I-methylene blue nokuguga kohlaka lwamathambo: kungani okutholwe kumaseli kungazange kubikezele imiphumela emathanjeni amagundane

Cabanga ngenhlanganisela yamakhemikhali ethulisa wona kanye amaseli adiliza amathambo, ihlolwe esitsheni selabhorethri futhi isebenze njengoba bekulindelwe. Ama-osteoclast anezinhlayiya eziyizinhloko eziningi aba mbalwa, kuba nokusabela okucacile okuhambisana nomthamo, futhi kube nencazelo ezwakalayo ephathelene nama-mitochondria. Bese yona le nhlanganisela inikezwa amagundane ingxenye enkulu yokuphila kwawo esekhulile, uhlaka lwawo lwamathambo luskenwe ngezinhlangothi ezintathu, kodwa kungabi nomehluko. Amathambo asekhulile abukeka esekhulile, kungakhathaliseki ukuthi anikezwe ukwelashwa noma cha.

Yilokho okwaboniswa ngokwethembeka ucwaningo olulodwa lwango-2024, futhi kubalulekile ukukuqonda ngokunembile, ngoba lusemngceleni lapho izimangalo eziningi ngezithasiselo zokudla namathambo zehluleka khona. Iphepha elikaPoudel nozakwabo, elashicilelwa ku-Aging ngoMashi 2024, futhi litholakala mahhala, ngakho zonke izilinganiso ezingezansi zingaqhathaniswa nomthombo. Okugqamile kulo umphumela ongabonisi mehluko. Imiphumela engabonisi mehluko ayikona ukwehluleka. Iyizimpendulo, futhi lona uphendula kahle umbuzo oqondile: ingabe ukwelashwa kwesikhathi eside nge-methylene blue noma nge-mitoquinone njengezivimbeli ze-oxidation ezisebenza emzimbeni wonke kulondoloza uhlaka lwamathambo egundane olugugayo? Cha.

Indlela ukuguga kwamathambo okubukeka ngayo ngezinombolo

Ukulahleka kwamathambo ngenxa yobudala akuyona into eyodwa, ngakho ucwaningo aluzange lulinganise into eyodwa kuphela. Besebenzisa i-micro-computed tomography, abaphenyi baskena izindawo ezintathu zohlaka lwamathambo: ungqimba lwethambo oluqinile phakathi kwengxenye ende yethambo lethanga (isiqu esiminyene), ithambo elinezikhala engxenyeni engezansi yethambo lethanga eseduze nelunga (inethiwekhi enjengesipontshi eduze kwelunga), kanye nethambo elinezikhala ethanjeni lesihlanu lomgogodla engxenyeni yesinqe. Endaweni ngayinye babhala izilinganiso ezijwayelekile zesakhiwo esincane samathambo. I-BV/TV iyisabelo somthamo esiyithambo kunokuba sibe yindawo yomnkantsha. I-BMD ukuminyana kwamaminerali. I-Tb.Th, i-Tb.N, ne-Tb.Sp zichaza ukujiya, inani, nezikhala phakathi kwemishayo emincane yethambo. Ukujiya kongqimba lwethambo oluqinile nezindawo zokunqamula isiqu zichaza isiqu.

Cabanga ngethambo elinezikhala njengohlaka lokusekela ngaphakathi kwendawo yokugcina impahla. Ukuguga kususa eminye imishayo ngokuphelele futhi kunciphise ukujiya kwesele, ngakho isabelo sendawo esigcwaliswe yisakhiwo siyehla, imishayo esele iqhelelana, futhi uhlaka lonke luthwala umthwalo omncane. Ungqimba lwethambo oluqinile lufana nodonga lwendawo yokugcina impahla uqobo, oluya ngokuya luncipha ukujiya. Ukwelashwa okuvikela uhlaka lwamathambo kufanele kubonakale kulezi zinombolo. Yilolo hlolo olwasetshenziswa ucwaningo.

Okwenzeka esitsheni selabhorethri

Umsebenzi wamaseli wasebenzisa umnkantsha othathwe emathanjeni amathanga nawemilenze engezansi amagundane e-UM-HET3 anezinyanga ezi-6 kuya kwezi-7 ubudala, uhlobo olunofuzo oluhlukahlukene olukhuliselwa ucwaningo lokuguga. Kwavela okutholakele okune, futhi kubhekisa ezindaweni ezihlukene, ngakho ngakunye kudinga owako umusho.

Okokuqala, amaseli esiqu e-mesenchymal, okuyiqoqo elihlanganisa amaseli azogcina akha amathambo, aqhubeka ephila emazingeni e-methylene blue nawe-mitoquinone asuka ku-0.125 kuya ku-0.5 micromolar phakathi kwamahora angu-48. Akubanga nobuthi, futhi akubanga nokwanda kokuphila kwawo. Okwesibili, lapho lawo maseli esiqu ekhuthazwa ukuba abe ama-osteoblast endaweni yokukhulisa amaseli ekhuthaza ukwakheka kwethambo futhi edaywa ukuze kuhlolwe i-alkaline phosphatase ngosuku lwe-18, ayikho kulezi nhlanganisela eyashintsha umphumela kunoma yiliphi izinga elahlolwa. Uhlangothi olwakha amathambo esitsheni aluzange lusabele.

Okwesithathu, uhlangothi oludiliza amathambo lwasabela. Amaseli angaphambi kokuvuthwa asemnkantsheni akhuthazwa nge-M-CSF ne-RANKL, okuyindlela ejwayelekile yokwakha ama-osteoclast, akha amaseli anezinhlayiya eziyizinhloko eziningi atholakala ene-TRAP ambalwa njengoba izinga lenhlanganisela lalikhuphuka. Ukuncipha okucacile kwavela ngaphezu kuka-0.25 micromolar, empeleni ezingeni eliphezulu elahlolwa elingu-0.5. Yilokhu okutholakele okudala izifinyezo eziningi ezigcwele ithemba, futhi kuyiqiniso ngaphakathi kwemingcele yako: ukuthintana okuqondile nokuqhubekayo kumaseli akhuliswe elabhorethri kwanciphisa ukwakheka kwama-osteoclast ngendlela ehambisana nomthamo.

Okwesine, ama-osteoblast avuthiwe athintana nalezi nhlanganisela amahora angu-48 futhi ahlolwa ngokuhlolwa kwengcindezi yama-mitochondria awazange abonise ushintsho olubalulekile ngokwezibalo ekusetshenzisweni komoya-mpilo okuyisisekelo noma okuphezulu kakhulu nganoma iyiphi inhlanganisela. Ku-0.5 micromolar zombili zabonakala zehlisa kancane ukuphefumula kwamaseli, kodwa ushintsho aluzange lufinyelele ezingeni elibalulekile ngokwezibalo. Ngakho esitsheni akubonakalanga kuthuthukiswa kokusebenza kwama-mitochondria kumaseli akha amathambo okungahambisana nomphumela wokunciphisa ama-osteoclast. Indlela yokusebenza eyayiphakanyisiwe, ukuthuthuka kokukhiqizwa kwamandla ngochungechunge lokudluliswa kwama-electron kuma-mitochondria, ayizange iveze ukwanda okulinganisekayo kokuphefumula kwamaseli lapho abaphenyi babekufuna khona.

Okwenzeka emagundaneni

Kwalandela ukuhlolwa okubili kwezilwane, ngezilinganiso ezihluke kakhulu.

Okuncane kwasebenzisa amagundane esifazane e-C57BL/6J akhuliselwe phakathi kozalo olufanayo, uhlobo olusetshenziswa kakhulu ocwaningweni lwezokwelapha nebhayoloji. Kusukela enezinyanga ezingu-18 ubudala, okucishe kube sekupheleni kobudala obuphakathi egundaneni, uhhafu wawo waphuza amanzi ane-methylene blue engu-250 micromolar izinyanga ezi-6 noma eziyi-12, aze afinyelela ezinyangeni ezingu-24 noma ezingu-30. Amagundane esifazane anezinyanga ezingu-18 angazange anikezwe ukwelashwa anikeza isilinganiso sokuqala, kanti anezinyanga ezi-5 abonisa indlela amathambo asemancane ayebukeka ngayo. Ukuguga kwenza lokho ukuguga okuvame ukukwenza. Ethanjeneni lomgogodla i-L5, isabelo somthamo esiyithambo sehla cishe ngamaphesenti angu-35, ukuminyana kwamaminerali cishe ngamaphesenti angu-18, nenani lemishayo emincane yethambo lehla cishe ngohhafu, kungakhathaliseki ukwelashwa. Ukujiya kongqimba lwethambo oluqinile kwehla cishe ngamaphesenti angu-10 phakathi kwezinyanga ezingu-18 nezingu-24 futhi cishe ngamaphesenti angu-30 lapho sekuyizinyanga ezingu-30. I-methylene blue ayizange iguqule noma iyiphi yalezi zindlela zoshintsho.

Ukuhlolwa okukhulu kwaqhutshwa ngaphansi kwe-Interventions Testing Program ye-National Institute on Aging, ehlola izinhlanganisela ezicatshangelwayo emagundaneni e-UM-HET3 obubili ubulili ezikhungweni ezintathu. I-methylene blue eyizingxenye ezingu-28 esigidini ngasinye ekudleni, ne-mitoquinone eyizingxenye ezingu-100 esigidini ngasinye, ngayinye yaqalwa ekuqaleni kobudala bokukhula futhi yaqhubeka kwaze kwaba cishe yizinyanga ezingu-22 kuya kwezingu-24 ubudala. Zona lezo zindawo ezintathu zohlaka lwamathambo zahlolwa nge-micro-CT. Ithebula elishicilelwe libika zonke izilinganiso ngokobulili nokwelashwa, kusetshenziswa izibalo ezilungisiwe, futhi ikholomu yokwelashwa isho okufanayo kuyo yonke indawo: akukho mphumela obalulekile ngokwezibalo, futhi akukho kuhlangana komphumela wokwelashwa nobulili, noma kuphi. Kungaba ungqimba oluqinile noma ithambo elinezikhala, ithambo lethanga noma umgogodla, owesilisa noma owesifazane, i-methylene blue noma i-mitoquinone. Uhlaka lwamathambo lwaguga ngesikhathi salo esijwayelekile.

Ithebula eliqhathanisayo elihlanganisa zombili izingxenye

Leli yithebula isihloko sonke esisekelwe kulo. Umugqa ngamunye uhlanganisa lokho okuboniswe esitsheni nalokho okuboniswe esilwaneni ngenqubo efanayo yebhayoloji, kusetshenziswa izilinganiso zocwaningo uqobo.

Uma lifundwa umugqa nomugqa, ithebula liqeda lokhu okubonakala njengokuphikisana. Yinye kuphela yezinqubo ezine zamaseli eyasabela, futhi yasabela kuphela eduze komkhawulo ophezulu wamazinga ahlolwa, ngaphansi kokuthintana okuqondile okuqhubekayo. Konke okunye esitsheni kwase kuvele kungabonisi mehluko. Umphumela wesilwane ube usubukeka ungaphansi kokuphikisana futhi usondele ekuqinisekiseni: imiphumela emithathu engabonisi mehluko nomphumela owodwa oncike ezimweni ezithile kumaseli akhuliswe elabhorethri kwaphenduka umphumela ongabonisi mehluko kukho konke esilwaneni.

Isakhiwo samathambo asisona isikhathi sokuphila

Kunokudideka okukodwa okudinga esako isigaba, ngoba i-methylene blue inawo umlando wemiphumela ephathelene nesikhathi sokuphila futhi kulula ukuwufunda ngephutha njengomlando wemiphumela yamathambo. Eqenjini langaphambilini le-Interventions Testing Program, i-methylene blue ngomthamo ofanayo ekudleni yandisa isikhathi sokuphila esisezingeni lamaphesenti angu-90 sokusatshalaliswa kwezikhathi zokuphila kwabesifazane be-UM-HET3 cishe ngamaphesenti angu-6. Lona kwakungumphumela wokusinda ekugcineni kokuphila, ngaphandle koshintsho esikhathini esimaphakathi sokuphila nangaphandle komphumela kwabesilisa. I-mitoquinone ayizange ibonise mphumela esikhathini sokuphila kunoma ibuphi ubulili. Lezo zinombolo zokusinda zivela emarekhodini ashicilelwe ohlelo aphathelene nobude bokuphila, hhayi ephepheni eliphathelene namathambo.

Igalelo lephepha lamathambo liwukugcina lezi zinhlobo zobufakazi zihlukene. Inhlanganisela ingashintsha ukusinda ekugcineni kokuphila kobulili obubodwa ngaphandle kokulondoloza noma isiphi isici esilinganisiwe sohlaka lwamathambo. Ukusinda kubala ukufa okuvela kuzo zonke izimbangela; i-micro-CT ibala imishayo emincane ethanjeni lomgogodla. Awukho umthetho othi ukungenelela kufanele kusebenze ngamathambo ukuze kwandise ukuphila, futhi lolu cwaningo luwubufakazi obuqondile bokuthi umphumela we-methylene blue ekugcineni kokuphila kwabesifazane wadlula ngenye indlela. Noma ubani ocaphuna okutholwe ngesikhathi sokuphila njengokwesekela i-methylene blue ne-osteoporosis usebenzisa ubufakazi obungahambisani naleyo ndaba. Indlela ejwayelekile yokugcina lezi zinhlobo zobufakazi zihlukene ichazwa kokuthi ungalufunda kanjani ucwaningo lwe-methylene blue.

Kungani isitsheni singazange sibikezele igundane

Izincazelo ezinhlanu zihambisana nedatha, futhi ukuba enye iyiqiniso akusho ukuthi ezinye azinakuba yiqiniso.

Ukuthintana nenhlanganisela kungokwokuqala. Amaseli akhuliswe elabhorethri ayecwiliswe ngokuqhubekayo emazingeni aziwayo. Amagundane adla ukudla okunemithamo izinyanga, futhi ucwaningo aluzange lulinganise amazinga omuthi emnkantsheni noma noma yiluphi uphawu lokulimala kwamathambo okubangelwa yi-oxidation. Izinga elinciphisa ama-osteoclast lapho lisetshenziswa ngokuqondile kungenzeka lingalokothi lifinyelele ngalelo zinga kumaseli andulela amanye emnkantsheni. Iphepha lahlola umthamo owodwa wenhlanganisela ngayinye ezilwaneni eziphilayo, ngakho ukusabela kwesilwane emithamweni ehlukene kusalokhu kungaziwa.

Ikhemistri ingeyesibili. Izinhlobo ezisebenzayo zomoya-mpilo azakhi iqoqo elilodwa elixubene kahle ngaphakathi kweseli. Zisebenza ezingxenyeni ezincane ngezikhathi ezimfushane, futhi izinhlelo ze-redox zama-mitochondria, ze-cytosol, nezolwelwesi zisebenza ngokwengxenye ngokuzimela. I-mitoquinone yakhelwe ukuba iqoqane kuma-mitochondria, kuyilapho izimpawu zama-oxidant eziqhuba ukwakheka kwama-osteoclast zingavela kwenye indawo. Ukunciphisa iqoqo elilodwa akudingeki kuphindaphinde lokho okufezwa ukucwiliswa okuqhubekayo kweseli lonke.

Ukuxhumana kwezinqubo kungokwesithathu. Ethanjeni eliphilayo, ukudilizwa nokwakheka kuxhumene: ama-osteoclast namaseli aseduze nawo akhipha izimpawu eziletha amaseli akha amathambo endaweni esanda kudilizwa. Ukunciphisa ukwakheka kwama-osteoclast akungezi ithambo elilingana nalokho okuncishisiwe, ngoba kungase futhi kususe izimpawu ezibiza abakhi. Isitsha esiqukethe amaseli andulela amanye kuphela asikwazi ukubonisa lokhu kubuyisana kwezinqubo. Uhlaka lwamathambo alukwazi ukukufihla.

Umthwalo ungowesine. Ama-osteocyte ezwa ukucindezelwa komshini futhi anqume ibhalansi yokuvuselelwa kwethambo lonke. Njengoba izilwane ziguga futhi zinyakaza kancane, uphawu lomthwalo womshini luyancipha, bese kulandela ukulahleka kungakhathaliseki umthelela omncane womuthi ohlotsheni olulodwa lweseli. Ababhali ngokwabo babiza ukuzivocavoca nokuthwala umthwalo okukhuthaza ukwakheka kwezicubu njengezindlela ezingase zibe nomthelela omkhulu, okuyindlela enomusa yokuthi uhlaka lwamathambo lulalela imithetho yefiziksi kakhulu kunekhemistri ye-redox.

Isikhathi sokuqala singesesihlanu, kodwa kunombandela. Ekuhlolweni kwe-C57BL/6J, ukwelashwa kwaqala ezinyangeni ezingu-18, lapho ukulahleka okukhulu kwesakhiwo kwase kwenzekile, futhi akukho okunciphisa ukudilizwa kwamathambo okungakha kabusha imishayo emincane yethambo esibhoboziwe futhi yasuswa ngaphandle kokukhuthazwa kokwakheka okuhambisana nakho. Kodwa isikhathi sokuqala asikwazi ukuthwala yonke incazelo, ngoba ukunikezwa kwemithamo kwe-Interventions Testing Program kwaqala ekuqaleni kobudala bokukhula futhi nalapho akuzange kushintshe lutho. Ukuqala sekwephuzile kuchaza ingxenye yokuhlolwa okukodwa. Akuchazi lutho ngokunye.

Lokho lokhu okukushoyo ngezimangalo ze-methylene blue ne-osteoporosis

Kumfundi ohlola i-methylene blue ngokuphathelene nempilo yamathambo noma ukuguga kohlaka lwamathambo, incazelo esebenzisekayo imfushane. Ubufakazi obuhle kakhulu obutholakalayo besikhathi eside emagundaneni bubika ukuthi ayikho inzuzo ohlakeni lwamathambo ngokusetshenziswa okuqhubekayo kwe-methylene blue noma kwe-mitoquinone, naphezu kophawu lwangempela lokunciphisa ama-osteoclast kumaseli akhuliswe elabhorethri. Imibhalo ephathelene nesikhathi sokuphila ibonisa umphumela omncane we-methylene blue ekugcineni kokuphila kwabesifazane kuphela, lolu cwaningo olubonisa ukuthi awudlulanga ngamathambo. Ayikho kulezi zinhlobo zobufakazi esekela ukusetshenziswa kubantu ngenxa ye-osteoporosis, futhi akukho datha yamathambo abantu engena kule ndaba nhlobo. Imibuzo ngemithamo nokuphepha kwemikhiqizo kubantu ingaphansi kwesihloko sokuphepha kwe-methylene blue nemiphumela engemihle, hhayi ngaphansi kwalolu hlaka lwamathambo egundane.

Isifundo esijulile siphathelene nokulandelana kobufakazi. Umphumela wamaseli uyisitatimende ngalokho inhlanganisela engakwenza ohlotsheni olulodwa lweseli ngaphansi kokuthintana okulawulwayo. Umphumela wamathambo uyisitatimende ngalokho okwenziwa izinyanga zokuthintana okusebenza emzimbeni wonke esithweni esixhumanisa izinhlobo ezimbili zamaseli ngaphansi kokulawulwa yimithwalo yomshini. Okokuqala yilapho kuzalwa khona imibono ehlolwayo. Okwesibili yilapho, ngokuphathelene nokuguga kohlaka lwamathambo nalezi zivimbeli ze-oxidation ezimbili, lo mbono waphela khona.