Yadda methylene blue ke aiki: canja wurin electron, da inda electron suke sauka

Kwalbar gwaji mai murfi a kan benci ita ce hanya mafi gajarta kuma mafi gaskiya ta gabatar da yadda methylene blue ke aiki. Ruwa, glucose, ɗan sodium hydroxide, da isasshen dye da zai mai da ruwan shuɗi mai zurfi wanda ba za a rikice da shi ba. Jira na ƴan mintuna, shuɗin zai gushe har kwalbar ta yi kama da kwalbar ruwa. Girgiza ta, launin zai dawo nan take. Daga nan ya sake dusashewa, kuma zai ci gaba da haka muddin akwai glucose da zai iya sakin electron da oxygen da zai iya karɓar su.

Babu abin da ya canza game da dye ɗin sai adadin electron da yake ɗauke da su. Idan yana oxidised, shuɗi ne. Idan yana reduced, ba shi da shuɗin. Kusan duk sauran bayanan da ake samu a rubuce-rubucen magani da bincike game da wannan molecule sun samo asali ne daga wannan mataki guda mai komawa baya, domin molecule da zai iya karɓar electron daga wani abokin hulɗa ya miƙa su ga wani shi ne relay, kuma relay ba ya yin komai shi kaɗai. Abin da yake yi ya dogara gaba ɗaya da abin da ke kowane ƙarshensa.

Siffofi biyu, da dalilin da ya sa haɗinsu yake da muhimmanci

Siffar oxidised tsarin zobe ne mai faɗi kuma mai positive charge. Wannan siffar mai faɗi da caji ce ta sa yake jeruwa kusa da nucleic acids, yake ɗaurewa da sialylated structures a kan cell membranes, kuma yake shan visible light sosai tsakanin kusan 600 and 700 nanometres, wato ɓangaren spectrum da idon ɗan adam ke gane shi a matsayin shuɗi.

Siffar reduced kuwa daban take. Ba ta ɗauke da charge, tana narkewa a cikin kitse maimakon ruwa, kuma ba ta da launi. Ionisation ɗinta yana da ƙasa sosai a pH na cikin jiki, kusan ƴan per cent. Saboda haka reduced molecule yana ketare cell membrane cikin sauƙi ta diffusion, kuma da zarar ya shiga ciki, inda yanayin ya fi oxidising, za a iya sake oxidise ɗinsa kuma a kama shi a can yadda ya kamata. Tsarin zoben oxidised yana da faɗi; reduced ɗin kuwa zai iya murɗewa. Waɗannan sakamakon tsarin, tare da sauyin launin kansa, suna cikin labarin siffofin biyu da kansu, kuma an yi bayani dalla-dalla a cikin kwatancin methylene blue da leucomethylene blue.

Karɓar electron

Oxidised dye zai karɓi electron daga masu bayarwa iri-iri. Glucose da ke cikin kwalbar gwajin nan misali ne guda. A cikin cell, reduced pyridine nucleotides da sassan mitochondrial electron transport chain su ma wasu ne. Lokacin da aka reduce methylene blue a tissue da ke ɗauke da oxygen, zagayen da yake haifarwa yana cinye reducing agent da oxygen duka, kuma yana samar da reactive oxygen species, musamman hydrogen peroxide. Dye ɗin yana fitowa daga zagayen ba tare da ya canza ba, a shirye ya sake maimaita zagayen.

Wannan batu na ƙarshe yana bayyana wani saɓani da in ba haka ba zai yi kama da sakaci. Ana bayyana methylene blue a cikin rubuce-rubuce a matsayin pro-oxidant da kuma antioxidant, kuma ana iya kare bayanin duka biyun. Zai iya rage samar da superoxide ta hanyar aiki a matsayin artificial electron acceptor, yana karkatar da kwararar electron daga enzyme sites inda in ba haka ba oxygen zai koma superoxide radicals. A liver cell cultures, an nuna cewa yana haifar da phase-2 antioxidant defence enzymes, ciki har da thioredoxin reductase 1 da NADPH quinone oxidoreductase. Wace fuska daga cikin biyun za ka gani ya dogara da wane ƙarshen relay ne yake da ƙarancin electron a wannan lokacin. Tambaya irin "shin methylene blue antioxidant ne" ba ta cika ba har sai an fayyace a wane system, a wane concentration, da me kuma ke ciki.

Miƙa electron gaba

Da zarar dye ɗin ya riƙe spare electron, abubuwa uku mabambanta sosai na iya faruwa daga nan, kuma sau da yawa ana haɗa su a ƙarƙashin take guda. Bai kamata a yi haka ba.

Zai iya miƙa electron zuwa metal centre. Misali mafi bayyana shi ne haemoglobin da aka oxidise daga ferrous zuwa ferric state. A wannan state ba zai iya ɗaukar oxygen ba. A cikin red cells, wani enzyme yana reduce dye ɗin, sannan reduced dye ɗin ya miƙa electron ga oxidised haem iron ta hanyar da ba enzymatic ba, yana maido da haemoglobin mai aiki kuma yana mayar da dye ɗin zuwa oxidised form. Ba a cinye dye ɗin a wannan aikin; yana zagayawa ne. Yanayin asibiti da ya dogara da wannan reaction batu ne na kulawar asibiti, kuma bayanin methylene blue a methemoglobinemia yana bayyana yadda ake amfani da shi da dalilin da ya sa ake kula da shi sosai.

Muhimmin bayanin gaskiya game da karatun: reviews biyu da aka gina wannan labarin a kansu ba su ambaci reducing cofactor iri ɗaya ba don matakin enzyme, ɗaya yana cewa NADH ɗayan kuma NADPH. Saɓanin gaskiya ne kuma ba a warware shi ba a cikin sources da ake da su a nan, don haka mafi aminci shi ne a bayyana rawar da kowane ɓangare yake takawa maimakon ɗaure bayanin ga suna guda na cofactor.

Zai iya miƙa energy, ko electron, zuwa oxygen, idan akwai haske. Wannan hanya tana samuwa ne kawai ƙarƙashin illumination. Excited dye ɗin yana kaiwa triplet state, kuma daga nan mechanisms biyu na iya faruwa. A na farko, excited dye ɗin yana hulɗa kai tsaye da neighbouring molecule, yana karɓar hydrogen ko yana miƙa electron, wanda ke samar da radicals; reaction da ruwa ta wannan hanya yana samar da hydroxyl radicals. A na biyu, ana miƙa excitation energy ga oxygen kansa, yana samar da singlet oxygen, wani highly electrophilic species da ke lalacewa cikin microseconds, kuma gaba ɗaya ana ɗaukarsa a matsayin babbar hanyar da illuminated methylene blue ke lalata microbial cells. Reactions da cell envelope lipids da peptides, da kuma guanine bases a DNA, suna cikin sakamakon da aka rubuta.

Sakamakon wannan hanya a aikace shi ne illumination yana canza lambobi da orders of magnitude. A bayanan da Wainwright da Crossley suka tattara, methylene blue a kan Pseudomonas aeruginosa yana da minimum bactericidal concentration kusan 500 micromolar a cikin duhu, kuma kusan 25 micromolar ƙarƙashin light dose na 12 joules per square centimetre. A kan strain guda na Staphylococcus aureus, values na duhu da illumination duka 5 micromolar ne. Wato, ko za a ɗauki molecule ɗin a matsayin antimicrobial ko a'a ba kaddara ce ta molecule kaɗai ba; kaddara ce ta molecule tare da haske.

Zai iya aiki kai tsaye a enzyme site, ba tare da relay ba. Methylene blue yana oxidise ferrous iron da ke ɗaure da nitric oxide synthase, wanda ke kashe aikin enzyme ɗin. Yana toshe soluble guanylate cyclase, kuma tsakanin 1 and 10 micromolar shi ne routine laboratory concentration da ake amfani da shi don wannan dalili. Yana inhibit monoamine oxidase type A da IC50 tsakanin 27 and 180 nanomolar, wanda shi ne mechanism da ke bayan sanannen interaction da serotonin reuptake inhibitors kuma dalilin da ya sa drug interactions na methylene blue suke da rukuni na kansu.

Waɗannan hanyoyi uku sun bambanta ta fuskar chemistry. Haɗa su duka cikin ikirari guda cewa methylene blue yana aiki ta redox cycling shi ne babban dalilin da ya sa yawancin rubutu game da shi yake karantawa kamar yana da cikakken tabbaci amma kuma ba ya fayyace komai sosai.

Bambance-bambance huɗu da ke tantance ko bayanin mechanism yana nufin wani abu

Wace siffa. Reduced dye ba za a iya musanya shi da oxidised dye ba. Acetylcholinesterase inhibition yana nuna wannan tarkon: a cikin dogon in vitro incubation, methylene blue yana ci gaba da raguwa zuwa siffarsa marar launi, kuma inhibition ɗin yana raguwa, domin reduced molecule ba ya inhibit wannan enzyme. Assay da aka yi tsawon lokacin da zai canza siffar zai nuna weaker effect kuma ya buɗe ƙofa ga bayanin da bai dace ba.

Wane concentration. Laboratory range don soluble guanylate cyclase inhibition shi ne 1 to 10 micromolar. Don kawar da glutamate-mediated transmission a rat hippocampal slices an buƙaci 5 to 50 micromolar. Ana bayar da rahoton toxic effects a cell culture sama da 100 micromolar. Waɗannan ba experiment ɗaya ba ne, kuma mechanism da aka tabbatar a 50 micromolar ba lallai ya samu ga patient wanda plasma concentration ɗinsa ƙasa yake sosai daga hakan ba.

Wace route. Absolute oral bioavailability yana kusan 72 per cent, kuma terminal plasma half-life yana 5 to 7 hours. Oral administration yana samar da concentrations mafi girma a intestine da liver; intravenous administration kuma yana samar da concentrations mafi girma a brain. Route ne ke tantance wane tissue zai haɗu da electron.

Wane model, kuma ko fitilu suna kunne. Mafi yawan mechanistic literature cell culture ne da aikin rodents, kuma in vitro results game da wannan molecule suna dogara da light exposure, membrane potential da redox state na cells, domin microscope lamp kaɗai ya isa ya photosensitise shi. Tsakanin 65 and 85 per cent na administered dose ana reduce shi zuwa siffar marar launi a red cells da peripheral tissue. Kowane ɗaya daga cikin waɗannan variables yana tsakanin mechanism mai tsabta a cikin dish da ikirari game da mutum.

Dalilin da ya sa tambayar purity tambayar chemistry ce

Illumination yana demethylate methylene blue. Samfurin farko shi ne Azure B, sai Azure A, sannan thionin, kuma higher light intensity yana samar da breakdown fiye da lower. Don haka molecule da amfaninsa ya ta'allaka kan iya karɓa da bayar da electron shi ma molecule ne da likely degradation products ɗinsa dangi ne na kusa ta fuskar chemistry waɗanda suke ci gaba da kasancewa cikin solution ɗaya. Abin da ke cikin container ba ƙaramin bayani na kwalliya ba ne.

Wannan ne dalilin da ya sa conformance to the USP specification shi ne gwaji mai ma'ana ga buyer. Product da bai cika shi ba dye ne, kuma dye yana ɗauke da impurities, ciki har da Azure B, heavy metals da residual solvents, waɗanda mutumin da zai haɗiye shi ba ya so. Ƙarin tarko shi ne seller da ke gwada heavy metals kawai sannan ya gabatar da sakamakon a matsayin full compliance. Blupreme ya yi haɗin gwiwa da pharmaceutical manufacturer na methylene blue kuma yana gwada full specification: identity, purity, organic impurities, residual solvents, elemental impurities, residue on ignition, microbial limits da bacterial endotoxins. Ana wallafa laboratory testing document da ke bayan kowane batch, kuma zaɓar supplier ya fi yawa zama batun duba ko wannan document yana nan da kuma abin da yake rufewa.

Reversible couple guda ɗaya. Hanyoyi da dama suna fita daga gare shi. Route ne ke tantance abin da ikirarin zai iya zama.