm rau txheeb xyuas seb EGCG puas muaj feem cuam tshuam rau qhov kev kawm spatial thiab nco kev ua haujlwm ntawm cov nas raug mob HAH, MWM xeem tau ua. Raws li pom nyob rau hauv daim duab 1A, tus naj npawb ntawm cov platform hla los ntawm pawg hypoxia txo qis (P< 0.001, vs. the n group). The treatment of EGCG did not affect the number of crossings of rats in either the normal altitude group (n group vs. n-EGCG group, P>{{0}}.05) lossis pawg HAH (hypoxia pawg vs. h-EGCG pawg, P> 0.05). Tom ntej no, kev khiav tawm latency raug xam raws li lub sij hawm coj los ntawm nas mus txog rau lub zais platform (Fig. 1B). Cov txiaj ntsig tau pom tias nyob rau hnub thib ob mus rau plaub hnub ntawm kev cob qhia, kev khiav tawm latency ntawm nas nyob rau hauv pawg hypoxia tau nce ntau heev (tag nrho cov txiaj ntsig P.<0.01, vs. the n group). Moreover, the h-EGCG group showed reduced escape latency than that in the hypoxia group (all P values<0.05). Also, the swimming speed and distance in MWM were measured. No significant difference was found among these four groups at different time points (All P values>0.05, Fig. 1C). Tom qab ntawd, peb pom tias kev ua luam dej nrawm ntawm cov nas hauv pawg hypoxia txo qis thaum piv nrog n pawg hnub ob thiab hnub peb (tag nrho cov txiaj ntsig P<0.01, Fig. 1D). Also, the swimming speed of rats in h-EGCG group was higher than that in hypoxia group (all P values<0.05, Fig. 1D).

DAB Enhanced Perls 'Staining

Tom qab 8 lub lis piam ntawm HAH raug mob ntev, cov hlau tau nce ntau hauv CA1 thiab CA3 thaj chaw ntawm hippocampus ntawm lub hlwb piv nrog cov pab pawg n (Txhua tus nqi P.<0.01). After EGCG intervention, iron accumulation in CA3 and CA1 of the hippocampus was reduced compared to that of the hypoxia group (P<0.0001, P<0.001, respectively; Fig. 2).

Sib nqus Susceptibility Hloov hauv Hippocampal Regions

Cov txiaj ntsig ntawm kev raug mob hauv hippocampus tau nce ntxiv tom qab HAH raug piv nrog cov pab pawg n (P<0.0001, Fig. 3A). After EGCG intervention, the values decreased compared to those of the hypoxia group (P<0. 0001, Fig. 3A).

Cov teebmeem ntawm EGCG ntawm Hippocampal Oxidative Stress, Hlau, thiab Fpn
Piv nrog rau cov pab pawg n, MDA thiab cov ntsiab lus hlau ntawm pawg hypoxia tau nce siab (P<0.05, Fig. 3B, C). EGCG treatment reduced the levels of MDA and iron, indicating that alleviation of oxidative stress may facilitate EGCG to play a protective role in chronic HAH-induced brain injury in rats. To understand the mechanisms by which brain iron contents were changed by HAH, we further examined the mRNA expression of hippocampal Fpn and found that the Fpn decreased in the hypoxia group and increased in the h-EGCG group (P < 0.01, P < 0.05, respectively, Fig. 3D).

Western Blotting Results

Peb tau soj ntsuam cov teebmeem ntawm EGCG ntawm Caspase {{0}} thiab BDNF theem. Western blotting (WB) kev tsom xam pom tias muaj kev qhia ntau dua ntawm Caspase-3 thiab qis dua ntawm BDNF hauv hippocampus ntawm pawg hypoxia ntau dua li hauv pawg n (tag nrho cov txiaj ntsig P <0.0001, Fig. 4). Ntxiv mus, EGCG kev kho mob txo qis cov theem ntawm Caspase-3 thiab nce qib BDNF (tag nrho cov txiaj ntsig P<0.0001, Fig. 4).

Neural Ultrastructural Hloov

Tom qab 2 lub hlis ntawm HAH raug mob ntev, mitochondria hauv hippocampal CA3 ntawm pawg hypoxia tau o vim qhov txo qis los yog tawg ntawm lub ridge. Tsis tas li ntawd, cov nas hypoxia pom pom condensed nuclear chromatin, nuclear shrinkage, qib siab ntawm endoplasmic reticulum o, nce lysosomes, thiab nce electron ceev. Tom qab kev kho mob nrog EGCG, mitochondrial o o, mitochondrial crystal dissolution, thiab electron ceev tau poob qis dua piv rau cov hauv pawg hypoxia (Fig. 5).

Kev sib tham

Hauv txoj kev tshawb fawb tam sim no, peb pom tias kev kho mob nrog EGCG tau txhim kho kev kawm, kev nco, thiab kev tshawb nrhiav spatial ntawm cov nas hypoxia pawg nas thiab txo cov hippocampal neural raug mob vim raug mob mus rau HAH ib puag ncig. Ntxiv mus, peb kuj pom tias, txawm tias muaj kev txhim kho tseem ceeb hauv txhua qhov ntsuas tom qab kho EGCG, cov ntsuas no tsis tshua rov qab mus rau qhov qub hypoxia.

Cov kev tshawb fawb ua ntej tau pom tias EGCG muaj qee qhov cuam tshuam rau ntau hom kev kawm thiab kev nco tsis zoo [28, 29]. Safar et al. pom tias EGCG tuaj yeem txhim kho kev nco ntawm morphine-kho nas [30]. Raws li lawv cov txiaj ntsig, hauv peb txoj kev tshawb fawb, tom qab kev cuam tshuam txhua hnub nrog 50 mg / kg EGCG hauv cov nas kis mus ntev rau HAH, kev kawm, kev nco, thiab kev tshawb nrhiav spatial ntawm cov kev kawm tau raug txhim kho, qhia tias EGCG tuaj yeem kho kev kawm thiab kev nco tsis zoo ntawm qhov siab.

Ntau cov kev tshawb fawb tau pom tias cov hlau ntau dhau ntawm lub hlwb muaj cov nyhuv neurotoxic vim nws ua rau oxidative puas tsuaj [31]. Nyob rau hauv txoj kev tshawb no, hlau tsub zuj zuj los ntawm MRI, immunohistochemistry staining, thiab biochemical ntsuam xyuas. Tsis tas li ntawd, FPN, raws li tib lub npe hu ua multitransmembrane hlau xa tawm protein nyob rau hauv mammalian hlwb, tau soj ntsuam nrog PCR. Cov txiaj ntsig zoo, cov hlwb zoo ntawm DAB txhim kho Perls 'staining thiab hippocampal hlau cov ntsiab lus nce hauv pawg hypoxia thiab txo qis hauv pawg h-EGCG thaum FPN qhia pom qhov sib txawv. Yog li, peb xaus lus tias HAH-induced hlau tsub zuj zuj yuav tshwm sim los ntawm inhibition ntawm hlau efflux los ntawm ib tug txo nyob rau hauv FPN qhia.

MDA yog cov khoom kawg ntawm lipid peroxidation [32, 33], thiab tuaj yeem siv los ua qhov taw qhia ntawm peroxidation. Cov kev tshawb fawb ua ntej tau pom tias oxidative kev nyuaj siab tuaj yeem nce nrog nce siab [34]. Peb txoj kev tshawb fawb kuj tau qhia tias MDA cov ntsiab lus tau nce ntau ntawm qhov siab. Tsis tas li ntawd, peb tau txheeb xyuas qhov kev qhia ntawm Caspase-3 thiab BDNF. Caspase-3 yog tus thawj coj tseem ceeb tshaj plaws thiab ua yeeb yam ntawm lub davhlau ya nyob twg cleavage enzymes thiab apoptosis [35], thaum BDNF, raws li neurotrophin, uas tuaj yeem tswj hwm kev muaj sia nyob thiab kev sib txawv nrog rau kev txhim kho synaptic kis tau tus mob [36], txhawb kev tsim dua tshiab. Lin et al. qhia tias hypoxia tuaj yeem nce qib Caspase-3 thiab ua rau apoptosis hauv hippocampal neurons [37]. Lee et al. tshaj tawm tias tom qab 3 min ntawm lub ntiaj teb ischemia hauv gerbil, EGCG tiv thaiv ischemia-induced hippocampal cell tuag [38]. Hauv peb txoj kev tshawb fawb, peb kuj pom tias EGCG txo qis qib ntawm cov protein proapoptotic, Caspase-3, thiab nce qhov kev qhia ntawm BDNF.

Mitochondrial kev puas tsuaj thiab kev ua haujlwm tsis zoo yog cov yam ntxwv tseem ceeb hauv lub hlwb hauv qab hypoxia [39]. Hauv peb txoj kev tshawb fawb, kev soj ntsuam ultrastructural kuaj pom muaj qhov sib sib zog nqus staining, mitochondrial o, thiab crista ploj hauv hippocampal neurons nyob rau hauv HAH, thaum hippocampal neurons tom qab EGCG kho tau zoo tiv thaiv los ntawm cov kev raug mob xws li karyopyknosis. Iwona Zwolak tau sau tseg tias EGCG tuaj yeem tiv thaiv cov hlau hnyav hnyav los ntawm kev khaws cia mitochondrial membrane muaj peev xwm thiab txhim kho mitochondrial antioxidant thiab ua pa ua haujlwm [40]. Peb xav tias cov txheej txheem no kuj tseem muaj nyob hauv lub luag haujlwm tiv thaiv EGCG ntawm lub hlwb raug rau HAH ib puag ncig. Hauv kev xaus, EGCG tuaj yeem txo cov hlau tsub zuj zuj, txo qis oxidative kev ntxhov siab, thiab apoptosis, thiab txhawb kev tsim kho neuronal, yog li ameliorating nas lub hlwb puas tsuaj los ntawm kev raug mob mus rau high-altitude hypoxia. Yog li ntawd, nws muaj peev xwm ua tau raws li cov tshuaj tshiab los kho thiab tiv thaiv kab mob hauv roob.

Kev lees paub

Peb xav ua tsaug rau peb lub siab dawb paug rau cov tub ntxhais kawm ntawm West China Tsev Kho Mob, Sichuan University, Chengdu, Tuam Tshoj, uas tau muab peb cov chaw thiab kev pab xws li kev sim qhib-thaj chaw thiab Morris dej maze xeem siv nyob rau hauv txoj kev tshawb no.

Sau Kev Pab Txhawb

CC thiab HC tau xeeb tub, tsim thiab ua qhov kev tshawb fawb no; CC tau ua tiav cov ntaub ntawv thiab sau cov ntawv sau; JC hloov kho cov ntawv sau; BL, BH, YW, DZ, thiab YQ tau pab ua qhov kev sim thiab FG yog lub luag haujlwm rau tag nrho cov txheej txheem.

Nyiaj txiag

Txoj kev tshawb no tau txhawb nqa los ntawm National Natural Science Foundation ntawm Tuam Tshoj (No. 81930046, 81771800, thiab 81829003).

Cov ntaub ntawv muaj

Cov ntaub ntawv tsim tawm thaum lub sijhawm thiab / lossis kev tshuaj xyuas thaum lub sijhawm kawm tam sim no muaj los ntawm tus kws sau ntawv raws li qhov kev thov tsim nyog.

Cov lus tshaj tawm

Kev tsis sib haum xeeb

Cov kws sau ntawv tsis muaj feem cuam tshuam txog nyiaj txiag lossis tsis yog nyiaj txiag los nthuav tawm.

Kev pom zoo rau kev ncaj ncees

Txoj kev tshawb no tau ua raws li cov hauv paus ntsiab lus ntawm Kev Tshaj Tawm ntawm Helsinki. Kev pom zoo tau tso cai los ntawm Pawg Neeg Tshawb Fawb Txog Kev Ncaj Ncees ntawm West China Tsev Kho Mob, Sichuan University, Chengdu, Suav.

Qhib Access

Kab lus no tau tso cai raws li Creative Commons Attribution 4.0 Daim ntawv tso cai thoob ntiaj teb, uas tso cai siv, sib koom, hloov kho, faib, thiab luam tawm hauv ib qho nruab nrab lossis hom ntawv, tsuav yog koj muab credit rau tus thawj sau ) thiab qhov chaw, muab qhov txuas mus rau Creative Commons daim ntawv tso cai, thiab qhia seb puas tau hloov pauv. Cov duab lossis lwm yam khoom siv thib peb hauv tsab xov xwm no suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai tshwj tsis yog tau qhia lwm yam hauv kab qiv nyiaj rau cov khoom siv. Yog tias cov ntaub ntawv tsis suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai thiab koj qhov kev npaj siv tsis raug tso cai los ntawm txoj cai lij choj lossis tshaj qhov kev tso cai siv, koj yuav tsum tau txais kev tso cai ncaj qha los ntawm tus tuav ntaub ntawv.

Cov ntaub ntawv

1. Jain V (2016) Lub hlwb noj zaub mov ntawm qhov siab. Adv Neurobiol 12: 307– 321. https://doi.org/10.1007/978-3-319-28383-8_16

2. Singh LC (2017) High altitude dermatology. Indian J Dermatol 62:59–65. https://doi.org/10.4103/0019-5154.198050

3. Shi J, Wang J, Zhang J, Li X, Tian X, Wang W, Wang P, Li M (2020) Polysaccharide muab rho tawm los ntawm Potentilla anserina L ameliorate acute hypobaric hypoxia-induced hlwb puas hlwb nyob rau hauv nas. Phytother Res 34:2397–2407. https://doi.org/10.1002/ptr.6691

4. Leon-Velarde F, Maggiorini M, Reeves JT, Aldashev A, Asmus I, Bernardi L, Ge RL, Hackett P, Kobayashi T, Moore LG, Penaloza D, Richalet JP, Roach R, Wu T, Vargas E, Zubieta -Castillo G, Zubieta-Calleja G (2005) Cov lus pom zoo ntawm cov kab mob ntev thiab subacute siab siab. High Alt Med Biol 6:147–157.https://doi.org/10.1089/ham.2005.6.147.

5. Villafuerte FC, Corante N (2016) Mob roob siab: kev kho mob, etiology, kev tswj hwm, thiab kev kho mob. High Alt Med Biol 17:61–69. https://doi.org/10.1089/ham.2016.0031

6. Germuska M, Chandler HL, Stickland RC, Foster C, Fasano F, Okell TW, Steventon J, Tomassini V, Murphy K, Wise RG (2019) Dual-calibrated fMRI kev ntsuas ntawm lub paj hlwb metabolic tus nqi ntawm oxygen noj thiab ua tau zoo oxygen diffusivity. Neuroimage 184: 717–728. https://doi.org/10.1016/j.neuroimage.2018. 09.035 Nws

7. Maiti P, Singh SB, Mallick B, Muthuraju S, Ilavazhagan G (2008) High altitude memory impairment is because of neuronal apoptosis in hippocampus, cortex, and striatum. J Chem Neuroanat 36:227–238. https://doi.org/10.1016/j.jchemneu.2008.07.003

8. Turner CE, Barker-Collo SL, Connell CJ, Gant N (2015) Cov pa roj ntsha hypoxic ua pa hnyav ua rau kev txawj ntse thiab kev kawm hauv tib neeg. Physiol Behav 142:104–110. https://doi.org/10. 1016/j.physbeh.2015.02.006

9. McMorris T, Hale BJ, Barwood M, Costello J, Corbett J (2017) Cov nyhuv ntawm kev mob hypoxia ntawm kev paub: kev tshuaj xyuas thiab kev tshuaj xyuas meta-regression. Neurosci Biobehav Rev 74:225–232.https://doi.org/10.1016/j.neubiorev.2017.01.019.

10. Wilson MH, Newman S, Imray CH (2009) Cov teebmeem cerebral ntawm ascent mus rau qhov chaw siab. Lancet Neurol 8:175–191. https://doi. org/10.1016/S1474-4422(09)70014-6

11. Hu S, Shi J, Xiong W, Li W, Fang L, Feng H (2017) Oxiracetam los yog fastigial nucleus stimulation txo kev paub raug mob ntawm qhov siab. Brain Behav 7:e00762. https://doi.org/10.1002/brb3.762

12. Zhang XY, Zhang XJ, Xv J, Jia W, Pu XY, Wang HY, Liang H, Zhuoma L, Lu DX (2018) Crocin attenuates acute hypobaric hypoxia-induced cognitive deficits ntawm nas. Eur J Pharmacol 818: 300–305. https://doi.org/10.1016/j.ejphar.2017.10.042

13. Graham HN (1992) Ntsuab tshuaj yej muaj pes tsawg leeg, noj, thiab polyphenol chemistry. Phau Ntawv Nkauj 21:334–350. https://doi.org/10. 1016/0091-7435(92)90041-f 14. Musial C, Kuban-Jankowska A, Gorska-Ponikowska M (2020) Cov khoom muaj txiaj ntsig ntawm cov tshuaj yej ntsuab catechins. Int J Mol Sci. https://doi.org/10.3390/ijms21051744

15. Eng QY, Thanikachalam PV, Ramamurthy S (2018) Molecular nkag siab ntawm Epigallocatechin gallate (EGCG) nyob rau hauv cov kab mob plawv thiab metabolic. J Ethnopharmacol 210:296–310.

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