Methylene blue nemasaini ekuzvimba: ma-inflammasome, IL-6, uye STAT3

Ngatitii marabhoritari matatu anoti chinhu chimwe chete chebhuruu chinoderedza kuzvimba. Imwe inoratidza IL-1beta shoma inobva mumacrophage. Imwe inoratidza enzyme ye-iNOS shoma mumhando imwe chete yemasero. Imwe inoratidza IL-6 shoma muropa uye kushanda kushoma kweSTAT3 muuropi. Zviri nyore kuda kubatanidza izvi zvitatu kuita chirevo chimwe. Kubatanidza ikoko kunovanza zvakawanikwa.

Mhedzisiro imwe neimwe ndeyenzira yakasiyana yekutumira masaini, ine chinokuchidzira chayo, danho rayo rekuunganidzwa, uye chiyero chayo. Methylene blue inobata nzira imwe neimwe panzvimbo yakasiyana. Kana izvi zvichivandudza mhedzisiro yeutano mubvunzo wakasiyana, unokurukurwa muongororo yedu yemethylene blue nekuzvimba. Peji rino rinongotarisa nzira dzacho.

Nzira 1: kuunganidzwa kwe-inflammasome necaspase-1

Inflammasome ihwaro hwemapuroteni hunovakwa pazvinenge zvichidiwa mukati mesero remyeloid. Zvikamu zvayo chinhu chinonzwa chinokuchidzira, chinobatanidza zvikamu chinonzi ASC, uye enzyme inoita basa rinonzi caspase-1. Kana zvikamu zvaungana, caspase-1 inocheka pro-IL-1beta nepro-IL-18 kuita zvimiro zvinobudiswa kunze kwesero, uye inogona kutanga pyroptosis, rudzi rwerufu rwesero runokonzera kuzvimba. Chinhu chinonzwa chinokuchidzira ndicho chinotsanangura mhando: NLRP3 inopindura kumakristaro nechepfu dzinogadzira maburi dzakadai senigericin neATP, NLRC4 inopindura kuflagellin yemabhakitiriya, AIM2 inopindura kuDNA ine tambo mbiri iri mucytosol, uye nzira isiri yechinyakare inopindura kulipopolysaccharide iri mukati mesero.

Kutanga kushanda kunoda masaini maviri. Saini yekutanga ndeyekugadzirira: ligand yereceptor yakaita seToll inokurudzira kunyorwa kwemagene eNLRP3 nepro-IL-1beta kunotsamira paNF-kB, kuitira kuti zvikamu zvivepo. Saini yechipiri ndeyekuunganidza: chimwe chinokuchidzira chinoita kuti ASC iungane kuita zvikwata zviduku, caspase-1 ichekwe kuita chidimbu chayo chep20, uye IL-1beta p17 yakura ibudiswe. Mhando dzeoxygen dzinokurumidza kupinda mumareaction dziri mumitochondria uye kutorwa kwemakristaro zvinozivikanwa kuti zvinobatsira pasaini yechipiri yeNLRP3.

Chidzidzo che2017 chemacrophage pamusoro pekudziviswa kwemhando dzakasiyana dzema-inflammasome chakaedza methylene blue pamasaini ose ari maviri. Mumacrophage emwongo wemapfupa emakonzo akanga agadzirirwa nelipopolysaccharide, nigericin, ATP, uye makristaro emonosodium urate chimwe nechimwe chakakurudzira kubudiswa kweIL-1beta necaspase-1 pamwe nekuumbwa kwezvikwata zviduku zveASC. Methylene blue pa20 kusvika 200 micromolar yakaderedza zvose zvitatu zvichienderana nehuwandu hwayo, isina kuuraya masero. Flagellin neSalmonella zveNLRC4, DNA ine tambo mbiri neListeria zveAIM2, uye lipopolysaccharide yakaiswa mukati memasero kana E. coli mhenyu zvenzira isiri yechinyakare zvakaratidza maitiro mamwe chete. Chinhu chacho choga hachina kukurudzira kubudiswa, uye mapureti ekuenzanisa haana kuratidza kudziviswa kwemabhakitiriya, saka mhedzisiro yacho iri pamhinduro yemuviri uri kutakura utachiona.

Pamadanho anotangira aya, zvinhu zvitatu zvakayerwa. Superoxide yemumitochondria yakanga yakurudzirwa nerotenone yakaderera nemethylene blue, uye kubudiswa kweIL-1beta kwakanga kwakurudzirwa nerotenone kwakadererawo. Kutorwa kwetubhora tunopenya kwakaderera, zvichienderana nekudzikira kwephagocytosis kusati kwanzwikwa makristaro. Chiyereso chinotungamirirwa nepromoter yeNLRP3 ine nzvimbo mbiri dzeNF-kB chakaratidza kushanda kwakaderera nemethylene blue, asi promoter yakanga yapfupikiswa isina nzvimbo idzodzo haina kupindura. Muchubhu isina masero, methylene blue yakaderedzawo zvakananga kushanda kwecaspase-1 yemunhu yakagadzirwa nenzira yerecombinant. Kuisa chinhu chacho pakugadzirira chete kana pakutanga kushanda chete kwakanga kuchiri kuderedza kubudiswa kweIL-1beta, IL-18, necaspase-1, saka matanho ose ari maviri anobatanidzwa.

Miedzo yemakonzo yakaenderana nekusarudzika uku. Huwandu hunouraya hwelipopolysaccharide hwakauraya makonzo ekuenzanisa mumaawa mashoma, asi methylene blue yakawedzera kupona; IL-1beta yemumhango yemudumbu yakaderera asi IL-6 iri mumvura imwe chete iyoyo haina kuderera. Mumodheru yeperitonitis yakakonzerwa neListeria, IL-1beta yemumhango yemudumbu yakaderera asi nhamba yose yemasero haina kuchinja. Monocyte dzemunhu dzeTHP-1 dzakaita semacrophage emakonzo mumhando dzose ina dzema-inflammasome. Mhedzisiro ye-inflammasome inotsanangura kukura kweIL-1beta neIL-18, kwete cytokine dzose. Ndokusakawo chimiro chemasero edziviriro muuropi mumiedzo yemicroglia neastrocyte chichipindura mubvunzo wakasiyana newekuunganidzwa kwehwaro.

Nzira 2: kunyorwa kwegene re-iNOS kuburikidza nekusunga kweNF-kB neSTAT1

Inducible nitric oxide synthase, kana kuti iNOS, inenge isipo kana sero rakazorora uye gene rayo rinonyorwa pazvinenge zvichidiwa. Lipopolysaccharide inonyanya kuikurudzira kuburikidza nekuparadzwa kweIkB-alpha, kuchiteverwa nekuwedzerwa kwephosphate paNF-kB uye kupinda kwayo munyukireyasi. Interferon-gamma inonyanya kuikurudzira kuburikidza nekuwedzerwa kwephosphate paSTAT1 neJanus kinase, kuchiteverwa nekupinda kweSTAT1 munyukireyasi. Protein imwe neimwe inobva yasunga panzvimbo yayo papromoter ye-iNOS. Kushanda kwe-enzyme nekukurudzirwa kwegene inzvimbo dzakasiyana dzekudzora, uye mushonga unogona kushanda pane imwe chete kana pazvose zviri zviviri.

Chidzidzo che2015 chekukurudzirwa kwe-iNOS mumacrophage nemakonzo ane endotoxin muropa chakaisa mhedzisiro padanho remashoko emRNA. Mumasero eRAW 264.7 nemacrophage akatorwa zvakananga mumakonzo, 1 micromolar yemethylene blue yakaiswa maminitsi 30 lipopolysaccharide, interferon-gamma, kana zvose zviri zviviri zvisati zvaiswa yakaderedza protein ye-iNOS nemRNA mukati memaawa 8 kusvika 24. Nitrite yaivawo shoma mumvura yaikuriswa masero, uye kugona kwemasero kurarama hakuna kuderera. Mumakonzo akapiwa 25 mg per kg yelipopolysaccharide, chipimo chimwe che5 mg per kg chakapiwa awa rimwe izvi zvisati zvaitwa chakaderedza protein ye-iNOS nemRNA mumapapu, chiropa, nemwoyo.

Chinhu chakasiyana pano ndechechisina kuchinja. Kuparadzwa kweIkB-alpha, kuwedzerwa kwephosphate paNF-kB, kuwedzerwa kwephosphate paSTAT1, uye kuungana kweprotein imwe neimwe munyukireyasi zvakaramba zvakadaro. Kuvhiringidzwa kwakauya padanho rakatevera: zvakabviswa munyukireyasi zvakaratidza kusunga kushoma kweNF-kB pachikamu chayo cheDNA mushure melipopolysaccharide uye kusunga kushoma kweSTAT1 mushure meinterferon-gamma. Chromatin immunoprecipitation yakasimbisa kuvapo kushoma kweprotein imwe neimwe papromoter ye-iNOS mukati memasero. Tumor necrosis factor alpha, ine promoter inoshandisawo NF-kB, yakadererawo. Mapuroteni aya akanga atanga kushanda uye asvika, asi haana kunyatsosunga DNA yaakanangana nayo.

Cherechedza musiyano nenzira 1. Izvi zvine chekuita nekusunga kweprotein kuDNA, zvinoshandisa STAT1 kwete STAT3, uye zvinoyera mRNA, protein, nenitrite panzvimbo pezvikwata zviduku zveASC nezvidimbu zvecaspase-1. Mepu yedu yezvinobatwa nemethylene blue mumamodheru emasero akasiyana inochengeta danho iri rekunyorwa kwemagene rakaparadzana nezviyero zvakananga zvema-enzyme nemitochondria.

Nzira 3: IL-6 muserum nekutanga kushanda kweSTAT3 mumatishu

IL-6 inotumira masaini kuburikidza nemhuri yemareceptor egp130 ichipinda munhevedzano yeJanus kinase kuenda kuSTAT3. STAT3 yakawedzerwa phosphate inobatana kuita mapeya, yopinda munyukireyasi, yokurudzira zvirongwa zvekuzvimba. IL-6 pachayo ndechimwe chezvinonangwa neNF-kB, saka zviviri izvi zvinoumba denderedzwa umo chimwe chinodzokorora kukurudzira chimwe, rinowanzonzi akisi yeIL-6 kuenda kuSTAT3. Zviyero zvinowanzoshandiswa iIL-6 yemuserum pamatanho ekutanga, reshiyo yeSTAT3 yakawedzerwa phosphate kuSTAT3 yose, uye ma-enzyme anoita basa akadai seiNOS neCOX2 pamatanho anotevera.

Chidzidzo che2023 chemakonzo pamusoro peIL-6 neSTAT3 mushure melipopolysaccharide chakayera akisi iyi munzvimbo dzakasiyana dzemuviri. Makonzo echirume eC57BL/6 akapiwa 1 mg per kg yelipopolysaccharide zuva nezuva kwemazuva matatu, pamwe ne5 kana 10 mg per kg yemethylene blue maminitsi 30 mushure mechipimo chimwe nechimwe. Pazuva rechitatu, IL-6 yemuserum yakanga yakaderera mumapoka ose ari maviri emethylene blue kupfuura muboka rakapiwa lipopolysaccharide chete, asi cytokine zhinji dzimwe dzakayerwa hadzina kuchinja nekuiswa kwemethylene blue. Reshiyo dzeSTAT3 yakawedzerwa phosphate kuSTAT3 yose dzakanga dzakaderera mucortex, hippocampus, uye ganda renzeve. Kupendwa kwemicroglia kunoratidza Iba-1 kwakanga kwaderera, iNOS neCOX2 zvakanga zvaderera, uye kurasikirwa nehuremu kwakanga kwadzikiswa zvishoma.

Pane zvinhu zviviri zvinoda kungwarirwa pano. IL-6 yakaderera muserum pamwe nekuwedzerwa kushoma kwephosphate paSTAT3 mumatishu zvinoenderana nekushandurwa kweakisi, asi hazvisarudzi nzvimbo huru inotanga kubatwa. Izvi zvinodaro nokuti cytokine shoma pamatanho ekutanga, masaini mashoma kubva kumareceptor, kana kushanda kwakachinja kwephosphatase chimwe nechimwe chinogona kubudisa reshiyo imwe chete. Uye huremu mumazuva matatu ekukuchidzirwa kwakanyanya chiyero chemhuka yose, kwete chibodzwa chechiratidzo chechirwere kana chekudzikira kwechirwere. Zvinowedzera mamiriro ekunzwisisa kupfuura mufananidzo weblot, zvisingashanduri mhedzisiro yemasaini kuita chirevo chekurapa. Mashandiro akasiyana ekemisitiri ekudzora redox mumitochondria anoitika pamwe chete netsananguro iyi yecytokine, uye haafaniri kubatanidzwa nayo.

Kuenzanisa nzira: nzvimbo dzinobatwa nezviyero zvemhedzisiro

Tafura iyi inodzivisa nzira nhatu dzisina kururama dzekuendesa mhedzisiro kune imwe nzira. Mhedzisiro yezvikwata zviduku zveASC hairatidzi kuti pane mhedzisiro yeSTAT3. Mhedzisiro yeSTAT1 papromoter ye-iNOS hairatidzi kuti pane mhedzisiro yeSTAT3, kunyange mapuroteni ose ari maviri ari emhuri yeSTAT. Mhedzisiro yeIL-6 yemuserum hairatidzi kuunganidzwa kwakaderera kwe-inflammasome: mumuedzo wemhango yemudumbu, IL-1beta yakaderera asi IL-6 haina kuderera. Mutsara mumwe nemumwe unoda chinokuchidzira chawo nechiyero chawo. Ndokusaka kuverenga zvidzidzo zvemethylene blue mumasero, mumhuka, nemuvanhu kuchida kuti chirevo chimwe nechimwe chienderane nedanho rakayerwa.

Hapana pane izvi chinosimbisa zvipimo, kuchengeteka, kana kubatsira muvanhu. Mamodheru elipopolysaccharide zvinhu zvinokuchidzira zvinodzorwa kuti paedzwe mashandiro enzira, kwete mifananidzo chaiyo yechirwere chisingaperi. Kupa chipimo pamwe chete nechinokuchidzira kunoedza kudzivirira shanduko inokonzerwa, kwete kudzorera shure chirwere chagara chiripo. Chengeta tsananguro iyi yemashandiro yakaparadzana nemhedzisiro yevarwere, uye ongorora chirevo chipi nechipi cheutano uchitarisa chidzidzo chevarwere chinotsigira chirevo chacho.