Chinyorwa
Methylene blue neferroptosis: simbi, peroxidation yemalipidhi, uye kufa kwemasero
Ferroptosis kufa kwesero kunokonzerwa nekukuvara kweganda raro. Mafuta epolyunsaturated ari mumaganda emasero anoitwa oxidation munhevedzano yezvinoitika zvinotevedzana, ganda resero rinotadza kushanda, uye sero rinofa. Simbi inodiwa kuti maitiro aya emakemikari aitike. Mutsara iwoyo ndiwo pfungwa yacho yose, uye zvose zviri munyaya ino zvinoedza chimwe chikamu chayo.
Mubvunzo uri pano wakamanikana kupfuura izvozvo: methylene blue inoitei kunhevedzano iyoyo muzviedzo zvemasero, uye mhedzisiro yacho inoratidzei chaizvo? Mhinduro pfupi ndeyekuti muzvidzidzo zviviri zvemurabhoritari, methylene blue iri pananomolar yakaderedza peroxidation yemalipidhi emitochondria uye yakachengeta masero ari mapenyu pashure pezvinhu zvinotanga maitiro aiwanzokonzera ferroptosis. Mhinduro yakareba ndiyo inonyanya kukosha, nokuti chinongedzo chimwe nechimwe munhevedzano yacho chakaedzwa nekupindira kwakasiyana, uye kupindira kumwe nekumwe kune zvakwakaganhurirwa.
Ferroptosis mundima imwe chete
Phospholipid yeganda resero ine muswe wefatty acid yepolyunsaturated ine nzvimbo isina kusimba: maatomu ehydrogen ari nyore kubvutwa. Kana radical yabvuta rimwe rawo, okisijeni inobatana nelipidhi, radical yeperoxyl yelipidhi inoumbwa, uye radical iyoyo inobvuta kubva kulipidhi rinotevera. Maitiro acho anopararira achitevedza ganda resero kunze kwekunge pane chinoamisa.
Chinhu chikuru chinomisa izvi ienzayimu inonzi GPX4, inoshandisa glutathione kushandura maperoxide emalipidhi kuita maalcohol emalipidhi asingakuvadzi. Zvinhu zvinotanga maitiro aya murabhoritari zvinorwisa dziviriro iyi panzvimbo dzakatarwa. Erastin inovhara kupinzwa kwecystine kuburikidza nesystem xc-minus, zvichiita kuti sero rishaye cysteine yekugadzira glutathione. Buthionine sulfoximine inovhara kugadzirwa kweglutathione zvakananga. RSL3 inodzivisa GPX4 pachayo. Zvose zviri zvitatu zvinosiya maperoxide asina kubviswa, asi zvinozviita pamatanho akasiyana, saka mhedzisiro inowanikwa nechinhu chimwe chinotanga maitiro aya haingonzi inoshanda kune zvimwe zvose.
Ferroptosis inosiyana neapoptosis pazvinhu zvinoitsanangura. Hapana nhevedzano yekushanda kwemacaspase uye hapana zvidimbu zvemasero zvinonzi apoptotic bodies. Zvinhu zvinosunga simbi zvakadai sedeferoxamine nezvinhu zvinobata maradical zvakadai seferrostatin-1 zvinodzvinyirira ferroptosis. Ichi ndicho chiyero chinoshandiswa nevatsvakurudzi kuti vati kufa kwesero nde kweferroptosis kwete kungova necrosis. Kutarisa nzira dzakasiyana-siyana dzekufa kwemasero kunodzorwa kunobatsira kunzwisisa musiyano uyu, uye mashandisiro anoita masero simbi inosununguka zviri nyore anotsanangura kuti nei kusungwa kwesimbi kuri nzira inokosha kudaro yekudzora zviedzo.
Panopinda mitochondria, uye nei pakanga paine gakava
Ongororo dzekutanga dzakashuma kuti masero asina DNA yemitochondria achiri kufa neferroptosis, zvichiratidza kuti mitochondria dzingasave dzinodiwa. Ongororo dzakazotevera dzakawana kuti kuderedza mitochondria, kana kuvhara kufema kwemasero, kunogona kudzvinyirira ferroptosis inobva pakushomeka kwecysteine asi kwete inobva pakudziviswa kweGPX4 zvakananga. Mhedzisiro yose iyi inogona kuva yechokwadi kana basa remitochondria richitsamira pachinhu chatanga maitiro acho, rudzi rwesero, uye nzvimbo inoitika peroxidation inosarudza kuti sero richafa here.
Chidzidzo chinotsigira nyaya ino chakaongorora gakava iri nepurobhu inoshanda mune chimwe chikamu chesero chete. MitoCLox ipurobhu inoratidza peroxidation yemalipidhi, yakagadzirwa kuti inange mitochondria: inoshandura fluorescence painosangana nemaradical eperoxyl emalipidhi mukati memitochondria. Mumafibroblast akaiswa erastin, oxidation yepurobhu yemitochondria yakaonekwa mumasero akanga achiri mapenyu asati afa, uye yaisafanana mumasero ose. Nguva iyoyo inokosha. Peroxidation inotangira kufa inogona kuva chikonzero chacho. Peroxidation inoyerwa mumasero atofa chete inongova masaririra. Gwaro rinofambidzana neiri pamusoro pezviyedzo zveperoxidation yemalipidhi nezvinoratidzwa nepurobhu imwe neimwe rinotsanangura kuti nei nzvimbo yepurobhu ichisarudza zvinorehwa nechiratidzo chayo.
Maitiro mamwe chete akaonekwa mune imwe modhi yechipiri. Mafibroblast emurwere ane Leber hereditary optic neuropathy, ane shanduko yegeneti mucomplex I uye kushushikana kweoxidation kwakakwirira kubva pakutanga, akafa neferroptosis pakaderedzwa glutathione, asi mafibroblast aishandiswa sekuenzanisa akaramba ari mapenyu. Oxidation yepurobhu yemitochondria yakatangazve kuonekwa. Tsanangudzo dzakadzama dziri muchidzidzo chemafibroblast pamusoro peperoxidation yemalipidhi emitochondria muferroptosis.
Zvakachinjwa nemethylene blue muzviedzo izvi
Methylene blue chinhu chinotakura maerekitironi mumaitiro eredox. Chimiro chayo chakaitwa oxidation chinogamuchira maerekitironi kubva mumaitiro emakemikari ane chokuita neNADH pacomplex I, uye chimiro chayo chakaitwa reduction chinoapa kucytochrome c, iyo inoendesa maerekitironi kucomplex IV. Izvi zvinogadzira nzira yepadivi inodarika chimwe chikamu chenhevedzano panowanzobuda maerekitironi achienda kuokisijeni, achiumba superoxide. Vaverengi vanoda kunzwisisa kufambiswa uku kwemaerekitironi zvizere vanofanira kutanga nemafambisirwo anoitwa maerekitironi nemethylene blue kuburikidza nemitochondria.
Mumafibroblast emapapu akaiswa erastin, methylene blue iri pa250 nanomolar yakachengetedza kurarama kwemasero pachiyero chekupona kwawo, pamwe chete nemaantioxidant anonanga mitochondria, SkQ1 neMitoTEMPO. Zvinhu zvakashandiswa kudzora zviedzo ndizvo zvinotsanangura zvinogona kunzi zvakaratidzwa. Ferrostatin-1, alpha-tocopherol, nedeferoxamine zvakadzivirirawo masero, zvichisimbisa kuti kufa kwacho kwaiva kweferroptosis. Chinhu chakafanana neSkQ1 asi chisina boka reantioxidant hachina kudzivirira masero, zvichinongedzera kumaitiro emakemikari eredox kwete kungoungana kusina kunanga chimwe chinhu. Dziviriro yakaderera pamusoro peanenge 200 nanomolar eSkQ1, zvichienderana nemaitiro anozivikanwa emaantioxidant equinone ekukurudzira oxidation kana awanda. Methylene blue inewo maitiro aya maviri anoenderana nehuwandu hwayo: inodzivirira pamazinga akaderera enanomolar mumidziyo iyi yekurimira masero, asi inogona kugadzira hydrogen peroxide kuburikidza nekutenderera kweredox kana huwandu hwayo hwakwidzwa kana ikavhenekerwa nechiedza chakasimba. Kukosha kwehuwandu hwechinhu mumamiriro akadai kunotsanangurwa mumarudzi eokisijeni anokurumidza kupinda mumaitiro emakemikari ari mumitochondria.
Zviedzo zvemitochondria dzakaparadzaniswa nemasero zvakazopatsanura kwaibva ROS. Paiva nesimbi yeferrous, zvinhu zvinopa complex I maerekitironi pamwe nerotenone zvakakurudzira oxidation yepurobhu. Rotenone inovhara nzira mushure menzvimbo yeflavin, saka zvinhu zvinotakura maerekitironi zviri pamberi payo zvinoramba zviri muchimiro chakaitwa reduction uye zvinoburitsa maerekitironi kuokisijeni kubva kuflavin yakatarisana nematrix. Zvinhu zvinopa complex III maerekitironi pamwe neantimycin A zvakagadzira hydrogen peroxide yakawanda asi hazvina kukonzera peroxidation yemalipidhi. Chiratidzo cheROS yose chaiva chakafanana, asi nzvimbo yaiva yakasiyana uye mhedzisiro yakasiyana. SkQ1 nemethylene blue imwe neimwe yakadzvinyirira peroxidation yaikonzerwa necomplex I pasina kudzvinyirira kugadzirwa kwehydrogen peroxide pachako. Izvi zvinoisa basa razvo mushure mekuumbwa kweROS, padanho reperoxidation yemalipidhi.
Chikamu chekuwandisa simbi uye chiratidzo chinobva kulipofuscin
Chidzidzo chechipiri chakabvunza mubvunzo uyu chichitarisa divi resimbi. Masero etsandanyama yemwoyo eH9c2 akaiswa ferric ammonium citrate iri pasi pemillimolar akatanga kuva neROS yemitochondria, peroxidation yemalipidhi emitochondria, ndokuzofa neferroptosis, zvichiteverwa nekuungana kwechinhu chine ruvara chakafanana nelipofuscin mukati menguva ye24 hours. Lipofuscin ndiwo masaririra ane simbi emalipidhi nemapuroteni akaitwa oxidation, anobudisa fluorescence pachawo uye anoungana pakukwegura. Kukurumidza kuoneka kwawo pano kunobatanidza peroxidation inokurumidza kukonzerwa nesimbi nechiratidzo chinozivikanwa chekukwegura. Methylene blue iri pananomolar neSkQ1 imwe neimwe yakadzivirira peroxidation, kufa kwemasero, uye kuungana kwechinhu chine ruvara. Mitochondria dzemwoyo dzakaparadzaniswa nemasero dzakasimbisa maitiro aya: rotenone pamwe nesimbi yeferrous zvakakonzera peroxidation nechiratidzo chakafanana nechechinhu chine ruvara, uye zvinhu zvose zviri zviviri zvakazvivhara. Nhevedzano yose inoshumwa muchidzidzo chekuwandisa simbi mumasero etsandanyama yemwoyo eH9c2. Divi renyaya rine chokuita nechinhu chine ruvara rinofambirana nelipofuscin nezviratidzo zvekukwegura kwemasero.
Zvisingaratidzwi nezviyedzo zvekupona kwemasero
Chiyero chekurarama kwemasero chakadai seCellTiterBlue chinoratidza kuti masero mangani ari mumudziyo anoramba achiita mabasa emetabolism pashure pekuiswa chinhu chinotanga maitiro acho pamwe nechinhu chiri kuedzwa kuti chingadzivirira here. Ichi chiyedzo chemashandiro mune hurongwa hwakatarwa. Haisi mhedzisiro yekurapwa. Hapana chimwe chezviedzo izvi chakapa murwere mushonga, chakayera mhedzisiro ine chokuita nechirwere, kana chakaratidza huwandu hunogona kubatsira pasina kukuvadza mumuviri.
Pane miganhu mina inofanira kutaurwa zvakajeka. Chekutanga, mhando dzemasero dzakashandiswa ishoma: mafibroblast emapapu akachinjwa, mafibroblast ekutanga akatorwa paganda kusanganisira rimwe boka rine chirwere chemitochondria, uye boka remasero etsandanyama yemwoyo yegonzo, pamwe nemitochondria dzakaparadzaniswa nemasero emwoyo yegonzo. Kukurumidza kukuvadzwa kwaisiyana neboka remasero, uye boka reLHON rakasarudzwa nokuti rainyanya kukuvadzwa. Chechipiri, zvinotanga maitiro aya zvinhu zvakanyanyisa zvemurabhoritari: kuvharwa kwakazara kwekutakurwa kwezvinhu kana kugadzirwa kwazvo, uye huwandu hwesimbi husingambosvikwi netishu munguva pfupi. Chechitatu, huwandu hwechinhu mumvura hahushandurwi kuita huwandu hwekupa munhu nekungowanza. Nanomolar mumvura yekurimira masero haisi huwandu hwekupa munhu, uye kupararira kwemethylene blue, kushandurwa kwayo kuita chimiro chisina ruvara muropa nemumatishu, kuvhenekerwa nechiedza, uye musiyano wemagetsi pamativi eganda resero zvose zvinopindira pakati pemudziyo wekurimira masero nemunhu. Chechina, mapurobhu anoratidza zviri kuitika muchikamu chawo chete. Oxidation yeMitoCLox inoratidza kuti maradical emalipidhi emitochondria akaumbwa; hairatidzi kuti kuputika kweganda rekunze resero, uko kungangova chiitiko chokupedzisira, kwakadzivirirwa zvakananga pane kungodziviswa nemaitiro akaitika pamberi pacho.
Saka zvinogona kutaurwa nokutendeseka ndeizvi: mumamodhi aya, ROS inobva kucomplex I pamwe nesimbi zvinokonzera peroxidation yemalipidhi emitochondria inotangira ferroptosis, uye methylene blue iri pahuwandu hwakaderera inodimbura chikamu ichocho. Chero chinopfuura chikamu ichocho chinoda zviedzo zvakasiyana mumhuka uye zvinozotevera muvanhu.