Inqaku
Imethylene blue nemultiple sclerosis: oko kwavavanywa luphando lwezilwanyana
Khangela imethylene blue ye-MS uze ufumane amaphepha ophando abonakala ephendula umbuzo omnye. Akunjalo. Elinye iphepha lanika iimpuku ezigulayo imethylene blue njengonyango. Elinye alizange linyange nasiphi na isilwanyana ngayo. Lasebenzisa imethylene blue njengedayi eluhlaza kwizilayi ezibhityileyo zomnqonqo ukuze ingcali yezifo zezicubu ikwazi ukubona imyelin ngemicroscope.
Le yeyona ngcamango isisiseko salo lonke eli nqaku. Le dayi ineendima ezimbini kwibhayoloji, yaye azinanto zifana ngayo ngaphandle kwebhotile. Xa ivavanywa njengechiza elinokusetyenziswa, inikwa isilwanyana esiphilayo ngomlinganiselo othile, yaye umbuzo ngowokuba isifo siyatshintsha na. Xa isetyenziswa njengedayi, ifakwa kwizicubu ezifileyo ezilondolozwe ngemichiza emva kovavanyo, yaye umbuzo ngowokuba izicubu zazikhangeleka njani. Ukuba awuwuboni lo mahluko, uya kufunda icandelo leendlela zohlolo lwezicubu ngokungathi sisiphumo sonyango.
Yintoni i-EAE, ngamazwi alula
I-encephalomyelitis yokuzihlasela kwamajoni omzimba ebangelwa kulingo, edla ngokufinyezwa ibe yi-EAE, yimodeli yezilwanyana eqhelekileyo esetyenziselwa ukujonga imiba ethile yemultiple sclerosis. Umphandi utofa impuku esesichengeni sokuchaphazeleka ngesiqwenga seproteni yemyelin kunye nesincedisi esinamandla sokuvuselela amajoni omzimba. Emva koko amajoni omzimba ahlasela inkqubo yemithambo-luvo esembindini yesilwanyana ngokwaso. Impuku iba nobuthathaka obunyuka busuka ezantsi, ekulula ukubulinganisa kwisikali sika-0 ukuya ku-5, ukusuka kumsila ojingayo ukuya kukukhubazeka kwemilenze yangasemva.
Oku kulula kokunika amanqaku kuchaza ukuba kutheni i-EAE isetyenziswa kakhulu, kwanokuba kutheni inokulahlekisa. I-MS ebantwini izivelela ngokwayo, ibandakanya uluhlu olubanzi lwezinto ezihlaselwa ngamajoni omzimba olungachazwanga ngokucacileyo kangako, yaye ibangela iimpawu ezichaphazela ukubona, ukuva ngemizwa, ukucinga, icerebellum, nokushukuma, ngokuhamba kwesifo okuphinda kuvele okanye okuqhubeka kusiba mandundu. I-EAE iphinda ivelise iinkqubo ezithile ezifana nokudumba, ukonakala kwemyelin, nokwenzakala kweeaxon phantsi kweemeko ezilawulwayo. Ayivelisi i-MS encinane. Abafundi abafuna umfanekiso obanzi banokuphonononga indlela ekucingelwa ukuba imethylene blue isebenza ngayo kwinqanaba leeseli ngaphambi kokuvavanya naliphi na ibango eliphathelele isifo esithile.
La maphepha mabini angezantsi asebenzisa neeantigen ezahlukeneyo ze-EAE, nto leyo ebalulekileyo. Isifundo sonyango sabangela isifo nge-MOG35-55, ipeptide emfutshane evela kwimyelin oligodendrocyte glycoprotein. Isifundo sokonakala kwezicubu sabangela isifo nge-MP4, umdibaniso wamacandelo emyelin basic protein neproteolipid protein. Ukhetho lomnombo weempuku nolweantigen lutshintsha indawo evela kuyo imimandla eyonakeleyo nendlela ukudumba okwenzeka ngayo, ngoko ezi zifundo zimbini azikokuphindwa kovavanyo olunye.
Isikhokelo: ukusuka kwimodeli ukuya kumbuzo nakwimida
Gcina le theyibhile ivuliwe. Yeyona ndawo isisiseko sokubhekisa kuyo kweli nqaku. Umqolo ngamnye uchaza umbuzo wonyango onokuphendulwa yimodeli, oko kwalinganiswa ngokwenene, nalapho ukudlulisela iziphumo ebantwini kuphelela khona.
| Isifundo sonyango (MOG EAE) | Isifundo sokonakala kwezicubu (MP4 EAE) | |
|---|---|---|
| Iphepha | Isifundo sonyango lwe-EAE ebangelwe yipeptide ye-MOG | Isifundo se-MP4 sokonakala kwezicubu ekuhambeni kwexesha |
| Indima yemethylene blue | Ungenelelo olunikwe iimpuku eziphilayo | Idayi efakwe kwizilayi zomnqonqo ezinobukhulu obuyi-1 micrometer, ezilondolozwe ngemichiza |
| Umbuzo wonyango enokuwuphendula | Ngaba ukuyinika kutshintsha ubunzima be-EAE nemiqondiso yenkqubo yamajoni omzimba | Ukonakala kwemyelin nokwenzakala kweeaxon kutshintsha njani kwiinyanga ezintandathu |
| Iziphumo ezilinganisiweyo | Amanqaku eempawu zemihla ngemihla, iiseli ezingene emnqonqweni, ukonakala kwemyelin, ukwenzakala kweeaxon, imiqondiso ye-AMPK ne-SIRT1, ukusebenza kwe-NF-kB, ulungelelwano lwe-Th17 ne-Treg | Ubungakanani bommandla owonakeleyo, amanqanaba okudumba nokufumba kolwelo, ubuninzi bemicu elahlekelwe yimyelin, i-g-ratio nge-electron microscopy, ukuqhekeka kweeaxon, ukudumba kweemitochondria, izithuba phakathi kweeneurofilament |
| Umda wokudlulisela iziphumo ebantwini | Umnombo omnye, isini esinye, ixesha elifutshane leentsuku ezingama-30, ukunxulumana kwezinto kungekhona ubungqina bendlela yokusebenza | Akukho nyango luvavanyiweyo, izicubu zithathwe kwizilwanyana ezahlukeneyo ngamaxesha athile, iindawo ezonakeleyo zithathwe ikakhulu kwinxalenye yomnqonqo esezantsi emqolo |
Nabani na ohlola isihloko esilandelayo angaphinda asebenzise ezi kholamu, encediswa sisikhokelo sendlela yokufunda izifundo zangaphambi kovavanyo ebantwini: buza ngohlobo lwesilwanyana neantigen, indima yekhompawundi, iziphumo ezilinganisiweyo, nokuba isiphumo sibonisa ukuhamba kwesifo okanye imeko yezicubu ngexesha elinye.
Isiphumo sonyango: oko kwatshintshwa kukunikwa kwechiza
Isifundo sonyango sango-2016 kwi-Journal of Neuroimmunology sasebenzisa iimpuku ezimazi ze-C57BL/6, ezineminyaka yobudala emalunga neeveki ezisi-8 ukuya kwezili-10, ezatofwa nge-250 micrograms ye-MOG35-55 kwimpuku nganye ukuvuselela amajoni omzimba. Emva kokubangela isifo, izilwanyana ezine-EAE zabelwa ngokungakhethiyo kwiqela lemethylene blue okanye kwiqela le-EAE elinganyangwanga, yaye amanqaku eempawu alandelwa de kube malunga nosuku lwama-30.
Amanqaku eempawu ayesezantsi kakhulu ngokweenkcukacha-manani kwiimpuku ezinyangiweyo kunakwiimpuku ze-EAE zeqela lothelekiso ukusuka kusuku lwe-15 ukuya kusuku lwama-30, nge-P engaphantsi kuka-0.01. Uhlalutyo lomnqonqo lwabonisa into efanayo, ngokungena kweeseli ezibangela ukudumba okuncinane, ukonakala kwemyelin okuncinane, nokulahleka kweeaxon okuncinane kwizilwanyana ezinyangiweyo.
Eyona nto yahlukileyo yayiluluhlu lwemiqondiso ehloliweyo. Unyango lwandisa ukuvuselelwa kwe-AMPK nokuveliswa kwe-SIRT1 lwaza lwacinezela ukuvuselelwa kwe-NF-kB, isilawuli esingundoqo sokudumba. Kwicala lamajoni omzimba aziqhelanisayo, lwanciphisa impendulo ye-Th17 lwaza lwomeleza impendulo ye-Treg. Ngoko ikhonkco elicetywayo lisuka kwimethylene blue liye kwi-AMPK ne-SIRT1, emva koko ekunciphiseni imiqondiso ye-NF-kB, liye ekutshintsheni ukusuka kwi-Th17 ebangela ukudumba ukuya kwi-Treg elawulayo, liye ekwenzakaleni kwezicubu okuncinane, lize liphele kumanqaku asezantsi.
Elo khonkco yindlela yokutolika iziphumo, kungekhona uthotho lwezizathu neziphumo olungqiniweyo. Isifundo sibonisa ukuba ichiza, utshintsho lwemiqondiso, nokuphucuka kwesifo zihamba kunye. Asingqini ukuba i-AMPK ne-SIRT1 zingunobangela walo lonke uncedo, yaye umbhalo ofakwe kwizalathiso ezifumaneka esidlangalaleni awuwuchazi ngokucace ngokwaneleyo umlinganiselo ochanekileyo wemilligram ngekilogram, indlela yokunika ichiza, usuku lokuqala, okanye ubude bexesha ukuze inkqubo iphinde yenziwe kusetyenziswa isishwankathelo kuphela. Thatha nawuphi na umlinganiselo wokunika ichiza ocatshulwe ngeli phepha kwiphepha lentengiso njengongaqinisekiswanga, ngaphandle kokuba ubhekisa kwiindlela ezichazwe kumbhalo opheleleyo. Ubungqina obubanzi ngemethylene blue ebantwini yindawo yokujonga ukuba lo mqondiso wokuqala wamajoni omzimba wakha wakhokelela kwidatha yovavanyo ebantwini na.
Isiphumo sokuhlola izicubu: oko kwaboniswa zizicubu kwiinyanga ezintandathu
Isifundo se-MP4 sango-2015 kwi-PLoS ONE sasebenzisa iimpuku ezimazi ze-C57BL/6 ezatofwa nge-200 micrograms ye-MP4 kwisixube esipheleleyo sika-Freund sokuvuselela amajoni omzimba, kunye netyhefu yepertussis ngosuku lokuvuselelwa kwamajoni omzimba, kwaphindwa emva kweeyure ezingama-48. Akukho silwanyana safumana imethylene blue njengechiza. Emva kokulondolozwa kwezicubu ngemichiza nokufakwa kwinto eziqinisa, izilayi ezinqumleza umnqonqo ezinobukhulu be-one micrometer zadaywa ngemethylene blue ukuze zihlolwe ngemicroscope yokukhanya, kwaza kwasetyenziswa i-electron microscopy kumzila ongaphambili osecaleni lomnqonqo ukuze kubonwe iinkcukacha ezincinane ngakumbi.
Izicubu zathathwa kwizigaba ezintathu: encotsheni yesigaba esibukhali ngeempuku ezisi-7, kwiinyanga ezintathu emva kokuqala ngeempuku ezi-5, nakwiinyanga ezintandathu emva kokuqala ngeempuku ezili-10. Amanqaku eempawu angumndilili kwezo zigaba ayekhangeleka ephantse afane, kufutshane no-1.7 ukuya ku-1.8 kwisikali sika-0 ukuya ku-5. Izicubu zabonisa into eyahlukileyo. Ukonakala kwemyelin kuphela kwenyuka ukusuka malunga nama-569,000 emicu nge-square millimeter encotsheni yesigaba esibukhali ukuya malunga ne-1,496,000 kwiinyanga ezintandathu, yaye umahluko phakathi kwesigaba esibukhali nexesha elide wawubalulekile ngokweenkcukacha-manani, nge-P elingana no-0.027. Ukonakala kwemyelin kunye neeaxon phantse kwaphindeka kabini kwiinyanga ezintandathu. Imicu eyayiphinda igqunywe yimyelin yahlala ingaphantsi kwepesenti enye kuzo zonke izigaba, nto leyo esisikhumbuzo esingqongqo malunga namathemba okulungiswa kwezicubu.
Ukwenzakala kweeaxon kwalandela ipateni emile okuka-U: kwakuphezulu encotsheni yesigaba esibukhali, kusezantsi kwiinyanga ezintathu, kuphezulu kakhulu kwiinyanga ezintandathu, yaye kwakunxulumene nokudumba kweemitochondria nokuncipha kwezithuba phakathi kweeneurofilament. Abahloli babengawazi amanqaku okanye isigaba xa behlola, nto leyo exhasa ngakumbi ukutolika ukuba amanqaku eempawu ayengawubonisi ngokupheleleyo umonakalo oqhubekayo kwisakhiwo sezicubu.
Oko kungahambelani yeyona nto iphambili esi sifundo siyinika umfundi onomdla kwi-MS. Amanqaku okukhubazeka angatshintshiyo ayengathethi ukuba izicubu azitshintshi. Kukwabonisa ukuba kutheni eli phepha lingenakuxhasa naliphi na ibango lonyango lwe-MS ngemethylene blue. Lavavanya ukuhamba kwexesha, hayi unyango.
Kutheni kungekho nasinye kwezi zifundo esingqina uncedo kwizigulana
Cinga ngephepha ngalinye njengeliphendula kakuhle umbuzo omxinwa, kodwa lingawuphenduli kwaphela umbuzo ophathelele izigulana.
Okokuqala, unobangela wahlukile. I-EAE ibangelwa ngabom ngeantigen echaziweyo nesincedisi sokuvuselela amajoni omzimba. I-MS ebantwini izivelela ngokwayo, ineenkqubo ezibanzi zamajoni omzimba ezibandakanya iiseli ze-T, iiseli ze-B, amajoni omzimba emvelo, neeseli zeglia ezihlala apho. Ikhompawundi etshintsha impendulo egxile kwi-MOG kwimpuku ayinyanzelekanga ukuba itshintshe loo mpendulo ibanzi ebantwini.
Okwesibini, indlela isifo esizibonakalisa ngayo yahlukile. Ubuthathaka obunyuka ukusuka emsileni ukuya kwimilenze yangasemva kwimpuku ngumlinganiselo olula. Buqukumbela ukwahlukahlukana kwe-MS ebantwini bube linani elinye elilinganisa ukushukuma. Amachiza adla ngokutshintsha amanqaku e-EAE exesha elifutshane ngaphandle kokutshintsha ukuhamba kwesifo ebantwini, yiyo loo nto i-EAE isisixhobo sokuhlola nokufunda indlela yokusebenza kunokuba ibe yikliniki encinane.
Okwesithathu, ukufikelela kwechiza emzimbeni nexesha lokulinika azikacaci. Ukunika ichiza ngokuqhubekayo kwangethuba kuvavanyo lwempuku lweentsuku ezingama-30 akufani nokusetyenziswa ngomlomo ixesha elide ngumntu onesifo esele sizinzile, yaye akukho nasinye isifundo esalinganisa amanqanaba echiza kwinkqubo yemithambo-luvo. Ngaphandle kwelo khonkco, akukho ndlela yokudibanisa ubungakanani bekhompawundi kwisitya sovavanyo okanye umlinganiselo onikwe impuku nenkqubo yokunika ichiza emntwini.
Okwesine, la maphepha mabini awaphindi uvavanyo olunye. Aneeantigen ezahlukeneyo, imibuzo eyahlukeneyo, iziphumo ezilinganisiweyo ezahlukeneyo, yaye kwelinye akukho nyango kwaphela. Ukuwacaphula kunye njengezifundo ezimbini ezibonisa iziphumo ezilungileyo kunyango lwe-MS yimpazamo yokwahlula iindidi.
Oku kuthetha ntoni kubafundi abavavanya ubungqina
Akukho nkqubo iqinisekisiweyo yokusebenzisa imethylene blue kwi-MS kula maphepha. Isifundo sonyango sixhasa uphando olungakumbi nge-AMPK, i-SIRT1, i-NF-kB, nolungelelwano lwe-Th17 ne-Treg kwi-MOG EAE. Isifundo sokonakala kwezicubu sisikhumbuzo sokuba ukwenzakala kwezicubu kunokuqhubeka ngoxa amanqaku ebonakala engatshintshi, kwanokuba imethylene blue kwicandelo leendlela ngamanye amaxesha ithetha ingqayi yedayi endaweni yekhabhathi yamayeza. Abafundi abalandela uphando oluqhubekayo ngemultiple sclerosis bafanele bawugcine engqondweni loo mahluko phakathi kwedayi nechiza. Imibuzo yokhuseleko, kuquka imingcipheko enxulumene negazi nokusebenzisana kwamachiza, ifanele ixoxwe nengcali yezonyango kunye nedatha ngokubanzi yokhuseleko lwemethylene blue ngaphambi kokuba nabani na athathe amanyathelo ngenxa yezihloko zeendaba zaselabhoratri.
Isigqibo sinokushwankathelwa kwisivakalisi esinye. Esinye isifundo esicatshuliweyo sanika iimpuku ezine-MOG EAE imethylene blue saza sanika ingxelo yesifo esingabukhali kangako kunye notshintsho lwamajoni omzimba olunciphisa ukudumba, ngoxa esinye isifundo esicatshuliweyo singazange sinike naliphi na ichiza saza sasebenzisa imethylene blue kuphela ukudaya umnqonqo ngoxa sasibhala ukwanda komonakalo wemyelin neeaxon kwiinyanga ezintandathu, ngoko ezi zifundo zimbini azingqini ukuba imethylene blue iyanceda kwi-MS ebantwini.