Methylene blue ndi mitochondria: kuyenda kwa maelekitironi ndi umboni

Yerekezerani zitsanzo ziwiri zokonzedwa kuti ziyesedwe, zili pambali pa zinzake. Zonse zili ndi mitochondria za mu ubongo zomwe zawonongeka ndi rotenone, mankhwala omwe amatseka Complex I ya unyolo wa kupuma. Mu chitsanzo chimodzi, woyesera akuikamo methylene blue yokhala ndi mlingo wa micromolar pogwiritsa ntchito pipette. Kugwiritsa ntchito oxygen kukubwerera pang'ono. Voteji ya nembanemba ikubwereranso pang'ono. Kupanga ATP kukuyambiranso pang'ono. Mu cuvette yoyandikana nayo, chipangizo choyezera peroxide chikufotokoza mbali ina ya nkhaniyi: kuchuluka kwa hydrogen peroxide yomwe ikutuluka kwawonjezeka kangapo, pa mlingo uliwonse womwe wayesedwa, ndi pa chakudya chilichonse cha kachitidwechi chomwe chayesedwa.

Zotsatira zonsezi ndi zolondola. Methylene blue ndi chonyamulira maelekitironi chomwe chingapitirizitse kuyenda kwawo podutsa pambali pa gawo lowonongeka la unyolo wa kupuma, koma chonyamulira chomwechi chimawonjezera peroxide pochita zimenezo. Mukakumbukira mfundo zonse ziwirizi nthawi imodzi, mabuku a kafukufuku okhudza ntchito ya methylene blue mu mitochondria amamveka bwino. Mukangoganizira imodzi yokha, mungawerenge nkhaniyi ngati yozizwitsa kapena yoopsa, malinga ndi pepala la kafukufuku lomwe mwatsegula poyamba.

Chonyamulira maelekitironi, chofotokozedwa mosavuta

Unyolo wa kupuma uli ngati mzere wopatsirana zinthu. Maelekitironi amalowa pa Complex I (kuchokera ku NADH) kapena pa Complex II ndi ma enzyme a flavin ogwirizana nayo (kuchokera ku succinate ndi zinthu zina zopereka maelekitironi), amadutsa mu Complex III kupita ku cytochrome c, chonyamulira chosunthika, kenako amadutsa mu Complex IV kupita ku oxygen. Kupatsirana kulikonse kumapopa mapulotoni kudutsa nembanemba yamkati, ndipo kusiyana kwa kuchuluka kwawo komwe kumabwera chifukwa cha zimenezi kumayendetsa kupanga ATP. Malo aliwonse akatsekedwa, mzere wa mbali isanakwane malowo umasiya kugwira ntchito.

Methylene blue imalowa ngati mthenga wodutsa m'njira ina yofanana ndi yoyamba. Mtundu wake wa oxidized umatenga maelekitironi kuchokera ku zonyamulira za mbali yoyambira, n'kusanduka mtundu wa reduced wopanda mtundu, leucomethylene blue. Mtundu umenewu umapereka maelekitironiwo ku mbali yotsatira n'kubwereranso kukhala wabuluu. Mitundu iwiriyi yafotokozedwa mu nkhani ya mitundu iwiri ya redox, ndipo mmene kupatsirana kumeneku kumagwirira ntchito mwachidziwitso chonse muli mu nkhani yofotokoza njira yonse yogwirira ntchito. Funso lomwe silinayankhidwebe ndi lakuti mthengayu amabwereranso pati mu mzerewu: m'thumba lomangira quinol la Complex III, lotchedwa malo a Qo, kapena mwachindunji pa cytochrome c. Njira yotulukira yomwe imagwiritsidwa ntchito kwambiri imadalira mmene zinthu zilili, monga momwe kuwunika njira yodutsa pambali pa Complex III kumafotokozera mwatsatanetsatane.

Zomwe kafukufuku atatu wa nyama zokutafuna anayezadi

Kafukufuku onse atatu anagwiritsa ntchito mitochondria zochotsedwa mu ubongo, kutanthauza tizigawo ta selo tomwe tachotsedwa m'selo n'kuikidwa m'madzi a buffer okonzedwa mwatsatanetsatane. Panalibe maselo athunthu, nyama zopatsidwa mankhwala, kapena anthu oyesedwa. Kuchuluka kwa mankhwala kumatanthauza kuchuluka kwake m'madzi ozungulira tizigawo ta selo, osati mlingo womwedwa ndi munthu. Mukasunga mfundo imeneyi m'maganizo, manambalawo amafotokoza zinthu momveka bwino.

Kafukufuku woyamba, wa mitochondria zochotsedwa mu ubongo wa guinea pig pa 100 nanomolar mpaka 2 micromolar, anasonyeza phindu ndi mtengo wake mu chitsanzo chimodzi. Mu mitochondria zathanzi, utotowo unawonjezera kugwiritsa ntchito oxygen pamene zinali zopuma koma sunasinthe kupuma kolimbikitsidwa ndi ADP, choncho kupanga ATP kogwirizana ndi kupumako sikunawonjezeke. Pamene Complex I inatsekedwa, 2 micromolar inabwezeretsa kupanga ATP kuchokera pa 46 kufika pa 93 nanomoles pa minute pa milligram, poyerekeza ndi liwiro labwinobwino la pafupifupi 1109. Pamene Complex III inatsekedwa, mlingo womwewo unabwezeretsa ATP kuchokera pa pafupifupi 46 kufika pa 63, poyerekeza ndi liwiro labwinobwino la pafupifupi 910. Voteji ndi kulowetsa calcium zinabwereranso pang'ono. Kutsekedwa kwa Complex IV ndi cyanide sikunathe kudutsidwa pambali, zomwe zimaika potulukira maelekitironi kothandiza patsogolo pa Complex IV yogwira ntchito. Kubwezeretsa ntchito kulikonse kunali kwenikweni malinga ndi ziwerengero, koma kunali kochepa: maperesenti a manambala amodzi okha a kupanga ATP kwabwinobwino.

Kafukufuku wachiwiri, wa mitochondria za mbali yakunja ya ubongo wa mbewa pa pafupifupi 1 micromolar, anatsimikizira njira yodutsa pambali pa Complex I ndipo anasonyeza malire pa Complex III. Rotenone inachepetsa kupuma kuchokera pa pafupifupi 121 kufika pansi pa 2 nanomoles za oxygen pa minute pa milligram, ndipo utotowo unakweza kufika pa pafupifupi 21, pafupifupi gawo limodzi mwa magawo asanu ndi limodzi a mulingo wabwinobwino, pamene voteji ya nembanemba inabwereranso pafupifupi yonse. Koma Complex III itatsekedwa, kupuma kapena voteji sizinabwerere pa 1 mpaka 5 micromolar, ndipo kutsekedwako kunachepetsa chizindikiro cha peroxide choyambitsidwa ndi utotowo. Olemba kafukufukuyu anapereka lingaliro lakuti utoto wa reduced umapereka maelekitironi ake pa malo a Qo a Complex III: malowo akakhala otsekedwa, mthengayu alibe malo operekera katundu wake.

Kafukufuku wachitatu, wa mitochondria za mu ubongo wa mbewa, makoswe ndi guinea pig, makamaka pa 2 micromolar, anayesa kusagwirizanaku mwachindunji ndipo anapeza kuti kutsekedwa kwa Complex III kungadutsidwe pambali. Pamene myxothiazol, yotsekereza malo a Qo, inachepetsa kupuma koyambira ndi 88 percent, utotowo unabwezeretsa kugwiritsa ntchito oxygen kufika pa pafupifupi 62 percent ya liwiro losatsekedwa. Poyesa mwachindunji, theka la micromolar linachititsa cytochrome c yowonjezeredwa kuchokera kunja kulandira maelekitironi ngakhale Complex III inali yotsekedwa. Izi zinathandiza lingaliro la njira yochokera ku NADH kudzera mu utotowo kupita ku cytochrome c, kenako ku Complex IV. Koma kupuma komwe kunabwezeretsedwako kunachita zinthu mosiyana ndi kwabwinobwino: ADP yowonjezeredwa inachepetsa kugwiritsa ntchito oxygen m'malo mowonjezera, ndipo kutseka kupanga ATP kunalimbikitsa kupuma. Kusiyana kwa voteji pa nembanemba kunangobwereranso pang'ono. Kugwiritsa ntchito oxygen kwambiri pamene kugwirizana kwake ndi kupanga ATP kwatembenuka sikupanga mphamvu kwabwinobwino.

Chifukwa chake ma labotale sakugwirizana, ndi zomwe zimathetsa kusagwirizanako

Kufotokoza kusiyanaku chifukwa cha mitundu ya nyama sikugwira ntchito. Gulu lachitatu linabwereza ntchitoyi mu mitundu itatu ya nyama zokutafuna ndipo linapeza njira yodutsa pambali yofanana m'mitundu yonse, kuphatikizapo mtundu wa mbewa womwe unanenedwa kale kuti unalibe njira imeneyi. M'malo mwake, zomwe zimasiyana ndi kuchuluka kwa mankhwala (1 poyerekeza ndi 2 micromolar), milingo ya zotsekereza, ma buffer, njira zochotsera mitochondria, ndipo koposa zonse, momwe kupanga mphamvu kunalili pa nthawi yoyerekezera: njira yodutsa pambali imaoneka bwino kwambiri ADP isanawonjezeredwe, pomwepo pamene kuyankha kodabwitsa kwa ADP kumaonekera. Choncho chithunzi cholondola chimasonyeza njira ziwiri zotulukira zomwe zimadalira mmene zinthu zilili, imodzi pa malo a Qo ndi ina pa cytochrome c. Mthengayu amapereka katundu pamene mmene makemikolo alili kumamulola kutero.

Mtengo wake wa peroxide

Mbali ya mtengo wake imasonyeza zotsatira zogwirizana mofanana ndi mbali ya kubwezeretsa ntchito. Mu kafukufuku wa guinea pig, utotowo unawonjezera kutulutsa peroxide pa mikhalidwe yonse yoyesedwa: popuma, popanga ATP, komanso ntchito itasokonezedwa ndi mankhwala, pa zakudya zonse zitatu za kachitidwechi. Pakupanga ATP, kuchuluka kotuluka kunakwera kuchokera pa pafupifupi 148 kufika kupitirira 1500 picomoles pa minute pa milligram pa 1 micromolar, ndipo ngakhale 100 nanomolar inawonjezera chizindikirocho kupitirira kanayi. Utotowo unachedwetsanso kuchotsa peroxide pafupifupi 2.6 fold, choncho chizindikirocho chimasonyeza kupanga mwachangu komanso kuchotsa pang'onopang'ono. Njira ya makemikolo yomwe yaperekedwa ngati lingaliro ndi yakuti utoto wa reduced umapereka maelekitironi mwachindunji ku oxygen popanda superoxide yooneka poyezera, ngakhale zimenezi zikadali kutanthauzira zotsatira, osati sitepe yotsimikiziridwa.

Kafukufuku wa mbewa amawonjezera mfundo zofunika zomwe zimapangitsa nkhaniyi kukhala yovuta kwambiri. Peroxide inakwera kwambiri pamene mlingo unkawonjezeka, kuchokera pa mulingo woyambira wa pafupifupi 76 kufika pafupifupi 2000 picomoles pa minute pa milligram pa 2 micromolar, koma kutsekedwa kwa Complex III kunachepetsa gawo loyambitsidwa ndi utotowo. Peroxide ikanachokera kokha ku utoto wa reduced wochita ndi oxygen, kutseka unyolowo sikukanakhudza kwambiri. Koma kumakhudza, choncho unyolowo umatenga nawo mbali. Kuwunika zochita zotsutsa ndi zolimbikitsa oxidation kumaika mbali ziwirizi m'chidziwitso chokulirapo, ndipo kuwunika oxygen poyerekeza ndi ATP kumafotokoza chifukwa chake manambala a oxygen okha sangathetse nkhani yonena za mphamvu.

Kubwezeretsa ntchito si kukonza

Mawu omwe ayenera kusiya kugwiritsidwa ntchito ndi akuti “kodi methylene blue imakonza mitochondria.” Kukonza kungatanthauze kusintha kokhalitsa: magulu a unyolowo omangidwanso, kugwirizana kwa kupuma ndi kupanga ATP komwe kumapitirira mankhwalawo atachoka, kubwerera kwa ntchito m'maselo athunthu, kenako kugwira ntchito bwino m'chamoyo chamoyo. Palibe mwa mapepala atatuwa lomwe limasonyeza zimenezo. Amasonyeza kusintha njira mwachangu mu tizigawo ta selo tomwe tangochotsedwa kumene, pamene njira yatsekedwa mwadala ndi mankhwala, kupanga ATP kuli m'magawo ochepa a mulingo wabwinobwino, ndipo nthawi zina kugwirizana kwa kupuma ndi kupanga ATP kumakhala kodabwitsa kwambiri. Rotenone ndi antimycin ndi zida za mu labotale, osati zitsanzo zoimira kukalamba kapena matenda a mitochondria mwa anthu, ndipo palibe pano chomwe chimatanthauza mphamvu zambiri kwa munthu wathanzi. Imeneyo ndi nkhani ina ya umboni yokhala ndi mabuku ake a kafukufuku wa anthu.

Malire atatu amathandiza kuti mfundo iliyonse yokhudza biohacking ikhale yoona. Choyamba, kuchuluka kwa mankhwala m'madzi oyezera si mlingo wopatsidwa: 0.1 mpaka 2 micromolar mozungulira tizigawo ta selo tochotsedwa sikunena chilichonse za kuchuluka komwe munthu angamwe kuti mankhwala aulere afike pa kuchuluka kumeneko pa mitochondrion ya mu ubongo wa munthu, makamaka popeza utotowo umasonkhana kapena kumasulidwa malinga ndi voteji ya nembanembayo. Chachiwiri, oxygen si mphamvu: utotowo ungawonjezere kugwiritsa ntchito oxygen kudzera m'kuzungulira kwa maelekitironi ndi kutuluka kwawo m'njira yosafunika, komanso kudzera m'phosphorylation yopindulitsa, choncho kuyeza kupuma popanda kuyezanso ATP ndi voteji sikungavomereze zonena za mphamvu. Chachitatu, phindu ndi mtengo wake ziyenera kutchulidwa m'chiganizo chimodzi: mawu aliwonse onena za voteji kapena ATP yobwezeretsedwa osatchula kuwonjezeka kwa peroxide komwe kwachitika nthawi yomweyo amafotokoza kuyeserako molakwika. Mfundo zina zoyenera kuganiziridwa ndi kulinganika kwa NAD redox, maonekedwe a kuyankha malinga ndi mlingo ndi chitsogozo cha kuyesa.

Mfundo imodzi yothandiza kuikumbukira: kunyamula maelekitironi kumeneku ndi khalidwe la molekyulu yeniyeniyo. Mamolekyulu achibale omwe achotsedwa magulu a methyl, monga Azure B, nawonso amagwira ntchito za redox, koma amachita mosiyana, choncho kuyesa madzi a mankhwala omwe ena ake awonongeka kale ndi kosiyana ndi kuyesa utoto wosasinthika. Ichi ndi chifukwa chimodzi cholimbikira kuti pakhale miyezo yonse ya buku lovomerezeka la mankhwala ndi chikalata chofalitsidwa cha gulu lopangidwa, m'malo mongoyezera zitsulo zokha. Zochita za makemikolo zoyendetsedwa ndi kuwala zimawonjezera chenjezo lina lofanana pakugwiritsa ntchito mankhwalawa mowunikira, lomwe lafotokozedwa pa kuphatikiza ndi kuwala kofiira.

Mfundo yochepayi imakhalabe yoona ngakhale zonsezi zitaganiziridwa, ndipo ndi yoyenera kuisunga: methylene blue ingakhale mthenga wonyamula maelekitironi malinga ndi mmene zinthu zilili mu mitochondria zowonongeka zomwe zachotsedwa m'maselo, kusunga kuyenda pang'ono koma ndi mtengo wowonjezera peroxide. Chilichonse chopitirira pamenepo, monga mphamvu, moyo wautali, kapena kukonza, chikudikira umboni womwe kafukufukuyu sanapangidwe kuti upereke.