Hemodialysis Raises Oxidative Stress Los Ntawm Carbon-centered Radicals Txawm tias Txhim Kho Biocompatibility

Leukocyte activation thiab qhov tshwm simoxidative kev nyuaj siabinduced los ntawmbioincompatible cov ntaub ntawvthaumhemodialysis cuam tshuamkev kwv yees ntawm cov neeg mob. Txawm tias muaj ntau yam kev nce qib hauvhemodialysis dialyzers, covprognosis ntawm hemodialysiscov neeg mob uas muaj teeb meem sib sib zog nqus rauoxidative kev nyuaj siab, zoo lintshav qab zib mellitus tseem tsis zoo. Yog li, peb rov ntsuam xyuas cov teebmeem ntawmhemodialysisntawm ntau hom reactive oxygen siv electron spin resonance-raws li txoj hauv kev rau kev txhim kho ntxiv ntawmbiocompatibilityhauvhemodialysis. Peb tau tso npe rau 31 tus neeg mob nyob rau hauv qhov chaw ruaj khov uas ua rau hemodialysis siv high-flux polysulfone dialyzers. Cov teebmeem ntawm hemodialysis ntawm cov pa oxygen reactive tau raug soj ntsuam los ntawm ob txoj hauv kev: MULtiS, uas ntsuas cov dej hauv cov dej ntws tawm tsam ntau hom hydrophilic reactive oxygen, thiab i-STrap, uas kuaj pom lipophilic carbon-center radicals. Zoo ib yam li cov kev tshawb fawb yav dhau los, peb pom tias cov ntshav hydroxyl radical scavenging ua haujlwm tau zoo dua tom qab hemodialysis. Tsis zoo li cov kev tshawb fawb yav dhau los, peb tau tshawb pom tias kev ua haujlwm pov tseg tawm tsam alkoxyl radicals tau txo qis tom qabhemodialysis. Tsis tas li ntawd, cov neeg mob ntshav qab zib mellitus tau pom qhov txo qis hauv cov tshuaj tua kab mob tawm tsam alkyl peroxyl radicals thiab nce lipophilic carbon-center radicals tom qab hemodialysis. Cov txiaj ntsig no qhia tau hais tias txawm tias muaj kev txhim kho dav hauv cov dialyzer membranes, cov ntaub ntawv ntawmreactive oxygen homuas tuaj yeem raug tshem tawm thaum lub sijhawm lim ntshav raug txwv, thiab ntau hom pa oxygen reactive tseem nyob ntawm qib nce thaum lub sijhawm hemodialysis.

Lo lus tseem ceeb:hemodialysis, biocompatibility, alkyl peroxyl radical electron spin resonance, MULTiS, i-STrap

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Mob raum mob(CKD) paub tias muaj feem nrogsiab oxidative kev nyuaj siab. (-4)Kev sib sau ntawm uremic toxins nrog pro-oxidative zog ua raumicroinflamationtshwm sim los ntawmnce ntawm pro-inflammatory cytokinesyog lub ntsiab feature ntawmCKD kab mob pathophysiology. (s-, Hemodialysis (HD), kev kho lub raum hloov pauv dav dav, ua haujlwm zoo li rab ntaj ob sab kom txo qis oxidative kev nyuaj siab hauv cov neeg mob CKD. Ua kom muaj cov kab mob polymorphonuclear leukocytes, vim yog kev sib cuag nrog daim nyias nyias lossis lwm qhov chaw dag hauv HD circuits. , yog lub ntsiab ua rau oxidative kev nyuaj siab induced los ntawm HD.s-) Conversely, kev tshem tawm ntawm pro-oxidative uremic toxins, tshwj xeeb tshaj yog tsis tshua muaj-molecular-hnyav hydrophilic tebchaw xws li trimethylamine-Noxideor guanidino tebchaw, yuav txhim kho lub antioxidative xwm ntawm HD, ua rau kev txhim kho mus sij hawm ntev ntawm kev ciaj sia thiab kev ua neej zoo nyob rau hauvcov neeg mob nrog CKD. (2,13) ​​Tsis tas li ntawd, kev kho cov acidosis ua rau rov ua haujlwm ntawm cov tshuaj tiv thaiv oxidative thiab txo cov ntim ntau dhau ua rau kev tiv thaiv cardio. Cov teeb meem vascular yog cov tshuaj tiv thaiv oxidative. (4 Yog li. yog qhov tsim nyog rau HD cov ntaub ntawv kom txo qis leukocyte ua kom muaj zog thiab ua kom muaj zog tiv thaiv oxidative. Kom kov yeej cov teeb meem no tiv thaiv oxidative thiab biocompatible HD cov ntaub ntawv ua piv txwv los ntawm vitamin E-coated dialyzers tau tsim. Qhov kev kwv yees ntawm HD cov neeg mob tseem tsis txaus siab, tshwj xeeb tshaj yog cov neeg mob ntshav qab zib mellitus (DM), ib tus kab mob uas txhawb nqa lub xeev oxidative heev.oxidative kev nyuaj siab cuam tshuamtsis tau raug soj ntsuam txaus. Yog li, txoj kev tshawb no tsom kom meej meej cov txiaj ntsig ntawm HD ntawm cov kab mob oxygen reactive (ROS) dynamics, raws li cov xwm txheej saum toj kawg nkaus ntawm oxidative kev ntxhov siab, raws li lub tswv yim uas txhua lub hom phiaj ntawm oxidative thiab tshuaj tiv thaiv antioxidant hauv lub cev yuav tsum tau piav qhia txog biochemically. nyob rau hauv ib lub sij hawm thiab qhov chaw. Ib qho kev piav qhia rau qhov no yog atsis muaj kev soj ntsuam ntxawsntawm sab sauv ntawm oxidative kev ntxhov siab cuam tshuam txog cov tshuaj tiv thaiv uas ua rau cov tshuaj tiv thaiv cellular. Kev txheeb xyuas ntawm ROS uas ua raws li stimulators ntawm oxidative kev ntxhov siab tshwm sim los yog kev sib cuam tshuam ntawm ROS los tsim oxidative stimulators yog ib qho nyuaj vim muaj cov tshuaj tiv thaiv siab thiab complex cov tshuaj tiv thaiv chains. Txij li thaum ROS tsis zoo ib yam thiab ib tus neeg ROS muaj cov yam ntxwv tshwj xeeb thaum muaj kev cuam tshuam vivo, yuav tsum muaj kev tshuaj xyuas ntau yam ROS. (9-2n Yog li, txhawm rau txhim kho biocompatibility raws li kev siv cov khoom siv tshuaj tiv thaiv oxidative thiab biocompatible, ib tug yuav tsum nkag siab txog cov kev hloov hauv ROS dynamics tshwm sim los ntawm HD.

Txhawm rau tshawb xyuas ROS dynamics thaum lub sijhawm HD, peb siv ob txoj kev tsim tshiab electron spin resonance (ESR) raws li txoj hauv kev. ntau yam hydrophilic dawb-radical scavenging assay (MULTIS) thiab lipophilic radical nrhiav kom tau ntawm cov ntshav tag nrho (i-STrap) Txawm hais tias kev tshawb fawb ntawm ROS scavenging kev ua los ntawm ESR tsis yog qhov dav, nws tseem yog tib txoj kev muaj rau kev txheeb xyuas hom thiab tshuaj xyuas cov dynamics. ntawm ROS. Txoj kev sib txuas ua ke ua kua dej chromatography hom kev ua haujlwm siab nrog ESR system thiab ntiav 5-(2,2-dimethyl-1}3-propoxycyclophosphoryl)-5- methyl-1-pyrrolineNoxide(CYPMPO) ua cov cuab yeej. MULTIS muab ntau dua rhiab heev thiab ruaj khov rau ROS scavenging kev ntsuas dua li cov kev ntsuam xyuas ESR ib txwm ua. Txawm li cas los xij, lawv tsis tau ntsuas qhov cuam tshuam ntawm HD.e2) Tsis tas li ntawd, txoj kev i-STrap yog ib txoj hauv kev ESRbased uas kuaj pom ROS hauv cov ntshav tag nrho siv 2-diphenylphosphinoyl-2-methyl-3,{ {20}}dihydro-2H-pyrrole N-oxideDPhPMPO) raws li nws cov ntxiab ntxiab thiab ntsuas ESR tom qab kev rho tawm cov organic, yog li cuam tshuam lipophilic ROS.2s.26 Hauv txoj kev tshawb no, peb pom tias theem ntawm ntau ROS yog nce thiab tsis raug tshem tawm los ntawm HD, qhia tau hais tias radical chain reactions tseem tsis tau tswj kom txaus raws li HD. Qhov no yuav cuam tshuam rauprognosis ntawm cov neeg mob nrog CKD.

Cov ntaub ntawv thiab cov txheej txheem

Cov ntsiab lus thiab kev tso cai pom zoo. Tag nrho cov txheej txheem ua rau tib neeg cov ntsiab lus tau ua tom qab tau txais kev tso cai sau ntawv rau tus kheej. Cov txheej txheem siv tau pom zoo los ntawm Tsukuba University of Technology Committee (AuthorizationNo. 201809). Qhov no yog ib qho kev tshawb fawb soj ntsuam; yog li ntawd, kev cuam tshuam tau ua rau cov neeg mob. Tag nrho ntawm 31 tus neeg mob HD ruaj khov tau suav nrog hauv txoj kev tshawb no. Peb tau txais thiab sau cov ntaub ntawv kho mob thiab pej xeem rau txhua tus neeg mob. Txhua tus neeg mob tau kho nrog high-flux polysulfone membrane dialyzers (Toray Medical Co., Ltd., Tokyo, Nyiv) nrog rau ntau qhov chaw ntawm daim nyias nyias, yoog raws lub cev kev cai ntawm txhua tus neeg. Cov neeg mob tau muab faib ua ob pawg nyob ntawm qhov ua rau lawv lub raum mob raum kawg: pab pawg uas tsis yog mob ntshav qab zib (non-DM pawg, n=17) thiab pab pawg ntshav qab zib, n=14).

Qauv sau. Cov ntshav kuaj tau txais thaum pib thiab qhov kawg ntawm kev kho ntshav lim ntshav los ntawm kev kuaj ntshav nyob rau sab hlab ntsha ntawm HD Circuit Court. Rau kev ntsuas MULTIS, sera tau muab cais thiab khaws cia rau hauv lub tub yees ntawm {{0}}} degree (txog thaum ntsuas tau ua tiav, rau kev ntsuas i-STrap. tag nrho cov ntshav kuaj tau sau rau hauv cov hlab ntshav heparin uas tau tso cai. Kev ntsuas ntawm ntau yam dawb radical scavenging kev ua si hauv ntshav. Hydrophilic ROS scavenging kev ua ub no tau ntsuas nrog MULTIS txoj kev siv cov txheej txheem piav qhia yav dhau los nrog cov kev hloov me me.2227) Kev khawb cov dej num tawm tsam tsib ROS, uas yog hydroxyl radical(OH) , superoxide(0,"), alkoxyl radical (RO"), alkyl peroxyl radical (ROO"), thiab singlet oxygen ('{16}},), tau ntsuas. Ib qho X-band ESR spectrometer( RR-Xl ESR: Radical Research Inc., Tokyo, Nyiv) siv 100 kHz teb kev hloov kho thiab WN-RAD ua haujlwm software (Radical Research Inc.) tau siv. ESR spin trapping reagents siv yog CYPMPO rau " OH, O, RO', thiab ROO' thiab 4-hydroxy-2,2,6,6-tetramethylpiperidine (TEMP) rau 'O. Cov chaw spectrometer raug raws li nram no: teb modulation width.0.1 mT; microwave zog, 10 mW; teb scan dav thiab tus nqi +7.5 mT/2 min; thiab lub sij hawm tas mus li, 0.1 s.Txhua ROS tau tsim los ntawm kev teeb pom kev hauv tsev nrog UV / pom lub teeb los ntawm lub teeb pom kev RUVF-203SR UV illuminator, Radical Research Inc.) nruab nrog 200W nruab nrab-siab mercury / xenon arc teeb thiab quartz teeb qhia, txuas nrog lub resonator kab noj hniav. Lub teeb qhov chaw, lub sij hawm illumination, precursors, thiab photosensitizers siv los tsim ROS yog summarized nyob rau hauv Table 1. ROS scavenging kev ua ub no raug xam raws li ib tug yav tas los piav txoj kev 22) thiab hloov mus rau hauv lub tsev sib npaug rau cov paub dawb huv scavengers: glutathione (GSH) rau OH thiab 'O, superoxide dismutase (SOD) rau O, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) rau RO', thiab a -lipoic acid rau ROO'MULTIS ntsuas rau txhua ROS tau ua hauv triplicate.

Kev ntsuas lipophilic

Ros scavenging kev ua si hauv cov ntshav tag nrho siv i-Strap. Lipophilic ROsscavenging kev ua haujlwm tau ntsuas siv i-STrap ESR ntsuas ki (Dojin-Glocal / Dojindo, Kumamoto, Nyiv) raws li cov chaw tsim khoom raws tu qauv thiab daim ntawv tshaj tawm dhau los.2s Txoj kev no yog ua raws li kev sib tw ntawm cov tshuaj tiv thaiv antioxidants hauv cov ntshav thiab DPhPMPO. nrog tert-butyl hydroperoxide (BuOOHradical. Tag nrho cov ntshav kuaj tau incubated nrog DPhPMPO(1{{10}}} mM) thiab kuaj-BuOOH (10 mM) rau 30 min ntawm chav tsev kub. Tom qab incubation, lub spin adducts nyob rau hauv cov organic theem tau muab rho tawm siv chloroform thiab methanol cov kev daws teeb meem; ESR kev ntsuas tau ua los ntawm cov organic cov qauv. Cov kev ntsuas tau zoo ib yam li rau MULTISmeasurement, tshwj tsis yog rau daim teb scan dav thiab tus nqi, teem rau ntawm +5 .0 mT / 2 min, Tsis muaj cov tshuaj tiv thaiv kab mob tshwj xeeb rau i-STrapmeasurement; yog li ntawd, kev ua haujlwm khawb tau nthuav tawm raws li tus nqi an10/-1, qhov "0" qhia txog lub teeb liab siv tsis muaj qauv thiab "' qhia txog lub teeb liab siv nrog ib tug qauv. (28)Reagents. CYPMPO tau txais los ntawm RR INC.(Nyiv); riboflavin, 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), text-BuOOH, dimethyl sulfoxide (DMSO), thiab rose Bengal tau yuav los ntawm Sigma-Aldrich Nyiv (Tokyo, Nyiv) thiab TEMP tau yuav los ntawm Tokyo Chemical Industry (Tokyo, Nyiv). Hydrogen peroxide thiab buffers tau txais los ntawm WakoChemical Co.(Osaka, Nyiv). Kev txheeb xyuas kev txheeb cais. "Statistical analysis tau ua los ntawm Prism 6 rau Mac OS X computer software (GraphPad SoftwareInc., La Jolla, CA). Paired t-test. Cov ntaub ntawv tau nthuav tawm raws li qhov tseem ceeb nrog 95% kev ntseeg siab ib ntus (95% CI.