Inqaku
I-methylene blue ne-ALS: kutheni iziphumo zahluka kwiimodeli zovavanyo
Xa ukhangela i-methylene blue ne-ALS, ufumana amabango amabini abonakala ephikisana. Elinye lithi idayi eluhlaza okwesibhakabhaka yakhusela iiseli zemithambo-luvo kwiimodeli ze-ALS. Elinye lithi ayizange yenze nto kwimpuku enemodeli ye-ALS. Omabini amabango acaphula amaphepha ophando okwenene, yaye omabini loo maphepha afanele athathelwe ingqalelo. Oku kuphikisana kuyaphela xa uchaza ingcamango esekelwe kuyo ilingelo ngalinye: imodeli yiyo emisela umda webango.
Isiphumo kwintshulube libango elimalunga nentshulube. Isiphumo kwintlanzi ekwinqanaba lombungu libango elimalunga nentlanzi ekwinqanaba lombungu. Akukho nasinye esilibango elimalunga nesigulana de inkqubela yesifo kwisilwanyana esanyisayo ixhase oko. Eli nqaku lithelekisa amaphepha amabini acatshulwa kakhulu, elinye ecaleni kwelinye, ukuze ubone ukuba kutheni iimpendulo zawo zahlukile.
Amalingelo amabini kwitheyibhile enye
Gcina le theyibhile ivuliwe xa ufunda. Wonke umahluko obalulekileyo phakathi kwezi zifundo uqulethwe kuyo.
| I-TDP-43 ne-FUS kwiintshulube neentlanzi | I-SOD1 kwiimpuku | |
|---|---|---|
| Iphepha lophando | UVaccaro noogxa bakhe, 2012, kwi-PLoS ONE | ULougheed noTurnbull, 2011, kwi-PLoS ONE |
| Isidalwa | Intshulube engqukuva i-C. elegans nemibungu ye-zebrafish | Impuku ye-congenic eneekopi ezininzi ze-SOD1 G93A yomntu |
| Imfuza ye-ALS emelwe yimodeli | I-TDP-43 yomntu eguqukileyo (A315T kwiintshulube, G348C kwiintlanzi) ne-FUS eguqukileyo (S57Δ kwiintshulube, R521H kwiintlanzi) | I-SOD1 G93A yomntu eguqukileyo, engunobangela we-ALS efumaneka kwiintsapho |
| Ukuvezwa kwichiza | Ukuvezwa ngokuntywiliselwa: 6 okanye 60 μM kwiipleyiti ukususela ekuqanduselweni, 600 μM kuvavanyo olufutshane kulwelo, 30 ukuya ku-60 μM ubusuku bonke kwiimbumba zeentlanzi | Malunga ne-25 mg/kg ngosuku ngomlomo kwijelatin enencasa, yonke imihla ukususela kubudala beentsuku eziyi-45 de kufike inqanaba lokugqibela |
| Iziphumo ezilinganisiweyo | Ukukhubazeka, ukonakala kwamasebe eeseli zemithambo-luvo ezilawula intshukumo, uvavanyo lweesinapsi, ukuqubha kweentlanzi nokumila kwee-aksoni, imilinganiselo yoxinzelelo lwe-oksideshini | Ixesha de kuqale iimpawu zemithambo-luvo, ubude bamaxesha amanqanaba esifo alandelelanayo, ukuphila de kufike inqanaba lokugqibela elichaziweyo, ubunzima bomzimba |
| Isiphumo esiphambili | Ukubuyiselwa kwentshukumo nokwakheka kweeseli zemithambo-luvo ngokubanzi | Akukho kulibaziseka kubalulekileyo ekuqaleni kwesifo xa sijongwa siphela, yaye akukho kwandiswa kubalulekileyo kokuphila |
Imfuza iyahluka, iintlobo zezilwanyana ziyahluka, indlela yokunika ichiza iyahluka, ixesha liyahluka, neziphumo ezilinganisiweyo ziyahluka. Uhlobo lwemfuza ngomnye kuphela wemahluko emihlanu, ngoko esi sibini sezifundo asinakungqina ukuba i-methylene blue isebenza ngokukhethekileyo kwindlela enye ye-ALS. Singqina into enomda omncinane kodwa eseluncedo: ukubuyiselwa okunamandla phantsi kokuvezwa ngokuthe ngqo kwangethuba kwiimodeli ezilula akuzange kudlulele kwinkqubela yesifo esingapheliyo kwisilwanyana esanyisayo esinikwa ichiza ngomlomo.
Isiphumo esihle: oko kwaboniswa luphando lweentshulube neentlanzi
Uphando lweentshulube neentlanzi luka-2012 lwabangela ukuba i-TDP-43 okanye i-FUS yomntu eguqukileyo iveliswe kwiiseli zemithambo-luvo ezilawula intshukumo kwiintshulube, lwaza lwatofa i-mRNA yomntu eguqukileyo kwiimbumba zeentlanzi. Ezi ziimodeli zetyhefu ebangelwa ziiproteni. Zivelisa iziphumo eziyityhefu zeproteni eguqukileyo kwintshukumo nakwimilo yeeseli zemithambo-luvo kwisithuba seentsuku, hayi inkqubela epheleleyo ye-ALS kumntu omdala kwiminyaka emininzi.
Unyango lwaqala kwangethuba kakhulu. Iintshulube zakhulela kwi-agar equlethe i-methylene blue ukususela ekuqanduselweni. Iimbumba zeentlanzi zahlala kwisisombululo sechiza ubusuku bonke, ukususela kwiiyure eziyi-24 emva kokuqhamiswa, kwaza kwavavanywa indlela eziziphatha ngayo malunga neentsuku ezimbini kamva. Umqobo ophakathi kwegazi nengqondo awuzange wahlule ichiza kwiseli yemithambo-luvo ngendlela owenza ngayo kwisilwanyana esanyisayo, yaye akukho nqanaba lechiza kwiplazma okanye engqondweni lalinganisiweyo.
Phantsi kwezo meko, iziphumo zazinkulu yaye zingaguquguquki. Zombini i-6 ne-60 μM zanciphisa ukukhubazeka okunxulumene nobudala kwiintshulube ze-TDP-43 ne-FUS. Ngosuku lwe-12 lobudala, malunga ne-80 ekhulwini leentshulube ze-TDP-43 ezinganyangwanga zazikhubazekile, xa kuthelekiswa malunga ne-10 ekhulwini leentshulube ezanyangwa ukususela ekuqanduselweni. Uvavanyo lweesinapsi lwakhomba kwicala elifanayo: iintshulube ze-TDP-43 zazinovakalelo olugqithisileyo kwi-aldicarb, ichiza elivavanya ukuhanjiswa kwemiqondiso phakathi kwemithambo-luvo nezihlunu, yaye i-methylene blue yayisondeza indlela ezisabela ngayo kwimeko eqhelekileyo. Inani lamasebe eeseli zemithambo-luvo ezilawula intshukumo awayesonakala lehla, isiphumo sibonakala ngokucace kakhulu kwi-60 μM. Kwiintlanzi, ukuqubha okubangelwa kukuchukunyiswa kwaphucuka kakhulu, yaye ii-aksoni ezilawula intshukumo ezazifutshane zakhula zaya kusondela kubude obuqhelekileyo kwi-30 okanye kwi-60 μM, nangona ukubuyiselwa kwakuyinxalenye kuphela kunokuba kuphelele.
Idatha yoxinzelelo lwe-oksideshini iyahambelana nenkcazelo yokusebenza njengento ethintela i-oksideshini. Iintshulube neentlanzi ezinemfuza eguqukileyo zabonisa imilinganiselo ephezulu yeentlobo zeoksijini ezisebenzayo, eyathi yehla ngonyango, yaye iintshulube eziqhelekileyo ezikwi-methylene blue zaphila ixesha elide kakhulu kunalezo zinganyangwanga xa zavezwa kwinto ekhuthaza i-oksideshini. Kodwa kukho imida emibini ebalulekileyo. Amanqanaba ewonke eeproteni ze-TDP-43 ne-FUS eziguqukileyo awazange ehle, yaye uphando aluzange lulinganise ngokuthe ngqo ukuqokelelana kweeproteni. Ngoko lubonisa ukubuyiselwa kokusebenza ngaphandle kokungqina ukususwa kweeproteni eziqokeleleneyo. Nabani na olandela indlela ezidla ngokuchazwa ngayo iingqikelelo ezimalunga ne-redox nemitokhondriya ufanele ayithathe le ndlela yokusebenza njengexhaswayo, kodwa engekaqinisekiswa ngokupheleleyo.
Ixesha lokuqalisa yayilelona nkcukacha lifundisayo. Xa ababhali balinda de kube lusuku lwe-5 lobudala ngaphambi kokuqala unyango, izinga lokukhubazeka ngosuku lwe-12 lalimalunga ne-55 ekhulwini, endaweni ye-10 ekhulwini xa unyango lwaqala ekuqanduselweni. Ichiza elifanayo, intshulube efanayo, utshintsho lwemfuza olufanayo, kodwa isiphumo esincinane kakhulu. Ukuvezwa kwangethuba ngenjongo yokuthintela kuko okwakubangela inxalenye enkulu yenzuzo ebonakalayo.
Isiphumo esingabonisi mahluko: oko kwaboniswa luphando lweempuku ze-SOD1
Uphando lweempuku luka-2011 lwabuza umbuzo onzima ngakumbi kwinkqubo engqongqo ngakumbi. Iimpuku ze-congenic ze-SOD1 G93A ziqhubela phambili ukusuka ekungcangcazeleni kwemilenze yangasemva, ukuya ekuphazamisekeni kokuhamba nasekulahlekelweni sisakhono sokuzibuyisela kwindawo yokuma, de zifike kwisiphelo esichazwe kwangaphambili. Unyango lwaqala ngaphambi kweempawu ngosuku lwe-45 lwaza lwaqhubeka yonke imihla, ababukeli benika amanqaku emeko yemithambo-luvo kwisikali esisuka ku-0 ukuya ku-6. Olu lilingelo lwenkqubela yesifo siphela, nto leyo elisondeza ngakumbi kwinto isigulana esiya kuyikhathalela kunayo nayiphi na igrafu yokukhubazeka kwintshulube.
Iziphumo eziphambili azizange zibonise mahluko ubalulekileyo. Ukuqala kwesifo xa sijongwa siphela kwafika kwiintsuku eziyi-121 kwiqela lolawulo, xa kuthelekiswa neentsuku eziyi-124 nge-methylene blue, umahluko owawungabalulekanga ngokwezibalo. Ukuphila de kufike inqanaba lokugqibela kwakuyiintsuku eziyi-167 xa kuthelekiswa neentsuku eziyi-170, nako okwakungabalulekanga ngokwezibalo. Ezinye iziphumo ezikwinqanaba lesibini zaxhasa unyango: iimazi zabonisa ukulibaziseka kokuqala kwesifo malunga neentsuku eziyi-7, yaye izilwanyana ezanyangwayo zachitha ixesha elithe chatha kumanqanaba athambileyo nelincinane kumanqanaba anzima. Ngenxa yamaqela amancinci ahlulwe ngokwesini nothelekiso oluninzi lwamanqanaba, ezi ziphumo zifanele zithathwe njengezivelisa iingqikelelo kunokuba zibubungqina. Akukho kubalwa kweeseli zemithambo-luvo, akukho vavanyo lokuqokelelana kwe-SOD1, yaye akukho zimpawu ze-oksideshini okanye zokudumba ezalinganisiweyo. Ngoko uphando alunakuxela nokuba ichiza lafikelela kwizinto elijolise kuzo laza lasilela, okanye alizange lisebenze kuzo kwaphela kule dosi. Kwavavanywa idosi enye kuphela.
Ababhali ngokwabo bawuphawula owona mda ubalulekileyo, osebenza kumacala omabini. Imodeli ye-SOD1 G93A eneekopi ezininzi imele kakuhle isifo esibangelwa yityhefu ye-SOD1, kodwa isifo se-SOD1 yinxalenye encinane ye-ALS efumaneka kwiintsapho, yaye ibhayoloji yaso yahlukile kuninzi lwezifo ezivela ngaphandle kwembali yosapho. Bacebisa ukuba amachiza anokuba luncedo aphinde avavanywe kwiimodeli zezilwanyana ezanyisayo ezisekelwe kwi-TDP-43. Elo cebiso liye lantsonkotha ngakumbi, kuba ababhali bophando lweentshulube neentlanzi baphawula ukuba olunye uphando lwe-methylene blue kwiimpuku ze-TDP-43 nalo aluzange luphucule iziphumo zentshukumo. Ngoko into ethile inokubonakala inamandla kwinkqubo enye ye-TDP-43 kodwa ingabonisi siphumo kwenye inkqubo ye-TDP-43. Inkqubo ibaluleke ngokulinganayo nemfuza.
Kutheni ukuphucuka kwintshulube okanye kwintlanzi kungenakungqina inzuzo kwizigulana
Cinga ngemodeli nganye njengophendula umbuzo owahlukileyo.
Okokuqala, ibhayoloji iyahluka. I-TDP-43 ne-FUS ziiproteni ezibophelela kwiiasidi zenyukliya, ezinonxibelelwano lwemfuza olufanayo kwezi nkqubo zilula, ngoxa ityhefu ye-SOD1 isebenza ngeendlela ezithe ngqo ezahlukileyo ngokuyinxenye. Ichiza elinciphisa uxinzelelo olubangelwa yiproteni ethile akuyomfuneko ukuba linciphise olo lubangelwa yenye. Impumelelo ekulweni ukuveliswa okugqithisileyo kweproteni kwimbumba ayiqinisekisi impumelelo ekulweni ukonakala okucothayo kwemithambo-luvo kubantu abadala, okuhamba nokudumba, ukubandakanyeka kweeseli ze-glia, nokwehla kokusebenza komzimba wonke.
Okwesibini, ukuvezwa kwichiza kuyahluka. Ukuvezwa ngokuqhubekayo ngokuntywiliselwa kwimixube yoxinaniso lwe-micromolar akufani nokunikwa ichiza ngomlomo. Ukufunxwa emathunjini, ukuguqulwa kwechiza ekuqaleni xa lidlula esibindini, ukubophelela kwiiproteni zeplazma, ukungena engqondweni, nokukhutshwa kwalo emzimbeni zonke ziphakathi kwedosi eginyiweyo neseli yomnqonqo elawula intshukumo. Akukho nasinye isifundo esalinganisa amanqanaba echiza kwinkqubo yemithambo-luvo, ngoko akukho nasinye esinokunxulumanisa uxinaniso lwaso nolwesinye.
Okwesithathu, ixesha liyahluka. Iinzuzo kwiintshulube neentlanzi zazixhomekeke kakhulu kunyango ngaphambi kokuba umonakalo wesifo uqokelelane. Unyango lweempuku nalo lwaqala ngaphambi kweempawu, nto leyo engamandla ophando. Kodwa unyango ebantwini beluya kuqala phantse ngalo lonke ixesha emva kokuqala kweempawu, ixesha elisemva nangakumbi. Umqondiso wokuthintela kuvavanyo lwembumba lweentsuku ezimbini sisiqalo sophando, hayi imodeli yokusetyenziswa kunyango lwezigulana.
Okwesine, iziphumo ezilinganisiweyo ziyahluka. Ukukhubazeka kwiintsuku eziyi-12, uvakalelo kwi-aldicarb, ubude bee-aksoni ngee-micrometer, nomgama wokuqubha ziimpawu ezisondele ngqo kumonakalo ovavanywayo. Ukuqala kwesifo nokuphila kwisilwanyana esanyisayo kudibanisa zonke iinkqubo zomzimba ezibuyekeza okanye ezenza imeko ibe nzima ngakumbi. Amachiza adla ngokutshintsha imilinganiselo ekufutshane nomonakalo ngaphandle kokutshintsha ukuphila, kungoko kukho imilinganiselo yexesha de kufike isiganeko kwizilwanyana ezanyisayo.
Abafundi abafuna uluhlu lokuhlola ibango eliphambili elilandelayo bangalusebenzisa olu lukwi-ndlela yokufunda izifundo zangaphambi kovavanyo ebantwini: buza ngohlobo lwesilwanyana, imfuza efakwe kuso, idosi nendlela yokuyinika, ukuba amanqanaba echiza alinganiswa na, ukuba unyango lwaqala nini xa kuthelekiswa nokuvela komonakalo wesifo, ukuba isiphumo esilinganisiweyo sasiyimpawu yexesha elifutshane okanye inkqubela yesifo, nokuba isiphumo saphindwa na kwilabhoratri yesibini. Ubungqina obubanzi obuvela ebantwini malunga ne-methylene blue buhlala buyindawo yokujonga ukuba nayiphi na kwezi zibonakaliso zangethuba iye yakhula yaba yidatha yonyango lwezigulana.
Kuthetha ukuthini oku kwizigulana neentsapho
La maphepha akamiseli nkqubo yonyango ye-methylene blue eqinisekisiweyo ye-ALS. Uphando olunesiphumo esihle luxhasa uphando olungakumbi ngeendlela ze-oksideshini nezesinapsi kwibhayoloji ye-TDP-43 ne-FUS. Uphando olungabonisanga mahluko luphikisana nokulindela ukutshintshwa kwenkqubela yesifo ngokunikwa ichiza ngomlomo ixesha elide kweli nqanaba lokuvezwa kwisifo sohlobo lwe-SOD1. Xa zidibene, ezi zifundo zichaza ukungangqinelani phakathi kweemodeli, hayi isindululo sonyango.
Ukhuseleko lufuna ukuchazwa ngobulumko obufanayo nokusebenza. Izifundo ze-ALS kwiimpuku azixeli nto ngeedosi ezikhuselekileyo ebantwini, yaye i-methylene blue ineengozi zokwenene zokutshatyalaliswa kweeseli ezibomvu zegazi xa kukho ukunqongophala kwe-G6PD, kunye netyhefu ye-serotonin xa idityaniswe namachiza achaphazela i-serotonin. Ukukrokrelwa kokunqongophala kwe-G6PD, ukusetyenziswa kwangoku kwamachiza okudakumba, nalo naluphi na utshintsho olucetywayo kumayeza kufuneka kuxoxwe ngazo nengcali yezonyango. Ngokunjalo, kufuneka kuphononongwe idatha ngokubanzi yokhuseleko lwe-methylene blue ngaphambi kokuba nabani na athathe inyathelo ngenxa yamabango aphambili avela elabhoratri.
Isigqibo esiphambili singangena kwisivakalisi esinye. I-methylene blue yabuyisela ngamandla iimpawu zentshulube neentlanzi ezikwinqanaba lombungu kwiimodeli ze-TDP-43 ne-FUS xa ukuvezwa kuyo kwaqala kwangethuba, kodwa ayizange ibonise nzuzo phantse kwaphela ekuqaleni kwesifo okanye ekuphileleni kuphando lweempuku ze-SOD1 ezaziyinikwa ngomlomo ixesha elide. Ngoko inkcazelo enyanisekileyo yeyokuba obu bubungqina obuxhomekeke kwimodeli, obungaxhasi ukusetyenziswa kwayo kwizigulana.