TUNEL Assay: Ib qho cuab yeej muaj zog rau kev ntsuam xyuas mob raum

Hu rau:joanna.jia@wecistanche.com/ WhatsApp: 008618081934791

Abstract:Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay yog ib qho kev soj ntsuam ntev uas siv los txheeb xyuas cov cell tuag-ssociated DNA fragmentation (3'-OH DNA termini) los ntawm endonucleases. Vim tias cov enzymes no tshwj xeeb tshaj yog nyob rau hauv lub raum, TUNEL tau siv dav los txheeb xyuas thiab txheeb xyuas DNA tawg thiab cell tuag hauv kab lis kev cai.raum cell thiab tsiaj thiab tib neeg lub raum uas tshwm sim los ntawm covmob toxic lossis hypoxic raug mob. Lub cim thaum ntxov ntawm TUNEL raws li kev ntsuam xyuas apoptotic tau ua rau ntau qhov kev txhais tsis raug ntawm cov txheej txheem ntawmlub raum cell raug mob.Txawm li cas los xij, TUNEL tau dhau los ua neeg nyiammob raumkev ntsuam xyuas vim tias nws tuaj yeem siv tau thoob plaws hauv cov kab lis kev cai thiab cov ntaub so ntswg thiab rau txhua lub tshuab ntawm kev tuag ntawm tes. Tsis tas li ntawd, nws yog rhiab heev, muaj tseeb, muaj nuj nqis, yooj yim txuas rau cov hlwb tshwj xeeb lossis cov ntaub so ntswg, thiab tuaj yeem ua ke nrog immunohistochemistry kom tso cai txheeb xyuas cov hom cell los yog cov txheej txheem ntawm cov cell tuag. Kev lig kev cai, TUNEL tsom xam tau txwv rau qhov muaj lossis tsis muaj TUNEL teeb liab. Txawm li cas los xij, cov ntaub ntawv ntxiv ntawm cov txheej txheem ntawm kev tuag ntawm tes tuaj yeem tau txais los ntawm kev tshuaj xyuas ntawm TUNEL cov qauv.

kev sib deev arousal raum mob kev kho mob nrog cistanche ntxiv hmoov

Taw qhia DNA yog tib lub molecule nyob rau hauv lub cell uas yuav kho tau, tab sis nws yuav tsis tau kiag li resynthesized tom qab kev puas tsuaj. DNA kev puas tsuaj yog ib qho ntawm cov theem kawg ntawm cov txheej txheem ntau-kauj ruam ntawm kev tuag, uas muaj peev xwm thim rov qab mus txog qhov kawg ntawm qhov kawg ntawm DNA tawg. Yog li ntawd, qhov tom kawg yog ib qho piv txwv thiab cov cuab yeej cuab tam ntawm irreversible cell tuag [1,2]. Cell death-associated DNA fragmentation feem ntau "visualized" los ntawm lub davhlau ya nyob twg deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, DNA laddering, los yog comet assay [3]. Pulse-field electrophoresis [4] thiab random oligonucleotide-primed synthesis (ROPS) assay [5] yog ob qho lwm yam tsis tshua siv cov kev ntsuam xyuas theem nrab. Kev siv cov comet assay tsuas yog siv rau hauv vitro (cov kab lis kev cai) nkaus xwb, thiab nws qhov ntau yog siv zog ua haujlwm [6]. Kev soj ntsuam comet nyob rau hauv cais cell nuclei yog ua tau, tab sis nws yog txuam nrog ntxiv confounding yam thaum lub sij hawm kev rho tawm nuclei. Qhov thib ob, DNA laddering assay, tsis yog ib txoj hauv kev kom muaj nuj nqis, thiab thawj qhov kev cia siab tias tus qauv ntaiv vs. smear yuav paub qhov txawv ntawm apoptosis thiab necrosis, ntsig txog, tau muab tshem tawm raws li txoj hauv kev no tau ua pov thawj tias tsis ntseeg tau [7]. Ntawm peb txoj hauv kev tseem ceeb, TUNEL yog qhov rhiab tshaj plaws, siv sijhawm tsawg kawg, thiab siv tau thoob plaws ntiaj teb. Nws tuaj yeem siv rau hauv kab lis kev cai thiab cov ntaub so ntswg thiab rau tag nrho cov cell tuag mechanisms. Nws yog qhov tseeb, muaj nuj nqis, yooj yim txuas rau cov hlwb tshwj xeeb lossis cov ntaub so ntswg, thiab tuaj yeem ua ke nrog immunohistochemistry kom tso cai txheeb xyuas cov hom cell lossis cov txheej txheem ntawm kev raug mob ntawm tes cuam tshuam nrog TUNEL-zoo teeb liab [2].

DNA rhuav tshem los ntawm TUNEL yog tsim los ntawm apoptotic endonucleases. Cov kab mob endonucleases feem ntau nyob hauv lub raum yog deoxyribonuclease 1 (DNase I) thiab endonuclease G (EndoG) [8–10]. DNase Kuv yog qhov nquag tshaj plaws thiab ntau tshaj apoptotic endonuclease hauv cov tsiaj txhu [11]. Lub raum, cov qog ua kua qaub, txiav, thiab cov hnyuv yog cov kabmob uas paub tias tsim cov DNase I. Epithelial cov hlwb hauv cov qog qaub ncaug, txiav, thiab cov hnyuv secrete DNase I raws li lub plab zom mov hauv txoj hnyuv. Qee cov cellular DNase kuv tawm tsam "txawv teb chaws" DNA invading host cells [12]. Tsis-secreted DNase kuv nthuav qhia qhov kev phom sij zais rau tus tswv-cell DNA, tab sis feem ntau DNases tsis ua rau tus tswv-cell DNA kom txog thaum raug mob lossis tuag ntawm tes. Nyob rau hauv lub raum, DNase I yog secreted los ntawm tubular epithelial hlwb, tej zaum yuav ua kom puas cov kab mob thiab cov kab mob nyob rau hauv cov zis. Qhov kev ua haujlwm no yuav tshwm sim nrog cov proteinases, tshwj xeeb, meprin, uas yog lub ntsiab proteinase hauv cov zis [13]. Txawm hais tias nws muaj kev tiv thaiv kab mob, qhov muaj DNase nquag hauv lub raum ua rau lub cev tsis muaj zog rau kev raug mob, qhov chaw ua haujlwm ntawm DNase paub tias yog cytotoxic rau cov tswv tsev. DNases txhawb kev tuag ntawm tes los ntawm cov tshuaj lom lossis hypoxic stimuli thiab rhuav tshem tag nrho cov kab mob hauv lub raum DNA tso tawm los ntawm kev tuag ntawm tes.

Lub raumyog lim cov kabmob uas tshem tawm cov tshuaj lom hauv lub cev. Kev ua haujlwm siab DNase I hauv ob lub raum ua rau lub raum hlwb nkag siab rau kev raug mob los ntawm cov tshuaj lom thiab lawv cov khoom lag luam metabolic. Qhov no ua rau TUNEL yog txoj kev tsim nyog tshaj plaws thiab siv tau los ntsuas kev raug mob rau lub raum. Hauv qhov kev tshuaj xyuas no, peb yuav tsom mus rau TUNEL raws li kev siv dav dav, kev qhia paub, thiab kev ntsuam xyuas meej rau kev ntsuas kev tuag ntawm tes thiab cov ntaub so ntswg raug mob hauv lub raum. Peb yuav sib tham txog yam ntawm TUNEL raws li nws siv rau cov kab mob hauv lub raum thiab kev raug mob thiab txheeb xyuas qhov zoo, cov tswv yim, thiab cov yam ntxwv tsis zoo ntawm TUNEL los txhawb kev tsim khoom thiab tsim nyog siv qhov kev ntsuam xyuas muaj zog hauv kev tshawb fawb, kev kuaj mob, thiab kev kho mob raum kab mob. Peb kuj tseem yuav tham txog ntau yam TUNEL cov qauv duab uas tuaj yeem txuas nrog cov txheej txheem kev tuag ntawm tes. Cov ntsiab lus uas tau hais tseg hauv qhov kev tshuaj xyuas no yog dav siv rau TUNEL siv rau hauv lub cev uas tsis yog lub raum.

Ntsiab lus:TUNEL; cell tuag; raum; DNases; endonucleases; cov ntaub so ntswg raug mob

TUNEL Cov CaiQhov kev ntsuam xyuas TUNEL tau tsim nyob rau xyoo 1992 los ntawm Gorczyca li al. [14] and Gavrieli et al. [15], siv fluorochrome thiab avidin-peroxidase labeling, feem. Thaum lub sijhawm ntawd, muaj qhov xav tau kev xav tau tshiab rau kev ntsuas apoptosis, thiab TUNEL ua tiav qhov sib txawv no. Pib ua lag luam raws li kev ntsuam xyuas rau DNA strand so thaum lub sij hawm los yog txuam nrog apoptosis [14,16], thaum tsis muaj kev soj ntsuam zoo dua, TUNEL tau dhau los ua tus qauv ntsuas rau apoptosis [17,18]. Txawm li cas los xij, nws tau lees paub yuav luag tam sim ntawd los ntawm cov kws tshawb fawb tias TUNEL tsis saib xyuas kev ntsuas DNA tawg, tsis yog ib qho cuam tshuam nrog apoptosis [19–21]. Txawm li cas los xij, qhov kev xav tau rau kev soj ntsuam apoptotic yog qhov zoo heev uas cov ntaub ntawv tsis tshua muaj ntawm TUNEL tsis yog tshwj xeeb rau apoptosis feem ntau tsis quav ntsej. Qhov no ua rau muaj ntau qhov kev ceeb toom tsis tseeb ntawm apoptosis nyob rau hauv lub raum thiab lwm yam kabmob, mus txog rau qib uas txog li 20 feem pua ​​​​ntawm tag nrho cov hlwb tau tshaj tawm tias apoptotic txawm tias tsis raug mob [22–25] thaum hloov pauv raws li me me li 0.01 feem pua ​​​​tuaj yeem ua pov thawj tseem ceeb ntawm cov ntaub so ntswg raug mob.

Qhov kev ntsuam xyuas TUNEL tau ntsib kev rov ua dua tshiab nyob rau xyoo tsis ntev los no txij li thaum kawg ntawm kev loj hlob hauv kev tshawb fawb apoptosis ob peb xyoos dhau los. Peb qhov kev tshawb nrhiav PubMed ntawm cov ntawv xov xwm hais txog "lub raum" thiab "TUNEL" cov ntsiab lus tau tsim ntau dua 1500 qhov txiaj ntsig los ntawm 1992 txog 2020. Hauv 2019, 170 cov ntawv tau tshaj tawm, tus lej siab tshaj plaws hauv ib xyoos dhau los 28 xyoo dhau los (Daim duab S1) thiab qhov twg yog feem ntau ntawm cov ntawv tshawb fawb rau qhov kev tshuaj xyuas no. Ntau cov kev tshawb fawb tau siv TUNEL-zoo li kev ntsuas thoob ntiaj teb ntawm DNA fragmentation-txuas nrog cell tuag nyob rau lwm hom cell tuag sib nrug ntawm apoptosis. Piv txwv li, ferroptosis, ib qho kev tshawb pom tshiab ntawm cov programmed, uas tsis yog-apoptotic cell tuag tshwm sim los ntawm oxidative puas tsuaj uas tshwm sim thaum lub raum ischemia-reperfusion (IR) hauv cov nas, tuaj yeem kuaj tau los ntawm TUNEL [26]. Pyroptosis, uas yog ib qho kev mob tshwm sim ntawm cov txheej txheem cell tuag uas txawv ntawm apoptosis thiab necrosis, tau pom tias muaj feem cuam tshuam nrog TUNEL-positive signals [27,28]. TUNEL-zoo tau pom thaum lub sij hawm necroptosis uas ua rau muaj kev loj hlob zuj zus ntawm lub raum tubule hlwb hauv cov nas raug rau subtotal nephrectomy [29]. Nyob rau hauv lub raum thiab lwm cov ntaub so ntswg, zoo TUNEL cov cim tau pom nyob rau hauv necrosis [30,31], dysregulated autophagy [32–35], anoikis [36,37], mitotic kev puas tsuaj [35,38], autolysis [39], Pautolysis [39], proptosis [40], thiab aponeurosis [30], ua kom pom TUNEL los ua ib qho kev ntsuam xyuas thoob ntiaj teb rau kev tuag ntawm tes tsis tuaj yeem.

Qhov kev ntsuam xyuas TUNEL yog raws li daim ntawv lo ntawm 3'OH xaus los ntawm 3'OH-kawg tshwj xeeb DNA enzyme, terminal deoxynucleotidyl transferase (TdT) (Daim duab). Kev siv DNA polymerase los yog Klenow fragment DNA polymerase nyob rau hauv qhov chaw ntawm TdT yuav pab txiav txim seb hom 3'OH xaus (nicks, khoob, los yog overhanging oligos) vim hais tias DNA polymerases xav tau ib tug template (qhov opposite DNA strand) thiab yog li yuav tsis sau ntawv. dai lossis protruding DNA xaus. Nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov tias kev tsim cov 3ZOH DNA xaus tsis yog tshwj xeeb rau apoptotic (DNA-degrading) endonucleases. 3'OH DNA xaus yog lub cim tseem ceeb "kev sib txuas lus", ib qho ntawm feem ntau ntawm DNA enzymes. Lawv tau tsim thiab siv los ntawm yuav luag txhua DNA enzymes hauv eukaryotes, suav nrog kho DNA (apurinic/apyrimidinic, AP) endonucleases, exonucleases, DNA polymerases, DNA ligases, DNA transferases, thiab topoisomerases. Tsis tas li ntawd, 3'Id 3z-qab zib, lossis 3z-protein conjugates tuaj yeem hloov mus rau 3'OH termini los ntawm phosphatase, glycosylases, thiab proteinases, feem. Qhov no yog vim li cas kev nkag siab meej ntawm qhov ntsuas yog qhov tsim nyog rau kev ua tiav ntawm TUNEL thiab kev txhais ntawm nws cov txiaj ntsig.

Piv nrog rau ob txoj hauv kev tseem ceeb ntawm kev ntsuas DNA fragmentation, DNA ntaiv thiab kev ntsuam xyuas comet, TUNEL muaj qhov zoo ntawm kev ua raws li kev txheeb xyuas ntawm DNA termini es tsis yog tawg (Daim duab 2). Raws li txoj cai, TUNEL shocld yog qhov rhiab heev thiab linear rau kev txheeb xyuas qhov pib (tsawg) DNA fragmentation dua DNA ntaiv los yog comet, ob qho tib si uas muaj lub sij hawm lag luam ntawm cov khoom me me ntawm cov khoom tawg yooj yim. Txawm li cas los xij, kev tshawb fawb ua ke nrog TUNEL nrog cov comet )41,42] lossis DNA ntaiv [43,44] muaj qee qhov zoo ntawm kev ntes DNA tawg thaum ntxov nrog qhov siab tshaj plaws.

Cov ntawv thov TUNELTUNEL yog siv los txheeb xyuas thiab ntsuas qhov kev raug mob raum hauv kev soj ntsuam thiab cov kev tshawb fawb toxi cological hauv ntau hom kev siv, suav nrog kev kho mob [45], ib puag ncig [46], kev ua liaj ua teb [41] thiab kev lag luam [47] raug, thiab tsiaj [48] thiab Food Sciences [49]. TUNEL siv tau rau txhua hom cell, kab mob, thiab hom uas muaj DNA thiab DNases, uas suav nrog txhua hom. TUNEL tau siv los txheeb xyuas thiab ntsuas lub raum raug mob hauv ntau hom tsiaj, suav nrog zebrafish [50], Japanese mov ntses [51], qaib [48], gerbils [52], nas [41], nas [45], rabb nws [53], mini- npua [54], thiab tib neeg [55]. Ib qho txiaj ntsig tseem ceeb ntawm TUNEL yog tias nws tuaj yeem siv rau hauv cov kab lis kev cai ruaj khov nrog rau cov ntaub so ntswg. Qhov no muab cov txheej txheem sib xws rau kev sib piv ntawm hauv vitro thiab hauv vivo cov txiaj ntsig hauv kev tshawb fawb hauv cov txheej txheem tuaj yeem tshawb xyuas hauv cov kab mob raum kab mob, thaum nyob hauv vivo cuam tshuam raug soj ntsuam hauv cov tsiaj lub raum. Txhua yam hauv nruab nrab ntawm tag nrho cov hlwb thiab cov ntaub so ntswg tuaj yeem siv tau zoo, suav nrog cov kab mob cell spheroids [56] thiab ex vivo raum slices [57,58].

Qhov kev siv ntawm TUNEL nyob rau hauv cov nqe lus ntawm lub teeb liab lub zog los yog tus naj npawb ntawm TUNEL- zoo hlwb yuav txawv ntawm hom nyob ntawm seb qhov kev ua ntawm DNases. Hauv peb qhov kev paub dhau los, nas lub raum muaj DNase I ntau dua li nas raum, thiab TUNEL-zoo teeb liab muaj zog cuam tshuam nrog qhov no. Thaum lub raum muab piv rau lwm yam kabmob, nws TUNEL-zoo teeb liab yog khaus heev thiab zoo ib yam li lwm cov kabmob uas muaj DNase I ua haujlwm siab, xws li cov hnyuv thiab cov qog qaub ncaug. Ntawm qhov kawg ntawm lub spectrum, qhov qis tshaj TUNEL-zoo teeb liab lub zog yog pom nyob rau hauv lub hlwb thiab qog / qog nqaij hlav hlwb, qhov twg DNase kuv qhia feem ntau yuav inactivated los ntawm lwm txoj kev splicing nyob rau hauv lub coding cheeb tsam [59]. TUNEL-zoo teeb liab tau pom nyob rau hauv txhua hom cell hauv lub raum. Qhov nruab nrab, cov tubular epithelium raug puas tsuaj ntau dua li glomeruli, tshwj xeeb tshaj yog nyob rau hauv kev raug mob hnyav (Daim duab 3A). Qhov no sib raug zoo nrog DNase I kev ua si uas muaj nyob rau hauv cov hom cell. Kev txiav txim los ntawm kev sib deev thiab hnub nyoog sib txawv uas tau pom hauv peb qhov kev sim, DNase I muaj ntau thiab nquag hauv cov txiv neej dua li poj niam thiab cov tsiaj nruab nrab hnub nyoog piv rau cov laus lossis cov menyuam yaus hnub nyoog qis dua. Txawm li cas los xij, tsis muaj qhov sib txawv ntawm TUNEL-qhov zoo yog qhov tshwm sim ntawm kev sib deev lossis hnub nyoog, tej zaum vim tias qhov tseem ceeb lossis muaj feem cuam tshuam los ntawm lwm yam. Hauv qee lub raum qauv (xws li ischemia-reperfusion), DNase I raug ntxias [9]; thaum nyob rau hauv lwm tus, nws tsis yog thiab tej zaum yuav txawm raug suppressed (piv txwv li, nyob rau hauv cisplatin toxicity) [10]. Kev ua kom endonucleases, yog tias tam sim no, tau pom tias tshwm sim hauv thawj 24 teev tom qab mob raum [9,60,61]. Yog li ntawd, nws tsis yog qhov xav tsis thoob tias tsawg kawg yog ib nrab ntawm TUNEL daim ntawv thov hauv kev tshawb fawb raum hauv qhov kev tshuaj xyuas no tau siv rau mob raum raug mob (AKI) txij li qhov kev ntsuam xyuas ntawm AKI feem ntau ua nyob rau hauv 24 mus rau 72 teev tom qab raug mob thaum endonucleases muaj zog tshaj plaws. . Cov npe ntawm AKI cov kev tshawb fawb ntev thiab suav nrog tshuaj lom, septic, hloov pauv, thiab kev raug mob hypoxic. Hauv txoj haujlwm no, peb tau soj ntsuam cov qauv, hom raug, kev kho mob, ncua sij hawm ntsuas, thiab hom cell cuam tshuam hauv cov kev tshawb fawb no. Cov kev tshawb fawb uas tau soj ntsuam tau hais txog qhov mob hnyav thiab sib xyaw ua ke ntawm cov mob raum mob thiab mob raum mob.

Txawm hais tias lub raum raug mob nrog TUNEL-zoo teeb liab pom nyob rau hauv tubules los yog lub glomerulus nyob ntawm seb hom kev puas tsuaj los yog kab mob. Piv txwv li, lupus nephritis nyob rau hauv tib neeg thiab nickel nanoparticle puas tsuaj tau piav raws li muaj nruj glomerular TUNEL-zoo signals [62,63]. Ntawm qhov tod tes, ischemia-reperfusion thiab streptozotocin-induced ntshav qab zib nephropathy nyob rau hauv nas tsim TUNEL-zoo hlwb nkaus xwb nyob rau hauv tubules [64,65]. Proximal tubules, Henle's voj, distal tubules, thiab cov ducts collective feem ntau yog TUNEL-zoo hauv cov qauv ischemia-reperfusion. TUNEL-zoo hlwb nyob rau hauv ob qho tib si glomeruli thiab tubules tau piav nyob rau hauv cov nas raug tshuaj xws li cyclosporine A [66] thiab doxorubicin [67] los yog cov khoom siv khoom siv xws li zinc oxide nanoparticles [68] thiab hauv cov qauv nas ntawm ntshav qab zib mellitus [ 69] ib. Septic shock-induced AKI hauv tib neeg kuj tseem cuam tshuam nrog ob lub glomerular thiab tubular TUNEL-zoo hlwb [70]. strangulated kaw voj me me plob tsis so tswj ua rau TUNEL-zoo nyob rau hauv lub raum in3h [87].

AKI tuaj yeem tshwm sim tom qab hloov pauv thiab raug mob hypoxic. Kev tsis lees paub Allograft hauv tib neeg feem ntau cuam tshuam nrog kev nce ntawm TUNEL-zoo hlwb [88,89]. Tom qab mob khaub thuas ischemia, TUNEL-zoo hlwb feem ntau pom nyob rau hauv 2-24 teev thaum lub sij hawm reperfusion [90], los yog ntau dua nqaim, ntawm 14 thiab 17 h tom qab ischemia [91], thiab qee zaum peb tseem pom lig li 7 hnub hauv Kev hloov pauv hloov pauv [92] Ib yam li ntawd, hauv cov qauv ischemia-reperfusion, TUNEL-zoo hlwb feem ntau pom tau 24 teev tom qab reperfusion [44,93,94], lossis nyob rau hauv thaj tsam ntawm 4 [95,96] thiab 72 h [97]. Hauv ob qho tib si kev hloov pauv thiab kev tshawb fawb AKI ischemic, TUNEL-zoo hlwb tau nyob rau thaj tsam ntawm cov kab mob hauv nruab nrab thiab cov tubular epithelial [44,93]. Hauv naslub raumTom qab ischemia-reperfusion raug mob, TUNEL-zoo hlwb tau piav qhia raws li cov ntsiab lus hauv corticomedullary junction, lub hom phiaj ntawm kev raug mob ischemic [98].

Kev raug mob hnyav-thiab-tshuaj, qhov uas raug tshuaj lom, kab mob hauv lub xeev, thiab / lossis kev soj ntsuam tau mob hnyav (piv txwv li, teev, hnub) thiab mob ntev (xws li, lub lis piam, hli, xyoo) hauv tib txoj kev tshawb fawb, tau pom hauv tib neeg kev hloov pauv. Kev tsis lees paub (biopsy 1 lub lis piam mus rau 3 xyoos posttransplant) [88], swine hepatitis E kab mob (7 thiab 14 hnub tom qab inoculation) [52], CO2 pneumoperitoneum-induced stress in hydronephroticlub raum(2 lub lis piam hydronephrosis, 2 hnub tom qab pneumoperitoneum) [53], mesangial proliferative glomerulonephritis induced los ntawm nab venom (1 mus rau 14 hnub tom qab txhaj tshuaj) [99], unilateral ureteral obstruction (1 mus rau 14 hnub ntawm obstruction) [100], uranyl acetate raug (1 mus rau 28 hnub ntawm raug) [101], aristocholic acid nephropathy (5 thiab 30-hnub raug txhua hnub) [102], thiab neonatal hyperoxia (kuaj 1 txog 60 hnub tom qab yug me nyuam, raug hyperoxia thawj 14 hnub) [103]. Kaum plaub hnub tom qab kev cuam tshuam yog lub sij hawm ntsuam xyuas ntau heev rau cov hom nomob raum[52,53,99,101,103]. Feem ntau, TUNEL-zoo hlwb tau nyob hauv cov tubules thiab cov ducts. Qhov tsuas yog qhov tshwj xeeb yog nab venom toxicity, uas feem ntau puas glomeruli [99].

Ntevraumkab mob (CKD), txhais tau tias yog muajraumkev puas tsuaj lossis txo qis glomerular filtration rate (GFR) hauv tib neeg ntev dua 3 lub hlis, kuj yog ib qho kev kawm rau TUNEL kev ntsuas, uas tuaj yeem txiav txim siab seb CKD puas cuam tshuam nrog kev loj hlob.raum puas.Feem ntau kawm CKD nyob rau hauv uas TUNEL siv yog mob ntshav qab zib nephropathy. Kev raug mob los ntawm TUNEL-zoo tshwm sim hauv tubules thiab podocytes [69] hauv cov neeg mob ntshav qab zib uas muaj ntshav qab zib mellitus ua ntej kuaj TUNEL txij li 7 txog 30 xyoo [104,105] thiab 2 mus rau 16 lub lis piam tom qab kho streptozotocin hauv nas [106,107]. Zoo sib xws TUNEL-zoo histopathology tau tshaj tawm hauv immunoglobulin A nephropathy [22]. Hauv lupus nephritis thiab membranoproliferative glomerulonephritis, TUNEL-zoo tau pom feem ntau hauv glomeruli [62,108,109]. Ntawm qhov tod tes, kev tshawb fawb txog kev nyab xeeb tshuaj ntawm cisplatin [110], chloroquine [45], doxorubicin [67], finasteride [23], tacrolimus [111], thiab levetiracetam [24] pom feem ntau ntawm tubular puas. Ib yam li ntawd, qhov kev raug mob txuas ntxiv raug txheeb xyuas los ntawm tubular TUNEL-zoo hlwb tau piav qhia hauv kev tshawb fawb ntawm uric acid nephropathy [112], methionine deficiency [48], raug rau ib puag ncig co toxins microcystin-LR [50], thiab cadmium [113], thiab hauv kev koom tes nrog calcium-oxalate inducedraumpob zeb [114] thiab calcifying nanoparticles [115]. Thaum kev txheeb xyuas qhov kev puas tsuaj rau lub raum thaum lub sij hawm CKD los ntawm kev siv TUNEL qhov kev ntsuam xyuas yog ib qho cuab yeej kuaj mob uas muaj txiaj ntsig zoo, kev sib koom ua ke ntawm TUNEL kev soj ntsuam nrog immunohistochemistry (IHC) tej zaum yuav muaj kev paub ntau ntxiv hauv kev txiav txim siab txog kev raug mob.

TUNEL Quantification thiab Colocalization Techniques

Cov txheej txheem ntawm TUNEL kom muaj nuj nqis sib txawv. Hauv feem ntau ntawm cov ntawv ceeb toom, TUNEL-zoo hlwb / nuclei raug suav tias yog feem pua ​​​​ntawm tag nrho cov hlwb / nuclei [83,116,117]. Qhov no zoo li yog qhov yuav tsum tau ua, txawm hais tias tus naj npawb ntawm TUNEL-zoo hlwb yuav tsis sib haum raws li tus naj npawb ntawm cov hlwb tuag. Qee lub hlwb yuav tsis tuaj yeem txheeb xyuas cov theem ntawm DNA fragmentation, thaum lwm tus tuaj yeem raug tshem tawm los ntawm macrophages. Kev ntsuas tus naj npawb ntawm TUNEL-zoo hlwb ib square unit (mm2) kuj tseem siv [118], tab sis txoj kev no zoo li tsis tshua muaj tseeb vim nws nyob ntawm qhov chaw thiab cov ntsiab lus ntawm tes ntawm thaj chaw. Cortical seem feem ntau muaj glomeruli, tubules, hlab ntsha, thiab medullary rays, thaum medullary seem feem ntau muaj loops ntawm Henle, sau ducts, thiab cov hlab ntsha. Quantification "ib Fifield" yeej nyob ntawm seb lub teb yog [72,119], thiab ntau npaum li cas "ib ntu" yog, txawm ntau, undefined [61,120]. Kev ntsuas tsis zoo, xws li "histological qhab nia" [121], "apoptotic index" [113], "apoptosis rate" [92], "feem pua ​​​​ntawm apoptosis" [122], lossis feem pua ​​​​ntawm kev tswj tsis tau kho [123,124], zoo li ua kom tsawg dua tsim nyog, thiab ua rau kev txhais lus nyuaj thiab tsis sib piv nrog feem ntau ntawm cov kev tshawb fawb.

Cell tuag hauv cov hlwb nyob sib ze pib tsis sib xws vim nws nyob ntawm vascularization, cell voj voog, thiab lwm yam hloov pauv. Qhov no feem ntau ua rau TUNEL tsis sib xws ntawm cov hlwb hauv cov ntaub so ntswg [45,96,125]. Kuj ceeb tias, qhov kev siv dav dav ntawm TUNEL-zoo teeb liab tsis siv rau kev ntsuas TUNEL, txawm tias nws ncaj qha nyob ntawm qib ntawm DNA fragmentation. Qhov no yog tej zaum vim hais tias cov hlwb raug suav hais tias tuag tsis muaj zog ntawm TUNEL teeb liab, vim tias lub xov tooj ntawm tes tsis tuaj yeem "tuag" ntau dua yog tias TUNEL staining hnyav dua. Txawm hais tias nws nyuaj rau kev sib piv cov kev tshawb fawb uas siv ntau txoj hauv kev los txheeb xyuas TUNEL cov txiaj ntsig, cov txiaj ntsig ntawm TUNEL-zoo hlwb hauv cov tsiaj tswj feem ntau yog qis dua 2 feem pua ​​[75,99,100,116,126]. Kev raug mob hnyav yog txuam nrog ~ 5–40 × nce hauv TUNEL-zoo [79,126,127]. Thaum lub sij hawm raug mob, qhov nce hauv TUNEL-zoo hlwb yog qis dua, ~ 2–{19}}fold, [29,43,65] piv rau kev tswj tsis tau kho.

Thawj TUNEL kev soj ntsuam raws li diaminobenzidine (DAB) staining yog tsim los rau lub teeb pom kev zoo [14,15]. Txoj kev cai no tseem siv nyob rau hauv ob feem peb ntawm cov kev tshawb fawb tshawb fawb hauv qhov kev tshuaj xyuas no thiab feem ntau ntawm cov kev tshawb fawb ntawm tib neeg biopsy. Qhov kev ntsuas yog ua los ntawm phau ntawv xov tooj suav thiab, thaum meej, nws siv sijhawm ntev. Feem ntau, kev ntsuas tsis tau siv tag nrho, thiab cov txiaj ntsig tau nthuav tawm siv cov duab kho mob thiab cov stained kuaj ua pov thawj yooj yim tias kev tuag ntawm tes tau tshwm sim. Automation yuav ua kom cov txheej txheem kom muaj nuj nqis [128,129]. Txawm li cas los xij, DAB tsis tuaj yeem ua ke nrog lwm cov staining hauv tib tus qauv, thiab yog li DAB-raws li TUNEL tsuas tuaj yeem muab cov cell tuag, tsis muaj kev txheeb xyuas ntawm hom cell (tshwj tsis yog pom pom) lossis cell tuag mechanism.

Thaum tib neeg kev tshawb fawb dhau los ntawm DAB-raws li TUNEL siv lub teeb microscopy, kwv yees li ib nrab ntawm cov kev tshawb fawb tsiaj siv FL fluorescent microscopy (Daim duab 3). Fluorescent TUNEL yog qhov pom kev zoo nkauj, txhais tau yooj yim, thiab muaj ntau yam zoo. Ntxiv nrog rau qhov tseeb kom muaj nuj nqis los ntawm kev txheeb xyuas ntawm TUNEL-positive nuclei lossis txhais tau tias siv cov xim, FL fluorescent TUNEL teeb liab tuaj yeem colocalized nrog lwm cov xim rau histochemistry (xws li, DAPI) (Daim duab 3A) lossis IHC (Daim duab 3B). Nyob rau yav tas los, kev siv cov cim cim protein tuaj yeem pab txheeb xyuas TUNEL-cov cim zoo cuam tshuam nrog qhov sib txawvraumcompartments los yog muab lus qhia txog hom cell tuag. Peb nrhiav tsis tau ib qho piv txwv ntawm TUNEL ua ke nrog ob chav IHC antibody staining. Txawm hais tias qhov kev sib xyaw ua ke no yuav qhia tau zoo heev, piv txwv li, rau kev txheeb xyuas qhov kev tuag ntawm tes hauv qee hom cell, cov teeb meem kev cuam tshuam nrog xws li staining yuav ntau dua cov txiaj ntsig.

Kev txheeb xyuas ntawmraumhom xovtooj ntawm tes yeej tsis muaj teeb meem rau ib tus neeg paub txograumkws kho mob [104]. Vim li no, kev siv cov cim ntxiv rau kev txheeb xyuas ntawm tes, piv txwv li, siv CD31 rau kev txheeb xyuas cov hlwb endothelial [130], tsis tshua muaj. Txawm li cas los xij, kev siv IHC cov cim cim rau cov cim ntawm cov ntaub so ntswg raug mob feem ntau siv. Feem ntau cov kev tshawb fawb siv TUNEL-positivity raws li ib qho pov thawj ntawm apoptosis. Hauv lwm cov kev tshawb fawb, kev lees paub tias TUNEL-zoo tsis yog tshwj xeeb rau apoptosis tau ua rau ntau yam morphological (xws li, hematoxylin thiab eosin lossis periodic acid-Schiff staining), biochemical, thiab IHC cov cim ua ke nrog TUNEL staining kom paub meej tias apoptosis. Feem ntau siv cov cim rau caspase-dependent apoptosis yog cleaved (active) caspase-3 los ntawm kev siv cov tshuaj tiv thaiv kab mob uas tsis paub txog qhov loj npaum caspase-3 [131,132]. Ob leeg TUNEL thiab cleaved caspase-3 kev soj ntsuam tau pom tsuas yog tubular, tsis glomerular, staining hauv nas ischemia-reperfusion [64]. Lwm cov kev tshawb fawb caspase siv caspase-1 [27], -9 [53], -11 [28], thiab -12 [75]. Lwm cov cim apoptotic suav nrog B-cell lymphoma 2 (Bcl2) tsev neeg ntawm cov proteins Bcl-2 [27], Bcl2L1 [43], Bcl-xl [71], Bax [99], Bak [49], thiab Phem [43]; apoptotic receptor Fas [52] thiab apoptotic receptor-ligand FasL [66]; forkhead box (FOX) protein transcription factor Fox01 [133] thiab Fox03 [27]; thiab cytochrome c [53]. Kev mob tshwm sim los ntawm IHC ntawm qog necrosis factor-alpha (TNF) [134]; qog loj hlob yam tseem ceeb TGF [66] thiab TGF- 1 [135]; serine/threonine kinase 1 (SGK1) [135]; electron-transfer flavoprotein, beta-subunit (ETF) [136]; Janus kinase-2 (JAK2) [137]; teeb liab transducer thiab activator ntawm transcription 3 (STAT3) [137]; nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB) [137] thiab nws cov inhibitor protein IκB [137]. RauraumKev soj ntsuam kev raug mob, lub raum raug mob molecule-1 (KIM-1) [24], aquaporin-1 channel (AQP-1) [45], hypoxia-inducible factor {{7} }alpha transcription factor (HIF-1) ​​[103], Asc-type amino acid transporter-1 (Asc-1) [121], mammalian phiaj ntawm rapamycin (mTOR) [131] kinase , thiab transglutaminase II [89] tau siv. Oxidative raum raug mob tau kawm siv cov cim xws li 8-hydroxyguanosine (8OHdG) [102], neutrophil gelatinase-associated lipocalin (NGAL) [102], NAD-dependent deacetylase sirtuin-1 (SIRT1) [138] , thiab heme oxygenase 1 (HO-1) [90]. Kinases tau kawm ntau ntxiv, suav nrog c-Jun N-terminal kinase (JNK) txoj hauv kev [101], phosphoinositide 3-kinase/protein kinase B/nuclear factor erythroid 2- cuam tshuam txog 2 qhov kev hloov pauv (PI3K/ Akt/Nrf2) pathway [46], pannexin-1 (PANX1) plasma membrane gap junction protein [26], thiab mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) [26]. Kev tshawb fawb ntawm apoptotic endonucleases koom nrog caspase-activated DNase (CAD) [139], DNase I [108], thiab EndoG [139], thiab DNA puas cov cim suav nrog qog protein p53 (p53) [99] thiab dynamin-related protein 1 (PARP1) Ib.) [66]. Qee cov kev tshawb fawb kuj pom tias qhov kev ntsuam xyuas ntawm endoplasmic reticulum stress-mediated apoptosis thiab cov ntaub so ntswg raug mob pab tau [75].

Muab ntau cov kev tshawb fawb txog kev tshawb fawb thiab qhov zoo ntawm kev sib txuas TUNEL nrog IHC cov cim rau kev txheeb xyuas cov txheej txheem kev tuag ntawm tes, ib tus yuav xav tias muaj ntau cov kev tshawb fawb siv colocalization ntawm ob. Kev tsom xam yuav ua tau los ntawm pixel-los-pixel colocalization ntawm TUNEL-zoo hlwb thiab IHC cov cim, IHC marker txhais tau hais tias siv hauv TUNEL-zoo, lossis TUNEL-zoo teeb liab txhais tau tias muaj zog hauv IHC marker-zoo thaj chaw. Hmoov tsis zoo, qhov zoo ntawm TUNEL no tau siv hauv ob peb qhov kev tshawb fawb xwb. TUNEL-zoo hlwb tau colocalized nrog cleaved caspase -3 nyob rau hauvraumtubules thaum lub sij hawm acetaminophen toxicity nyob rau hauv nas [74]. IHC colocalization nrog TUNEL-zoo teeb liab tso cai rau kev tuag ntawm tes ntawm mesangial hlwb thaum Habu nephritis [99]. Nyob rau hauv lub teeb microscopy, colocalization nrog mechanical markers ntawm toxicity tau ua nyob rau hauv serial seem, raws li qhia los ntawm Ott li al. [89].

TUNEL Teeb meem thiab kev txwv

Ntau qhov teeb meem muaj peev xwm tuaj yeem tshwm sim thiab yuav tsum xav tau thaum ua haujlwm nrog TUNEL. Ib qho yog tias 30OH DNA termini tshwm sim tsis tau tsuas yog thaum lub sijhawm tuag ntawm tes los ntawm kev ua haujlwm endonuclease tab sis kuj yog ib qho nruab nrab metabolite hauv yuav luag tag nrho cov tshuaj tiv thaiv enzymatic nrog DNA. Ib qho tseem ceeb ntawm tes ua haujlwm uas ua rau 30OH termini yog DNA synthesis, uas ntau Okazaki fragments tej zaum yuav theoretically tsim ib tug cuav-zoo TUNEL teeb liab, tshwj xeeb tshaj yog nyob rau hauv heev-rhiab heev kev hloov kho ntawm qhov kev ntsuam xyuas. Thaummob raum,Kev kho DNA kuj tseem tuaj yeem ua rau TUNEL-cov cim tsis zoo raws li qhov tshwm sim ntawm AP endonuclease. Loj oxidative raum raug mob tshwm sim los ntawm tshuaj lom neeg (hydrogen peroxide, bleomycin) los yog lub cev (gamma irradiation) raug mob yuav ntxiv rau tus naj npawb ntawm 30OH kawg, ua rau cov ntawv nyeem tsis tseeb-zoo TUNEL sib nrug los ntawm endonuclease mediated DNA fragmentation.

Ntau zaus thiab ntau dua, teeb tsa kev sim tsis raug ntawm TdT cov tshuaj tiv thaiv (lub sijhawm, qhov kub thiab txias, kev ua kom muaj zog, lossis cov cofactors) tuaj yeem ua rau muaj qhov tsis zoo ntawm TUNEL-zoo teeb liab keeb kwm yav dhau. Txawm hais tias nws tseem tsis tau hais meej txog qhov uas suav tias yog lub hauv paus ib txwm rau kev kuaj mob raum tsis zoo, nws zoo li qhov tseeb tias nyob ib ncig ntawm 1 feem pua ​​​​ntawm tag nrho cov hlwb yuav yog tus lej tsim nyog. Muaj tseeb tiag, 10 feem pua ​​​​lossis siab dua keeb kwm yav dhau los yog cov pov thawj tseeb ntawm qhov tsis zoo thiab tsis raug tshuaj xyuas los yog qhov sib txawv ntawm cov qauv khaws cia tsim nyog.

Lwm qhov teeb meem uas xav tau yog qhov ua haujlwm DNase I qis hauv qee hom cell (piv txwv li, endothelium),raumcompartments (piv txwv li, glomeruli), los yog cov ntaub so ntswg pathological (xws li, qog). Cov hlwb no yuav tsum tau ua kom pom qhov tsis zoo ntawm TUNEL, tsis yog vim tias lawv raug mob, tab sis vim tias tsis muaj cov haujlwm endonuclease txaus los tsim cov teeb liab ntsuas hauv TUNEL. Hauv peb qhov kev paub dhau los, txawm tias qhov hloov pauv ntawm {{0}}.01 feem pua ​​​​thiab 0.1 feem pua ​​​​ntawm TUNEL cov hlwb zoo tuaj yeem ua pov thawj tseem ceeb thiab siv los ua pov thawj ntawm lub raum (ntawm lwm cov ntaub so ntswg) raug mob.

Thaum kawg, ib qho teeb meem nrog rau qhov muaj pes tsawg leeg ntawm TUNEL staining tshwm sim thaum tus nqi ntawm DNA fragmentation siab dhau lawm, piv txwv li, thaum lub sij hawm lig ntawm cell tuag los yog nyob rau hauv high-kev raug mob, qhov uas seem DNA tsis tuaj yeem raug counterstained nrog DAPI vim disassembly ntawm lub ob chav helix (Daim duab 3D). Nyob rau hauv cov ntaub ntawv no, TUNEL-zoo teeb liab yog "dai hauv huab cua", thiab zoo li tsis cuam tshuam nrog lub nucleus. Nws dhau los ua lub luag haujlwm ntawm tus kws kho mob los txhais cov no raws li TUNEL-zoo hlwb es tsis yog khoom cuav. Nrog fluorescein-labeled TUNEL, muaj qhov txaus ntshai ntawm kev txheeb xyuas keeb kwm yav dhau autofluorescence hauv cov ntaub so ntswg li TUNEL-zoo [140]. Hauv cov xwm txheej no, hloov mus rau liab spectrum TUNEL [51,54] raug pom zoo.

TUNEL Cov Qauv raws li qhov chaw ntawm cov ntaub ntawv ntxiv

Ntau tus qauv TUNEL tuaj yeem ua kom pom tseeb thiab siv los txiav txim siab txog qhov muaj peev xwm thiab qib kev raug mob (Table 1, Daim duab 3). Feem ntau, ib hom nuclei nkaus xwb ntawm TUNEL qauv yog tsim los ntawm kev raug mob me me, thaum muaj kev raug mob hnyav dua (piv txwv li, kev raug mob siab) ua rau muaj zog staining thiab disintegration ntawm nuclei lossis hlwb. Ib tug nruj apoptotic TUNEL qauv yog nuclear fragmentation raws li ib tug tshwm sim ntawm apoptotic lub cev tsim (Daim duab 3E). Cov tawg me me ntawm TUNEL-zoo nuclei, uas zoo li lub cev apoptotic, tau pom hauv nas ischemia-reperfusion ntawm 24-72 h [97]. Apoptotic lub cev kuj tau pom nrog TUNEL sau npe ntawm nuclei hauv lub raum tubular epithelium thiab interstitium 14 hnub tom qab ischemia-reperfusion raug mob hauv nas [141]. Kev nce koob tshuaj ntawm cov roj lavender txo cov nuclei-tsuas yog TUNEL cov qauv thiab hloov mus rau necrosis zoo li TUNEL nrog cytoplasmic to [94]. Cov tom kawg yog tshwm sim los ntawm kev puas tsuaj ntawm lub hnab ntawv nuclear vim qhov kev ua ntawm cov proteinases thiab lipases hauv necrosis. Cytoplasmic TUNEL (Daim duab 3F) pom raws li TUNEL-zoo cytoplasm tau pom nyob rau hauv lub teeb microscopy nyob rau hauv lub rhabdomyolysis qauv nyob rau hauv nas tubules [142] thiab nyob rau hauv podocytes kab lis kev cai nyob rau hauv lub xub ntiag ntawm siab qabzib thiab nyob rau hauv mob ntshav qab zib nephropathy nyob rau hauv nas [135]. Cytoplasmic TUNEL kuj tau piav qhia hauv lub raum ischemia-reperfusion hauv cov nas uas siv FL fluorescent TUNEL [98] thiab hauv ischemia-reperfusion hauv nas siv lub teeb microscopy [143]. Mercuric chloride-induced profound thiab tshwj xeeb tubular puas tsuaj nrog muaj zog cytoplasmic TUNEL staining qhia txog nuclear membrane puas [60]. Yog tias qhov kev raug mob txuas ntxiv los yog qhov koob tshuaj ntawm cov neeg ua kom puas tsuaj nce ntxiv, cytoplasmic TUNEL tuaj yeem hloov mus rau ib daim duab "ntsiab" ntawm TUNEL dispersed (Daim duab 3G) vim yog cov theem ntawm kev puas tsuaj ntawm tes vim necrosis thiab spillage ntawm cellular cytoplasm thiab khib nyiab. Piv txwv li, ob qho tib si cytoplasmic thiab dispersed TUNEL tau pom nyob rau hauv cisplatin-kho nas [71,72] thiab nas [144], thiab hauv podocyte raug mob hauv streptozotocin-induced diabetic nas [145]. Dispersed necrotic TUNEL kuj tau pom thaum lub sij hawm uranyl acetate-induced nephrotoxicity hauv nas [101]. Thaum kawg, lwm TUNEL teeb liab sib txawv los ua tib zoo saib yog qhov kev siv ntawm TUNEL-zoo cov cim qhia, uas tuaj yeem sib txawv ntawm lub xov tooj mus rau ntawm tes nyob ntawm seb muaj pes tsawg tus so (degree ntawm kev raug mob) txhua qhov kev paub ntawm tes, raws li sau tseg los ntawm qee cov kev tshawb fawb [45,96,125 ]. Rau cov qauv uas sib txawv ntawm TUNEL teeb liab siv tau pom, kev txiav txim siab los ntawm ob qho tib si TUNEL-zoo khoom thiab qhov nruab nrab ntawm qhov staining yog pom zoo.

Cov lus xaus

Vim muaj kev ua haujlwm siab ntawm apoptotic endonucleases hauv lub raum, tshwj xeeb yog DNase I thiab EndoG, kev siv TUNEL rauraum tshawb nrhiav thiab kuaj mobyog cov ntaub ntawv heev. Nws yuav dhau los ua qhov tseem ceeb dua yog tias TUNEL tau siv dhau ntawm nws qhov kev siv thawj zaug ua ib txoj hauv kev los txheeb xyuas apoptosis. Kev thov TUNEL nkaus xwb rau kev tshawb fawb apoptosis yog ib qho piv txwv zoo ntawm kev tshawb fawb yuav ua li cas ua yuam kev los ntawm txoj kev siv tsis tsim nyog. Txawm hais tias nws siv los ua qhov kev ntsuam xyuas raug mob feem ntau, TUNEL tuaj yeem siv tau zoo los ntsuas ob qho tib si AKI thiab CKD. Txawm li cas los xij, hauv CKD, qhov degree ntawm TUNEL-zoo yuav tsum xav tias yuav qis dua qhov raug mob hnyav. Vim tias qhov kev ua ntawm endonucleases (tsuas yog, DNase I) ntau dua hauv tubular epithelium dua li hauv lwm lub raum, kev raug mob rau lub raum tubules feem ntau pom tias yog qhov tseem ceeb. Txawm li cas los xij, kev raug mob glomerular lossis vascular raug mob kuj tuaj yeem kawm siv TUNEL yog tias qhov kev raug mob tshwm sim hauv cov khoom no. Lub colocalization ntawm FL fluorescent TUNEL nrog IHC cov cim tso cai rau kev txheeb xyuas thiab kev koom tes ntawm cov xov tooj ntawm tes thiab cov txheej txheem kev tuag ntawm tes nrog TUNEL cov cim. TUNEL kom muaj nuj nqis tuaj yeem pab txhawb cov ntaub ntawv tseem ceeb ntawm qib ntawm kev tuag ntawm tes, thiab siv cov duab tsom xam software tuaj yeem pab tau zoo hauv qhov no. Xav txog ntau yam ntawm TUNEL cov qauv, cov duab sawv cev tau pom zoo los tsim thiab ua qauv qhia

TUNEL cov cim qhia uas suav tias yog qhov zoo. Thaum kawg, TUNEL cov qauv yuav tsum tau siv los ua qhov chaw ntawm cov ntaub ntawv ntxiv, xws li cov txheej txheem ntawm kev tuag ntawm tes thiab qib ntawm toxicity. Piv txwv li, qhov sib txawv ntawm TUNEL-zoo apoptotic lub cev, muaj cov cim cytoplasmic TUNEL, lossis cov qauv TUNEL tawg tuaj yeem pab txheeb xyuas cov kab mob apoptotic vs. necrotic TUNEL-zoo hlwb rau kev txhais lus meej ntawm kev tshawb fawb thiab kev kuaj mob.

kev sib deev arousal raum kev kho mob