Isihloko
I-methylene blue ne-multiple sclerosis: lokho ucwaningo lwezilwane olukuhlolile
Uma usesha i-methylene blue ye-MS, uthola amaphepha ocwaningo abukeka sengathi aphendula umbuzo ofanayo. Akunjalo. Elinye iphepha lanikeza amagundane agulayo i-methylene blue njengokwelapha. Elinye alizange lelaphise noma yisiphi isilwane ngayo. Lasebenzisa i-methylene blue njengodayi oluhlaza okwesibhakabhaka ezingcezwini ezincane zomgogodla ukuze uchwepheshe wezifo abone i-myelin ngaphansi kwesibonakhulu.
Lona ngumqondo oyisisekelo salo lonke lesi sihloko. Lo dayi unezindima ezimbili kwisayensi yezinto eziphilayo, futhi lezo zindima azihlanganyeli ngalutho ngaphandle kwebhodlela. Uma uhlolwa njengomuthi ongase usetshenziswe, unikezwa isilwane esiphilayo ngomthamo, bese kubuzwa ukuthi isifo siyashintsha yini. Uma usetshenziswa njengodayi, ufakwa ezicutshini ezingasaphili ezilondoloziwe ngemva kokuhlolwa, bese kubuzwa ukuthi izicubu zazibukeka kanjani. Uma ungawuboni lowo mehluko, uzofunda isigaba sezindlela zokuhlola izicubu sengathi siwumphumela wokwelapha.
Iyini i-EAE, ngamagama alula
I-encephalomyelitis yokuhlola ebangelwa amasosha omzimba ahlasela umzimba uqobo, evame ukufushaniswa ngokuthi i-EAE, iyimodeli yezilwane ejwayelekile esetshenziselwa ukutadisha izici ze-multiple sclerosis. Umcwaningi uvusa ukusabela kwamasosha omzimba egundaneni elisengozini yokuba nalesi simo esebenzisa ingxenye yephrotheni ye-myelin kanye nesikhuthazi esinamandla samasosha omzimba. Amasosha omzimba abe esehlasela isimiso sezinzwa esimaphakathi saleso silwane. Igundane liba nobuthakathaka obukhuphuka emzimbeni, okulula ukubulinganisa esikalini esingu-0 kuya ku-5, kusukela emsileni olengayo kuye ekukhubazekeni kwemilenze yangemuva.
Ukuba lula kwalokho kunikeza amaphuzu kuchaza ukuthi kungani i-EAE ithandwa, nokuthi kungani ingadukisa. I-MS kubantu iqala ngokwayo, ihilela izinhlobonhlobo ezibanzi nezingacacisiwe kahle zezinto ezihlaselwa amasosha omzimba, futhi ibangela izimpawu ezithinta ukubona, ukuzwa, ukucabanga, i-cerebellum, nokunyakaza, ngokuhamba kwesifo okuphindaphindayo noma okuqhubekayo. I-EAE ilingisa izinqubo ezithile ezikhethiwe, njengokuvuvukala, ukulahleka kwe-myelin, nokulimala kwama-axon, ngaphansi kwezimo ezilawulwayo. Ayiyona i-MS eyenziwe yaba ncane. Abafundi abafuna ukuqonda isisekelo esibanzi bangabuyekeza ukuthi kucatshangwa ukuthi i-methylene blue isebenza kanjani ezingeni lamangqamuzana ngaphambi kokwahlulela noma yisiphi isimangalo ngesifo esithile.
Amaphepha amabili angezansi aphinde asebenzisa ama-antigen ahlukene e-EAE, futhi lokho kubalulekile. Isifundo sokwelapha sabangela isifo nge-MOG35-55, i-peptide emfushane evela ku-myelin oligodendrocyte glycoprotein. Isifundo sezinguquko zezicubu sabangela isifo nge-MP4, inhlanganisela yezingxenye ze-myelin basic protein ne-proteolipid protein. Ukukhethwa kohlobo lofuzo lwamagundane ne-antigen kushintsha lapho kuvela khona izindawo ezilimele nendlela ukuvuvukala okuziphatha ngayo, ngakho lezi zifundo ezimbili azikona ukuphindwa kabili kokuhlolwa okukodwa.
Imephu: imodeli, umbuzo, nemikhawulo
Gcina leli thebula livuliwe. Liyinkomba eyisisekelo yalesi sihloko. Umugqa ngamunye uchaza ukuthi imodeli ingaphendula muphi umbuzo wezokwelapha, yini eyalinganiswa ngempela, nokuthi ukudlulisela okutholakele kubantu kuphelela kuphi.
| Isifundo sokwelapha (MOG EAE) | Isifundo sezinguquko zezicubu (MP4 EAE) | |
|---|---|---|
| Iphepha locwaningo | Isifundo sokwelapha i-EAE ebangelwe yi-peptide ye-MOG | Isifundo se-MP4 sezinguquko zezicubu ngokuhamba kwesikhathi |
| Indima ye-methylene blue | Ukungenelela okunikezwe amagundane aphilayo | Udayi ofakwe ezingcezwini zomgogodla ezilondoloziwe ezingu-1 micrometer |
| Umbuzo wezokwelapha esingawuphendula | Ingabe ukunikeza imithamo kushintsha ubukhulu be-EAE nokudluliswa kwezimpawu zamasosha omzimba | Ukulahleka kwe-myelin nokulimala kwama-axon kushintsha kanjani phakathi nezinyanga eziyisithupha |
| Imiphumela elinganisiwe | Amaphuzu ezimpawu nsuku zonke, ukungena kwamangqamuzana okuvuvukala emgogodleni, ukulahleka kwe-myelin, ukulimala kwama-axon, ukudluliswa kwezimpawu kwe-AMPK ne-SIRT1, ukusebenza kwe-NF-kB, ibhalansi ye-Th17 ne-Treg | Ubukhulu bendawo elimele, amazinga okuvuvukala nokunqwabelana koketshezi, ukuminyana kwemicu elahlekelwe yi-myelin, i-g-ratio ngesibonakhulu sama-electron, ukubhidlika kwama-axon, ukuvuvukala kwama-mitochondria, izikhala phakathi kwama-neurofilament |
| Umkhawulo wokudlulisela okutholakele kubantu | Uhlobo lofuzo olulodwa, ubulili obubodwa, isikhathi esifushane sezinsuku ezingu-30, ukuhlobana hhayi ubufakazi benqubo yokusebenza | Akukho kwelapha okuhloliwe, amasampula ezicubu ezilwane ezihlukene ngezikhathi ezithile, izindawo ezilimele ezithathwe ikakhulukazi engxenyeni yomgogodla esezansi emhlane |
Noma ubani ohlola isihloko sezindaba esilandelayo angaphinde asebenzise amakholomu afanayo ngosizo lwe-indlela yokufunda izifundo zangaphambi kokuhlolwa kubantu: buza ngohlobo lwesilwane ne-antigen, indima yento ehlolwayo, imiphumela elinganisiwe, nokuthi umphumela uchaza ukuhamba kwesifo noma isimo sezicubu ngesikhathi esithile.
Umphumela wokwelapha: lokho okwashintshwa ukunikeza imithamo
Isifundo sokwelapha sango-2016 ku-Journal of Neuroimmunology sasebenzisa amagundane esifazane ohlobo lwe-C57BL/6, cishe anamasonto angu-8 kuya ku-10 ubudala, avuselwe ukusabela kwamasosha omzimba nge-250 micrograms ye-MOG35-55 ngegundane ngalinye. Ngemva kokubangelwa kwesifo, izilwane ezine-EAE zabelwa ngokungahleliwe eqenjini le-methylene blue noma eqenjini le-EAE elingelashwanga, futhi amaphuzu ezimpawu alandelelwa kuze kube cishe usuku lwama-30.
Amaphuzu ezimpawu ayengaphansi ngokuphawulekayo ngokwezibalo emagundaneni elashwayo kunasemagundaneni eqembu lokuqhathanisa eline-EAE kusukela ngosuku lwe-15 kuya osukwini lwama-30, u-P engaphansi kuka-0.01. Ukuhlaziywa komgogodla kwakhomba ohlangothini olufanayo: ukungena okuncane kwamangqamuzana okuvuvukala, ukulahleka okuncane kwe-myelin, nokulahleka okuncane kwama-axon ezilwaneni ezelashwayo.
Ingxenye eyayihlukile kwakuwuchungechunge lokuhlolwa kokudluliswa kwezimpawu. Ukwelapha kwandisa ukwenziwa kusebenze kwe-AMPK nokukhiqizwa kwe-SIRT1, futhi kwacindezela ukwenziwa kusebenze kwe-NF-kB, okuyisilawuli esiyinhloko sokuvuvukala. Ohlangothini lwamasosha omzimba afunda ukubona izinto ezithile, kwanciphisa ukusabela kwe-Th17 futhi kwaqinisa ukusabela kwe-Treg. Ngakho uchungechunge oluphakanyisiwe lusuka ku-methylene blue luye ku-AMPK ne-SIRT1, luye ekuncipheni kokudluliswa kwezimpawu kwe-NF-kB, luye ekusukeni ku-Th17 ekhuthaza ukuvuvukala kuye ku-Treg elawulayo, luye ekulimaleni okuncane kwezicubu, bese luya emaphuzwini aphansi.
Lolo chungechunge luyincazelo, hhayi ukulandelana kwembangela nomphumela okufakazelwe. Isifundo sibonisa ukuthi umuthi, izinguquko ezimpawini ezidluliswa emangqamuzaneni, nokuba ngcono kwesifo kuhamba ndawonye. Asifakazeli ukuthi i-AMPK ne-SIRT1 yizona eziletha yonke inzuzo, futhi umbhalo ofakwe ezinkombeni zomphakathi awuvezi ngokucace ngokwanele umthamo oqondile we-milligram per kilogram, indlela yokuwunikeza, usuku lokuqala, noma ubude besikhathi ukuze indlela yokwelapha iphinde yenziwe kusetshenziswa isifinyezo kuphela. Thatha noma iyiphi inombolo yomthamo ecashunwe ngaleli phepha ekhasini lokuthengisa njengengakaqinisekiswa, ngaphandle uma ibhekisela ezindleleni ezisembhalweni ogcwele. Ubufakazi obubanzi babantu nge-methylene blue yindawo yokuhlola ukuthi lokhu kutholakele kwasekuqaleni ngamasosha omzimba kwagcina yini sekuba idatha yezokwelapha kubantu.
Umphumela wezicubu: lokho okwaboniswa izinyanga eziyisithupha
Isifundo se-MP4 sango-2015 ku-PLoS ONE sasebenzisa amagundane esifazane ohlobo lwe-C57BL/6, avuselwe ukusabela kwamasosha omzimba nge-200 micrograms ye-MP4 ekwi-complete Freund adjuvant kanye ne-pertussis toxin ngosuku lokuvuswa kwalokho kusabela, kwaphinde kwanikezwa i-pertussis toxin ngemva kwamahora angu-48. Asikho isilwane esanikezwa i-methylene blue njengomuthi. Ngemva kokulondolozwa kwezicubu nokushumekwa kwazo entweni ezisekelayo, izingcezu ezinqamula umgogodla ezingu-one micrometer zadaywa nge-methylene blue ukuze zihlolwe ngesibonakhulu sokukhanya, kwasetshenziswa nesibonakhulu sama-electron ukuhlola umgudu ongaphambili naseceleni ukuze kubonakale imininingwane emincane.
Izicubu zathathwa ngezigaba ezintathu: esicongweni sesigaba esibukhali kusetshenziswa amagundane angu-7, ezinyangeni ezintathu ngemva kokuqala kwesifo kusetshenziswa amagundane angu-5, nasezinyangeni eziyisithupha ngemva kokuqala kwesifo kusetshenziswa amagundane angu-10. Amaphuzu amaphakathi ezimpawu kulezo zigaba ayebukeka ecishe afane, eseduze kuka-1.7 kuya ku-1.8 esikalini esingu-0 kuya ku-5. Izicubu zaveza indaba ehlukile. Ukulimala kwe-myelin kuphela kwakhuphuka kusuka cishe emicwini engu-569,000 per square millimeter esicongweni sesigaba esibukhali kuya cishe ku-1,496,000 ezinyangeni eziyisithupha, futhi ukuqhathanisa isigaba esibukhali nesikhathi eside kwaba okuphawulekayo ngokwezibalo, u-P elingana no-0.027. Ukulimala okuhlanganisa i-myelin nama-axon kwacishe kwaphindeka kabili ezinyangeni eziyisithupha. Imicu eyayiphinde yakha i-myelin yahlala ingaphansi kwephesenti elilodwa kuzo zonke izigaba, okuwumniningwane odambisa amathemba okulungiseka kwezicubu.
Ukulimala kwama-axon kwalandela iphethini efana no-U: kwakuphezulu esicongweni sesigaba esibukhali, kwehla ezinyangeni ezintathu, kwase kuba phezulu kakhulu ezinyangeni eziyisithupha. Kwakuhlobene nokuvuvukala kwama-mitochondria nokuncipha kwezikhala phakathi kwama-neurofilament. Abahloli babengawazi amaphuzu ezimpawu noma isigaba sesifo ngesikhathi behlola, okuqinisa incazelo yokuthi amaphuzu ezimpawu ayengabubonisi ngokugcwele ubukhulu bokulimala kwesakhiwo sezicubu okwakuqhubeka.
Lokho kungahambisani yikona okufaka isandla ngempela kulesi sifundo kumfundi ophathelene ne-MS. Amaphuzu okukhubazeka angashintshi ayengasho ukuthi izicubu nazo azishintshi. Lokhu kuphinde kubonise ukuthi kungani leli phepha lingakwazi ukusekela noma yisiphi isimangalo sokwelapha i-MS nge-methylene blue. Lahlola ukuhamba kwesikhathi, hhayi ukwelapha.
Kungani kungekho sifundo esifakazela inzuzo ezigulini
Cabanga ngephepha ngalinye njengeliphendula kahle umbuzo omncane othile, kodwa elingawuphenduli nhlobo umbuzo ophathelene neziguli.
Okokuqala, imbangela ihlukile. I-EAE ibangelwa ngamabomu nge-antigen echaziwe nesikhuthazi samasosha omzimba. I-MS kubantu iqala ngokwayo futhi ihilela izinhlobonhlobo ezibanzi zamasosha omzimba, okuhlanganisa amangqamuzana e-T, amangqamuzana e-B, amasosha omzimba emvelo, namangqamuzana e-glia ahlala ezicutshini. Into eshintsha ukusabela okugxile ku-MOG egundaneni akusho ukuthi izoshintsha nalokho kusabela okubanzi kubantu.
Okwesibili, indlela isifo esiziveza ngayo ihlukile. Ubuthakathaka obukhuphuka busuka emsileni buye emilenzeni yangemuva egundaneni buyisilinganiso esilula. Bufingqa ukuhlukahluka kwe-MS kubantu kube yinombolo eyodwa ephathelene nokunyakaza. Imithi ivame ukushintsha amaphuzu e-EAE esikhathini esifushane ngaphandle kokushintsha ukuhamba kwesifo kubantu. Yingakho i-EAE iyithuluzi lokuhlunga izinto ezingase zisebenze nokutadisha izinqubo zokusebenza, hhayi umtholampilo omncane.
Okwesithathu, ukuchayeka emuthini nesikhathi sokuwunikeza akukacaciswa. Ukunikeza imithamo ngokuqhubekayo kusenesikhathi ekuhlolweni kwamagundane kwezinsuku ezingu-30 akufani nokusebenzisa umuthi ngomlomo isikhathi eside kumuntu osenesifo esizinzile, futhi akukho kulezi zifundo okwalinganisa amazinga omuthi esimisweni sezinzwa. Ngaphandle kwalokho kuhlobana, akukho bhuloho elisuka ekugxileni kwento esitsheni sokuhlola noma emthamo osegundaneni liye ohlelweni lokuwusebenzisa kumuntu.
Okwesine, la maphepha amabili awaphindani. Ama-antigen ahlukene, imibuzo ehlukene, imiphumela elinganisiwe ehlukene, futhi kwesinye isifundo akukho kwelapha nhlobo. Ukuwacaphuna ndawonye njengezifundo ezimbili ezinemiphumela emihle yokwelapha i-MS kuyiphutha lokuhlanganisa izinto zezigaba ezingafani.
Lokhu kusho ukuthini kubafundi abahlaziya ubufakazi
Ayikho indlela yokusebenzisa i-methylene blue ye-MS esunguliwe kula maphepha. Isifundo sokwelapha sisekela ucwaningo olwengeziwe nge-AMPK, i-SIRT1, i-NF-kB, nebhalansi ye-Th17 ne-Treg ku-MOG EAE. Isifundo sezinguquko zezicubu sikhumbuza ukuthi ukulimala kwezicubu kungaqhubeka lapho amaphuzu ebukeka engashintshi, nokuthi i-methylene blue esigabeni sezindlela zokuhlola ngezinye izikhathi isho imbiza kadayi esikhundleni sekhabethe lemithi. Abafundi abalandela ucwaningo oluqhubekayo nge-multiple sclerosis kufanele bawugcine engqondweni lowo mehluko phakathi kukadayi nomuthi. Imibuzo yokuphepha, okuhlanganisa izingozi eziphathelene negazi nokusebenzisana neminye imithi, kufanele ixoxwe nochwepheshe wezokwelapha kanye nedatha evamile yokuphepha kwe-methylene blue ngaphambi kokuba noma ubani athathe isinyathelo ngenxa yezihloko zezindaba zemiphumela yaselabhorethri.
Isiphetho singangena emshweni owodwa. Esinye isifundo esicashuniwe sanikeza amagundane ane-MOG EAE imithamo ye-methylene blue futhi sabika isifo esithambile kanye noshintsho lwamasosha omzimba olunciphisa ukuvuvukala, kuyilapho esinye isifundo esicashuniwe singazange sinikeze mithamo yokwelapha futhi sasebenzisa i-methylene blue kuphela ukudaya umgogodla ngenkathi sibhala phansi ukunqwabelana kokulimala kwe-myelin nama-axon ezinyangeni eziyisithupha, ngakho lezi zifundo ezimbili azifakazeli ukuthi i-methylene blue isiza i-MS kubantu.