Shuɗin methylene da siginar kumburi: inflammasome, IL-6, da STAT3

A ce dakunan gwaje-gwaje uku sun ba da rahoton cewa sinadarin shuɗi ɗaya yana rage kumburi. Ɗaya ya nuna ƙarancin IL-1beta daga macrophage. Ɗaya ya nuna ƙarancin enzyme na iNOS a irin waɗannan ƙwayoyin halitta. Ɗaya ya nuna ƙarancin IL-6 a jini da ƙarancin aikin STAT3 a kwakwalwa. Ana iya son haɗa waɗannan sakamako uku a matsayin da'awa guda ɗaya. Wannan haɗawa tana ɓoye ainihin abin da aka gano.

Kowane sakamako yana da alaƙa da wata hanyar sigina daban, mai abin tayar da ita, matakin haɗuwa, da ma'aunin sakamako na kanta. Shuɗin methylene yana shafar kowace hanya a wani mataki daban. Ko ɗaya daga cikin waɗannan yana inganta sakamakon lafiya, tambaya ce dabam, wadda muka tattauna a bayaninmu na gaba ɗaya kan shuɗin methylene da kumburi. Wannan shafi yana mai da hankali kan hanyoyin sigina.

Hanya 1: haɗuwar inflammasome da caspase-1

Inflammasome wani tsarin furotin ne da ake ginawa bisa buƙata a cikin ƙwayar myeloid. Abubuwan da suka haɗa shi su ne mai gane abu, mai haɗawa da ake kira ASC, da enzyme mai aiwatarwa da ake kira caspase-1. Bayan sun haɗu, caspase-1 yana yanke pro-IL-1beta da pro-IL-18 zuwa siffofinsu da ake fitarwa, kuma yana iya tayar da pyroptosis, wani nau'in mutuwar ƙwayar halitta da ke haifar da kumburi. Mai gane abin ne ke ƙayyade nau'in: NLRP3 yana amsawa ga lu'ulu'u da gubobin da ke buɗe ramuka, kamar nigericin da ATP; NLRC4 yana amsawa ga flagellin na ƙwayoyin bacteria; AIM2 yana amsawa ga DNA mai zare biyu a cytosol; hanyar da ba ta gargajiya ba kuma tana amsawa ga lipopolysaccharide da ke cikin ƙwayar halitta.

Kunna wannan tsarin yana buƙatar sigina biyu. Sigina na farko shi ne shiryawa: wani abu da ke ɗaure da mai karɓar Toll-like yana ƙarfafa rubuta bayanan NLRP3 da pro-IL-1beta zuwa RNA, ta hanyar da ta dogara da NF-kB, domin abubuwan haɗin su kasance. Sigina na biyu shi ne haɗuwa: wani abin tayarwa na biyu yana sa ASC ya haɗu zuwa tarin oligomer a matsayin ɗigon taruwa, a yanke caspase-1 zuwa gutsurensa na p20, sannan a fitar da balagaggen IL-1beta p17. An san nau'o'in oxygen masu saurin yin martanin sinadarai daga mitochondria da shigar lu'ulu'u cikin ƙwayar halitta suna taimakawa wajen sigina na biyu na NLRP3.

Binciken macrophage na 2017 kan hana nau'o'in inflammasome da dama ya gwada shuɗin methylene a duka siginonin biyu. A macrophage na ɓargon ƙasusuwan beraye da aka shirya da lipopolysaccharide, nigericin, ATP, da lu'ulu'un monosodium urate kowannensu ya tayar da fitar IL-1beta da caspase-1 tare da samar da ɗigon taruwar ASC. Shuɗin methylene a 20 zuwa 200 micromolar ya rage duka ukun gwargwadon adadin da aka yi amfani da shi, ba tare da kashe ƙwayoyin halittar ba. Flagellin da Salmonella don NLRC4, DNA mai zare biyu da Listeria don AIM2, da lipopolysaccharide da aka shigar cikin ƙwayoyin halitta ko E. coli mai rai don hanyar da ba ta gargajiya ba, duk sun nuna irin wannan tsarin sakamako. Sinadarin shi kaɗai bai tayar da fitarwa ba, kuma farantin gwaji na kwatantawa bai nuna tasirin kashe bacteria ba. Saboda haka, sakamakon yana da alaƙa da amsawar jikin mai ɗauke da su.

A matakan da ke gabata, an auna tasiri guda uku. Superoxide na mitochondria da rotenone ya tayar ya ragu da shuɗin methylene, kuma fitar IL-1beta da rotenone ya tayar ita ma ta ragu. Shigar ƙananan ƙwallaye masu hasken fluorescence cikin ƙwayoyin halitta ya ragu, abin da ya dace da raguwar phagocytosis kafin gane lu'ulu'u. Tsarin rahoton aiki da yankin fara rubuta bayanan NLRP3 mai wuraren NF-kB guda biyu ke sarrafawa ya nuna ƙarancin aiki da shuɗin methylene, amma gajeriyar sigar wannan yankin da ba ta da waɗannan wuraren ba ta amsa ba. A bututun gwaji marar ƙwayoyin halitta, shuɗin methylene ya kuma rage aikin caspase-1 na ɗan Adam da aka samar ta fasahar haɗa bayanan gado kai tsaye. Ƙara sinadarin a lokacin shiryawa kaɗai ko a lokacin kunnawa kaɗai har yanzu ya rage fitar IL-1beta, IL-18, da caspase-1, don haka matakan biyu suna da hannu.

Gwaje-gwajen beraye sun yi daidai da wannan takamaiman tasiri. Adadin lipopolysaccharide mai kisa ya kashe berayen kwatantawa cikin sa'o'i, amma shuɗin methylene ya inganta tsira; IL-1beta a ramin ciki ya ragu, amma IL-6 a wannan ruwan bai ragu ba. A samfurin kumburin murfin ramin ciki da Listeria ta haifar, IL-1beta a ramin ciki ya ragu, yayin da jimillar ƙwayoyin halitta ba ta canza ba. Monocyte na ɗan Adam na THP-1 sun nuna hali irin na macrophage na beraye a duk nau'o'in inflammasome huɗu. Sakamakon inflammasome yana bayyana balagar IL-1beta da IL-18, ba kowane cytokine ba. Wannan kuma shi ne dalilin da ya sa siffar ƙwayoyin kariyar kwakwalwa a gwaje-gwajen microglia da astrocyte ke amsa wata tambaya dabam da ta haɗuwar tsarin furotin.

Hanya 2: rubuta bayanan iNOS zuwa RNA ta ɗaurin NF-kB da STAT1

Enzyme na inducible nitric oxide synthase, ko iNOS, yana kasancewa kaɗan ƙwarai a yanayin hutawa, kuma ana rubuta bayanansa zuwa RNA bisa buƙata. Lipopolysaccharide yana ƙarfafa wannan musamman ta lalacewar IkB-alpha, wadda ƙara rukunin phosphate ga NF-kB da shigarsa cikin tsakiya ke biyo baya. Interferon-gamma yana ƙarfafa shi musamman ta ƙara rukunin phosphate ga STAT1 ta Janus kinase, wadda shigar STAT1 cikin tsakiya ke biyo baya. Daga nan kowanne furotin mai sarrafawa yana ɗaure da wurinsa a yankin fara rubuta bayanan iNOS. Aikin enzyme da tayar da aikin kwayar gado matakan sarrafawa ne daban, kuma magani na iya yin tasiri a ɗaya ko duka biyun.

Binciken 2015 kan tayar da iNOS a macrophage da beraye masu endotoxin a jini ya gano tasirin a matakin saƙon RNA. A ƙwayoyin RAW 264.7 da macrophage na farko da aka ɗauko daga beraye, 1 micromolar na shuɗin methylene da aka bayar mintuna 30 kafin lipopolysaccharide, interferon-gamma, ko duka biyun ya rage furotin da mRNA na iNOS tsakanin sa'o'i 8 zuwa 24. Nitrite a ruwan renon ƙwayoyin ya ragu, ba tare da raguwar iya rayuwar ƙwayoyin halittar ba. A berayen da aka ba 25 mg per kg na lipopolysaccharide, adadi ɗaya na 5 mg per kg sa'a ɗaya kafin hakan ya rage furotin da mRNA na iNOS a huhu, hanta, da zuciya.

Abin da ya bambanta shi ne abin da bai canza ba. Lalacewar IkB-alpha, ƙara rukunin phosphate ga NF-kB, ƙara rukunin phosphate ga STAT1, da taruwar kowanne daga cikin furotin masu sarrafawar a tsakiya duk sun ci gaba kamar yadda suke. Katsewar ta faru mataki ɗaya bayan haka: abubuwan da aka fitar daga tsakiya sun nuna ƙarancin ɗaurin NF-kB da sashen DNA ɗinsa bayan lipopolysaccharide, da ƙarancin ɗaurin STAT1 bayan interferon-gamma. Hanyar chromatin immunoprecipitation ta tabbatar da ƙarancin kasancewar kowanne daga cikin furotin masu sarrafawar a yankin fara rubuta bayanan iNOS a cikin ƙwayoyin halitta. Tumor necrosis factor alpha, wanda yankin fara rubuta bayanansa shi ma yake amfani da NF-kB, ya ragu. An kunna furotin masu sarrafawar kuma sun isa wurin, amma ba su ɗaure da DNA da suke nufi sosai ba.

A lura da bambancin da hanyar 1. Wannan ya shafi ɗaurin furotin mai sarrafawa da DNA, yana amfani da STAT1 maimakon STAT3, kuma yana auna mRNA, furotin, da nitrite maimakon ɗigon taruwar ASC da gutsuren caspase-1. Taswirarmu ta wuraren da shuɗin methylene ke shafa a samfurorin ƙwayoyin halitta tana ware wannan matakin rubuta bayanai zuwa RNA daga ma'aunin aikin enzyme kai tsaye da ma'aunin mitochondria.

Hanya 3: IL-6 a serum da kunna STAT3 a nama

IL-6 yana aika sigina ta dangin masu karɓar gp130 zuwa jerin matakan Janus kinase zuwa STAT3. STAT3 da aka ƙara wa rukunin phosphate yana haɗuwa bibbiyu, ya shiga tsakiya, sannan ya ƙarfafa shirye-shiryen kumburi. Shi kansa IL-6 kuma abin da NF-kB ke sarrafawa ne, don haka su biyun suna samar da zagayen amsawa da ake yawan kira tsarin IL-6 zuwa STAT3. Ma'aunan da aka saba amfani da su su ne IL-6 a serum a matakin farko, rabo tsakanin STAT3 da aka ƙara wa rukunin phosphate da jimillar STAT3, da enzymes masu aiwatarwa kamar iNOS da COX2 a matakan baya.

Binciken beraye na 2023 kan IL-6 da STAT3 bayan lipopolysaccharide ya auna wannan tsarin a sassa daban-daban. Berayen C57BL/6 maza sun karɓi 1 mg per kg na lipopolysaccharide kullum na tsawon kwana uku, tare da 5 ko 10 mg per kg na shuɗin methylene mintuna 30 bayan kowane adadi. A rana ta uku, IL-6 a serum ya yi ƙasa a rukunonin shuɗin methylene biyu fiye da rukunin lipopolysaccharide kaɗai, yayin da mafi yawan sauran cytokine da aka auna ba su canza da abin da aka ba su ba. Rabo tsakanin STAT3 da aka ƙara wa rukunin phosphate da jimillar STAT3 ya yi ƙasa a cortex, hippocampus, da fatar kunne. Haka kuma, alamar Iba-1 da aka nuna ta rina microglia ta ragu, iNOS da COX2 sun ragu, kuma raguwar nauyi ta yi sauƙi zuwa wani mataki.

Akwai abubuwa biyu da ya kamata a yi taka-tsantsan da su a nan. Ƙarancin IL-6 a serum tare da ƙarancin ƙara rukunin phosphate ga STAT3 a nama ya dace da sauya wannan tsarin, amma bai ware ainihin matakin da aka fara shafa ba. Wannan saboda ƙarancin cytokine a matakin farko, ƙarancin siginar mai karɓa, ko canjin aikin phosphatase kowannensu zai iya haifar da irin wannan rabo. Kuma nauyi a tsawon kwana uku na tayar da martani mai tsanani ma'auni ne na dabbar gaba ɗaya, ba ma'aunin alamomin rashin lafiya ko lafar cuta ba. Yana ƙara mahalli fiye da hoton gwajin blot, ba tare da mai da sakamakon sigina da'awar magani ba. Hanyar sinadarai dabam ta sarrafa redox a mitochondria tana gudana a layi ɗaya da wannan bayanin cytokine, kuma bai kamata a haɗa su ba.

Kwatanta hanyoyin: wuraren tasiri da ma'aunan sakamako

Teburin yana hana amfani da sakamako ta hanyoyi uku marasa inganci. Sakamakon ɗigon taruwar ASC ba ya tabbatar da sakamakon STAT3. Sakamakon ɗaurin STAT1 da yankin fara rubuta bayanan iNOS ba ya tabbatar da sakamakon STAT3, ko da yake furotin biyun duka 'yan dangin STAT ne. Sakamakon IL-6 a serum ba ya tabbatar da raguwar haɗuwar inflammasome: a gwajin ramin ciki, IL-1beta ya ragu amma IL-6 bai ragu ba. Kowane layi yana buƙatar abin tayarwa da ma'aunin sakamako na kansa. Wannan shi ne dalilin da ya sa karanta binciken shuɗin methylene na ƙwayoyin halitta, dabbobi, da mutane ke buƙatar a daidaita kowace da'awa da matakin da aka auna.

Babu ɗaya daga cikin waɗannan da ke tabbatar da adadin amfani, aminci, ko fa'ida ga mutane. Samfurorin lipopolysaccharide abubuwan tayarwa ne da aka sarrafa don gwada hanyar aiki, ba kwafin rashin lafiya mai ɗorewa ba. Bayar da sinadari tare da abin tayarwa kuma yana gwada hana canjin da aka jawo, maimakon mayar da cutar da ta riga ta kafu baya. A ware wannan bayanin hanyar aiki daga sakamakon marasa lafiya, kuma a tantance kowace da'awar lafiya bisa binciken marasa lafiya da ke goyon bayanta.