Inqaku
I-methylene blue neeseli ezixhasa ingqondo: ii-astrocyte, i-microglia, nokudumba
Masithi wenza iimvavanyo ezimbini ngedayi enye eluhlaza okwesibhakabhaka uze ufumane iziphumo ezimbini ezikhuthazayo. Kwesinye isitya, iiseli ezixhasayo ezimile okweenkwenkwezi ziyasinda ekususweni kweoksijini neglucose. Kwesinye, iiseli zokhuselo lomzimba ezisengqondweni zigcina ukumila kwazo okuzolileyo okunamasebe nangona kukho ityhefu yebhaktheriya. Kulula ukufuna ukudibanisa ezi ziphumo zibe libango elinye: idayi ithomalalisa ukudumba engqondweni ize ikhusele ingqondo. Kanye kolu dibaniso apho abafundi abanenyameko kufuneka banqumame khona.
Ingqondo inamaqela amabini eeseli ezixhasayo anemisebenzi eyahlukeneyo, yaye i-methylene blue idibana neqela ngalinye ngento eyahlukileyo ebangela impendulo kuvavanyo. Ii-astrocyte ziiseli ezixhasa imetabolism. Zihlala phakathi kwemithambo yegazi nee-neuron, zisusa iglutamate egqithisileyo, zilungelelanisa ipotassium, zigcina iglycogen, zize zidlulisele umthombo wamandla kwii-neuron xa imfuno inyuka ngesiquphe. I-microglia ziiseli zokhuselo lomzimba ezihlala apho, zihlola imeko. Izandiso zazo ezibhityileyo zihlola izicubu, zibhaqe imiqondiso yeebhaktheriya neyomonakalo, zize xa kukho isilumkiso zirhoxe zibe nokumila okungqukuva okufana neamoeba, okulungiselelwe ukuhamba nokugubungela izinto, ngoxa zikhupha iimolekyuli ezidlulisa imiqondiso. Isiphumo kwelinye iqela asidluleli kwelinye, yaye umlinganiselo osetyenziswa kwimeko nganye ngowona umisela into eboniswe ngokwenene.
Ii-astrocyte phantsi koxinzelelo lwemetabolism: ixesha lilo isiphumo
Uvavanyo lwee-astrocyte luqala ngokuzivimba izinto ezizidingayo. Abaphandi bakhulisa ii-astrocyte ezivela kwi-cortex yeempuku ezisandul’ ukuzalwa, basusa iglucose phantse nayo yonke ioksijini kangangeeyure ezintandathu, baza babuyisela zombini kangangeeyure ezingamashumi amabini anesine. Le yimodeli yaselabhoratri yestroke elandelwa kukubuyela kokuhamba kwegazi. Xa i-methylene blue yayikho kwisigaba sokuvinjwa kweoksijini neglucose, ukusinda kweeseli kwaba kubi ngakumbi. Xa yafakwa kuphela kwisigaba sokubuyiselwa kweoksijini, kwi-0.1 nakwi-1 micromolar, ukusinda kwaphucuka ngokuphawulekayo ngokovavanyo lokuphila kweeseli olusebenzisa icalcein. Uphononongo lwee-astrocyte zeempuku luchaza lo mahluko wexesha kunye nedatha yokuphefumla kweeseli.
Loo mahluko ngowona uphambili kwiziphumo. Ngexesha lokuvinjwa, phantse akukho oksijini inokwamkela iielectron, ngoko indlela yokudlulisa iielectron ayinandawo yokuzisa kuyo. Xa ioksijini ibuyiselwa, iinkqubo zeseli ziqala kwakhona yaye idayi inomsebenzi enokuwenza. Ii-astrocyte ezifumene idayi zasebenzisa ioksijini engakumbi, zangenisa iglucose engakumbi, zavelisa i-ATP engakumbi, zaza zakha kwakhona oovimba beglycogen, kunye nokwanda komsebenzi wehexokinase ngaphandle kokunyuka kwesuperoxide kwiimitochondria. Ingcaciso ecetywayo yileyo iqhelekileyo kwingcaciso ye-methylene blue njengomthwali wee-electron kwiimitochondria: idlulisa iielectron ngaphaya kweendawo ezivalekileyo kwikhonkco lokuphefumla kweeseli ukuze ukuphefumla kuqale kwakhona kakuhle, endaweni yokuvuza kwee-electron ezivelisa iimolekyuli ezisebenza kakhulu ngokwekhemikhali.
Qaphela ukuba olu luhlobo luni lobungqina. Sisiphumo sokusinda nemetabolism kuhlobo olunye lweeseli ezahlulwe kwezinye, esinefestile yexesha ecacisiweyo apho idayi isebenza khona — ngexesha lokuchacha, kungekhona ngexesha lokuvinjwa — kunye noluhlu olucacisiweyo lokuxinana kwayo. Asitsho nto malunga nokudluliselwa kwemiqondiso yokhuselo lomzimba, yaye asitsho nto malunga nokuziphatha.
I-microglia phantsi kwesilumkiso sebhaktheriya: imetabolism ehlengahlengisiweyo
Iimvavanyo ze-microglia ziqala ngento eyahlukileyo ebangela impendulo: asikokulamba, kodwa lusulelo olungeyonyani. Ilipopolysaccharide, isiqwenga senwebu yangaphandle yebhaktheriya, ibophelela kwizivamvo zokhuselo lomzimba lwendalo ize yenze i-microglia ibone ngathi iintsholongwane ezibangela izifo zifikile, nangona kungekho bhaktheriya ziphilayo. Kwiimpuku ezinikwe ilipopolysaccharide nakumgca weeseli ze-microglia okhuliswe elabhoratri, isiphumo yinkqubo yokudumba eqhelekileyo: izinto ezikhuthaza ukudumba ziyanda, i-microglia itshintshela kwimeko ye-M1 ehlaselayo, ii-neuron ezikufutshane ziyafa, yaye izilwanyana ziqhuba kakubi ngakumbi kwimisebenzi yokufunda nokukhumbula.
Uphononongo lwale mpendulo kwiimpuku luyilandela ukuya kwindlela yokuziva ioksijini: ilipopolysaccharide ibangela ukuveliswa okugqithisileyo kwe-PHD3, enye yee-enzyme ezintathu zeprolyl hydroxylase, ngokusebenzisa ikhonkco le-Siah2/Morg1 elingaphambi kwayo kule ndlela. Kwangaxeshanye, iiseli zihlengahlengisa ukusetyenziswa kwamandla zize zitshintshele kwi-aerobic glycolysis, kunye nokwanda kweemolekyuli zeoksijini ezisebenza kakhulu ngokwekhemikhali kwiimitochondria. Ababhali balulinganisa ngokuthe ngqo olu tshintsho ngeemvavanyo zeSeahorse zemetabolism ezilinganisa ukusetyenziswa kweoksijini nokwanda kobumuncu ngaphandle kweeseli. I-methylene blue yarhoxisa zombini utshintsho lwemetabolism nokuveliswa okugqithisileyo kwe-PHD3, yanciphisa izinto ezikhuthaza ukudumba, yanciphisa ukufa kwee-neuron, yaza yaphucula amanqaku eemvavanyo zokuziphatha. I-positron emission tomography yongeza ubungqina bemifanekiso: ilipopolysaccharide yandisa ukungeniswa kweglucose kwihippocampus njengoko kubonwa kwi-FDG-PET, yaye unyango lwayibuyisela ezantsi, nto leyo ehambelana nokuthomalala kwemetabolism yokudumba engqondweni ephilayo.
Le yingqokelela yemilinganiselo ebanzi ngakumbi kuneyophononongo lwee-astrocyte — iicytokine, imetabolism, imifanekiso, nokuziphatha kwizilwanyana — kodwa yeyento eyahlukileyo ebangela impendulo nakwiseli eyahlukileyo. Abafundi abathelekisa ezi zifundo zimbini kufuneka bagcine izinto ezibangela iimpendulo zahlukene: olunye uphononongo luhlangula iiseli ezixhasayo ezilambileyo ngexesha lokuchacha, kanti olunye luthomalalisa isilumkiso sosulelo olungeyonyani kwiiseli zokhuselo lomzimba.
I-microglia phantsi kwemikroskopu: ukumila njengomlinganiselo
Uvavanyo lwesithathu lujonga isilumkiso esifanayo sebhaktheriya ngokumila kuphela. Kwiiseli zeglia ezikhuliswe zivela kwi-cortex yeempuku ezinkulu, iiseli ze-microglia zeqela lolawulo ezidaywe ukubonisa uphawu lwe-IBA1 zibonisa izandiso ezininzi ezibhityileyo, oko kukuthi ukumila okunamasebe okulungiselelwe ukuhlola. Emva kweeyure ezingamashumi amabini anesine zelipopolysaccharide kwi-10 micrograms nge-milliliter, iiseli ziba mcaba zize zithathe ukumila okufana neamoeba. Ummandla womzimba weseli uyakhula ngoxa umjikelezo wawo uhlala uphantse ungatshintshi, nto leyo eluphawu lwejometri lokuba ngqukuva. I-methylene blue kwi-1 micromolar, enikwe kunye netyhefu, yagcina izandiso ezinamasebe yaza yagcina ummandla nomjikelezo womzimba weseli kufutshane namaxabiso eqela lolawulo. Uphononongo lweeseli ezikhuliswe zivela kwiimpuku ezinkulu lulinganisa oku kugcinwa kokumila ngemilinganiselo yommandla nomjikelezo womzimba weseli.
Iinkcukacha ezimbini zenza olu phononongo lube nexabiso elingaphezu komfanekiso nje. Okokuqala, ikhompawundi yokuthelekisa, imenadione, eyenye imolekyuli esebenzayo kwiinkqubo ze-redox kuxinano olufanayo, ayizange ihlangule ukumila, yaye yona ngokwayo yatyhalela iiseli ekumileni kwemeko yokusebenza. Ngoko ke esi siphumo asiyonto eqhelekileyo kuzo zonke iimolekyuli ezijikeleza phakathi kweemeko ze-redox. Okwesibini, ababhali banxulumanisa isiphumo sedayi nokuvuselelwa kwe-Nrf2, inkqubo yento elawula ukukhutshelwa kolwazi lwemfuza ehambisana nemfuza yokulwa ioksideshini neyokulwa ukudumba, kungekhona nje nokususwa ngokuthe ngqo kweemolekyuli ezisebenza kakhulu ngokwekhemikhali. Ingxoxo enxulumene ne-redox nokudluliselwa kwemiqondiso yokudumba ibeka olu nxulumano lwe-Nrf2 kumxholo walo.
Nangona kunjalo, ukumila kokona kunobungqina obuncinane kule milinganiselo mithathu. Iseli enamasebe idla ngokuba yiseli ehlolayo, yaye iseli engqukuva idla ngokuba kwimeko yokusebenza, kodwa ukumila sisalathisi esingathanga ngqo, asiyongxelo ethe ngqo yemisebenzi yeseli. Ezi seli zikhuliswe elabhoratri zazingenazo iiseli zokhuselo lomzimba ezifika ngegazi, zingenazo iisekethe zemithambo-luvo, yaye zingenaziphumo zokuziphatha.
Ukufunda ezi ziphumo zintathu ecaleni kwenye
| Into ebangela impendulo | Uhlobo lweseli | Oko kwalinganiswayo | Oko kwenziwa yi-methylene blue |
|---|---|---|---|
| Ioksijini neglucose zisusiwe, zaza zabuyiselwa | Ii-astrocyte zeempuku | Ukuphila kweeseli, ukusetyenziswa kweoksijini, ukungeniswa kweglucose, i-ATP, iglycogen, isuperoxide | Yakhusela kuphela xa inikwe ngexesha lokubuyiselwa kweoksijini; yandisa ukuphefumla kweeseli noovimba ngaphandle kokwandisa isuperoxide |
| Isiqwenga setyhefu yebhaktheriya | I-microglia yeempuku, kwiiseli ezikhuliswe elabhoratri nakwiimpuku eziphilayo | Iicytokine, iimpawu ze-M1, imetabolism yeSeahorse, iiproteni zendlela ye-PHD3, i-FDG-PET, imisebenzi yokukhumbula | Yarhoxisa ukuhlengahlengiswa kwe-glycolysis nokuveliswa okugqithisileyo kwe-PHD3; yanciphisa ukudumba; yaphucula ukuziphatha |
| Isiqwenga setyhefu yebhaktheriya | I-microglia yeempuku ezinkulu kumxube weeseli zeglia ezikhuliswe elabhoratri | Ummandla womzimba weseli ophawulwe nge-IBA1, umjikelezo, ukwenziwa kwamasebe ezandisweni | Yagcina ukumila okunamasebe kwemeko yokuphumla; imenadione ayizange ikwenze oko |
Into efanayo kule miqolo kukuba into nganye ebangela impendulo inomlinganiselo wayo nempendulo yayo. Ukuhlangulwa kwemetabolism kuphendula umbuzo wemetabolism. Idatha yeecytokine neyokuziphatha iphendula umbuzo wokudumba. Ukumila kuphendula umbuzo wokumila. Akukho nasinye kwezi zintathu esiphendula omnye umbuzo ngokwaso.
Oko kungangqinisiswa yi-microglia ebonakala izolile ngakumbi
Esi sisivakalisi esifunekayo kuso sonke esi sihloko: i-microglia ebonakala ingavuselelwanga kangako, okanye imetabolism ye-microglia etshintshileyo, ayikungqinisisi ngokwayo ukuphucuka kokusebenza kwengqondo.
Ukumila kunokutshintsha ngaphandle kokuba umsebenzi ulandele. Iseli inokugcina amasebe ayo ngoxa isakhupha iicytokine, okanye ibe ngqukuva ngoxa isenza umsebenzi oluncedo wokususa inkunkuma yeeseli. Imilinganiselo yemetabolism inomda ofanayo: umkhondo weSeahorse okanye iskeni se-PET sibonisa indlela aphathwa ngayo amandla, kungekhona amandla okuqonda. Uphononongo lweempuku olwadibanisa imilinganiselo yemetabolism nefanayo neyokumila kunye neemvavanyo zokuziphatha lunamandla ngakumbi kanye ngenxa yokuba lwadibanisa amanqanaba obungqina endaweni yokufuna umlinganiselo omnye uthwale isigqibo sonke. Nalapho, imida eqhelekileyo isekhona, kuba ilipopolysaccharide ingumzekelo wesilumkiso esingenalo usulelo lokwenene, iimpuku ayingobantu, yaye ababhali ngokwabo bayawuphawula umahluko phakathi kweentlobo zezilwanyana. Nabani na oceba umsebenzi olandelayo kufuneka aqwalasele indlela iziphumo ze-microglia ezisebenza ngayo kwiimodeli ezahlukeneyo ngaphambi kokucinga ukuba le ndlela iziphatha ngokufanayo kwizicubu zabantu.
Oku kukhokelela kwisigqibo esisebenzayo malunga nexesha. Isiphumo see-astrocyte sibonisa ukuba ukunika idayi kwangethuba kwenza imeko ibe mandundu, kodwa ukuyinika ngexesha lokuchacha kuzisa inzuzo. Iziphumo ze-microglia zivela ekunikezweni kwedayi malunga nexesha lento ebangela ukudumba. Ukwenzakala kwengqondo kubomi bokwenene kudibanisa zombini ezi zigaba ngaxeshanye: ukusilela kwamandla kunye nesilumkiso sokhuselo lomzimba, eziguqukayo ekuhambeni kweeyure. Uvavanyo olwahlula isigaba esinye alunakumisela ishedyuli yemeko edibanisa zombini, yiyo loo nto iingxoxo ngamaxesha okunika idayi zigcina ixesha lokuvinjwa kweoksijini neglucose nelokudumba zahlukene.
Isishwankathelo esinyanisekileyo singena kumhlathi omnye. Kwii-astrocyte ezahlulwe kwezinye iiseli, i-methylene blue enikwe ngexesha lokubuyiselwa kweoksijini iqala kwakhona ukuphefumla kweeseli ize izisindise. Kwi-microglia evuselelwe ngesiqwenga sebhaktheriya, irhoxisa inkqubo yemetabolism yokudumba ngendlela ethile ye-PHD3 ize igcine ukumila kwemeko yokuphumla. Isiphumo ngasinye siyinyani ngaphakathi kwemeko yovavanyo lwaso, yaye akukho nasinye esisisigqibo ngokubanzi malunga nokudumba engqondweni. Ukuthelekisa iiseli zeglia kuluncedo kanye kuba kuthintela ezi ngxelo zimbini ekuxubeni zibe yinye.