Isihloko
I-methylene blue namaseli asekela ubuchopho: ama-astrocyte, ama-microglia, nokuvuvukala
Ake sithi wenza izivivinyo ezimbili ngodayi ofanayo oluhlaza okwesibhakabhaka bese uthola imiphumela emibili ekhuthazayo. Esitsheni esisodwa, amaseli asekela amanye amaseli, amise okwezinkanyezi, ayasinda lapho elahlekelwa umoya-mpilo ne-glucose. Kwesinye, amaseli okuvikela ubuchopho agcina ukuma kwawo okunamagatsha, okukhombisa ukuzola, naphezu kobuthi bamagciwane. Kulula ukulingeka uhlanganise lokhu kube isimangalo esisodwa: udayi udambisa ukuvuvukala kobuchopho futhi uvikele ubuchopho. Yilapho kanye abafundi abacophelelayo kufanele bame bacabangisise khona.
Ubuchopho bunamaqembu amabili amaseli asekelayo anemisebenzi ehlukene, futhi i-methylene blue ihlangana neqembu ngalinye ngesikhuthazi esihlukile esivivinyweni. Ama-astrocyte angamaseli asekela i-metabolism. Atholakala phakathi kwemithambo yegazi nama-neuron, asusa i-glutamate eyeqile, alawula amazinga e-potassium, agcina i-glycogen, futhi adlulisela izinto ezinikeza amandla kuma-neuron lapho isidingo sikhuphuka kakhulu. Ama-microglia angamaseli okuvikela ahlala khona aqapha isimo. Izandiso zawo ezincane zigada izicubu, zithole izimpawu zamagciwane nezokulimala, bese kuthi lapho kuba nesexwayiso zihoxe zibe ukuma okuyindilinga okufana ne-amoeba, okulungele ukuhamba nokugwinya izinto, kuyilapho zikhipha ama-molecule okusayina. Umphumela eqenjini elilodwa awudluleli kwelinye, futhi isilinganiso esisetshenziswe esimweni ngasinye sinquma lokho okuboniswe ngempela.
Ama-astrocyte ngaphansi kokucindezeleka kwe-metabolism: isikhathi siwumphumela
Isivivinyo sama-astrocyte siqala ngokuncishwa izinto azidingayo. Abacwaningi bakhulisa ama-astrocyte avela engxenyeni engaphandle yobuchopho bamagundane amancane asanda kuzalwa, basusa i-glucose nomoya-mpilo cishe wonke amahora ayisithupha, base bebuyisela kokubili amahora angamashumi amabili nane. Lokhu kuyimodeli yaselabhorethri yokushaywa unhlangothi okulandelwa ukubuyela kokugeleza kwegazi. Lapho i-methylene blue ikhona esigabeni sokuncishwa, ukusinda kwamaseli kwaba kubi kakhulu. Lapho ifakwa kuphela esigabeni sokubuyiselwa komoya-mpilo, ku-0.1 no-1 micromolar, ukusinda kwathuthuka ngokuphawulekayo ngokwesivivinyo se-calcein sokuhlola ukuphila kwamaseli. Ucwaningo lwama-astrocyte amagundane amancane lubika lo mehluko wesikhathi kanye nedatha yokuphefumula kwamaseli.
Lowo mehluko yiwona mphumela obaluleke kakhulu. Ngesikhathi sokuncishwa cishe awukho umoya-mpilo wokwamukela ama-electron, ngakho into edlulisa ama-electron ayinandawo yokuwahambisa. Ngesikhathi sokubuyiselwa komoya-mpilo, izinqubo zamaseli ziqala futhi, bese udayi uba nomsebenzi ongawenza. Ama-astrocyte afakwe udayi asebenzisa umoya-mpilo omningi, amunca i-glucose eningi, akhiqiza i-ATP eningi, futhi akha kabusha izinqolobane ze-glycogen. Umsebenzi we-hexokinase wakhuphuka, kodwa i-superoxide kuma-mitochondria ayizange yande. Incazelo ehlongozwayo yileyo ejwayelekile evela encazelweni ye-methylene blue njengomthwali wama-electron kuma-mitochondria: idlulisa ama-electron adlule ezindaweni ezivinjiwe ochungechungeni lokuphefumula kwamaseli, ukuze ukuphefumula kuqale futhi ngendlela ehlelekile kunokuba ama-electron avuze aye ekwakheni izinhlobo zamakhemikhali ezisebenza kakhulu.
Qaphela ukuthi lobu ubufakazi baluphi uhlobo. Kungumphumela wokusinda ne-metabolism ohlotsheni olulodwa lwamaseli oluhlukanisiwe, onesigaba esishiwo lapho kusebenza khona kahle — ukululama, hhayi ukuncishwa — kanye nobubanzi bokugxila obushiwo. Awusho lutho ngokusayina kwamasosha omzimba, futhi awusho lutho ngokuziphatha.
Ama-microglia ngaphansi kwesexwayiso samagciwane: i-metabolism ihlelwa kabusha
Izivivinyo zama-microglia ziqala ngesikhuthazi esiphambene: hhayi ukuswela ukudla, kodwa isexwayiso esingamanga sokutheleleka. I-lipopolysaccharide, ingxenye yolwelwesi lwangaphandle lwamagciwane, ibophezela ezinzwa zamasosha omzimba azalwa nawo futhi yenze ama-microglia acabange ukuthi sekufike amagciwane abangela izifo, nakuba kungekho magciwane aphilayo. Emagundaneni amancane anikezwe i-lipopolysaccharide nasohlwini lwamaseli ama-microglia akhuliswa elabhorethri, umphumela uyinqubo ejwayelekile yokuvuvukala: izinto ezikhuthaza ukuvuvukala ziyanda, ama-microglia ashintshela esimweni se-M1 esinolaka, ama-neuron angomakhelwane ayafa, futhi izilwane zenza kabi kakhulu emisebenzini yokufunda nokukhumbula.
Ucwaningo lwamagundane amancane ngalokhu kusabela lululandelela luye endleleni ezwa umoya-mpilo: i-lipopolysaccharide ikhuthaza ukukhiqizwa ngokweqile kwe-PHD3, enye yama-enzyme amathathu e-prolyl hydroxylase, ngochungechunge lwe-Siah2/Morg1 olungaphambi kwayo endleleni yokusayina. Ngesikhathi esifanayo, amaseli ahlela kabusha ukusebenzisa kwawo amandla aqonde ku-glycolysis lapho kukhona umoya-mpilo, kuhambisane nokwanda kwezinhlobo zomoya-mpilo ezisebenza kakhulu kuma-mitochondria. Ababhali balinganisa lolu shintsho ngokuqondile ngezivivinyo ze-Seahorse ze-metabolism ezikala ukusetshenziswa komoya-mpilo nokwanda kobumuncu ngaphandle kwamaseli. I-methylene blue yabuyisela emuva kokubili ushintsho lwe-metabolism nokukhiqizwa ngokweqile kwe-PHD3, yehlisa izinto ezibangela ukuvuvukala, yanciphisa ukufa kwama-neuron, futhi yathuthukisa amaphuzu okuziphatha. Ukuthwebula nge-positron emission tomography kwengeza ubufakazi bezithombe: i-lipopolysaccharide yandisa ukumuncwa kwe-glucose ku-hippocampus njengoba kubonwe ku-FDG-PET, kanti ukufakwa kwe-methylene blue kwakwehlisa futhi, okuhambisana ne-metabolism yokuvuvukala ethule kakhudlwana ebuchosheni obuphilayo.
Leli iqoqo lezilinganiso elibanzi kunelocwaningo lwama-astrocyte — ama-cytokine, i-metabolism, izithombe, nokuziphatha kwezilwane — kodwa lihlobene nesikhuthazi esihlukile neseli elihlukile. Abafundi abaqhathanisa lokhu okubili kufanele bahlukanise izikhuthazi: ucwaningo olulodwa lusindisa amaseli asekelayo aswele ukudla ngesikhathi sokululama, olunye ludambisa isexwayiso esingamanga sokutheleleka kumaseli okuvikela.
Ama-microglia ngaphansi kwesibonakhulu: ukuma njengenkomba
Isivivinyo sesithathu sibheka isexwayiso esifanayo samagciwane ngokuma kwamaseli kuphela. Kumaseli e-glia akhuliswa elabhorethri avela engxenyeni engaphandle yobuchopho bamagundane amakhulu, ama-microglia eqembu lokuqhathanisa afakwe umbala ukuze kubonakale umaka i-IBA1 abonisa izandiso eziningi ezincane, okuwukuma okunamagatsha kokugada. Ngemva kwamahora angamashumi amabili nane e-lipopolysaccharide engu-10 micrograms per milliliter, amaseli aba isicaba athathe ukuma okufana ne-amoeba, futhi indawo yemizimba yawo iyakhula kuyilapho ubude bemingcele buhlala cishe bufana. Lokhu kuwuphawu lwejiyomethri lokuba yindilinga. I-methylene blue engu-1 micromolar, efakwe kanye nobuthi, yagcina izandiso ezinamagatsha futhi yagcina indawo yomzimba nobude bomngcele kusondele kumanani eqembu lokuqhathanisa. Ucwaningo lwamaseli amagundane amakhulu akhuliswa elabhorethri lulinganisa lokhu kugcinwa kokuma ngezilinganiso zendawo yomzimba weseli nobude bomngcele.
Imininingwane emibili yenza lolu cwaningo lube ngaphezu kwesithombe esilula. Okokuqala, i-menadione eyasetshenziswa ukuqhathanisa, enye i-molecule esebenza nge-redox ekugxileni okufanayo, ayizange ibuyisele ukuma, futhi iyodwa yaphushela amaseli esimweni esivuselelwe. Ngakho lo mphumela awuyona into ejwayelekile kuzo zonke izinhlanganisela ezijikeleza phakathi kwezimo ze-redox. Okwesibili, ababhali baxhumanisa umphumela kadayi nokuvuselelwa kwe-Nrf2, uhlelo lwesici esilawula ukulotshwa kwezakhi zofuzo ezingemuva kokulwa nokonakala nge-oxidation nokulwa nokuvuvukala, kunokuwuxhumanisa kuphela nokubamba ngokuqondile izinhlobo zamakhemikhali ezisebenza kakhulu. Izingxoxo ezihlobene ne-redox nokusayina kokuvuvukala zibeka lokhu kuxhumana kwe-Nrf2 kumongo wakho.
Noma kunjalo, ukuma kuyinkomba enobufakazi obuncane kunazo zonke kulezi ezintathu. Iseli elinamagatsha ngokuvamile liyiseli eligadayo, futhi iseli eliyindilinga ngokuvamile liyiseli elivuselelwe, kodwa ukuma kuyinkomba engathathi indawo yokubona konke okwenzekayo, hhayi umbhalo oqondile wako. Leli qoqo lamaseli lalingenawo amaseli okuvikela avela egazini, lingenamanethiwekhi ezinzwa, futhi lingenamphumela wokuziphatha.
Ukufunda imiphumela emithathu ihambisana
| Isikhuthazi | Uhlobo lweseli | Okwalinganiswa | Okwenziwa yi-methylene blue |
|---|---|---|---|
| Umoya-mpilo ne-glucose kwasuswa, kwase kubuyiselwa | Ama-astrocyte amagundane amancane | Ukuphila kwamaseli, ukusetshenziswa komoya-mpilo, ukumuncwa kwe-glucose, i-ATP, i-glycogen, i-superoxide | Yavikela kuphela lapho ifakwa ngesikhathi sokubuyiselwa komoya-mpilo; yandisa ukuphefumula kwamaseli nezinqolobane ngaphandle kokwandisa i-superoxide |
| Ingxenye yobuthi bamagciwane | Ama-microglia amagundane amancane, akhuliswa elabhorethri nasemagundaneni aphilayo | Ama-cytokine, omaka be-M1, i-metabolism ye-Seahorse, amaprotheni endlela ye-PHD3, i-FDG-PET, imisebenzi yokukhumbula | Yabuyisela emuva ukuhlelwa kabusha kwe-glycolysis nokukhiqizwa ngokweqile kwe-PHD3; yehlisa ukuvuvukala; yathuthukisa ukuziphatha |
| Ingxenye yobuthi bamagciwane | Ama-microglia amagundane amakhulu eqoqweni elixubile lamaseli e-glia akhuliswa elabhorethri | Indawo yomzimba emakwe nge-IBA1, ubude bomngcele, ukuba namagatsha kwezandiso | Yagcina ukuma okunamagatsha kwesimo sokuphumula; i-menadione ayikwenzanga lokho |
Iphethini kuyo yonke imigqa ukuthi isikhuthazi ngasinye sinesilinganiso saso nempendulo yaso. Ukusindiswa kwe-metabolism kuphendula umbuzo we-metabolism. Idatha yama-cytokine nokuziphatha iphendula umbuzo wokuvuvukala. Ukuma kuphendula umbuzo wokuma. Akukho kulokhu okuthathu okudlulela komunye umkhakha ngokwakho.
Lokho ama-microglia abukeka ezolile angakufakazeli
Lona umusho odingwa yilesi sihloko sonke: ama-microglia abukeka engavuselelwe kakhulu, noma i-metabolism yama-microglia eshintshile, akufakazeli ngokwakho ukuthi ukusebenza kobuchopho kuthuthukile.
Ukuma kungashintsha ngaphandle kokuba ukusebenza kulandele. Iseli lingagcina amagatsha alo kodwa lisakhipha ama-cytokine, noma libe yindilinga kuyilapho lenza umsebenzi owusizo wokususa izinsalela. Izilinganiso ze-metabolism zinomkhawulo ofanayo: igrafu ye-Seahorse noma isithombe se-PET sibonisa ukuphathwa kwamandla, hhayi amandla okucabanga. Ucwaningo lwamagundane amancane oluhlanganise izilinganiso ze-metabolism nezinjengokuma kwamaseli kanye nokuhlolwa kokuziphatha lunobufakazi obuqinile ngenxa yokuthi luhlanganise amazinga ahlukene obufakazi, kunokulindela ukuba isilinganiso esisodwa sithwale isiphetho. Nalapho, imikhawulo evamile iyasebenza, ngoba i-lipopolysaccharide imelela isexwayiso esingenamagciwane aphilayo kunokutheleleka kwangempela, amagundane akubona abantu, futhi ababhali ngokwabo baqokomisa umehluko phakathi kwezinhlobo zezinto eziphilayo. Noma ubani ohlela ucwaningo olulandelayo kufanele ahlole ukuthi okutholakele ngama-microglia kudlulela kanjani phakathi kwamamodeli ngaphambi kokucabanga ukuthi le ndlela isebenza ngokufanayo ezicutshini zabantu.
Kulokhu kulandela iphuzu elisebenzayo mayelana nesikhathi. Umphumela wama-astrocyte ubonisa umphumela omubi wokufaka kusenesikhathi nomphumela omuhle wokufaka ngesikhathi sokululama. Imiphumela yama-microglia ivela ekufakweni ngesikhathi esiseduze nesikhuthazi sokuvuvukala. Ukulimala kobuchopho empilweni yangempela kuhlanganisa zombili lezi zigaba ngesikhathi esisodwa: ukuntuleka kwamandla kanye nesexwayiso samasosha omzimba, okushintsha phakathi namahora. Isivivinyo esihlukanisa isigaba esisodwa asikwazi ukunquma uhlelo lwezikhathi lwesimo esihlanganisiwe. Yingakho izingxoxo zezikhathi zokufaka zigcina imigqa yesikhathi yokuncishwa neyokuvuvukala ihlukene.
Isifinyezo esithembekile singena esigabeni esisodwa. Kuma-astrocyte ahlukanisiwe, i-methylene blue efakwa ngesikhathi sokubuyiselwa komoya-mpilo iqala kabusha ukuphefumula kwamaseli futhi isindise amaseli. Kuma-microglia abhekene nengxenye yegciwane, ibuyisela emuva uhlelo lwe-metabolism yokuvuvukala ngendlela ethile ye-PHD3 futhi igcine ukuma kwesimo sokuphumula. Umphumela ngamunye uyiqiniso ngaphakathi kwezimo zesivivinyo sawo, futhi akukho kuwo okuyisinqumo esijwayelekile ngokuvuvukala kobuchopho. Ukuqhathanisa amaseli e-glia kuwusizo ngoba kuvimbela lezi zindaba ezimbili ukuba zifiphale zihlangane zibe yindaba eyodwa.