Methylene blue nenitric oxide: NOS, cGMP, uye kutumirana masaini mutsinga

Mubvunzo unonzwika seuri nyore: methylene blue inodzivisa nitric oxide here? Murabhoritari, mhinduro inopatsanuka kuita mibvunzo mina yakasiyana, uye umwe neumwe une muyedzo wakasiyana. Kuvhiringidza mibvunzo iyi ndicho chikonzero chikuru chinoita kuti zvinyorwa pamusoro pemethylene blue nenitric oxide zvionekwe sezvine chivimbo asi zvisina kujeka.

Funga nezvesaini yenitric oxide secheni ine zvikamu zvina zvakabatana. Mashoko ari mugene anokopwa kuti pave nekugadzirwa kwenitric oxide synthase. Enzyme inoshandura arginine kuita nitric oxide. Gasi rinopararira richienda kwarakananga risati raita shanduko nekusangana nechimwe chinhu. Soluble guanylate cyclase inoriona yoita cyclic GMP. Pane zvakabudiswa zvinoratidza kuti methylene blue ine zvainoita pachikamu chimwe nechimwe, asi humbowo hunobva mumagadzirirwo emiedzo akasiyana, huwandu hwayo hwakasiyana, uye maisoform akasiyana. Mhedzisiro pachikamu chimwe hairatidzi kuti iri kushandawo pane chimwe.

Cheni inowanzozivikanwa, yakatsanangurwa zviri nyore

Mumadziro etsinga yeropa, zvinowanzoitika zvinotevedzana seizvi. Chinhu chinokurudzira chinoshanda pasero reendothelium. Endothelial nitric oxide synthase, inonzi eNOS kana NOS3, inogadzira nitric oxide shoma kwenguva pfupi. Gasi rinopararira richipinda musero retsandanyama yakatsetseka riri pedyo, robatana neheme ine iron yakaderedzwa yesoluble guanylate cyclase, uye rokurumidzisa kushandurwa kweGTP kuita cyclic GMP. Cyclic GMP inomutsa protein kinase G, mabatirwo ecalcium anoshanduka, uye tsandanyama inoregedzeka.

Pane imwe cheni inofanira kuramba ichionekwa yakasiyana. Panguva yeutachiona kana masero asangana nemacytokine, masero anokopa mashoko ekugadzira imwe enzyme, inducible nitric oxide synthase kana iNOS, inonziwo NOS2. Inobudisa nitric oxide yakawanda uye kwenguva refu kupfuura eNOS. Isoform yechitatu, neuronal NOS kana nNOS, inoshanda mune mamwe mamiriro akasiyana. Kana bepa richiti nitric oxide synthase risingadudzi isoform yacho, mira utarise kuti ndeipi yakayerwa. Tsananguro yokufambisa kunoita methylene blue maelectron inoburitsa pfungwa imwe chete pamusoro pezvinonangwa neredox kazhinji: mufambisi haana chaanoita ari oga, uye chinhu chaanoshanda nacho kumucheto umwe neumwe ndicho chinosarudza mhedzisiro.

Cyclic GMP yakayerwa chiratidzo chinosanganisa zviri kuitika zvakawanda, kwete muyedzo weenzyme wakananga. Huwandu hwayo hunoratidza inogadzirwa necyclase kuchibviswa inoputswa nemaphosphodiesterase, kunyanya PDE5 mutsandanyama yakatsetseka yetsinga. Saka kuderera kwecyclic GMP kunoreva kuti saini shoma yasvika kuchinhu chinoiona. Hakuratidzi pachako kuti ndechipi chikamu chakundikana.

Chikamu 1: iNOS enzyme shoma inogadzirwa

Mumamacrophage emakonzo madiki akakurudzirwa nelipopolysaccharide yebhakitiriya, interferon gamma, kana zvose zviri zviviri, methylene blue yakadzivisa kuwedzera kwaitarisirwa kweprotein yeiNOS uye yakaderedza zvikuru messenger RNA yeiNOS mumiedzo yekukopwa kwemashoko emagenes mumamacrophage. Chinonyanya kupa ruzivo ndechezvinhu zvisina kushanduka. Kuputswa kwechinhu chinodzivisa chinonzi I kappa B alpha, kuwedzerwa kwephosphate paNF kappa B neSTAT1, uye kufamba kwezvinhu izvozvo zvichipinda munucleus zvakanyanya kuramba sezvazvaiva. Chakaderera kwaiva kubatana kwazvo neDNA, zvakasimbiswa nechromatin immunoprecipitation yakaratidza kuderera kwekuvapo kwazvo papromoter yeiNOS.

Izvozvo zvinokonzerwa nekudzorwa kwegene kunoitika huwandu hweenzyme husati hwasarudzwa, kwete kuvharwa nokukurumidza kweenzyme iripo. Zviri mukutumirana masaini pakuzvimba, apo zvinhu zvinotungamirira kukopwa kwemashoko emagenes zvinofanira kubatana nepromoter protein isati yavapo. Hazvitauri zvakananga nezveeNOS inogara iripo mutsinga isina kukurudzirwa, uye zvinofanira kutsanangurwa maererano nemiganhu iyoyo.

Chikamu 2: NOS enzyme iripo inoshanda zvishoma

Ongororo yakaitwa pamakonzo makuru yakaedza rimwe danho. Makonzo makuru echirume aiva akasvinura akapiwa lipopolysaccharide kuburikidza netsinga, uye mushure memaawa matatu akapiwa mvura ine munyu kana methylene blue pa3 mg per kg kuburikidza netsinga. Kushanda kwenitric oxide synthase mumedial basal hypothalamus kwakayerwa kuburikidza nekushandurwa kwearginine yakaiswa chiratidzo kuita citrulline muongororo yekushanda kweNOS muhypothalamus. Kushushikana pamwe chete nemvura ine munyu chete kwakamutsa kumwe kushanda kusingatsamiri pacalcium, uye lipopolysaccharide yakakuwedzera kusvika kunenge kakapetwa kagumi pachiyero cheboka rekuenzanisa. Methylene blue yakabvisa zvose zviri zviviri zvakanga zvamutswa. Kushanda kunogara kuripo kunotsamira pacalcium hakuna kushanduka munyama iyoyo pahuwandu hwacho uye panguva iyoyo.

Verenga mhedzisiro iyoyo sezvayakanyorwa chaizvo: kushanda kwakayerwa kwerudzi rweiNOS kwakaderedzwa mumamiriro iwayo. Haisi kuyerwa kwekuratidzwa kwegene, uye haisi humbowo hwekuti methylene blue inogara ichisarudza iNOS kupfuura eNOS kana nNOS. Ongororo dzeenzyme yakacheneswa dzinowedzera kukosha kwekungwarira uku. Mumiedzo yakananga yenitric oxide synthase yakacheneswa, methylene blue yakavhara zvachose kuumbwa kwecitrulline pa30 micromolar, nehuwandu hunodzivisa kushanda nehafu hwaiva pedyo ne5.3 micromolar pasina superoxide dismutase uye 9.2 micromolar iinayo, sezvakataurwa mukuenzaniswa kweNOS yakacheneswa neguanylate cyclase. Imwe ongororo yenNOS yakacheneswa yakaratidza kuti oxidation yeheme yakabatana neenzyme yaibatanidzwa. Chidzidzo chinoshanda ndechekuti kudziviswa kweNOS zvakananga kunoitika zvechokwadi mumasisitimu akaparadzaniswa, asi mhedzisiro yehypothalamus haigoni kutambanudzwa kuita chirevo chakazara chekuti methylene blue haikanganisi NOS yose inogara iripo munzvimbo dzose.

Chikamu 3: nitric oxide inorasika isati yasvika

Kuratidzwa kwakajeka zvikuru kwekuti chimwe chinhu chisiri kuvharwa kweenzyme chinogona kutsanangura kuderera kwecyclic GMP kunobva kumasero etsandanyama yakatsetseka yetsinga dzemapapu etsuro akarerwa murabhoritari. Magadzirirwo emuyedzo akapfuura NOS inogadzirwa nemasero pachawo zvachose nekuwedzera zvinhu zvinobudisa nitric oxide zvakananga, ndokubvunza kana methylene blue yaizoramba ichideredza kupindura kwecyclic GMP muongororo yesuperoxide mutsandanyama yakatsetseka yetsinga.

Zvinhu zvitatu zvinobudisa nitric oxide zvakaedzwa kubva pa1 kusvika pa100 micromolar: S nitroso L cysteine, sodium nitroprusside, uye chinhu chakafanana nazvo asi chakabviswa boka reamine uye chakagadzikana, chinonzi MPANO. Zvose zvakawedzera cyclic GMP kubva pakapetwa kanenge 1.5 kusvika pakapetwa ka12. Methylene blue pa10 micromolar yakadzivisa zviviri zvekutanga ne51 kusvika ku100 muzana uye yakasiya kupindura kweMPANO kusina kushanduka. Dai methylene blue yaingovhara nzvimbo yecyclase inoita shanduko yemakemikari, kusarudza zvinhu zvinobudisa nitric oxide kwakadaro kwaizonetsa kutsanangura, nokuti zvose zviri zvitatu zvinopedzisira zvasvika paenzyme imwe chete iri pamberi munzira yacho.

Miedzo mitatu yekuenzanisa yakazoratidza kuti nzira yacho yaibatanidza superoxide. Kutanga, kugadzira superoxide kunze kwemasero nexanthine pamwe chete nexanthine oxidase kwakabudisa chimiro chimwe chete: CYSNO nenitroprusside zvakavharwa, MPANO isina kukanganiswa. Chechipiri, methylene blue yakaita kuti masero etsandanyama yakatsetseka abudise superoxide zvichienderana nehuwandu hwayo uye nenguva, zvakayerwa kuburikidza nekuderedzwa kwecytochrome c. Chechitatu, superoxide dismutase yakadzivisa zvachose kuvharwa uku asi catalase isina kudaro, zvichinongedzera kusuperoxide kwete hydrogen peroxide sechinhu chaikonzera izvi zvakananga mumagadzirirwo iwayo.

Rudzi rwemuyedzo rwakabatanidza kurera masero emhando dzakasiyana pamwe chete runofanira kutsanangurwa nokuti runoratidza nzvimbo yakaitikira zvacho. Masero eendothelium aiva pazvimagirazi zvekufukidza akarerwa kwenguva pfupi pamwe chete nemasero etsandanyama yakatsetseka, zvokuti chinhu chaibudiswa neendothelium pasina kumutswa chakawedzera cyclic GMP yemutsandanyama yakatsetseka zvakapetwa katatu kusvika kasere. Kutanga kubata rimwe nerimwe remapoka emasero nemethylene blue kwakaderedza saini iyoyo, uye superoxide dismutase yakadzorera chimwe chikamu chemhedzisiro yakanga yaderedzwa nekutanga kubatwa kweendothelium. Nemamwe mashoko, methylene blue haifaniri kushanda mukati mesero retsandanyama yakatsetseka rinoona saini. Inogona kuderedza saini nekubvisa mutumwa paanenge achifamba. Gwaro rekuverenga tsvakurudzo yemethylene blue rinotora izvi semutemo unoshanda kazhinji: doma nzvimbo yakaitwa shanduko usati wadoma chinangwa chemamorekuru.

Kudana izvi kuti kudziviswa kweNOS kunenge kusiri iko. Hapana danho resynthase raidiwa kuti zvinhu zvinobudisa nitric oxide zvishande, asi saini yakangodzikira.

Chikamu 4: chinoona saini pachacho chinopindura zvishoma

Methylene blue ine zvainoita zvakananga pasoluble guanylate cyclase, asi huwandu hwayo hwakakosha. Mukuenzaniswa kweenzyme yakacheneswa kwataurwa pamusoro, cyclase yakamutswa nenitric oxide yaida methylene blue inenge 60 micromolar kuti idziviswe nehafu, zvichienzaniswa nekudziviswa kweNOS nehafu pedyo ne5 kusvika ku9 micromolar, uye kunyange 1 millimolar yakangoivhara zvishoma, kwete zvachose. Iwo musiyano wakapetwa kanenge katanhatu kusvika kagumi nerimwe. Saka muyedzo wemasero une huwandu hushoma huri muma micromolar unoderedza cyclic GMP haugoni kunzi kudziviswa kwecyclase kwakasarudzwa zvichibva panhamba yecyclic GMP chete.

Imwe ongororo yeredox yakazouya yakawedzera chimwe chikonzero chekungwarira: methylene blue yakadzima kushanda kwecyclase yakanga yamutswa nenitric oxide nenzira yakasiyana neoxidation yakajairika yeiron yeheme inokonzerwa nemhepo. Saka chinoona saini chinogona kukanganiswa, asi mashandiro emakemikari acho haasati anyatsojekeswa, uye musiyano wehuwandu hunodiwa unoreva kuti zvinokonzerwa nesuperoxide neNOS zvinofanira kutanga zvabviswa sezvikonzero kana huwandu hwacho huri hushoma muma micromolar. Apa ndipo panofanirawo kuitwa muyedzo wekudzora phosphodiesterase. Kana kuputswa kusina kuyerwa kana kuvharwa, kuderera kwekuungana kwecyclic GMP kunogona kuenderana nekugadzirwa kushoma, kuputswa kunokurumidza, kana zvose zviri zviviri.

Nei izvi zvisingagoni kupfupikiswa kuita chirevo chimwe pamusoro peBP

Chikamu chimwe nechimwe chakasimbiswa musisitimu yakasiyana. Kukopwa kwemashoko emagenes mumamacrophage hakusi kushanda kweenzyme muhypothalamus. Kushanda muhypothalamus mushure melipopolysaccharide hakusi mamiriro eendothelium isina kukurudzirwa. Chinhu chinobudiswa neendothelium pasina kumutswa kana masero akarerwa pamwe chete hachisi kuwedzera kwetsinga kunomutswa neagonist. Masero etsinga dzemapapu akarerwa murabhoritari ane methylene blue pa10 micromolar haasi kufamba kweropa mumunhu. Kuzvibatanidza kuita chirevo chakadai sekuti methylene blue inokwidza BP nekuvharira nitric oxide kunorasa misiyano chaiyo iyo miedzo yekuenzanisa yakagadzirirwa kuchengetedza.

Kuongorora zvinhu zvitatu kunoita kuti tsananguro irambe yakatendeseka. Kutanga, doma isoform: eNOS pamamiriro etsinga dzisina kukurudzirwa, iNOS pakubudiswa kwakawanda kunomutswa nekuzvimba, nNOS mumamiriro emasero etsinga dzinotakura mashoko uye mimwe miedzo yeenzyme yakacheneswa. Chechipiri, doma kuti saini yenitric oxide yakaunzwa sei: kugadzirwa mukati memasero, kubudiswa neendothelium pasina kumutswa, kana chinhu chakatarwa chinobudisa nitric oxide chakadai seCYSNO, nitroprusside, kana MPANO, nokuti chinhu chakagadzikana hachina kukanganiswa nemethylene blue asi zviya zvisina kugadzikana zvakakanganiswa. Chechitatu, doma huwandu uye chakadzorera kushanda: kudzorerwa nesuperoxide dismutase kunoratidza kuti superoxide yaibatanidzwa, kutadza kwecatalase kunopikisa hydrogen peroxide sechikonzero mumagadzirirwo iwayo, kutevedzera kunoitwa nexanthine oxidase kunoratidza kuti izvi zvakakwana kubudisa mhedzisiro yacho, uye miedzo yeenzyme yakacheneswa inoparadzanisa zvinokonzerwa nekushandura makemikari zvakananga nezvinorasika saini ichiri kufamba.

Pfupiso yacho ndeiyi. Zviri nani kutora methylene blue sechinhu chinokanganisa nzvimbo dzakawanda pakutumirana masaini kubva kunitric oxide kuenda kucyclic GMP pane kuitora semuvhariri wecyclase akasarudzika. Zvichienderana nemagadzirirwo emuyedzo uye huwandu hwayo, inogona kuderedza kubatana kwezvinhu zvinotungamirira kukopwa kwemashoko egene reiNOS, kudzivisa maenzyme eNOS zvakananga, kuderedza kuwanikwa kwenitric oxide kuburikidza nekugadzirwa kwesuperoxide, uye kukanganisa cyclase yakamutswa nenitric oxide. Basa rekutumirana masaini kwecyclic GMP muongororo dzetsinga uye kuenzaniswa kwezvinhu zvinobudisa nitric oxide zvinoshandiswa mumiedzo yemasero chimwe nechimwe chinotsanangura rimwe remapazi iwayo zvakadzama, uye nzira dzinoshandiswa kuparadzanisa zvinokonzerwa neredox nezvinokonzerwa pakushanda kwemaenzyme dzinotsanangura nei superoxide dismutase pamwe necatalase pamwe nemuyedzo wekuenzanisa unoshandisa chinhu chakagadzikana chinobudisa nitric oxide zvichiwanzowanikwa muzvinyorwa izvi.

Huwandu hushoma hwecyclic GMP mushure memethylene blue hunoratidza cheni yakadambuka. Miedzo yekuenzanisa chete ndiyo inotaura kuti ndechipi chikamu chakadambuka.