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Meta-analysis Ntawm NAD(P)(H) Quantifcation Results Exhibits Variability Across Mammalian Tissues Ⅱ

Jun 01, 2023

Qhov cuam tshuam ntawm premortem piv rau cov ntaub so ntswg tom qab sau ntawm NAD(P)(H) qib.

Peb kwv yees tias tej zaum yuav muaj qee qhov sib txawv ntawm NAD(P)(H) redox lub xeev uas yog nyob ntawm cov txheej txheem sau cov ntaub so ntswg. Txhawm rau teb cov lus nug no, peb tau tshuaj xyuas cov txiaj ntsig ntawm cov txheej txheem sau cov ntaub so ntswg ua ntej ntawm NAD(P)(H) redox. Rau qhov kev tshuaj ntsuam no, peb piv cov txiaj ntsig los ntawm cov ntaub so ntswg nas uas tau muab ntau dua ntawm cov kev tshawb fawb qhia txog lawv cov txheej txheem kev txi. Thaum nthuav tawm, tag nrho cov kev tshawb fawb soj ntsuam tau hais tias cov qauv ntaub so ntswg tau khaws cia txias thiab muab rho tawm tam sim ntawm cov dej khov lossis khov ntawm - 80 degree ua ntej NAD(P)(H) ntsuas. Qhov kev tshuaj xyuas no tau txiav txim siab tias tsis muaj qhov sib txawv tseem ceeb hauv NAD ntxiv rau qhov muaj nyob hauv nas daim siab (Fig. 6a) ntawm cov ntaub so ntswg muab rho tawm ua ntej lossis tom qab euthanasia. Txawm li cas los xij, qhov qhia txog NADH qib hauv nas daim siab yog qhov siab dua hauv cov qauv tom qab tuag (Fig. 6b). Tsis muaj qhov sib txawv ntawm tag nrho NAD(H) qib (Fig. 6c) tau pom tab sis qhov NAD ntxiv / NADH piv tau qis dua hauv cov ntaub so ntswg uas tau sau tseg tom qab euthanasia (Fig. 6d), uas yog ib qho zoo sib xws nrog cov kev hloov pauv hauv NADH qib. NADP ntxiv , NADPH, thiab NADP ntxiv / NADPH piv tau tshwm sim hauv nas daim siab pab pawg los ntawm cov ntaub so ntswg sau lub sij hawm taw tes qhia nyob rau hauv Ntxiv Fig. 9; Txawm li cas los xij, tus lej tsis txaus ntawm kev kuaj mob tom qab tsis tso cai rau peb txhais cov txiaj ntsig ntawm cov ntaub so ntswg sau lub sijhawm ntawm NADP(H) qib. Hauv nas lub siab, tsis muaj qhov sib txawv ntawm qhov ua ntej thiab tom qab-mortem NAD ntxiv rau qib, txawm li cas los xij, tus naj npawb ntawm cov kev tshawb fawb qhia tau txwv (Daim duab 6e). Tsis muajNADH,NADP plus, los yogNADPHpost-mortem data nyob rau hauvmus cevtwb muaj rau kev sib piv.

Flavonoid (2)

Nyem qhov no kom tau txais Hebrs Cistanche


Kev sib tham

Ntau qhov kev tshawb fawb tsis ntev los no tau pom tias NAD(P)(H) kev tswj hwm yog qhov tseem ceeb rau cellular homeostasis thiab redox xwm txheej. Cov ntaub ntawv tau los ntawm cov nas tsuag tau coj mus rau kev tshawb fawb soj ntsuam tib neeg cov ntshav NAD ntxiv rau qib, raws li qhov tsis tshua muaj kev cuam tshuam ntawm tag nrho lub cev NAD ntxiv rau qib. Cov kev tshawb fawb soj ntsuam tsis ntev los no tau pom tias NAD ntxiv rau qib tau txo qis hauv cov kab mob hauv xeev 23,47 thiab nce ntxiv tom qab NAD ntxiv rau kev txhawb nqa cov tswv yim23,44,48,49. Peb tau ua ib qho kev soj ntsuam ntawm txhua qhov kev tshawb fawb luam tawm thaum xyoo 1946 thiab Lub Rau Hli 20, 2021, uas muaj cov ntaub ntawv ntau rau NAD(P)(H) qib hauv hom tsiaj nrog rau kev tsom tshwj xeeb rau nas, nas, thiab tib neeg vim lawv sawv cev rau feem ntau. tau kawm txog hom kab mob hauv biomedical. Qhov kev tshuaj xyuas no tau ua tiav hauv ib feem los txiav txim siab qhov nruab nrab qib ntawm NAD(P) (H) cov concentrations hauv cov ntaub so ntswg ntawm tus tsiaj ib txwm muab qhov paub zoo sib txawv hauv cov kev ntsuas no thoob plaws kev tshawb fawb. Peb cia siab tias cov ntaub ntawv no tuaj yeem siv los txhawb cov txheej txheem kev cai hauv thaj tsam ntawm NAD ntxiv rau kev tshawb fawb thiab txhawb nqa kev siv NAD(P)(H) qib thiab redox piv raws li kev ntseeg siab biomarkers rau cov kab mob thiab kev kho mob.

Txawm hais tias muaj ntau qhov sib txawv hauv kev ntsuas thoob plaws cov ntaub so ntswg, qhov kev tshuaj ntsuam meta no tau pom zoo li NAD ntxiv rau qib thoob cov ntaub so ntswg nrog qee qhov kev zam. Interestingly, nas skeletal leeg nthuav tawm qis nruab nrab NAD ntxiv rau qib (Fig. 2a) dua li lwm cov ntaub so ntswg uas muaj metabolic (xws li, daim siab, lub raum, lub plawv, thiab lub hlwb). Qhov no yuav qhia tau tias txawv NAD(H) redox cov xwm txheej hauv cov leeg pob txha lossis tej zaum yuav taw tes rau cov teeb meem loj dua nrog cov metabolite rho tawm hauv cov ntaub so ntswg. Txawm li cas los xij, cov kev soj ntsuam no tsis tuaj yeem txheeb xyuas hauv tib neeg vim tias tsis muaj cov ntaub so ntswg ntau dhau ntawm cov ntshav thiab cov leeg.

Flavonoid-1

Muaj ntau yam tseem ceeb uas tuaj yeem cuam tshuam qhov tseeb ntawm physiological NAD(P)(H) kev ntsuas hauv cov ntaub so ntswg. Thaum tshaj tawm, txhua qhov kev tshawb fawb tau piav qhia cov ntaub so ntswg uas tau sau thiab ncaj qha rau hauv cov kua nitrogen, los yog khaws cia rau hauv dej khov ua ntej khov lossis ua tiav txhawm rau txhawm rau khaws cov khoom kub-sensitive (NAD ntxiv thiab NADP ntxiv). Cov nyhuv ntawm pH ntawm ntau yam NAD(P)(H) feem, xws li cov kua qaub-labile qhov txo qis NAD(P)H cov ntaub ntawv, tau coj mus rau kev siv cov pH-tsis nruab nrab extraction kuab tshuaj los ntawm feem ntau ntawm cov kev tshawb fawb. . Txawm li cas los xij, muab qhov kev hloov pauv sai ntawm kev txo qis thiab oxidized metabolites thiab cov txiaj ntsig ntawm anoxia ntawm NAD(H) 50-53, nws tseem ceeb heev kom paub tseeb tias cov ntaub so ntswg metabolites sai sai thiab cov txheej txheem tsim nyog rau quenching cellular NAD(P)(H. )-redox / noj enzymes, xws li los ntawm deproteinization. Raws li cov kab no, peb qhov kev soj ntsuam ntawm kev sau cov ntaub so ntswg ua ntej-mortem qhia tau hais tias kev soj ntsuam tom qab kev tuag nyiam qhov txo qis ntawm NAD ntxiv. Kev txo qis ntawm cov ntaub so ntswg NAD ntxiv tau piav qhia yav dhau los hauv vivo hauv lub hlwb, lub siab, thiab daim siab ntawm cov nas kis mus rau qhov txuas ntxiv 2.5-h hypoxic cua54, ib puag ncig uas tej zaum yuav rov ua dua ib nrab los ntawm kev ncua sij hawm tom qab- txi cov ntaub so ntswg ua ntej snap khov. Qhov no yuav qhia tau tias kev rho tawm cov ntaub so ntswg thaum nyob rau hauv tshuaj loog los yog qhov tseem ceeb ntawm kev sau tam sim ntawm cov ntaub so ntswg npaj los siv rau kev tshuaj ntsuam metabolite tom qab kev txi, yuav tsum tshwm sim kom tau txais cov neeg sawv cev NAD(P)(H) ntsuas.

Kuj tseem muaj peev xwm rau ntau txoj hauv kev kom muaj nuj nqis, txhua qhov muaj kev txwv sib txawv, los pab txhawb rau tag nrho cov kev kawm sib txawv ntawm NAD(P)(H) kev ntsuas. Hauv ob lub xyoo dhau los, cov kev siv ntau tshaj plaws yog enzyme cycling assays, LC-MS, thiab HPLC. Txhua yam ntawm cov txheej txheem no cuam tshuam los ntawm cov txheej txheem metabolite extraction, ntsuas qhov ntsuas, thiab kev siv cov kev tswj kom zoo. Siv LC-MS, Lu et al. tau pom tias qhov kev sib hloov ntawm cov ntaub ntawv txo thiab oxidized ntawm NAD(P)(H) metabolites tshwm sim ntawm cov nqi sib txawv nyob rau hauv ntau yam extraction buffers los yog cov kuab tshuaj nrog cov acetonitrile: methanol: dej nrog 0.1 M formic acid sib tov yielding lub zoo tshaj plaws rov qab nrog qhov tsawg kawg interconversions31. Txawm li cas los xij, thaum siv LC-MS qhov kev hloov pauv no tuaj yeem saib xyuas ntawm tus kheej cov qauv ntawm kev tshawb fawb los ntawm kev sib tw nrog kev tswj hwm sab hauv xws li NAD(P)(H) isotopes, uas yog qhov zoo ntawm LC-MS cov txheej txheem dhau ntawm HPLC thiab enzyme. 31, 33, 55. Txawm li cas los xij, txawm tias muaj kev tswj xyuas zoo LC-MS cov txheej txheem, ntau yam tuaj yeem cuam tshuam nrog cov teeb liab ntsuas, suav nrog cov teebmeem matrix thiab kev hloov pauv hauv ionization efficiency. Piv txwv li, qee qhov kev tshawb fawb siv 13C-labeled poov xab rho tawm raws li cov qauv hauv LC-MS-based metabolomics. Txawm li cas los xij, spiking cov qauv rho tawm cov qauv metabolite matrix nrog 13C-labeled poov xab extract metabolite matrix tuaj yeem muaj ntau yam tshwm sim, xws li ion suppression, uas txo cov teeb liab ntawm cov ntawv sau thiab / lossis cov metabolites tsis muaj npe, thiab tuaj yeem ua rau muaj qhov yuam kev hauv qhov kom muaj nuj nqis yog tias tsis yog. kuaj pom los ntawm kev ntsuam xyuas zoo 56. Tsis tas li ntawd, txawm hais tias LC-MS cov tswv yim theoretically tso cai rau kev ntsuas ntawm ntau yam NAD(P)(H) metabolites hauv ib qho kev sim, tseem muaj kev txwv vim tias qhov kev pom zoo yuav tsum tau khiav yog tias cov metabolites ntawm kev txaus siab muaj qhov sib txawv loj hauv qhov ntau. Yog li, txawm hais tias lawv qhov zoo dua li qhov kev ntsuam xyuas yooj yim enzyme cycling, qhov nyuaj ntawm LC-MS txoj kev ua rau qhov tsim nyog rau kev ua kom zoo dua qub. Tsis ntev los no cov kev tshuaj ntsuam tshiab, xws li, NAD ntxiv rau biosensors thiab imaging-based mass spectrometry, tau tsim tab sis tseem muaj cov ntaub ntawv tsis txaus tsim los ntawm cov tswv yim no los suav nrog hauv kev tshuaj ntsuam meta. Txawm hais tias, ib txoj kev tshawb fawb suav nrog hauv peb qhov kev tshuaj ntsuam meta siv cov ntawv-raws li bioluminescent biosensor los ntsuas NAD ntxiv rau qib hauv nas lub siab thiab lwm yam qauv 57. Txoj kev tshawb no tau suav nrog hauv pab pawg kuaj bioluminescent vim yog txoj kev tshawb pom zoo sib xws (Fig. 1a).


Flavonoid

Rau lub hom phiaj ntawm qhov kev tshuaj ntsuam no, cov ntaub ntawv los ntawm 677 kev tshawb fawb tau raug cais tawm. Qhov no suav nrog cov kev tshawb fawb uas muab cov txheeb ze NAD(P)(H) cov txiaj ntsig (51 feem pua) es tsis yog qhov ntau, nrog rau cov kev tshawb fawb uas NAD(P)(H) concentratetsis tau normalizedcov ntaub so ntswg hnyav, cov ntsiab lus protein, los yogntshav ntim(13 feem pua). 36 feem pua ​​​​ntawm cov kev tshawb fawb tsis suav nrog tau ua rau cov qauv tsis tsim nyog (xws li, yam tsis yog tsiaj txhu, kab lis kev cai ntawm tes lossis cov ntaub so ntswg). Tshaj li cov kev tshawb fawb tsis suav nrog, cov kev txwv loj hauv qhov kev ntsuam xyuas meta no suav nrog qhov kev hloov pauv ntawm, lossis tsis muaj cov ntaub ntawv qhia txog, cov txheej txheem ua ntej kev tshuaj ntsuam (ua ntej lossis tom qab kev tuag los yog rho tawm cov ntaub ntawv tsis txaus) thiab / lossis cov txheej txheem tshuaj xyuas, nrog rau nrog lub hnub nyoog, poj niam txiv neej, keeb kwm caj ces, kev noj haus, thiab kev noj zaub mov ntawm kev txi rau nas.
Txawm hais tias peb qhov kev ntsuam xyuas meta qhia txog qhov sib txawv ntawm kev kawm sib txawv thiab qhov tsim nyog los ua qauv NAD(P) (H) kev ntsuas ntau, peb qhov kev soj ntsuam tsis ntsuas lossis sib cav txog qhovvalidity ntawm cov txiaj ntsig sib pivhauv ib qho kev kawm ntawm tus kheej. Muab qhov cuam tshuam ntawm ntau yam NAD(P)(H) metabolites ua biomarkers ntawmkev noj qab haus huv nyob rau hauv tib neeg, tus txheej txheem los yog kev kawm kom zoo optimization ntawm pre-analytical thiabcov txheej txheem analyticalyuav pub raukev sib piv zoo dua ntawm cov txiaj ntsig ntawm kev tshawb fawb. Qhov no ua qhov tseem ceeb tshaj nrog kev nce ntxivTus naj npawb ntawm cov kev tshawb fawb soj ntsuam kuaj cov nyhuv ntawm cov tshuaj los yog cov tshuaj siv los hloov cov ntaub so ntswg NAD(P)(H) mus rautxhim kho kev noj qab haus huv ntawm tib neeg.

Flavonoid (11)

Cov txheej txheem

Nrhiav cov ntaub ntawv. "Epub Ua Ntej Luam Ntawv", "In-Process & Other Non-Indexed Citations", "Versions", "PubMed-Not-MEDLINE", "Txhua hnub hloov tshiab", "Pem ntej ntu hloov tshiab txhua lub limtiam", "Rov qab los ntawm 1946 los pib. ntawm pem hauv ntejntu "citations ntawm 1946 thiab Lub Rau Hli 20, 2021, hauv MEDLINE ALL (Ovid) database tau tshawb nrhiav thiabtshuaj xyuas rau cov khoom uas muaj cov ntaub ntawv ntau ntawm NAD(P)(H) metabolite theem hauv cov ntaub so ntswg mammalian li tawmnyob rau hauv Daim Ntawv Ntxiv S1. Koj tshawb nrhiav cov tswv yim yielded 3377 kab lus. Kev tshawb nrhiav ntshav ntxiv yogua tau siv tib lub database thiab yielded 1513 tsab xov xwm (Appendix S2).
Kev tshuaj ntsuam xyuas. Cov lus piav qhia thiab cov npe qhia txog NAD(P)(H) metabolite kom muaj nuj nqis nyob rau hauv cov ntaub so ntswg thiab ntshav tau soj ntsuam tag nrho cov ntawv nyeem. Tag nrho cov kev ntsuam xyuas paub daws teeb meem tau txheeb xyuas nrog kev tshuaj xyuas thib obthiab tej kev tsis sib haum xeeb raug daws. Tom qab kev tshuaj ntsuam xyuas paub daws teeb meem, 643 cov ntawv tshaj tawm raug xaiv rau kev tshuaj xyuas tag nrho cov ntawv nyeem raucov ntaub so ntswg tsom rau kev tshawb nrhiav thiab 272 rau kev tshawb nrhiav ntshav (Ntxiv daim duab 10). Kev tshawb fawb uas tsuas yog ua ntejxa cov txheeb ze NAD(P)(H) metabolite cov ntaub ntawv raug tshem tawm thaum lub sij hawm soj ntsuam tag nrho cov ntawv, nrog rau kev zam ntawmkev tshawb fawb qhia NADntxiv/ NADH ratios. Cov txiaj ntsig tau nruj me ntsis los ntawm tssues, hlwb, cais cov organelles khaws cianyob rau hauv xov xwm, los yog los ntawm cov ntshav eluent los ntawm perfusion raug cais tawm. Qhov kev tshuaj xyuas tag nrho cov ntawv no coj mus rau kev cais tawm ntawm 667(457 ntxiv rau 210) kab lus vim feem ntau tsis muaj cov ntaub ntawv NAD metabolite (50.5 feem pua), cov ntaub ntawv muab los ntawm tsis tsim nyogcov qauv (36.3 feem pua ​​​​) los yog cov ntaub ntawv nthuav tawm raws li cov txheeb ze / cov neeg txiav txim siab (13.2 feem pua). Thaum kawg, 241 kab lus tau suav nrogqhov zoo meta-analysis thiab 205 nyob rau hauv lub quantitative meta-analysis (Supplementary Fig. 10). Txawm tias pebKev tshawb nrhiav thawj zaug suav nrog cov kev tshawb fawb luam tawm los ntawm 1946 txog rau Lub Rau Hli 20, 2021. Raws li tau hais los saum no, qhov kev kawm qub tshaj plaws nrogquantitative NAD(P)(H) cov ntaub ntawv uas ua tau raws li peb qhov kev lees paub tau luam tawm xyoo 1961


Kev rho tawm cov ntaub ntawv.

Cov ntaub ntawv naj npawb rau NAD(P)(H) cov metabolites tau ncaj qha los ntawm cov khoom lossis muab rho tawm los ntawm cov ntawv xov xwm siv cov ntaub ntawv ib nrab tsis siv neeg rho tawm software (WebPlotDigitizer58). Tsis tas li ntawd, qhov twg siv tau, cov ntaub so ntswg sampling lub sij hawm cov ntsiab lus txheeb ze rau kev txi thiab / lossis cov ntaub so ntswg / cov qauv ntshav (piv txwv li txoj kev siv tshuaj loog thiab / lossis euthanasia, cov ntaub so ntswg tuav, thiab qhov kub thiab txias) tau sau tseg. Rau cov qauv tsiaj, hom, tus yam ntxwv (xws li hom, hom kab mob, hnub nyoog, thiab qhov hnyav), thiab ib puag ncig (xws li kev pw tsaug zog, hom kev noj zaub mov, kev pub noj ntau zaus, thiab kev noj zaub mov hauv xeev txheeb ze rau cov ntaub so ntswg sampling) tau sau tseg. Tsis tas li ntawd, rau tag nrho cov kev tshawb fawb, kev kho mob (xws li kev kho tshuaj, kev phais mob, irradiation, qog induction, thiab lwm yam txheej txheem siv rau cov kev kawm) thiab tag nrho cov ntaub ntawv kawm pab pawg (xws li kev kho mob lossis kab mob thiab kev tswj hwm sib raug) raug muab rho tawm. Thaum kawg, cov txheej txheem ntawm NAD(P)(H) quantification method (xws li enzymatic assays, mass spectrometry-based, HPLC, NMR, bioluminescence) thiab cov ntaub ntawv tshaj tawm (xws li cov ntawv tshaj tawm, cov cai siv [DOI, Medline UI, PMID] lossis hyperlink thiab xyoo ntawm kev tshaj tawm) tau sau tseg (Cov Khoom Ntxiv 2).



Cov ntaub ntawv tsom xam. Ua ntej kev soj ntsuam, tag nrho cov concentrations tau hloov mus rau nmol / g ntawm cov ntaub so ntswg los yog nmol / g ntawm cov proteins rau cov ntaub so ntswg los yog nmol / ml rau cov ntshav feem. Vim qhov tsawg ntawm cov txiaj ntsig normalized rau cov ntsiab lus protein, peb tsuas yog qhia cov txiaj ntsig normalized rau cov nqaij mos qhov hnyav thiab cov ntshav ntim. Kev txheeb xyuas txheeb cais tau ua nrog GraphPad Prism version 9.3.1 (GraphPad Sofware Inc., San Diego, California, USA,Cov ntaub ntawv muajTag nrho cov ntaub ntawv tau muab rho tawm thiab tshuaj xyuas thaum lub sijhawm kawm no suav nrog hauv tsab xov xwm tshaj tawm no thiab nws cov khoom ntxivlus fles.Tau txais: 29 Plaub Hlis 2022; Tau Txais: 7 Lub Ob Hlis 2023



Cov ntaub ntawv

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