Oxidative Stress, Dysfunctional Energy Metabolism, Thiab Destabilizing Neurotransmitters Hloov Cov Cerebral Metabolic Profile nyob rau hauv ib tus nas qauv ntawm Simulated Heliox Saturation Diving rau 4.0 MPa

Abstract

Lub hom phiaj tseem ceeb ntawm txoj kev tshawb fawb tam sim no yog txhawm rau txiav txim siab txog kev hloov pauv ntawm metabolic hauv lub hlwb ntawm cov nas tom qab simulated heliox-saturated diving (HSD) mus rau 400 meters dej hiav txwv piv rau cov kev tswj tsis pub muaj. Kev hloov pauv hauv cov polar metabolome nyob rau hauv nas hlwb vim HSD tau tshawb xyuas hauv covcortex, hippocampus, thiabstriatum cov ntaub so ntswgkuaj los ntawm kev siv NMR-raws li metabolomic txoj hauv kev ua ke nrog kev tshawb nrhiav biochemical hauv lub cortex. Covtxo glutathione thiab taurine ntau ntautej zaum yuav hypotheticallytxhawb kev tiv thaiv antioxidantthaum lub sij hawm saturation dhia dej, uas kuj proven los ntawm covnce qib malondialdehyde, covtxo superoxide dismutase, thiab covtxo glutathione peroxidasenyob rau hauv lub cortex. Kev txo qis hauv aerobic thiab anaerobic metabolic txoj hauv kev suav nrog downregulated zog metabolism, uas kuj tau ua pov thawj los ntawm biochemical quantification ntawm metabolic enzymes Na-K ATPase thiab LDH nyob rau hauv cerebral cortex cov ntaub so ntswg. Covtseem ceeb metabolic abnormalities ntawm amino acid neurotransmitters, xws li GABA, glycine, thiab aspartate, txo cov aromatic amino acids, suav nrogtyrosinethiab phenylalanine, ob qho tib si uas muaj feem cuam tshuam nrogmetabolism ntawm dopaminethiab noradrenaline, uas yog downregulated nyob rau hauv lub cortex. Qhov tseem ceeb, kev poob qis hauv N-acetyl aspartate yog txuam nrog kev puas hlwb neuronal. Hauv cov ntsiab lus, hyperbaric decompression ntawm 400 msw HSD cuam tshuam rauhlwb metabolomeHauv cov qauv nas, muaj peev xwm suav nrog ntau yam kev cuam tshuam ntawm amino acid homeostasis, metabolites cuam tshuam nrog oxidative kev nyuaj siab thiab lub zog metabolism, thiab destabilizing neurotransmitter Cheebtsam. Cov nokev cuam tshuamtuaj yeem pab txhawb rauneurochemicalthiabNeurological phenotypes ntawm HSD.

Nyem qhov no kom tau txais Anti-Oxidative Cistanche Supplements

Taw qhia

Kev kub siab siab tshaj 1.3 MPa (ntawm kwv yees li 120- meter seawater) yog induced nyob rau hauv tib neeg thiab tsiaj txhu muaj kev pheej hmoo ntawm central nervous system (CNS) hloov [1, 2]. CNS tej zaum yuav yog ib lub hom phiaj rhiab tshaj plaws nyob rau hauv huab cua ntau dhau, cov pa npuas hauv lub cev, thiab kev mob nkeeg (DCS) tshwm sim los ntawm kev dhia dej hiav txwv tob. Nyob rau hauv cov xwm txheej zoo li no, cov kev xav ntawm psychomotor thiab kev paub txog kev paub yog qhov nyuaj heev, nrog rau qhov distal thiab proximal tremors, electroencephalographic abnormalities, fasciculations, myoclonus, pw tsis tsaug zog, xeev siab, mob taub hau, kiv taub hau, thiab txo kev ua tau zoo ntawm kev ntsuam xyuas kev txawj ntse [2-4]. Moen et al. ntsuas lub cheeb tsam neurological txawv txav los ntawm diffusion- thiab perfusion-weighted magnetic resonance imaging (MRI). Perfusion deficits nyob rau hauv cerebral microvascular muaj nuj nqi nrog arterial microemboli [5] tau pom nyob rau hauv North Sea divers, proven los ntawm txo lub sij hawm hloov pauv vim txo complexity ntawm lub microvascular los yog capillary system. Alvhild Alette Bjørkum et al. pom kev cuam tshuam cov protein homeostasis, piv txwv li, hauv synaptic vesicles, thiab destabilizing cytoskeletal Cheebtsam tom qab heliox saturation dhia dej hauv tus qauv nas. Txawm li cas los xij, Arvid Hope et al. qhia tias tsis pom CNS raug mob ntawm kev hloov pauv ntawm morphological nyob rau hauv MRI scan tau pom nyob rau hauv cov nas nrog cov tsos mob ntawm lub paj hlwb loj ntawm decompression mob tom qab heliox saturation decompression [6]. Yog li, peb xav tias muaj peev xwm hloov pauv ntawm cov molecular profile tom qab qhov tseem ceeb tab sis qhov tsis meej ntawm lub cev tsis meej pem yuav raug pom hauv CNS cov ntaub so ntswg tom qab heliox saturation dhia dej.

Kev txheeb xyuas ntawm biomarkers rau kev saib xyuas cov xwm txheej ntawm cellular physiological mechanisms yog qhov tseem ceeb rau kev nkag siab txog cov xwm txheej biochemical. Nws yog ib txwm muaj kev sib tw los txiav txim siab txog qhov nyuaj thiab ntau haiv neeg ntawm txoj hauv kev molecular koom nrog hauv cov lus teb ntawm cov kab mob lom neeg rau ntau yam xwm txheej hauv lub sijhawm tshwj xeeb lossis xwm txheej. Cov kev sim yav dhau los txuas tus neeg biomarkers nrog kev ua haujlwm CNS perturbation tau muab qee qhov kev pom rau oxidative puas thiab lub zog metabolism tom qab dhia dej [3, 4, 7]. Ib yam li ntawd, ntau qhov kev tshawb fawb tau muab piv rau cov amino acid neurotransmitter profiles ntawm cov nas uas muaj cov kab mob siab ntsws rau cov neeg noj qab haus huv [2]. Kev piav qhia txog tus ntiv tes ntawm lub paj hlwb metabolic ntawm CNS kev lom zem tuaj yeem nyiam cov kev xav etiological [8, 9], kev tiv thaiv [10], thiab kev kho mob [11]. Txawm li cas los xij, kev sib koom ua ke ntawm neurological metabolic perturbation induced los ntawm qhov tob ntawm heliox saturation diving tsis tau tshawb xyuas. Xws li cov ntaub ntawv tso tawm los ntawm cov txheej txheem lom neeg muaj peev xwm tuaj yeem sau tau sai vim tias kev nce qib hauv kev siv thev naus laus zis [12]. Daim ntawv thov ntawm high-resolution nuclear magnetic resonance (NMR) spectroscopy tuaj yeem muab ntau qhov nyuaj tab sis txhais tau thiab muaj zog metabolic profiling cov ntaub ntawv. Cov ntaub ntawv spectral ntawm biofluids thiab cov ntaub so ntswg metabolite rho tawm tuaj yeem tau txais los ntawm cov txheej txheem chemometric thiab bioinformatic los qhia txog cov ntaub ntawv ntawm lub cev lossis kab mob. NMR spectroscopy muab cov ntaub ntawv zoo thiab muaj nuj nqis tam sim ntawm kwv yees li 102 qhov sib txawv me me uas muaj nyob hauv cov qauv lom neeg. NMR nrhiav tau ua rau muaj kev sib cav dav dav yam tsis muaj kev xaiv ua ntej ntawm cov txheej txheem biochemical tshwj xeeb. Tsis tas li ntawd, NMR tso cai rau kev soj ntsuam siab dhau los thiab ua kom rov ua tau zoo, thiab nws yog cov txheej txheem ntau yam ntawm cov khoom siv dav dav vim yog cov lus teb ntawm NMR cov teeb liab hauv qhov concentration. Cov thev naus laus zis no tau ua tiav rau cov kab mob neurological xws li cerebellar ataxia [13], Huntington's disease [14, 15], thiab Alzheimer's disease [16, 17] hauv ob qho tib si kev tshawb fawb thiab kev kho mob.

Txawm hais tias muaj txiaj ntsig zoo ntawm NMR-raws li metabolomics, nws daim ntawv thov los ntsuas qhov nruab nrab ntawm cov hlab ntsha hauv nruab nrab ntawm cov hlab ntsha metabolic perturbation cuam tshuam ntawm heliox saturation diving tsis tau tshaj tawm. Lub hom phiaj ntawm txoj kev tshawb fawb tam sim no yog txhawm rau tshawb xyuas cov kev hloov pauv hauv cov kab mob sib txawv ntawm lub paj hlwb (cortex, hippocampus, thiab striatum) thiab biochemical index theem hloov hauv lub cortex hauv tus qauv nas tom qab simulated 400 meters ntawm seawater (MSW) heliox saturation dhia dej. , raws li qhia los ntawm schematic sim tsim ntawm kev tshawb fawb tam sim no nyob rau hauv daim duab 1. Cov duab qhia txog cov metabolic ntiv tes ntawm lub hom phiaj ntawm lub cev yuav muab cov txheej txheem biomarkers muaj uas tuaj yeem pab txhim kho cov txheej txheem dhia dej.

Fig 1. Schematic sim tsim ntawm kev tshawb fawb tam sim no

Cov txheej txheem sim

Peb lees paub tias peb tau nyeem Phau Ntawv Xov Xwm txoj haujlwm ntawm cov teeb meem cuam tshuam nrog kev tshaj tawm kev ncaj ncees thiab lees paub tias daim ntawv tshaj tawm no ua raws li cov lus qhia.

Reagents thiab cov ntaub ntawv Analytical-qib sodium chloride, DMSO, NaN3, NaH2PO4•2H2O, thiab Na2HPO4•12H2O tau yuav los ntawm Sinopharm Chemical Reagent Co. Ltd. (Shanghai, Tuam Tshoj). HPLC-qib CHCl3 thiab CH3OH tau txais los ntawm Merck (Darmstadt, Lub Tebchaws Yelemees). D2O (99.9 feem pua ​​​​hauv D) muaj sodium 3-(trimethyl-silyl) propionate-2, 2, 3, 3, d4 (TSP) raws li tus qauv sab hauv rau kev siv tshuaj hloov pauv tau muab los ntawm Sigma-Aldrich (MO, USA). Ib lub tshuab tsis muaj 0.2 M Na2HPO4/NaH2PO4 hauv D2O ntawm pH 7.4 tau npaj los tiv thaiv pH cuam tshuam rau cov tshuaj hloov pauv ntawm cov metabolites ntawm ntau qhov sib txawv. Cov khoom siv tshuaj ntsuam xyuas rau kev txiav txim siab ntawm sodium-potassium ATPase (Na-K-ATPase), cholinesterase (AChE) thiab lactate dehydrogenase (LDH) tau yuav los ntawm Abcam (USA). Cov khoom siv tshuaj ntsuam xyuas rau dopamine (DA) tau yuav los ntawm RD, Tebchaws Asmeskas, cov khoom siv tshuaj ntsuam rau epinephrine (E) thiab norepinephrine (NE) yog los ntawm Abnova, Taiwan, 5-hydroxytryptamine (5HT) cov khoom siv tshuaj ntsuam xyuas yog los ntawm BioSource, thiab Cov khoom siv tshuaj gamma-aminobutyric acid (GABA) yog los ntawm Santa Cruz, Tebchaws Asmeskas. Cov khoom siv tshuaj ntsuam xyuas ntawm superoxide dismutase (SOD), malondialdehyde (MDA), thiab glutathione peroxidase (GPx) tau yuav los ntawm Cayman, Tebchaws Asmeskas.

Kev pom zoo ntawm Ethics

Tag nrho cov txheej txheem kev sim tau pom zoo los ntawm Pawg Saib Xyuas Kev Ncaj Ncees Tsiaj ntawm Naval Medical Center ntawm PLA, Naval Medical University (pom zoo tsis yog: SYXK(Shanghai)2017-0019, Pom zoo xyoo: 2020). Txhua tus tsiaj tau txais kev saib xyuas zoo raws li Phau Ntawv Qhia Txog Kev Saib Xyuas thiab Kev Siv Cov Tsiaj Laboratory ntawm Naval Medical University. Peb lees paub tias txhua txoj hauv kev tau tshaj tawm raws li ARRIVE cov lus qhia (https://arriveguidelines.org) rau kev tshaj tawm txog kev sim tsiaj. Cov neeg koom nrog tshawb fawb hauv qhov kev tshawb fawb no yog nas, tsis yog tib neeg; yog li ntawd, tsis muaj kev tso cai los koom.

Tsiaj txhu thiab pab pawg tsim

Cov txiv neej laus Sprague-Dawley nas uas muaj hnub nyoog nruab nrab ntawm 8 lub lis piam thiab hnyav 200–220 g tau suav nrog hauv qhov kev tshawb fawb no. Tsiaj txhu tau yuav los ntawm Shanghai Slac Laboratory Tsiaj Co., Ltd. (Shanghai, Suav). Kev siv zog siab tshaj plaws tau ua los txo qis kev txom nyem tsiaj thiab cov tsiaj uas tsim nyog los ntes cov ntaub ntawv ntseeg tau. Tus qauv nas chow thiab dej haus muaj rau txhua tus tsiaj ad libitum. Tsiaj txhu nrog 3 nas rau ib lub tawb tau nyob hauv ib chav tsiaj tsis muaj kab mob (SPF) (kub, 22–24˚C, av noo, 45– 55 feem pua) thiab tswj cov teeb pom kev zoo (12/12 teev lub teeb-dub voj voog) rau ib lub lim tiam ua ntej qhov kev sim. Ua ntej pib qhov kev sim dhia dej, nas tau ua kom zoo rau tsib hnub hauv qhov chaw kuaj mob, suav nrog chav ua haujlwm hyperbaric. Kaum rau tus nas raug hnyav, sau npe, txwv tsis pub muab tso rau hauv kev tswj hwm (qhov raug rau huab cua normobaric tab sis ua rau lub teeb thiab suab nrov zoo sib xws, npe hu ua CON pawg, n=8) pab pawg thiab kev sim (raug simulated heliox saturation dhia mus rau 4.0 MPa (400 msw), hu ua HSD pawg, n=8).

Stimulated saturation dhia dej ntawm 400 msw

Cov nas nyob hauv pawg HSD tau raug thawb hauv helium-oxygen gas hauv lub chav tsev hyperbaric. Hauv luv luv, plaub theem ntawm compression, cia, decompression, thiab khoov saib tau suav nrog

lub sij hawm saturation. Tus nas compression tus nqi yog 1 msw / min mus txog 10 msw nrog 20 feem pua ​​​​Nws O roj sib tov thiab 3.54 msw / min los ntawm 10 msw rau 400 msw nrog ntshiab Nws. Lub sij hawm compression los ntawm qhov chaw mus rau 400 msw siv li ntawm 120 feeb. Cov theem hauv qab ntawm 400 msw yog 120 feeb. Cov pa oxygen concentration tau ntxiv nrog cov pa ntshiab kom tswj tau cov pa ib feem siab ntawm 35 ~ 50 kPa thaum lub sij hawm compression txheej txheem thiab qhov tob cia. Lub sijhawm decompression khoov saib yog 40 min ntawm 300 msw, 45 min ntawm 200 msw, 50 min ntawm 105 msw, 60 min ntawm 45 msw, 75 min ntawm 10 msw, 75 min ntawm 3 msw, thiab tom qab ntawd ntawm qhov chaw. Tus ascent tus nqi siv nyob rau hauv no decompression qauv yog kwv yees li 10 meters (33 ft) ib min. Cov pa oxygen ib feem siab tau khaws cia ntawm 38 ~ 67 kPa thaum lub sijhawm decompression. Tom qab decompression mus rau 10 m, cov pa oxygen concentration tau khaws cia ntawm 20 ~ 24 feem pua. Lub sijhawm saturation diving timeline overview is show in Fig 2. Rats in the air control group (Group CON, hu ua pab pawg tswj, nrog yim nas) tau bred nyob rau hauv atmospheric ib puag ncig nyob rau hauv tib lub chaw soj nstuam nrog tib chamber raws li HSD pawg. Pawg tswj hwm tsis tau txais cov txheej txheem compressing lossis decompressing tab sis ua rau lub suab nrov zoo sib xws thiab lub teeb zoo li HSD pawg.

Qauv sau

Tom qab lub sij hawm decompression, nas raug tshuaj loog nrog pentobarbital sodium ({{0}}}.3 feem pua, 1.0 ml / kg nas nyhav) intraperitoneally, tom qab ntawd tshem tawm ntawm lub hlwb. Sab laug cerebral hemispheres tau txiav sai sai. Hauv ob pawg tsiaj, lub cortex, hippocampus, thiab striatum cov ntaub so ntswg tau muab faib ua ntu zus raws li HSDC, HSDH, HSDS, CONC, CONH, thiab CONS. Raws li cov ntaub so ntswg loj tshaj plaws ntawm peb qhov chaw, txhua tus qauv cortex tau txiav rau hauv ob daim nrog kwv yees li qhov hnyav sib npaug (ib feem ntawm cov qauv rau kev txheeb xyuas metabolomics, lwm daim qauv rau kev ntsuam xyuas biochemical). Tag nrho cov ntaub so ntswg tau snap-kua hauv cov kua nitrogen thiab khaws cia ntawm −80˚C kom txog thaum kev soj ntsuam ntxiv.

Nug ntxiv:

Email: wallence.suen@wecistanche.com

Whatsapp / Tel: ntxiv rau 86 15292862950