Chinyorwa
Methylene blue nekuchembera kwemasero: zvinoyerwa nemiedzo yefibroblast
Kusanzwisisa kunowanzoitika kwakadai. Pepa retsvakurudzo rinotaura kuti methylene blue yakawedzera kanopfuura 20 nhamba yenguva idzo masero efibroblast anorimwa murabhoritari akawedzera kaviri, saka muverengi anobva afunga kuti inobvisa masero achembera kana kurebesa upenyu hwevanhu nechiyero chakafanana. Mhedziso yacho inoita seinonzwisisika. Asi yakakanganisawo pazvinhu zviviri zvakasiyana.
Dambudziko guru riri pamanzwisisiro epfungwa, kwete panhamba. Kuwedzera kwekaviri kwenhamba yemasero, zviratidzo zvekuchembera kwemasero, uye zviyero zvetelomere chimwe nechimwe zvinopindura mubvunzo wakasiyana. Tsvakurudzo yemethylene blue pafibroblast inonyatsonzwisiswa sechidzidzo chekunonotsa kupinda mukuchembera kwerudzi rumwe rwemasero ari mundiro yekurimiramo, ichitsigirwa nezviyero zvemitochondria. Izvi zvakasiyana nekubvisa zvakanangwa masero anenge atochembera, uye zvakasiyanawo nekuchembera kwechisikwa chose. Tsananguro yakafara yemusiyano uyu chikamu chenzira iyo kutakurwa kwemaerekitironi mumitochondria kunoumba simba remasero uye nzira yekududzira zvinotaurwa nezvetsvakurudzo yemethylene blue.
Kuwedzera kwekaviri kwenhamba yemasero kunoverenga zvakaitika, kwete hudiki hwawo
Mizhinji yemigumisiro mikuru inobva kumasero eIMR90, mutsara wemasero efibroblast akajairika anobva mumapapu emwana wemunhu asati aberekwa, ane muganhu wekugona kupatsanuka. Muchidzidzo chekutanga chemethylene blue nenguva yekugona kupatsanuka kwemasero efibroblast eIMR90, masero airimwa aichinjirwa mumidziyo mitsva maererano nehurongwa, uye nhamba yakaunganidzwa yenguva dzaaiwedzera kaviri yainyorwa kusvikira amira kupatsanuka. Kushandisa methylene blue pamwero wenanomolar, kunyanya pa100 nM, kwakabvumira masero kuunganidza kanopfuura 20 dzimwe nguva dzekuwedzera kaviri asati amira.
Kuwedzera kwekaviri kwenhamba yemasero chikamu chekuverenga. Kunonyora kuti boka remasero rakawedzera kanenge kaviri kangani. Hakutauri zvakananga kana sero rimwe nerimwe rachembera, rakangozorora pakupatsanuka, rashanduka kuita sero rine basa rakati, kana rafa. Masero ari kurimwa anogona kuwedzera kaviri kakawanda nekuti anoramba achikwanisa kupatsanuka kwenguva yakareba, nekuti mashoma anomira kupatsanuka nekukurumidza pasi pekushushikana, kana nekuti mamiriro acho anosarudza boka diki remasero rinokwanisa kuramba richiwanda. Kuverenga kwacho chete hakugoni kusiyanisa nzira idzi.
Ndokusaka nhamba dzenguva dzekuwedzera kaviri dzichibatanidzwa nezviratidzo. Nhamba yacho inotaura kuti kuwanda kwemasero kwakaramba kuchiitika kwenguva yakareba sei. Zviratidzo zvinotaura mamiriro akapindwa nemasero akamira kupatsanuka kana akasara ari mapenyu.
Gwaro rezviyero zvekupedzisira: chinotaurwa nechiratidzo chimwe nechimwe
Kuchembera kwemasero anorimwa murabhoritari kunozivikanwa nezvimiro zvinobva pazviyero zvakati wandei zvakazvimirira. Hapana dhayi rimwe chete kana puroteni imwe chete inozviratidza zvechokwadi.
Kumira kukura pamwe nekurasikirwa nekugadzirwa kweDNA. Kumira kunoramba kuripo kunoongororwa nekudzikira kwekubatanidzwa kweBrdU kana EdU, chimiro chakati sandara, uye kutadza kukura zvakare kana masero adyarwa mumudziyo mutsva. Izvi zvinosiyanisa kumira kunokonzerwa nekuchembera kwemasero nekuzorora kwechinguvana pakupatsanuka.
Beta galactosidase ine chekuita nekuchembera kwemasero. SA-beta-gal inoratidza basa remalysosome nedhayi pedyo nepH 6.0. Masero efibroblast achembera anowanzobata dhayi sezvo huwandu hwemalysosome huchiwedzera. Haisi chiratidzo chekuchembera chete, saka inodudzirwa pamwe nezviratidzo zvekuwanda kwemasero nezvemamorekuru.
Nzira dzep16 nep21. p16 inoshanda kuburikidza neCDK4 neCDK6 uye nzira yeRB. p21 inowanzokurudzirwa kuburikidza nep53. Kuwedzera kwehuwandu hwadzo kunotsigira humbowo hwekumira kupatsanuka, asi hapana pamapuroteni aya ine chekuita nekuchembera kwemasero chete.
Mamiriro ekuburitsa zvinhu. Chimiro chekuburitsa zvinhu chine chekuita nekuchembera kwemasero, kana kuti SASP, chinoreva kuchinja kwekuburitswa kwemacytokine, machemokine, zvinhu zvinokurudzira kukura, nemaprotease. Miedzo yeRNA kana mapuroteni yezvinhu zveSASP inotsanangura kuti masero anotumira sei mashoko kumasero ari pedyo. Haiyeri kugona kupatsanuka zvakananga.
Zvinhu zvinokonzera oxidation nezviyero zvemitochondria. Mapepa emethylene blue anowedzera zviyero zvemitochondria: huwandu nebasa recomplex IV, kushandiswa kweokisijeni, kugadzirwa kweheme, huwandu hwemembrane, musiyano wemagetsi pamembrane, ATP, uye zviratidzo zvezvinhu zvinokonzera oxidation. Izvi zvinotsanangura mamiriro ekushandurwa kwezvinhu nesimba anoenderana nekunonoka kumira kupatsanuka. Hazvitsananguri kuchembera kwemasero zviri zvoga, asi zvinoedza nzira inofungidzirwa kuti ndiyo inoshandiswa. Muenzaniso wekutenderera kweredox, umo NAD(P)H inopa methylene blue maerekitironi uye cytochrome c yoibvisa maerekitironi zvakare, unotsanangurwa muchidzidzo chimwe chete cheIMR90 chinobatanidza kutenderera kweredox nekudzikira kwekugadzirwa kwezvinhu zvinokonzera oxidation. Tsananguro ine chekuita nehuwandu hunoshandiswa nekupararira kwayo inobatwa muongororo yemashandiro emethylene blue mumuviri uye kusvika kwayo munyama dzemuviri.
Mwero wekupfupika kwetelomere. Chikamu chakakodzera chekuyera inhamba yemapeya emabhesi anorasika pakuwedzera kwekaviri kwega kwega mumavhiki akati wandei, kwete urefu hwacho panguva imwe chete. Kupfupika kunononoka kunoreva kurasikirwa neurefu hushoma pakupatsanuka kwega kwega. Hazvirevi kureba, uye kushushikana kunogona kumisa kupatsanuka kwemasero eIMR90 pasina shanduko huru yetelomere.
Kuenzanisa marongerwo matatu emiedzo yefibroblast
Chinhu chepakutanga chakagadzirirwa peji iri kuenzanisa kuri pazasi. Kunoisa rudzi rwemasero, kusangana nechinoshandiswa, zviyero, nemiganhu munzvimbo imwe chete.
| Chidzidzo | Masero nekusangana nechinoshandiswa | Zvakayerwa | Zvachisingagoni kuratidza |
|---|---|---|---|
| Atamna nevamwe, FASEB J 2008 | Masero efibroblast eIMR90 akajairika, methylene blue pamwero wenanomolar kwenguva refu, 100 nM semamiriro akajairwa | Nhamba yakaunganidzwa yenguva dzekuwedzera kaviri, huwandu hwecomplex IV, kushandiswa kweokisijeni kwakakwira ne37 kusvika 70 muzana, kugadzirwa kweheme, kudzivirirwa pakuchembera kwemasero nguva isati yakwana kunokonzerwa nehydrogen peroxide kana cadmium, maenzyme echikamu 2 mumasero eHepG2 | Hapana muedzo wekubviswa kwemasero achembera. Hapana chiyero chekureba kweupenyu hwemhuka kana vanhu. Nguva yekugona kupatsanuka pano inoreva kuwedzera kwekaviri kuri mundiro yekurimiramo. |
| Atamna nevamwe, Redox Biology 2015, zvinotsanangurwa muchidzidzo chakazotevera pamusoro penzira dzemashandiro eAMPK nekuumbwa kwecomplex IV | Masero efibroblast eIMR90, zvikurukuru 100 nM methylene blue, ichienzaniswa neAICAR sechinhu chinomutsa AMPK kwenguva refu | Shanduko nekufamba kwenguva yechiyero cheNAD kuNADH, kubatanidzwa kwemapoka ephosphate kuAMPK kwechinguvana, kukurudzirwa kwePGC-1alpha neSURF1, kukwira kwebasa recomplex IV neanopfuura 100 muzana, kudzikira kwezvinhu zvinokonzera oxidation ne28 muzana, kunonoka kwekupfupika kwetelomere, dzimwe nguva dzekuwedzera kaviri dzinenge 18 zvichienzaniswa nedzinenge 2 neAICAR, hapana shanduko pakutenderera kwekupatsanuka kwesero | Kuenzanisa neAICAR kunosiyana pahuwandu hunoshandiswa nekanenge 1000 uye pazvinhu zvinonyanya kunangwa, saka kunoedza nzira yekumutswa kwacho kwete simba rekushanda rinoenzanirana. Mugumisiro wetelomere une chekuita nemasero airamba achipatsanuka mumavhiki akati wandei. |
| Xiong nevamwe, Aging Cell 2016, zvinotaurwa muchidzidzo chefibroblast dzeprogeria pamusoro pekuvandudzwa kwemitochondria nenyukireyasi | Masero efibroblast eganda akatorwa zvakananga kuvanhu vane Hutchinson-Gilford progeria pamwe nemasero akajairika akafananidzwa nawo kuti zvienzaniswe, zvikurukuru 100 nM kwemavhiki 5 kusvika 12 | Chimiro chemitochondria chakaonekwa nemaikorosikopu efluorescence neemaerekitironi, kufamba kwadzo nekuona masero ari mapenyu, superoxide, musiyano wemagetsi pamembrane, ATP, PGC-1alpha, zviratidzo zvekuwanda nekuchembera kwemasero, pamwe nemapundu panyukireyasi, nzvimbo inowanikwa progerin, heterochromatin, uye kushanda kwemajini | Muenzaniso wekuchembera nguva isati yakwana une chekuita nechirwere chakati, kwete kuchembera kwakajairika. Kuvandudzwa kwechimiro nezviratidzo hakusi kubviswa kwemasero achembera. Hapana chiyero chekushanda mumhuka kana chekuchipatara. |
| Ongororo yaXue nevamwe, Cells 2021, yakapfupikiswa muongororo yemethylene blue sechinhu chinogona kurwisa kuchembera | Kubatanidzwa kwezvinyorwa pamusoro pekuchembera kweuropi, kuparara kwemasero etsinga, ganda, kupora kwemaronda, neprogeria | Inotsanangura methylene blue sechinhu chinokurudzira kutenderera kweredox chinogona kunzvenga zvimwe zvikamu zvekufamba kwemaerekitironi mucomplex I neIII, ichienzaniswa nedzidziso dzekuchembera dzinobva pamafree radical | Kubatanidzwa kwezvinyorwa kwetsananguro chete. Kunoronga pfungwa dzinofungidzirwa uye kunoratidza panoramba humbowo hwezvidzidzo zvemuvanhu husina mhedziso yakajeka. |
Maitiro anoonekwa mumitsara yose anokosha kupfuura mutsara mumwe chete. Methylene blue inoshandiswa muhuwandu hudiki inoramba ichibatanidzwa nenguva yakareba yekuwanda kwemasero, kuwedzera kwebasa recomplex IV, kudzikira kwezviratidzo zvezvinhu zvinokonzera oxidation, nekuvandudzwa kwechimiro. Marongerwo emiedzo anoramba achitevera masero nekufamba kwenguva: kutanga nemasero anopatsanuka, kushandisa chinhu panguva yekuwanda kwawo, uye kunyora kuti kupatsanuka kunoramba kuchiitika kwenguva yakareba sei. Marongerwo aya anogona kutsigira chirevo chekunonotsa. Haagoni kutsigira chirevo chekubvisa pasina marongerwo akasiyana.
Kunonotsa kuchembera kwemasero hakusi kubvisa masero achembera
Kunonotsa kunoreva kuti masero ari kupatsanuka anoramba achipatsanuka kwenguva yakareba asati amira. Kubvisa, kunowanzonzi kushanda kwesenolytic, kunoreva kuti masero anenge atochembera anourayiwa kana kuderedzwa zvakanangwa uku masero ari pedyo ari kuwanda achiramba ari mapenyu.
Zvinyorwa zvemethylene blue pafibroblast zvinoedza marongerwo ekutanga. Masero anotanga achiwanda, opiwa chinhu panguva yekuramba achichinjirwa mumidziyo mitsva, uye ozomira kupatsanuka pave paya. Naizvozvo zviratidzo zvakaita seSA-beta-gal, p16, p21, nezvinhu zveSASP zvinoonekwa pave paya kana muhuwandu hushoma pane imwe nhanho yekuchinjirwa mumudziyo mutsva. Maitiro aya anotarisirwa pose panosundirwa nguva yekutanga kuchembera mberi. Haaratidzi kuti chero sero rakatochembera rakabviswa.
Chirevo chekushanda kwesenolytic chingada marongerwo akasiyana: kutanga kusvitsa masero ari kurimwa pakuchembera kwakavimbiswa, vozoshandisa chinhu chacho, vozoratidza kurasika kwakanangwa kwemasero achembera zvichienzaniswa nemasero ari kuwanda anoshandiswa pakuenzanisa, pamwe nekuyera zvakananga kurarama nekufa kwemasero. Kudzikira kwechiratidzo chete hakuna kukwana nekuti kuburitswa kwacho kunogona kuchinja pasina kurasika kwemasero, uye masero akasara achiwanda anogona kuzadza ndiro yekurimiramo. Hapana pamanyuko mana ataurwa anoshandisa marongerwo aya ekuderedza masero zvakanangwa pamethylene blue, saka tsananguro yakarurama inoramba iri kunonotsa kana kurwisa kuchembera kwemasero murabhoritari, kwete kushanda kwesenolytic. Chikamu chemusiyano uyu chine chekuita nekushushikana kunokonzerwa neoxidation chinotsanangurwa zvakare munzira iyo masero anodzora nayo reactive oxygen species.
Mugumisiro wekutumirwa kwezviratidzo kwechinguvana unojekesa pfungwa iyi. Mubasa reIMR90 ra2015, methylene blue yakaita kuti chiyero cheNAD kuNADH chikwire kwenguva pfupi uye kuti mapoka ephosphate abatanidzwe kuAMPK kwenguva pfupi, zvichiteverwa nekukurudzirwa kwePGC-1alpha neSURF1, zvose zviri zviviri zvine chekuita nekuumbwa kwecomplex IV. Kumutswa kweAMPK kwenguva refu neAICAR kwakava nemigumisiro yakaderera zvikuru pakuwedzera dzimwe nguva dzekuwedzera kaviri.
Nguva yekugona kupatsanuka kwemasero haisi kureba kweupenyu hwevanhu
Nguva yekugona kupatsanuka kwesero inoverenga nguva dzekuwedzera kaviri kusvikira ramira, mundiro inodzorwa ine okisijeni, zvinovaka masero, uye nguva dzekuchinjirwa mumidziyo mitsva zvakatarwa. Kureba kweupenyu hwevanhu kunobatanidza makumi emakore ekukura, kudzivirirwa kwemuviri, tsinga dzeropa, nzvimbo dzinogara masero anokwanisa kubereka mamwe marudzi emasero, maitiro, nezvirwere. Chinhu chinogona kurebesa yekutanga chisingachinji yechipiri, uye zvakawanda zvinoita saizvozvo.
Masero efibroblast eprogeria ane shanduko yakati muLMNA inogadzira progerin, aine mitochondria dzakapatsanuka kuita zvidimbu uye dzisingafambi zvakanyanya, ATP yakaderera, manyukireyasi ane chimiro chisina kujairika, uye kurasika kweheterochromatin. Methylene blue pamwero wenanomolar yakavandudza zviyero zvemitochondria uye yakavandudza zvakanyanya zviyero zvenyukireyasi, kusanganisira kusunungurwa kweprogerin kubva pamembrane. Uku kuvandudzwa kunoshamisa padanho remasero. Kuchiri kugumira kumugumisiro wefibroblast, uye vanyori vakaisa kuedza mumhuka sebasa remangwana. Muenzaniso wechirwere wacho unokurukurwa zvakare mumethylene blue mumienzaniso yeprogeria nekuchembera nguva isati yakwana.
Izvi zvinopa rondedzero inoshanda yezvinofanira kubvunzwa pakuverenga.
- Bvunza kana kushandisa chinhu kwakatanga masero asati amira kupatsanuka kana mushure mekunge kumira kwacho kwasimbiswa. Kushandisa chinhu asati amira kunotsigira kunonotsa. Kushandisa chinhu mushure mekunge amira, pamwe nedata rekufa kwakanangwa, zvinodiwa kuratidza kubvisa.
- Bvunza kuti mutsetse wegirafu wekuwedzera kwekaviri kwemasero ekuenzanisa wakafamba sei. Pasina nhamba yakaunganidzwa yenguva dzekuwedzera kaviri, shanduko dzezviratidzo dzinoshaya chinodzibatanidza nezvakaitika.
- Kumbira mapoka maviri kana anopfuura ezviratidzo, akadai sechiratidzo chekuwanda kwemasero pamwe nep16 kana p21 pamwe neSA-beta-gal, pane dhayi rimwe chete.
- Bvunza kuti matelomere akatsanangurwa sei. Mwero pakuwedzera kwekaviri kwega kwega nekufamba kwenguva une zvinorehwa nawo. Chiyero chimwe chete cheurefu panguva imwe chete hachina.
- Bvunza kuti chii chakachengetwa chisina kuchinja. Kuwanda kwemasero pakuchinjirwa mumudziyo mutsva, okisijeni, kuchinjwa kwezvinovaka masero, uye methylene blue ichangobva kugadzirwa kana yagara kwenguva, zvose zvinoshandura kumira kupatsanuka kwefibroblast.
Kana zvichiverengwa nenzira iyi, zvinyorwa zvefibroblast zvinopa methylene blue basa rinoenderana asi rine muganhu: chinhu chinoshanda muredox muhuwandu hudiki, chinononotsa kuchembera kunoenderana nekupatsanuka kwemasero efibroblast emunhu anorimwa murabhoritari uku chichivandudza zviyero zvemitochondria, chine tsananguro yakasimba zvikuru yemashandiro ine chekuita nekutumirwa kwezviratidzo zvesimba kwechinguvana nekuumbwa kwecomplex IV, uye pasina kuratidzwa mumapepa aya kwekubviswa kwemasero achembera kana kurebeswa kweupenyu hwevanhu.