Methylene blue nemitochondria: kutakurwa kwemaerekitironi nehumbowo

Fungidzira sampuli mbiri dziri padivi nepadivi. Dzose dzine mitochondria dzeuropi dzakaiswa muchetura werotenone, kemikari inovhara Complex I yeketani yekufema. Mune imwe, muongorori anoshandisa pipette kuisa methylene blue pachiyero chemicromolar. Kushandiswa kweokisijeni kunodzoka zvishoma. Musiyano wemagetsi pamembrane unodzokawo zvishoma. Kugadzirwa kweATP kunotangazve pamwero mudiki. Mucuvette iri padivi, sensor yeperoxide inoratidza rumwe rutivi rwenyaya: kugadzirwa kwehydrogen peroxide kwawedzera kakawanda, pachiyero chose chakaedzwa, uye pasubstrate yese yakaedzwa.

Zviyero zvose zviri zviviri ndezvechokwadi. Methylene blue inotakura maerekitironi ichikwanisa kuita kuti kuyerera kwemagetsi kurambe kuchiitika ichipfuura chikamu chakakuvadzwa cheketani yekufema, asi kutakura ikoko kunouyawo nemutengo weperoxide. Kana ukachengeta chokwadi chacho chiri zviviri pamwe chete, zvinyorwa zvesainzi pamusoro pemashandiro emethylene blue mumitochondria zvinonzwisisika. Kana ukabata chimwe chete, zvinoita sechishamiso kana sengozi, zvichienderana nepepa rawakatanga kuverenga.

Kutakurwa kwemaerekitironi, kwakatsanangurwa zviri nyore

Ketani yekufema mutsara wekupfuudzana. Maerekitironi anopinda paComplex I (achibva kuNADH) kana paComplex II nemaenzayimu eflavin ane hukama nayo (achibva kusuccinate nezvimwe zvinopa maerekitironi zvakafanana), opfuura nepaComplex III achienda kumutakuri anofamba anonzi cytochrome c, ozopfuura nepaComplex IV achienda kuokisijeni. Kupfuudzana kumwe nokumwe kunopomba mapurotoni achiyambuka membrane yemukati, uye musiyano unovapo ndiwo unosunda kugadzirwa kweATP. Kana nzvimbo ipi zvayo ikavharwa, chikamu chemutsara chisati chasvika ipapo chinomira.

Methylene blue inopinda semumwe mutakuri anoshanda parutivi rwemutsara uyu. Chimiro chayo chine oxidation chinotora maerekitironi kubva kuvatakuri vari kumusoro kweketani, chova chimiro chisina ruvara chakaitwa reduction, chinonzi leucomethylene blue. Chimiro ichocho chinobva chapfuudza maerekitironi mberi neketani, chodzokera pakuva chebhuruu. Zvimiro zviviri izvi zvinotsanangurwa mutsananguro yezviviri zvinopindurana muredox, uye mafambiro acho ekupfuudzana anotsanangurwa mutsananguro yemashandiro acho zvachose. Mubvunzo usati wapindurwa ndewekuti mutakuri uyu anodzokera papi mumutsara: muhomwe inosungirira quinol yeComplex III, inonzi nzvimbo yeQo, kana kuti akananga kucytochrome c. Nzira yekubuda inonyanya kushandiswa inoenderana nemamiriro acho, sezvinotsanangurwa zvakadzama muongororo yekupfuura Complex III.

Zvakayerwa chaizvo nezvidzidzo zvitatu zvemakonzo

Zvidzidzo zvose zvitatu zvakashandisa mitochondria dzeuropi dzakaparadzaniswa, kureva maorganelles akabviswa musero akaiswa mubuffer yakatarwa. Hapana masero akazara, hapana mhuka dzakapihwa mushonga, uye hapana vanhu vakapinda muzvidzidzo izvi. Zviyero zveconcentration zvinoreva mvura yakakomberedza maorganelles, kwete chiyero chemushonga chinonwiwa nemunhu. Kana ukachengeta mamiriro aya mupfungwa, nhamba dzacho dzinotaura zvakajeka.

Chidzidzo chekutanga, chakaitwa mumitochondria dzeuropi hweguinea pig dzakaparadzaniswa, pa100 nanomolar kusvika pa2 micromolar, chakaratidza zvose zvakanakira nezvinodhura musampuli imwe chete. Mumitochondria dzine utano, dhayi yakawedzera kushandiswa kweokisijeni pakuzorora asi haina kuchinja kufema kunokurudzirwa neADP, saka kugadzirwa kweATP kwakabatana nekufema hakuna kuvandudzika. Pakavharwa Complex I, 2 micromolar yakadzorera kugadzirwa kweATP kubva pa46 kusvika pa93 nanomoles paminiti pamilligram, zvichienzaniswa nemwero wakajairika uri pedyo ne1109. Pakavharwa Complex III, chiyero chimwe chetecho chakadzorera ATP kubva panenge pa46 kusvika pa63, zvichienzaniswa nemwero wakajairika uri pedyo ne910. Musiyano wemagetsi nekutorwa kwecalcium zvakadzoka zvishoma. Kuvharwa kweComplex IV necyanide kwakanga kusingagoni kupfuura, zvichiratidza kuti maerekitironi anobuda panzvimbo inobatsira iri pamberi peComplex IV ichiri kushanda. Kudzorerwa kwose kwekushanda kwaiva kwechokwadi maererano nenhamba asi kuri kudiki: zvikamu zveATP yakajairika zvaiva muzvikamu kubva muzana zvine digit rimwe chete.

Chidzidzo chechipiri, chakaitwa mumitochondria dzecortex yeuropi hwembeva paanenge 1 micromolar, chakasimbisa kupfuura Complex I uye chakaratidza muganhu paComplex III. Rotenone yakaderedza kufema kubva panenge pa121 kusvika pasi pe2 nanomoles dzeokisijeni paminiti pamilligram, uye dhayi yakakusimudza kusvika panenge pa21, chinenge chikamu chimwe pazvitanhatu zvemwero wakajairika, ukuwo musiyano wemagetsi pamembrane wakadzoka unenge wakazara. Asi mushure mekuvharwa kweComplex III, kufema kana musiyano wemagetsi hazvina kudzoka pazviyero zve1 kusvika 5 micromolar, uye kuvharwa kwacho kwakadzvinyirira chiratidzo cheperoxide chaikonzerwa nedhayi. Vanyori vakakurudzira kuti dhayi yakaitwa reduction inopa maerekitironi ayo panzvimbo yeQo yeComplex III: kana nzvimbo iyoyo yakagarwa, mutakuri wacho haana pekuaendesa.

Chidzidzo chechitatu, chakaitwa mumitochondria dzeuropi hwembeva, makonzo neguinea pig, kazhinji pa2 micromolar, chakaedza zvakananga kusawirirana uku ndokuona kuti kuvharwa kweComplex III kunogona kupfuura. Myxothiazol, inovhara nzvimbo yeQo, payakaderedza kufema kwekutanga ne88 percent, dhayi yakadzorera kushandiswa kweokisijeni kusvika panenge pa62 percent yemwero usina kuvharwa. Mumuedzo wakananga, hafu ye micromolar yakaita reduction yecytochrome c yakawedzerwa kubva kunze kunyange Complex III yakanga yakavharwa, zvichitsigira nzira kubva kuNADH ichipfuura nemudhayi ichienda kucytochrome c yozopfuurira kuComplex IV. Asi kufema kwakadzorerwa kwakaita zvisina kujairika: ADP yakawedzerwa yakaderedza kushandiswa kweokisijeni pane kukukwidza, uye kuvhara kugadzirwa kweATP kwakakurudzira kufema. Musiyano wemagetsi pamembrane wakadzoka zvishoma chete. Kushandiswa kwakawanda kweokisijeni uku kubatanidzwa kwemaitiro kwakapinduka hakusi kugadzirwa kwesimba kwakajairika.

Nei marabhoritari asingawirirani, uye chii chinogadzirisa kusawirirana kwacho

Tsananguro yekuti mhando dzemhuka ndidzo dzinokonzera kusiyana haitsigirwi. Boka rechitatu rakadzokorora basa racho mumhando nhatu dzemakonzo rikawana kupfuura kweketani kwakafanana mune dzose, kusanganisira rudzi rwembeva rwakanga rwambotaurwa kuti kupfuura uku kwakanga kusipo. Zvinosiyana pachinzvimbo chazvo iconcentration (1 ichienzaniswa ne2 micromolar), zviyero zvezvinovhara ketani, mabuffer, nzira dzekuparadzanisa mitochondria, uye kupfuura zvose mamiriro esimba anenge achienzaniswa: kupfuura ketani kunonyatsooneka ADP isati yawedzerwa, panguva chaiyo panoonekwa mhinduro yeADP inopesana nezvaitarisirwa. Naizvozvo dhiyagiramu inoratidza chokwadi inofanira kuva nenzira mbiri dzekubuda dzinoenderana nemamiriro acho, imwe panzvimbo yeQo uye imwe pacytochrome c. Mutakuri anoendesa maerekitironi panzvimbo inobvumirwa nekemisitiri yemunzvimbo iyoyo.

Mutengo weperoxide

Rutivi rwemutengo runoramba rwakafanana sezvinoita rutivi rwekudzorerwa kwekushanda. Mubasa reguinea pig, dhayi yakawedzera kuburitswa kweperoxide mumamiriro ose akaedzwa: pakuzorora, pakugadzirwa kweATP, uye pakukanganiswa nemakemikari, pasubstrate dzose nhatu. Panguva yekugadzirwa kweATP, kuburitswa kwakakwira kubva panenge pa148 kusvika pamusoro pe1500 picomoles paminiti pamilligram pa1 micromolar, uye kunyange 100 nanomolar yakawedzera chiratidzo ichi kanopfuura kana. Dhayi yakanonotsawo kubviswa kweperoxide nekanenge 2.6, saka chiratidzo chacho chinoratidza kugadzirwa kuri kukurumidza pamwe nekubviswa kuri kunonoka. Kemisitiri yakakurudzirwa ndeyekuti dhayi yakaitwa reduction inoita reduction yeokisijeni zvakananga pasina superoxide inoonekwa, kunyange izvi zvichiri dudziro kwete danho rakaratidzwa kuti nderechokwadi.

Basa rembeva rinowedzera kunetsa kwenyaya nenzira inobatsira. Peroxide yakakwira zvakanyanya nechiyero, kubva pamwero wekutanga uri pedyo ne76 kusvika panenge pa2000 picomoles paminiti pamilligram pa2 micromolar, asi kuvharwa kweComplex III kwakadzvinyirira chikamu chaikonzerwa nedhayi. Dai peroxide yaingobva pakudyidzana kwedhayi yakaitwa reduction neokisijeni chete, kuvhara ketani kwaisafanira kuva nesimba rakakura kudaro. Asi kune simba, saka ketani yacho inobatanidzwawo. Ongororo yekudzivirira nekukurudzira oxidation inoisa mativi maviri aya mumamiriro akafara, uye ongororo yeokisijeni ichienzaniswa neATP inotsanangura nei nhamba dzeokisijeni dzoga dzisingambokwanisi kupedza mubvunzo pamusoro pesimba.

Kudzorerwa kwekushanda hakusi kugadziriswa

Mutsara unofanira kuregedzwa ndewekuti “methylene blue inogadzirisa mitochondria here.” Kugadziriswa kwaizoreva kururamiswa kunogara: macomplex akavakwazve, kubatanidzwa kwemaitiro kunoramba kuripo mushure mekunge mushonga wabva, kudzoka kwekushanda mumasero akazara, uyezve kushanda kwakavandudzika muchipenyu. Hapana pamapepa matatu aya panoratidza izvozvo. Anoratidza kuchinjwa kwenzira pakarepo mumaorganelles achangobva kuparadzaniswa, panguva yekuvharwa nemakemikari kwakagadzirwa murabhoritari, kugadzirwa kweATP kuri pazvikamu zvidiki zvemwero wakajairika uye dzimwe nguva kubatanidzwa kwemaitiro kusingawanzoitiki zvachose. Rotenone neantimycin maturusi emurabhoritari, kwete mamodheru ekukwegura kana echirwere chemitochondria muvanhu, uye hapana pano chinoreva simba rakawanda kumunhu ane utano. Izvozvo mubvunzo wehumbowo wakasiyana une zvinyorwa zvawo zvezvidzidzo zvevanhu.

Miganhu mitatu inoita kuti mhedziso ipi zvayo yebiohacking irambe yakatendeseka. Chekutanga, concentration iri mumvura yakakomberedza maorganelles haisi chiyero chemushonga: 0.1 kusvika 2 micromolar yakakomberedza maorganelles akaparadzaniswa haitauri kuti chiyero chipi chinonwiwa chinoita kuti concentration yemushonga wakasununguka yakadaro isvike pamitochondrion yeuropi hwemunhu, kunyanya sezvo dhayi ichiungana kana kuburitswa zvichienderana nemusiyano wemagetsi pamembrane pachawo. Chechipiri, okisijeni haisi simba: dhayi inogona kuwedzera kushandiswa kweokisijeni kuburikidza nekutenderera nekudonha kwemaerekitironi uyewo kuburikidza nephosphorylation inogadzira ATP, saka kuyera kufema pasina zviyero zveATP nemusiyano wemagetsi hakugoni kupa hwaro hwekutaura nezvesimba. Chechitatu, kubatsira nemutoro zvinofanira kutaurwa mumutsara mumwe chete: kutaura kupi zvako nezvekudzorerwa kwemusiyano wemagetsi kana kweATP kusingatauri kukwira kweperoxide kunoitika panguva imwe chete kunomiririra muyedzo zvisiri izvo. Zvimwe zvinofanira kufungwa nezvazvo zvinosanganisira kuenzana kweNAD muredox, chimiro chemhinduro inoenderana nechiyero uye gwaro remiedzo yekuyera.

Chimwe cherechedzo chinoshanda: kutakura uku hunhu hwemorekuru iroro chairo. Mamorekuru ane hukama naro akabviswa mapoka emethyl, akadai seAzure B, anoshandawo muredox asi ane maitiro akasiyana, saka mushonga wakanyungudutswa wakatoparara zvishoma unoita muyedzo wakasiyana newedhayi isina kuparara. Ichi ndicho chimwe chikonzero chekusimbirira pazvinodiwa zvose zvepharmacopeia negwaro rebatch rakaburitswa, pane ongororo yesimbi chete. Kemisitiri inokurudzirwa nechiedza inowedzera imwe yambiro pakushandiswa kune mwenje, inotsanangurwa pasi pekubatanidzwa nechiedza chitsvuku.

Pfungwa yakaganhurirwa iyi inoramba ichitsigirwa nezvose izvi uye yakakodzera kuchengetwa: methylene blue inogona kushanda semutakuri wemaerekitironi zvinoenderana nemamiriro acho mumitochondria dzakakuvadzwa uye dzakaparadzaniswa, ichichengetedza kuyerera zvishoma asi nemutengo weperoxide. Zvose zvinopfuura ipapo, simba, kurarama kwenguva refu, kugadziriswa, zvakamirira humbowo uhwo zvidzidzo izvi zvisina kumbogadzirirwa kupa.