Methylene blue, Nrf2, uye kuumbwa kwemitochondria mitsva: zvakayerwa nezvichiri kufungidzirwa

Mutsara mumwe unowanzoonekwa muzvinyorwa pamusoro pemethylene blue: inomutsa Nrf2 uye inovaka mitochondria mitsva. Mutsara uyu unoita sokunge unotaura chinhu chimwe chete, asi une pfungwa mbiri dzakasiyana, dzakadzidzwa mumasisitimu akasiyana, dzine marudzi akasiyana ehumbowo. Yokutanga inzira yokutumirana masaini, chinonzwa shanduko chomutsa majini okudzivirira. Yechipiri chirongwa chokuvaka, sero richigadzira zvimwe zvikamu zvemitochondria. Saini inogona kutanga kuvaka, asi kuratidza kuti saini yatumirwa hakuna kufanana nokuratidza kuti chivako chavakwa, uye kuratidza zvinhu zvakawanda zvokuvakisa hakuna kufanana nokuratidza kuti simba riri kugoverwa zviri nani. Nyaya ino inochengeta zvinhu zvitatu izvi zvakaparadzana.

Pfungwa yeNrf2: chinonzwa shanduko, kwete nhoo

Nrf2 (nuclear factor erythroid 2-related factor 2) ipuroteni inodzora kunyorwa kwemajini, ine basa rokumutsa majini. Kana pasina zvinokanganisa sero, inobatwa mucytoplasm nepuroteni inonzwa shanduko inonzi Keap1, iyo inoiendesa kumidziyo yesero yokurasa zvinhu kuti irambe ichiparadzwa. Kana zvinhu zvinokonzera oxidation kana maelectrophile zvachinja macysteine anokwanisa kuita makemikari paKeap1, kuraswa uku kunomira. Nrf2 ichangogadzirwa inoungana, yopinda munucleus, yobatana nemapuroteni madiki eMaf, yobva yanamatira panhevedzano dzeDNA dzinonzi antioxidant response elements. Kunamatira ikoko kunomutsa boka remajini okudzivirira.

Zviratidzo zvinowanzoshandiswa kuyera nzira iyi zvakanyatsotsanangurwa. NQO1 (NAD(P)H quinone dehydrogenase 1) neTrxR1 (thioredoxin reductase 1) zviri pakati pemajini anotarisirwa neNrf2 anovimbika zvikuru. HO-1 (heme oxygenase-1) inogona kumutswa zvikuru asi inodzorwawo nezvimwe zvinhu. Maenzyme akaita seSOD necatalase chikamu chemidziyo inodzivirira oxidation, asi huwandu hwawo hunodzorwa nemapuroteni akawanda anodzora kunyorwa kwemajini, saka kuwedzera kweSOD hakuratidzi kwoga kuti Nrf2 ndiyo yakakonzera izvi. Humbowo huri pedyo zvikuru nechiitiko chokutumirwa kwesaini ndiko kuungana kweNrf2 munucleus, kumutswa kwechiratidzi cheARE, kana kumutswa kwemajini anozivikanwa kuti anotarisirwa neNrf2 kunonyangarika kana Nrf2 yabviswa. Zvimwe zvose, kuderera kwezvinokonzera oxidation, kurarama zviri nani pakushushikana kwesero, kuwanda kweglutathione, migumisiro inotevera inogona kukonzerwa nenzira dzakawanda.

Mugumisiro wokutanga wemethylene blue munyaya iyi uri pedyo nedivi rehumbowo rakanangana nenzira iyi. Mafibroblast akaiswa mumethylene blue iri pahuwandu hwenanomolar akaratidza kunonoka kusvika pakuchembera kwesero, pamwe chete nekuwanda kwecomplex IV, kushandiswa kweoxygen kwakawanda, uye kumutswa kwemaenzyme echikamu-2 TrxR1 neNQO1, sezvakataurwa muongororo yakatanga kubatanidza methylene blue nekuvandudzwa kwemitochondria uye kunonoka kuchembera kwemasero. TrxR1 neNQO1 ndiwo majini anozivikanwa kuti anotarisirwa neNrf2 ayo muverengi anongwarira anoda kuona. Chisina kuratidzwa neongororo iyi chiitiko chokutumirwa kwesaini pachacho: hapana kuyerwa kweNrf2 munucleus, hapana chiratidzi cheARE, hapana kuenzanisa nemasero akabviswa Nrf2. Saka kuverenga kwakarurama ndekwekuti methylene blue inomutsa majini anozivikanwa kuti anotarisirwa neNrf2 musisitimu iyi, izvo zvinoenderana nokumutswa kweNrf2 asi hazviratidzi Nrf2 yoga sechikonzero. Kumutswa kwejini rinotarisirwa idanho rimwe rinotevera chinhu chinoribatidza.

Pfungwa yokuumbwa kwemitochondria mitsva: chirongwa chine nhanho nhatu dzokuongorora

Kuumbwa kwemitochondria mitsva ndicho chirongwa chinoita kuti sero riwedzere kugona kwemitochondria yaro. Kutevedzana kwacho kunotsanangurwa mumabhuku kunopfuura nenhanho nhatu dzokuongorora. Kutanga, puroteni inobatsira kumutsa kunyorwa kwemajini, PGC-1alpha, inotanga chirongwa chacho. Chechipiri, mapuroteni emunucleus anodzora kunyorwa kwemajini (chinovhiringidza ndechokuti imwe inonzi NRF1, nuclear respiratory factor 1, iri puroteni yakasiyana neNrf2) anokurudzira kushanda kwemajini emitochondria, kusanganisira TFAM, iyo inochengetedza uye inokopa DNA yemitochondria. Chechitatu, sero rinopedzisira rava nemakopi akawanda egenome yemitochondria uye nemidziyo yakawanda yokufema kwemasero, zvinoyerwa sehuwandu hwemakopi emtDNA, kushanda kwecitrate synthase, kana kuwanda nekushanda kwezvikwata zvemapuroteni zvokufema.

Nhanho imwe neimwe inopa humbowo husina kusimba kana hwakasimba kupfuura hwakapfuura. Kuwanda kweRNA inotakura mashoko ePGC-1alpha kana TFAM kunoratidza kuti chirongwa chakumbirwa, kwete kuti chapera. Kuwanda kwemtDNA kana kwepuroteni yecomplex IV kunoratidza kuwanda kwezvinhu, asi mitochondria dzakagadziridzwa dzinogona kuratidzawo saizvozvo. Kufema kuri nani, kushandiswa kweoxygen kwakawanda kana kushanda kwezvikwata zvemapuroteni kwakawedzera, kunoratidza kuti kushanda kwachinja, asi kushanda kunogona kuvandudzika pasina kana mitochondria itsva. Kutaura kwakasimba pamusoro pokuumbwa kwemitochondria mitsva kunobatanidza zvose zvitatu: chinodzora, huwandu hwezvikamu, uye kushanda.

Ongororo yakatevera yemafibroblast yakapfuura nepakawanda pakutevedzana uku. Mumasero evanhu eIMR90 akaiswa mu100 nanomolar yemethylene blue, reshiyo yeNAD kuNADH yakakwira kwenguva pfupi mukati me15 minutes ikadzokera paya payakatangira mukati mezuva rimwe. Saini iyoyo yenguva pfupi ine chokuita nesimba yakateverwa nekuwedzerwa kweboka rephosphate kwenguva pfupi paAMPK, puroteni inonzwa mamiriro esimba, ndokuzoteverwa nokumutswa kwePGC-1alpha neSURF1, puroteni inodiwa pakuunganidza complex IV inoshanda. Kushanda kwecomplex IV kwakawedzera kusvika panenge kaviri, zvinokonzera oxidation musero zvakaderera neinenge 28 percent, uye matelomere akapfupika zvishoma nezvishoma kupfuura kare. Kutevedzana kwacho kwose kunotsanangurwa muchinyorwa chinobatanidza kushanda kwemethylene blue muredox neAMPK uye nekuumbwa kwemitochondria mitsva.

Zvinhu zviviri zviri muchinyorwa ichocho zvinoda kucherechedzwa nokuti ndizvo zvinowanzosiyiwa. Kutanga, maitirwo ekumutswa ainyanya kukosha kupfuura kungovapo kwawo: methylene blue yakamutsa AMPK kwenguva pfupi, ukuwo mushonga wekuenzanisa AICAR wakaimutsa kwenguva refu, uye chinhu chaiimutsa kwenguva pfupi chakanonotsa kuchembera kwemasero zvakapetwa kanenge kasere. Saka kutaura kwokuti “methylene blue inomutsa AMPK” hakuna kukwana; chinokosha ndechokuti inoimutsa kwenguva pfupi, uye kuenzanisa uku kunopa pfungwa yokuti kumutswa kwenguva refu hakuiti basa rimwe chete. Chechipiri, methylene blue haina kuchinja kutenderera kwematanho esero, izvo zvinobvisa imwe tsananguro iri nyore zvikuru, yokuti masero aingopatsanuka zvishoma nezvishoma obva aita sokunge achiri madiki nokuda kwaizvozvo.

Bazi rembeva: maitiro, magenome, uye chinotanga shanduko chinoita sechinopesana nemaune

Basa rakaitwa pamasero akakuriswa rakawedzerwa kumhuka neboka raidzidza mbeva dzaiva pakati peupenyu hwadzo. Pashure pe60 days dzichipiwa methylene blue nemuromo, mbeva dzakapiwa iyi dzakachengetedza kwakawanda kwekutsungirira kwadzo pakuita basa remuviri uye maitiro okuongorora zvakadzipoteredza, uye dzakaratidza maitiro mashoma akafanana nekuzvidya mwoyo kupfuura mbeva dzakwegura dzisina kupiwa iyi. Munyama yeuropi, majini akanga aderedzwa kushanda nokuchembera, NRF1, TFAM, SOD2, uye chikamu checomplex IV chinonzi COX1, akadzorerwa achiswedera pahuwandu hunowanikwa mumbeva dzakura dzisati dzakwegura, uye huwandu hwemakopi emtDNA hwakawedzera. Migumisiro iyi, maitiro pamwe chete nechinodzora pamwe chete nekuwanda kwegenome, yakataurwa muongororo yembeva dziri pakati peupenyu pamusoro pemethylene blue uye kuumbwa kwemitochondria mitsva muuropi.

Chinhu chinofungidzirwa kutanga shanduko ichi ndicho chinonyanya kufadza, nokuti chinopesana nenyaya iri nyore yokuti methylene blue inodzivirira oxidation. Mbeva dzakapiwa iyi dzakaratidza kuwedzera kwekugadzirwa kwemhando dzeoxygen dzinokurumidza kuita makemikari mumitochondria yeuropi, zvikurukuru hydrogen peroxide. Fungidziro yevanyori ndeyokuti saini shoma yeperoxide ndiyo inomutsa nzira yeNrf2, iyo inobva yatungamirira chirongwa chokuumbwa kwemitochondria mitsva. Mumuenzaniso uyu methylene blue inokurudzira oxidation padanho rokutanga shanduko uye yoidzivirira padanho remugumisiro, maitiro anonzi nenyanzvi dzemishonga hormesis, apo kushushikana kudiki kunomutsa dziviriro yakakura kupfuura kushushikana kwacho. Kuti kuwedzera ikoko kweperoxide kunonyatsokonzera kumutswa kweNrf2 here hakuna kuyedzwa zvakananga; uku ndiko kubatana kunofungidzirwa, kwete kubatana kwakayerwa.

Imwe ongororo yakafambidzana neiyi yakaitwa neboka rimwe chete yakaongorora zvakasimba rutivi rwekuchengeteka rwefungidziro iyoyo. Mumitochondria yeuropi yakaparadzaniswa kubva kune zvimwe zvikamu, methylene blue yakawedzera kugadzirwa kwehydrogen peroxide pahuwandu hwakaderera kusvika pa50 nanomolar, asi huwandu hunoshandiswa pakurapa hahuna kukonzera kukuvara kunoonekwa kuDNA yemitochondria. Pa100 micromolar chete, iri kure zvikuru pamusoro pehuwandu hunoshandiswa pakurapa, ndipo pakaratidza kukuvara kwezvimwe zvikamu zvemtDNA zvakasarudzwa, uye kubatanidza 20 micromolar yedhayi nechiedza chitsvuku kwakagadzira singlet oxygen yakakuvadza mtDNA munzvimbo dzakawanda, zvinova zvinozivikanwa kuti dhayi iyi inoita painomutswa nechiedza, kwete kushanda kwayo semushonga murima. Kupiwa kwayo pamberi kwakatochengetedza mitochondria yechikamu chepazasi cheuropi hwepakati pakukuvara kwemtDNA kunokonzerwa nerotenone. Kuchinja kwemugumisiro zvichienderana nehuwandu uye bvunzo dzokuenzanisa kukuvara kweDNA zvinotsanangurwa muongororo yokugadzirwa kweperoxide nemethylene blue kusingakuvadzi zvinhu zvemajini.

Zvakayerwa nezvinofungidzirwa: kuverenga mufananidzo

Mufananidzo wenzira uri pazasi wakagadzirirwa nyaya ino kuti usimbise musiyano wanga uchitaurwa munyaya yacho. Mabhokisi ane miganhu isina kudamburwa ndiwo nhanho dzine humbowo hwakananga hwekuedza muzvinyorwa zvemethylene blue: kutenderera kweredox kunochinja NAD/NADH, kuwedzerwa kweboka rephosphate kwenguva pfupi paAMPK, kumutswa kwePGC-1alpha neSURF1, kuwedzera kwekushanda kwecomplex IV, kumutswa kweNQO1 neTrxR1, kudzorerwa kwekushanda kwemajini okuumbwa kwemitochondria mitsva uye huwandu hwemakopi emtDNA muuropi hwembeva, uye kuwedzera kweperoxide pasina kukuvara kwemtDNA pahuwandu hunoshandiswa pakurapa. Mabhokisi ane miganhu yakadamburwa ndiwo mabatanidzo anofungidzirwa: Keap1 ichinzwa saini yeperoxide, kuungana kweNrf2 munucleus kuchitungamirira majini okudzivirira, uye museve unoratidza kuti kumutswa kweNrf2 kunokonzera kutanga kwechirongwa chokuumbwa kwemitochondria mitsva. Nhanho idzodzo dzine miganhu yakadamburwa dzine musoro, dzinoenderana nezvakawanikwa, uye ibhayoroji inowanzoonekwa mumabhuku, asi muzvinyorwa izvi hadzina kuyerwa pachishandiswa bvunzo dzokuenzanisa (Nrf2 yemunucleus, zviratidzi zveARE, kubviswa kweNrf2) dzaizodziita mabhokisi ane miganhu isina kudamburwa.

Zvinhu zvitatu zvinovhiringidza zvinoonekwa kana mufananidzo wava pamberi pako. Kutanga, NRF1 neNrf2 mapuroteni akasiyana ane mazita anenge akafanana; ongororo yembeva yakayera nuclear respiratory factor 1, chinodzora kuumbwa kwemitochondria mitsva, kwete puroteni inodzora kunyorwa kwemajini anodzivirira oxidation, uye kusanganisa zviviri izvi kunoita kuti mufananidzo wose uite sokunge unongova nepfungwa imwe. Chechipiri, kushanda kwemajini ndiko kutanga kwenyaya, kwete kuguma kwayo; kudzorerwa kwemashoko eTFAM kana COX1 kunotsigira kuti kuumbwa kwemitochondria mitsva kwakumbirwa, ukuwo huwandu hwemakopi emtDNA uye kushanda kwecomplex IV zviri humbowo hwokuti chikumbiro chacho chazadziswa. Chechitatu, mugumisiro wokudzivirira oxidation hauratidzi kuti nzira yeNrf2 ndiyo yashandiswa; kuderera kwe28 percent kwezvinokonzera oxidation mumafibroblast uye kudzivirirwa parotenone mumbeva migumisiro chaiyo inotevera, asi nzira dzakawanda dzinosvika pairi.

Chii chingashandura bhokisi rine muganhu wakadamburwa kuva nerisina kudamburwa

Kuedza kusipo ndiko kunowanzoshandiswa. Kuyera puroteni yeNrf2 munucleus pashure pokunge masero aiswa mumethylene blue, panguva dzinoenderana nesaini yenguva pfupi yeAMPK, kwaizoedza kuona kana mapazi maviri aya achinyatsomutswa pamwe chete, sezvinotaurwa nefungidziro yokumutswa panguva imwe chete. Chiratidzi chinodzorwa neARE chaizosimbisa kushanda kwekunyorwa kwemajini panzvimbo pokungozvifungidzira kubva pamajini anotarisirwa. Kudzokorora zviyero zvokuumbwa kwemitochondria mitsva kana Nrf2 yaderedzwa kana kubviswa kwaizoedza zvakananga museve unoratidza chikonzero: kana kumutswa kwecomplex IV uye kuwanda kwemtDNA zvikaitika pasina Nrf2, kubatana kunofungidzirwa kunenge kusina kururama uye bazi reAMPK ndiro rinenge richikonzera mugumisiro roga. Kudivi remhuka, ongororo yokupa mushonga kwenguva refu ine nhanho nhatu dzimwe chete dzokuongorora, chinodzora, huwandu hwezvikamu, kushanda, yaizoedza kuona kana chirongwa chichiramba chichishanda kana kuti chichichinjika. Kusvikira ipapo, kumutswa panguva imwe chete kwenzira dzesimba nedzokudzivirira kunoramba kuri izvo zvakadanwa nevanyori vacho: chinhu chikuru chinogona kuva chinokosha, kwete mashandiro akasimbiswa.

Vaverengi vanobva paruzivo rwakafara rwemashandiro emishonga vangada mamwe mashoko akapoteredza nyaya iyi: kuti methylene blue inotamisa sei maelectron mumitochondria, zvimwe zvainoita kumitochondria nesimba remasero, nzvimbo yezvakawanikwa pamusoro pokuchembera kwemasero mukutsvakurudza kuchembera, kuti zvinotaurwa pamusoro pokuumbwa kwemitochondria mitsva zvinobatana sei nekugona kufunga, uye humbowo hwekuchengeteka hunoisa miganhu pakukurukurirana kwose kwehuwandu hunopiwa. Chimwe nechimwe cheizvi ipfungwa yakazvimirira ine humbowo hwayo, uye chimwe nechimwe chinoda kupatsanurwa kwezvakayerwa nezvinofungidzirwa sezvaitwa neNrf2 munyaya ino.