I-methylene blue uma iqhathaniswa nezindlela ze-NAD+: izindlela zokusebenza, ubufakazi, nezimangalo zokuzihlanganisa

I-methylene blue nezithasiselo ezithengiswa njengezikhuphuli ze-NAD+ kokubili kuchazwa ngamagama athi ama-mitochondria namandla amaseli. Ayikho kulezi zincazelo ekutshela ukuthi umkhiqizo wenzani ngempela.

Lezi zindlela ezimbili zithinta izici ezihlukene. Enye izama ukushintsha ukuthi amaseli akho aqukethe i-NAD+ engakanani. Enye ishintsha isilinganiso phakathi kwezinhlobo zayo ezimbili. Lezi akuzona izindlela ezimbili eziya endaweni efanayo.

Izilinganiso ezimbili ezihlale zididaniswa

I-NAD+ iyi-coenzyme. Yamukela ama-electron amabili, esesimweni se-ion ye-hydride, bese iba yi-NADH. Lapho i-NADH idlulisela lawo ma-electron phambili, iphinde ibe yi-NAD+. Amaseli asebenzisa lo mjikelezo njalo: yindlela amandla akhishwe ekudleni afinyelela ngayo emishinini yamaprotheni eyakha i-ATP.

Njengoba i-NAD+ ne-NADH kuyizimo ezimbili ze-molecule efanayo, kunamanani amabili ahlukene okufanele alandelwe eselini. Elokuqala yinqolobane ephelele: ukuthi ingakanani i-NAD+ ne-NADH uma kuhlanganisiwe. Elesibili yisilinganiso phakathi kwazo, esichaza ukuthi izithwali zama-electron zilayishwe kangakanani ngaleso sikhathi.

Ichibi ledamu lenza lo mehluko ucace. Umthamo wamanzi agcinwe ngemuva kwedamu uyinqolobane. Ukwahlukaniswa kwamanzi agcinwe ngenhla kwedamu nalawo akhishelwe ngezansi yisilinganiso. Ungasikhuphula leso silinganiso ngaphandle kokwengeza ilitha, ngokuvula isango. Lokho akusona isingathekiso salokho okwenziwa yi-methylene blue. Kusondelene nencazelo engokoqobo.

Indlela yokwengeza izinto zokwakha

Izithasiselo eziyizanduleli ze-NAD+, okuhlanganisa i-nicotinamide riboside, i-nicotinamide mononucleotide, ne-nicotinamide, ziyizinto zokwakha. Amaseli akha i-NAD+ ngomzila wokuphinda asebenzise izinto obizwa ngokuthi umzila wokubuyisa, futhi izanduleli zingena kulo mzila. Inhloso ukukhulisa inqolobane.

Ezilwaneni, lokhu kuqinisekiswe kahle: i-NAD+ ezicutshini iyancipha ngokukhula kweminyaka nangezifo zokushintshaniswa kwezinto emzimbeni kumamodeli amaningi, futhi ukunikezwa kwezanduleli kuvame ukuyikhuphula. Kubantu, ubufakazi buncane kakhulu. Izivivinyo kubantu ziyabonisa ukuthi lezi zinhlanganisela zikhuphula amazinga e-NAD+ alinganiswa egazini. Lokho ezingakakuvezi ukuthuthuka okungaguquguquki emiphumeleni yomtholampilo, okusho izinto umuntu angazibona kuye noma eziklanyelwe ukulinganiswa esivivinyweni.

“Amazinga e-NAD+ akhuphukile” kulinganisa okuthile ngaphakathi emzimbeni. Akusona isiphetho esimayelana nokuthi umuntu wazizwa kanjani, wanyakaza kanjani, noma walulama kanjani.

Lokho okwenziwa yi-methylene blue esikhundleni salokho

I-methylene blue ayisona isanduleli. Ingudayi we-redox, osekuyisikhathi eside kangangokuba ususetshenziswe kwezokwelapha iminyaka engaphezu kwekhulu, futhi ukusetshenziswa kwayo okugunyaziwe kuncane kunalokhu okusaphenywayo.

Ngaphakathi kweseli, ama-enzyme ancike ku-flavin ayinciphisa ngokuyinikeza ama-electron ibe yisimo esingenambala, i-leucomethylene blue, esibe sesiphinde senziwe i-oxidation. Ama-electron abezodlula ku-complex I, isiteshi sokuqala sochungechunge lokuphefumula angakhishwa ku-NADH nge-methylene blue esikhundleni salokho. I-NADH iba yi-NAD+.

Isilinganiso siyakhuphuka. Inqolobane ayikhuli.

Ucwaningo lwamagundane enhliziyweni enesifo sikashukela lwalinganisa lokhu ngokuqondile kuma-mitochondria enhliziyo ayehlukanisiwe. Ukwengeza i-methylene blue emazingeni okusuka ku-6 kuya ku-18 micromolar ngesikhathi sokwenziwa kwe-oxidation kwama-asidi anamafutha kwabangela ukwehla kwe-NADH nesilinganiso se-NADH ne-NAD+ okuncike enanini elengeziwe. Inani le-NAD+ ngokwalo alishintshanga kakhulu njenge-NADH, okuyilokho ongakulindela enhlanganiselweni eguqula uhlobo olulodwa lube olunye kunokuba ikhiqize olunye olwengeziwe.

Kubalulekile ukuthi lawo ma-electron agcina ephi. Kuma-mitochondria obuchopho ayehlukanisiwe, i-methylene blue emazingeni okusuka ku-100 nanomolar kuya ku-1 micromolar yathuthukisa ukukhiqizwa kwamandla lapho uchungechunge lokuphefumula luvinjelwe, futhi ngesikhathi esifanayo yandisa ukukhiqizwa kwe-hydrogen peroxide kuyilapho inciphisa ukususwa kwayo, kuzo zonke izimo ezahlolwa. Ababhali bathola ukuthi le hydrogen peroxide yayivela ekuncishisweni komoya-mpilo ngokuqondile yi-methylene blue esithole ama-electron. Ama-electron aphuma ku-NADH ngalo mzila awanyamalali. Agcina esemoyeni-mpilo.

Ucwaningo oluhlukile ezicutshini zomsipha i-Achilles luchaza leli phuzu ngokusobala. I-methylene blue yabuyisela ibhalansi ye-NAD+/NADH futhi yakhuphula ukukhiqizwa kwe-ATP kumaseli omsipha alimele nakumodeli yamagundane yokuphazamiseka kwemisipha, futhi iphepha libika ukuthi inqolobane ephelele ye-NAD ayizange ibonise ushintsho olubalulekile.

Lokho okuqinisekiswa izivivinyo zezicubu nezilwane

Izivivinyo ezine ezenziwe emashumini amane eminyaka zilandela umqondo ofanayo futhi ziveza umkhawulo wawo ngokuphindaphindiwe.

Ekuphuzisweni kwamagundane utshwala isikhathi eside, i-ethanol yehlisa izilinganiso ze-NAD+/NADH kokubili ku-cytosol nakuma-mitochondria esibindini. Ukwengeza i-methylene blue ekudleni kwavimbela kakhulu lolo shintsho. Akuzange kunciphise ukunqwabelana kwamafutha esibindini okwabangelwa yi-ethanol. Ukulungisa isilinganiso akuzange kulungise umphumela. Lowo mphumela wango-1985 kufanele ukhunjulwe ngaphambi kokwamukela noma yisiphi isimangalo sokuthi ukushintsha kwe-redox kuyindlela ebangela inzuzo.

Kumaseli esibindi nasemagundaneni adla ukudla okunamafutha amaningi, i-methylene blue yakhuphula isilinganiso se-NAD+/NADH, yandisa i-SIRT1, yanciphisa ukunamathiselwa kwamaqembu e-acetyl ku-PGC-1alpha, yavula ukusebenza kwe-AMPK, futhi yanciphisa ukunqwabelana kwamafutha esibindini. Lapha umphumela washintsha kanye nesilinganiso. Lona leli phepha libonisa ukuthi umphumela awuqhubeki ukhula ohlangothini olulodwa: ama-milligram amathathu ngekhilogremu ngosuku andisa inani le-DNA yama-mitochondria kangcono kunayishumi, kuyilapho ukusetshenziswa komoya-mpilo nokudluliswa kwezimpawu kwe-AMPK kwakhuphuka ngokwanda kwenani.

Enhliziyweni enesifo sikashukela, i-methylene blue eyayinikezwa emanzini okuphuza amasonto ayishumi nanye yehlisa isilinganiso se-NADH ne-NAD+ kuma-mitochondria enhliziyo futhi yakhuphula ukusebenza kwe-sirtuin 3. Ukunamathiselwa kwe-acetyl ku-lysine kwehla ezindaweni ezingama-83 emaprothenini angama-34, ikakhulu kulezo eziphathelene nokuthuthwa nokwenziwa kwe-oxidation kwama-asidi anamafutha. Ingxenye yegazi ekhishwa yinhliziyo lapho ifinyela yathuthuka. Kodwa ukuphefumula okusekelwa yi-complex I akuzange kulungiswe. Futhi isivivinyo sokucisha isakhi sofuzo sahlukanisa imiphumela emibili okulula ukuyididanisa: ukushintsha kwesilinganiso kwenzeka kakhulu ngaphandle kwe-sirtuin 3, kuyilapho ukususwa kwe-acetyl kwakuncike kuyo.

Ocwaningweni lomsipha, i-methylene blue yasetshenziswa ngesikhathi i-complex I ivinjwe ngamakhemikhali nge-rotenone. Yayisabuyisela ibhalansi ye-NAD+, futhi yayisahluleka ukuvimbela ukucindezeleka okubangelwa yi-oxidation, ukuwohloka kwe-matrix, nokulahleka kwamandla amaseli okuhlala ephila. Ukubuyisela isilinganiso kwakunganele ukuze izicubu zilulame.

Iphepha eliphrintekayo elinezinkomba zocwaningo eziphelele lisebenzisa imigqa efanayo.

Kungani ama-sirtuin efakwa kule mpikiswano

Ama-sirtuin asusa amaqembu e-acetyl emaprothenini, futhi asebenzisa i-NAD+ ukwenza lokho. Ukusebenza kwawo kusabela ekutholakaleni kwe-NAD+, ngakho noma iyiphi inhlanganisela eshintsha ukutholakala kwayo ishintsha ukusebenza kwama-sirtuin okulandelayo.

Yilokhu ukuthengisa okuningi okuthi “i-methylene blue ye-NAD+” okuncike kukho. Yilapho futhi ukucabanga kuphambuka khona. Ukushintsha isilinganiso kungakhuphula ukusebenza kwama-sirtuin. Lokho akusho ukuthi umkhiqizo unikeza i-NAD+, futhi akusho ukuthi umphumela uyafana nokuthatha isanduleli. Ukuchazwa kwalezi zilinganiso ngokwe-redox kunezihibe zako okufanele ufunde ngazo ngaphambi kokwamukela noma iyiphi yalezi zincazelo.

Lokho osekuhloliwe kubantu

I-methylene blue isisetshenziswe emitholampilo amashumi eminyaka ezimweni eziphuthumayo eziqashwe ngokucophelela: i-methemoglobinemia, ubuthi obuthinta isimiso sezinzwa obuhlobene ne-ifosfamide, kanye nomfutho wegazi ophansi kakhulu ngesikhathi sokuhlinzwa. Lokhu ukusetshenziswa okulinganiselwe nokuqondiswayo, hhayi ubufakazi bokuyithatha ngomlomo nsuku zonke njengento ethuthukisa ukusebenza.

Ukusesha okwenziwe ngalesi sihloko akutholanga ucwaningo kubantu olulinganisa imikhiqizo yokushintshaniswa kwe-NAD+ emzimbeni ngemva kwe-methylene blue kumavolontiya aphilile. Okutholakele okungenhla kuvela kuma-mitochondria ayehlukanisiwe, kumaseli akhuliselwe elebhu, ezicutshini zamagundane, nasezicutshini ezikhishwe emzimbeni womuntu. Umsipha okhishwe emzimbeni wagcinwa uphila esitsheni uyisinyathelo esiwusizo esidlula amaseli akhuliselwe elebhu. Nokho awuyena umuntu.

Imibhalo yocwaningo lwezanduleli ngempela ihlanganisa abantu, futhi imiphumela yayo incane. I-NAD+ egazini iyakhuphuka. Imiphumela yomtholampilo ixubile futhi ngokuvamile incane.

Ngakho isimangalo sokuzihlanganisa siwumbuzo ofanele onempendulo engagculisi: alukho ucwaningo olushicilelwe, isivivinyo, noma uchungechunge lwamacala olutholakele ngalesi sihloko oluhlole i-methylene blue kanye ne-nicotinamide riboside, i-nicotinamide mononucleotide, noma ukufakwa kwe-NAD+ ngomthambo. Noma ubani ochaza le nhlanganisela njengokusekelwa ubufakazi, noma njengokusebenza ngokuhlanganyela ukuze kwandiswe umphumela, welula iziphetho ezivela emaqoqweni amabili obufakazi ahlukene.

Ukwelula iziphetho kungaphambuka ngezindlela zombili. Kungaba nethemba elikhulu ngokweqile, uma inqolobane ibingeyona into ebeka umkhawulo lapho isilinganiso sesibuyiselwe. Kungabuye kube lula ngokweqile, ngoba ukusabela enanini le-methylene blue kuyafika endaweni lapho kushintsha khona indlela. Abacwaningi bayo bayichaza njengenhlanganisela enomphumela we-hormesis: amazinga aphansi avuna ukuncishiswa ngokwamukela ama-electron, kuyilapho amazinga aphezulu ephambukisa ama-electron awayise kude nochungechunge lokuphefumula futhi engase akhuthaze i-oxidation. Ukwengeza izinto eziningi ezisetshenziswa kule nqubo akulenzi lelo jika liphephe kakhulu.

Kukhona futhi ukukhathazeka ngokusebenzisana nezinye izinto okungahlangene ne-NAD+. I-methylene blue ivimbela i-monoamine oxidase A, yingakho inobungozi obuhlobene ne-serotonin lapho ihambisana nemithi ethile yokucindezeleka ngokwengqondo; ukubuyekezwa kwemibiko yobuthi obuthinta isimiso sezinzwa esimaphakathi kwathola ukuthi amacala angu-13 kwangu-14 ahlangabezana nemigomo ka-Hunter yobuthi be-serotonin. Ingabangela futhi i-methemoglobinemia lapho umuntu echayeka kuyo ngezinga eliphezulu. Uma uthatha imithi kadokotela, umbuzo wokuhlanganisa okufanele uwubuze uchwepheshe wakho wezempilo umayelana naleyo mizila, hhayi ibhalansi ye-redox. Ukuqapha okufanayo kuyasebenza nasezinhlanganiselweni ezivamile: ezinye izithasiselo ezithathwa kanye ne-methylene blue zineqoqo lazo lobufakazi, ngokuyinhloko obungakahlolwa.

Kungani ukuhlanzeka kunquma ukuthi yisiphi isivivinyo osenzayo ngempela

Umkhiqizo we-methylene blue uyingxube kuze kube yilapho ilabhorethri ifakazela okwehlukile. Ukungcola okubalulekile lapha yi-Azure B, i-phenothiazine ekhishwe iqembu le-methyl futhi engahlali ingasebenzi. Ingudayi osebenza ku-redox onesisekelo esifanayo sokusebenza kwamakhemikhali kagesi, ngakho umkhiqizo one-Azure B enanini elibalulekile awulona nje uhlobo oluhlanzeke kakhulu noma olungcole kakhulu lwento efanayo. Uyingxube ehlukile esebenza emzileni ofanayo, ngesilinganiso esingakachazwa muntu.

Izinga le-USP liyindlela umthengi afinyelela ngayo entweni echazwe ngezindinganiso. Umkhiqizo ongekho ezingeni le-USP cishe ungudayi wezindwangu, futhi amadayi ezinga lezindwangu aqukethe ukungcola okuwenza angafaneleki ukudliwa, okuhlanganisa izinsimbi ezisindayo nezinsalela zezincibilikisi.

Igebe alikho phakathi kwamazinga kuphela. Abathengisi abaningi abathi bathobela i-USP bahlola izinsimbi ezisindayo kuphela, bese bethula lelo qoqo elilodwa lokuhlola njengokuthobela okuphelele. Imininingwane yezidingo ephelele ihlanganisa ukuhlonzwa kwento, ukuhlanzeka, ukungcola kwezinto eziphilayo ngokwesakhiwo samakhemikhali, izinsalela zezincibilikisi, ukungcola kwezakhi, insalela ngemva kokushiswa, imikhawulo yamagciwane, kanye nama-endotoxin amagciwane. Iqoqo lokuhlola elingaphelele alikwazi ukufakazela izici elizishiyayo, futhi okushiywayo kuhlanganisa izinto ezihlobene nezinsalela zezincibilikisi.

Blupreme ithenga kumkhiqizi we-methylene blue wemithi futhi ishicilela isitifiketi sokuhlaziywa esihlanganisa yonke imininingwane yezidingo, ukuze into esebhodleleni kube yiyona echazwa ucwaningo. Uma ufuna ukuzihlolela ukucabanga ngaphambi kokuthenga, indlela yokufunda ucwaningo lwe-methylene blue ichaza lokho uhlobo ngalunye locwaningo olungakubonisa nolungenakukubonisa.

Umehluko ohlala uqinisekile lapho uhlolwa ngokucophelela

Ukwengeza isanduleli se-NAD+ kushintsha ukuthi ingakanani i-NAD+ ekhona. I-methylene blue ishintsha lokho okwenziwa yi-NAD+ ekhona. Okokuqala kuwumbuzo wokunikezwa kwezinto, okwesibili kuwumbuzo wokugeleza kwazo, futhi izivivinyo ezisekela okukodwa zisho okuncane ngokunye.

Okusekela i-methylene blue yindlela yokusebenza ehambisanayo, eqinisekiswe kuma-mitochondria ayehlukanisiwe nasezicutshini zezilwane, enomkhawulo ocacile ophindaphindiwe: ukushintsha isilinganiso akufani nokulungisa izicubu. Okushodayo yizivivinyo kubantu ezilinganisa umphumela we-NAD+ kubantu. Okungekho kunoma yiliphi iqembu ukuhlolwa okuqondile kwalokhu okubili lapho kuthathwa ndawonye.

Womabili amaqembu afanelwe indlela efanayo yokuwahlola. Buza ukuthi yisiphi isici umkhiqizo othi uyasishintsha, bese ubuza ukuthi yiziphi izilinganiso ezikhona zaleso sici kubantu. Uma impendulo iwucwaningo lwamaseli, igundane, noma umzila ongase usebenze, yilokho okuyikho. Akuyona inhlanganisela yezithasiselo efakazelwe.