Methylene blue ndi maselo othandiza ubongo: astrocytes, microglia, ndi kutupa

Tiyerekeze kuti mukuchita mayesero awiri pogwiritsa ntchito utoto wabuluu womwewo ndipo mukupeza zotsatira ziwiri zolimbikitsa. M'mbale imodzi, maselo othandiza ooneka ngati nyenyezi amapulumuka pamene mpweya wa oxygen ndi glucose zasowa. M'mbale ina, maselo a chitetezo cha m'thupi omwe ali mu ubongo amasungabe mawonekedwe awo abata okhala ndi nthambi ngakhale pali poizoni wa mabakiteriya. N'zosavuta kufuna kuphatikiza zotsatira ziwirizi n'kunena kuti utotowo umachepetsa kutupa mu ubongo ndiponso umateteza ubongo. Kuphatikiza kumeneku ndi pamene owerenga mosamala ayenera kuima kaye ndi kuganizira.

Ubongo uli ndi magulu awiri a maselo othandiza omwe amagwira ntchito zosiyana, ndipo methylene blue imayesedwa pa gulu lililonse ndi choyambitsa chosiyana. Maselo a astrocyte amathandiza kagayidwe ka zinthu. Amakhala pakati pa mitsempha ya magazi ndi maselo a minyewa, amachotsa glutamate yochuluka, amasunga potassium pa mlingo woyenera, amasunga glycogen, ndipo amatumiza zinthu zopangira mphamvu ku maselo a minyewa pamene kufunika kwake kwakwera mwadzidzidzi. Maselo a microglia ndi alonda a chitetezo cha m'thupi omwe amakhala mu ubongo. Nthambi zawo zopyapyala zimayang'anira minofu, zimazindikira zizindikiro za mabakiteriya ndi kuwonongeka, ndipo pakakhala chenjezo zimabwerera mkati n'kupanga mawonekedwe ozungulira ngati amoeba, oyenera kusuntha ndi kumeza zinthu, pamene zimatulutsa mamolekyulu otumiza zizindikiro. Zotsatira za gulu limodzi sizingotengedwa n'kuikidwa pa lina, ndipo muyeso wogwiritsidwa ntchito pa mayesero aliwonse ndi umene umasankha zimene zasonyezedwadi.

Maselo a astrocyte pamene kagayidwe kakusokonekera: nthawi ndiyo mfundo yaikulu ya zotsatira

Mayesero a astrocyte amayamba ndi kuchotsa zinthu zofunika. Ofufuza anakulitsa mu labotale maselo a astrocyte ochokera ku kotekisi ya mbewa zobadwa kumene, anachotsa glucose ndi pafupifupi oxygen yonse kwa maola asanu ndi limodzi, kenako anabwezeretsa zonse ziwirizo kwa maola makumi awiri ndi anayi. Ichi ndi chitsanzo cha mu labotale cha sitiroko yotsatiridwa ndi kubwerera kwa magazi. Pamene methylene blue inalipo pa nthawi yosowa zinthuzi, kupulumuka kwa maselo kunachepa. Pamene inaikidwa pa nthawi yobwezeretsa oxygen yokha, pa 0.1 ndi 1 micromolar, kupulumuka kunakwera kwambiri malinga ndi mayeso a calcein oyeza ngati maselo akadali amoyo. Kafukufuku wa maselo a astrocyte a mbewa akufotokoza kusiyana kumeneku kwa nthawi limodzi ndi deta ya kupuma kwa maselo.

Kusiyana kumeneku ndiko kofunika kwambiri pa zimene anapeza. Pa nthawi yosowa zinthuzi, pafupifupi palibe oxygen yolandira ma elekitironi, choncho njira yopatsira ma elekitironi ilibe poti iwafikitse. Pa nthawi yobwezeretsa oxygen, njira za m'selo zimayambiranso ndipo utotowo umakhala ndi ntchito yoti ugwire. Maselo a astrocyte omwe analandira utotowo anagwiritsa ntchito oxygen yambiri, anatenga glucose yambiri, anapanga ATP yambiri, ndipo anabwezeretsa nkhokwe za glycogen. Ntchito ya hexokinase inakwera, koma superoxide ya m'mitochondria sinakwere. Kufotokozera komwe akuganiza kuti kungakhale kolondola ndi komwe kumadziwika kuchokera ku nkhani ya methylene blue ngati chonyamulira ma elekitironi m'mitochondria: imanyamula ma elekitironi n'kuwadutsitsa malo otsekeka m'unyolo wa kupuma kwa maselo, kuti kupuma kuyambenso bwino m'malo motayikira ma elekitironi n'kupanga zinthu zogwira ntchito kwambiri pa mankhwala.

Dziwani mtundu wa umboni umenewu. Ndi zotsatira zokhudza kupulumuka ndi kagayidwe ka zinthu mu mtundu umodzi wa maselo olekanitsidwa, zomwe zili ndi nthawi yeniyeni imene zinathandiza, ndiyo nthawi yobwerera bwino osati nthawi yosowa zinthu, ndiponso milingo yeniyeni ya kuchuluka kwa utotowo. Sizinena chilichonse chokhudza kutumizana zizindikiro za chitetezo cha m'thupi, kapena khalidwe.

Maselo a microglia pa chenjezo la mabakiteriya: kagayidwe kakusinthidwa

Mayesero a microglia amayamba ndi choyambitsa chosiyana: osati kusowa chakudya, koma chenjezo labodza la matenda opatsirana. Lipopolysaccharide, chidutswa cha nembanemba yakunja ya bakiteriya, imamangirira ku zolandirira zizindikiro za chitetezo chobadwa nacho ndipo imachititsa maselo a microglia kuganiza kuti tizilombo toyambitsa matenda tafika, ngakhale palibe mabakiteriya amoyo. M'mbewa zopatsidwa lipopolysaccharide ndiponso mu mzere wa maselo a microglia okulitsidwa mu labotale, pamachitika ndondomeko yodziwika ya kutupa: zinthu zolimbikitsa kutupa zimakwera, maselo a microglia amasinthira ku mkhalidwe waukali wa M1, maselo a minyewa oyandikana nawo amafa, ndipo nyama zimachita moipa kwambiri pa ntchito zoyesa kuphunzira ndi kukumbukira.

Kafukufuku wa mbewa wokhudza zimenezi akulondola chiyambi chake ku njira yozindikira oxygen: lipopolysaccharide imachititsa kupanga PHD3 yochuluka kwambiri, imodzi mwa ma enzyme atatu a prolyl hydroxylase, kudzera mu unyolo wa Siah2/Morg1 womwe uli koyambirira kwa njirayi. Pa nthawi yomweyo, maselo amasintha mmene amagwiritsira ntchito mphamvu n'kudalira kwambiri aerobic glycolysis, pamodzi ndi kuwonjezeka kwa mitundu ya oxygen yogwira ntchito kwambiri m'mitochondria. Olembawo anayeza kusintha kumeneku mwachindunji pogwiritsa ntchito mayeso a Seahorse a kagayidwe, omwe amayeza kugwiritsidwa ntchito kwa oxygen ndi kuwonjezeka kwa asidi kunja kwa maselo. Methylene blue inabwezeretsa kusintha kwa kagayidwe ndi kupanga PHD3 kochuluka kwambiri, inachepetsa zinthu zoyambitsa kutupa, inachepetsa kufa kwa maselo a minyewa, ndipo inakweza zotsatira za mayeso a khalidwe. Kujambula ndi positron emission tomography kunawonjezera umboni wa zithunzi: lipopolysaccharide inakweza kutengedwa kwa glucose mu hippocampus pa FDG-PET, ndipo chithandizo chinachepetsanso kutengedwako. Izi zinagwirizana ndi kuchepa kwa kagayidwe kogwirizana ndi kutupa mu ubongo wamoyo.

Miyeso imeneyi ndi yochuluka kuposa ya kafukufuku wa astrocyte: ma cytokine, kagayidwe, zithunzi, ndi khalidwe la nyama. Komabe, imakhudza choyambitsa china ndi mtundu wina wa maselo. Owerenga oyerekezera kafukufuku awiriwa ayenera kusiyanitsa zoyambitsa zake: kafukufuku wina amathandiza maselo othandiza omwe akusowa chakudya pa nthawi yobwerera bwino, pamene winayo amachepetsa chenjezo labodza la matenda opatsirana m'maselo a chitetezo cha m'thupi.

Maselo a microglia pa maikulosikopu: mawonekedwe ngati muyeso

Mayesero achitatu amaona chenjezo lomwelo la mabakiteriya kudzera m'mawonekedwe okha. M'maselo a glia okulitsidwa kuchokera ku kotekisi ya makoswe, maselo a microglia a gulu loyerekezera, opakidwa utoto wozindikira chizindikiro cha IBA1, amasonyeza nthambi zambiri zopyapyala, mawonekedwe anthambi a maselo oyang'anira. Atakhala kwa maola makumi awiri ndi anayi mu lipopolysaccharide ya 10 micrograms pa milliliter, maselo amakhala athyathyathya n'kutenga mawonekedwe ngati amoeba. Malo okutidwa ndi thupi la selo amakula pamene utali wozungulira m'mphepete mwake umakhala pafupifupi womwewo, chomwe ndi chizindikiro cha masamu chosonyeza kuti selo likuzungulira. Methylene blue ya 1 micromolar, yoperekedwa limodzi ndi poizoniyo, inasunga nthambizo ndipo inasunga malo okutidwa ndi thupi la selo ndi utali wozungulira m'mphepete mwake pafupi ndi miyeso ya gulu loyerekezera. Kafukufuku wa maselo okulitsidwa a makoswe akuyeza kusungidwa kwa mawonekedwe kumeneku pogwiritsa ntchito malo okutidwa ndi thupi la selo ndi utali wozungulira m'mphepete mwake.

Mfundo ziwiri zimapangitsa kafukufukuyu kukhala wofunika kuposa chithunzi chokha. Choyamba, menadione, chinthu choyerekezera chomwenso ndi molekyulu yogwira ntchito pa redox pa kuchuluka komweko, sinasunge mawonekedwe ndipo payokha inakankhira maselo ku mawonekedwe a mkhalidwe woyatsidwa. Choncho zotsatirazi si khalidwe la zinthu zonse zomwe zimazungulira pakati pa mikhalidwe ya redox. Chachiwiri, olembawo amagwirizanitsa zotsatira za utotowo ndi kuyatsidwa kwa Nrf2, puloteni yolamulira kulembedwa kwa majini yomwe imayendetsa ndondomeko ya majini oteteza ku oxidation ndi ochepetsa kutupa, osati kungochotsa mwachindunji zinthu zogwira ntchito kwambiri. Nkhani zina zokhudza redox ndi kutumizana zizindikiro za kutupa zimathandiza kumvetsa kugwirizana kumeneku kwa Nrf2.

Komabe, mawonekedwe ndi muyeso wokhala ndi umboni wochepa kwambiri pa miyeso itatuyi. Selo lokhala ndi nthambi nthawi zambiri limakhala selo loyang'anira, ndipo selo lozungulira nthawi zambiri limakhala loyatsidwa, koma mawonekedwe ndi chizindikiro choloŵa m'malo mwa zomwe zikuchitika, osati zolemba zake zonse. Maselo okulitsidwa amenewa analibe maselo a chitetezo cha m'thupi ochokera m'magazi, analibe maukonde a minyewa, ndipo analibe zotsatira za khalidwe.

Kuwerenga zotsatira zitatuzi moyerekezera

Mfundo yofanana m'mizereyi ndi yakuti choyambitsa chilichonse chili ndi muyeso wake ndi yankho lake. Kupulumutsa kagayidwe kumayankha funso la kagayidwe. Deta ya ma cytokine ndi khalidwe imayankha funso la kutupa. Mawonekedwe amayankha funso la mawonekedwe. Palibe mwa zitatuzi chomwe chokha chingayankhe mafunso a zinazo.

Zimene maselo a microglia ooneka abata sizitsimikizira

Ichi ndi chiganizo chofunikira pa nkhani yonseyi: maselo a microglia ooneka ngati sayatsidwa kwambiri, kapena kagayidwe ka microglia kosinthidwa, paokha sizitsimikizira kuti ntchito ya ubongo yasintha kukhala yabwinopo.

Mawonekedwe angasinthe popanda ntchito kusintha motsatira. Selo lingasunge nthambi zake pamene likutulutsabe ma cytokine, kapena lingazungulire pamene likugwira ntchito yothandiza yochotsa zotsalira za maselo. Miyeso ya kagayidwe ilinso ndi malire omwewo: zotsatira za Seahorse kapena chithunzi cha PET zimasonyeza mmene mphamvu zikugwiritsidwira ntchito, osati kuganiza ndi kuzindikira. Kafukufuku wa mbewa amene anaphatikiza miyeso ya kagayidwe ndi yofanana ndi ya mawonekedwe pamodzi ndi mayeso a khalidwe ali ndi umboni wolimba kwambiri chifukwa anaunjika mitundu yosiyanasiyana ya umboni m'malo modalira muyeso umodzi kuti uthandize mfundo yomaliza. Ngakhale pamenepo, malire anthawi zonse amakhalabe, chifukwa lipopolysaccharide imapanga chitsanzo cha chenjezo lopanda tizilombo tamoyo m'malo mwa matenda enieni opatsirana, mbewa si anthu, ndipo olembawo eni ake amachenjeza za kusiyana kwa mitundu ya zamoyo. Aliyense amene akukonzekera kafukufuku wotsatira ayenera kuganizira mmene zomwe zapezeka pa microglia zingagwirire ntchito m'mitundu yosiyanasiyana ya mayesero asanaganize kuti njirayi imagwira ntchito mofanana mu minofu ya anthu.

Mfundo yothandiza yokhudza nthawi imatsatira pamenepa. Zotsatira za astrocyte zimasonyeza kuipa kopereka utotowo msanga ndi ubwino woupereka pa nthawi yobwerera bwino. Zotsatira za microglia zimachokera pa kupereka utotowo nthawi yoyandikana ndi choyambitsa kutupa. Kuvulala kwa ubongo m'moyo weniweni kumasakaniza magawo onse awiri nthawi imodzi: kulephera kupeza mphamvu pamodzi ndi chenjezo la chitetezo cha m'thupi, zomwe zimasintha m'maola angapo. Mayesero olekanitsa gawo limodzi sangathe kukhazikitsa nthawi yoperekera zinthu pamene magawowo aphatikizana. N'chifukwa chake zokambirana za nthawi yoyenera yopereka zinthu zimasiyanitsa nthawi ya kusowa zinthu ndi nthawi ya kutupa.

Chidule chosakokomeza chingakwane m'ndime imodzi. M'maselo a astrocyte olekanitsidwa, methylene blue yoperekedwa pa nthawi yobwezeretsa oxygen imayambitsanso kupuma kwa maselo ndipo imapulumutsa maselo. M'maselo a microglia oikidwa pa chenjezo ndi chidutswa cha bakiteriya, imabwezeretsa ndondomeko ya kagayidwe yoyambitsa kutupa kudzera mu njira yeniyeni ya PHD3 ndipo imasunga mawonekedwe a mkhalidwe wopuma. Zotsatira zilizonse ndi zenizeni malinga ndi mmene mayeserowo anakhazikitsidwira, ndipo palibe chimodzi mwa izo chomwe chili chigamulo cha kutupa konse kwa mu ubongo. Kuyerekezera maselo a glia kumathandiza makamaka chifukwa kumaletsa nkhani ziwirizi kusanganikirana n'kukhala imodzi.