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Hydrogen sulfide (H2S) yana da tasirin ilimin halittar jiki da na cututtuka da yawa a jikin ɗan adam. Ana amfani da Sodium hydrosulfide (NaHS) sosai a matsayin kayan aikin magani don tantance tasirin H2S a cikin gwaje-gwajen halittu. Duk da cewa asarar H2S daga maganin NaHS yana ɗaukar mintuna kaɗan kawai, an yi amfani da maganin NaHS a matsayin mahaɗan masu ba da gudummawa ga H2S a cikin ruwan sha a wasu nazarin dabbobi. Wannan binciken ya binciki ko shan ruwa mai yawan NaHS na 30 μM da aka shirya a cikin kwalaben bera/linzami zai iya kasancewa mai karko na akalla awanni 12-24, kamar yadda wasu marubuta suka ba da shawara. Shirya maganin NaHS (30 μM) a cikin ruwan sha kuma nan da nan a zuba shi a cikin kwalaben ruwan bera/linzami. An tattara samfurori daga ƙarshen da cikin kwalbar ruwa a awanni 0, 1, 2, 3, 4, 5, 6, 12 da 24 don auna abubuwan da ke cikin sulfide ta amfani da hanyar methylene blue. Bugu da ƙari, an yi wa beraye maza da mata allurar NaHS (30 μM) na tsawon makonni biyu kuma ana auna yawan sinadarin sulfide a cikin jini kowace rana a cikin makon farko da kuma ƙarshen mako na biyu. Maganin NaHS a cikin samfurin da aka samu daga ƙarshen kwalbar ruwa bai yi ƙarfi ba; ya ragu da kashi 72% da 75% bayan awanni 12 da 24, bi da bi. A cikin samfuran da aka samu daga cikin kwalaben ruwa, raguwar NaHS ba ta yi yawa ba cikin awanni 2; duk da haka, ya ragu da kashi 47% da 72% bayan awanni 12 da 24, bi da bi. Allurar NaHS ba ta shafi matakin sinadarin sulfide na jini na beraye maza da mata ba. A ƙarshe, bai kamata a yi amfani da maganin NaHS da aka shirya daga ruwan sha don bayar da gudummawar H2S ba saboda maganin ba shi da ƙarfi. Wannan hanyar bayarwa za ta fallasa dabbobi ga adadin NaHS marasa tsari da ƙanƙanta fiye da yadda ake tsammani.
An yi amfani da sinadarin hydrogen sulfide (H2S) a matsayin guba tun daga shekarar 1700; duk da haka, Abe da Kimura sun bayyana yiwuwar rawar da yake takawa a matsayin kwayar halittar biosignaling ta ciki a shekarar 1996. A cikin shekaru talatin da suka gabata, an fayyace ayyuka da yawa na H2S a cikin tsarin ɗan adam daban-daban, wanda ya haifar da fahimtar cewa ƙwayoyin masu ba da gudummawar H2S na iya samun aikace-aikacen asibiti a cikin magani ko kula da wasu cututtuka; duba Chirino et al. don sake dubawa kwanan nan.
An yi amfani da Sodium hydrosulfide (NaHS) sosai a matsayin kayan aikin magani don tantance tasirin H2S a cikin al'adun ƙwayoyin halitta da nazarin dabbobi da yawa5,6,7,8. Duk da haka, NaHS ba shine mafi kyawun mai ba da gudummawar H2S ba saboda ana canza shi cikin sauri zuwa maganin H2S/HS-in, yana gurɓata shi da polysulfides cikin sauƙi, kuma yana da sauƙin oxidize da volatilization4,9. A cikin gwaje-gwajen halittu da yawa, NaHS yana narkewa a cikin ruwa, wanda ke haifar da volatilization mara aiki da asarar H2S10,11,12, oxidation na H2S11,12,13, da photolysis14. Sulfide a cikin maganin asali yana ɓacewa da sauri saboda volatilization na H2S11. A cikin akwati buɗe, rabin rai (t1/2) na H2S yana kusan mintuna 5, kuma yawansa yana raguwa da kusan 13% a minti ɗaya10. Duk da cewa asarar hydrogen sulfide daga ruwan NaHS yana ɗaukar mintuna kaɗan ne kawai, wasu nazarin dabbobi sun yi amfani da ruwan NaHS a matsayin tushen hydrogen sulfide a cikin ruwan sha na tsawon makonni 1-21, suna maye gurbin ruwan da ke ɗauke da NaHS a kowace awa 12-24.15,16,17,18,19,20,21,22,23,24,25,26 Wannan aikin bai yi daidai da ƙa'idodin binciken kimiyya ba, tunda ya kamata a yi amfani da yawan magunguna bisa ga amfani da su a wasu nau'ikan halittu, musamman mutane.27
Binciken da aka yi kafin a fara aikin likitanci a fannin likitanci yana da nufin inganta ingancin kulawar marasa lafiya ko sakamakon magani. Duk da haka, sakamakon yawancin nazarin dabbobi ba a fassara shi zuwa ga mutane ba tukuna28,29,30. Ɗaya daga cikin dalilan da suka haifar da wannan gazawar fassara shine rashin kulawa ga ingancin hanyoyin nazarin dabbobi30. Saboda haka, manufar wannan binciken ita ce a binciki ko maganin NaHS 30 μM da aka shirya a cikin kwalaben ruwan bera/linzami zai iya kasancewa a cikin ruwan sha na tsawon awanni 12-24, kamar yadda aka yi iƙirari ko aka ba da shawara a wasu nazarin.
An gudanar da dukkan gwaje-gwajen da aka yi a cikin wannan binciken bisa ga ka'idojin da aka buga don kulawa da amfani da dabbobin dakin gwaje-gwaje a Iran31. Duk rahotannin gwaji a cikin wannan binciken sun kuma bi ka'idojin ARRIVE32. Kwamitin Ɗabi'a na Cibiyar Kimiyyar Endocrine, Jami'ar Kimiyyar Lafiya ta Shahid Beheshti, ya amince da duk hanyoyin gwaji a cikin wannan binciken.
An sayi sinadarin zinc acetate dihydrate (CAS: 5970-45-6) da kuma sinadarin anhydrous ferric chloride (CAS: 7705-08-0) daga Biochem, Chemopahrama (Cosne-sur-Loire, Faransa). An sayi sinadarin sodium hydrosulfide hydrate (CAS: 207683-19-0) da kuma N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) daga Sigma-Aldrich (St. Louis, MO, Amurka). An sayi sinadarin Isoflurane daga Piramal (Bethlehem, PA, Amurka). An sayi sinadarin hydrochloric acid (HCl) daga Merck (Darmstadt, Jamus).
Shirya maganin NaHS (30 μM) a cikin ruwan sha sannan a zuba shi nan da nan a cikin kwalaben ruwan bera/linza. An zaɓi wannan yawan dangane da wallafe-wallafe da yawa ta amfani da NaHS a matsayin tushen H2S; duba sashen Tattaunawa. NaHS kwayar halitta ce mai ruwa-ruwa wacce za ta iya ƙunsar ruwa mai ruwa-ruwa daban-daban (watau, NaHS•xH2O); a cewar masana'anta, kashi na NaHS da aka yi amfani da shi a bincikenmu shine 70.7% (watau, NaHS•1.3 H2O), kuma mun yi la'akari da wannan ƙimar a cikin lissafinmu, inda muka yi amfani da nauyin kwayoyin halitta na 56.06 g/mol, wanda shine nauyin kwayoyin halitta na NaHS mai ruwa-ruwa. Ruwan ruwa (wanda kuma ake kira ruwan lu'ulu'u) shine kwayoyin ruwa waɗanda suka samar da tsarin lu'ulu'u33. Hydrates suna da halaye na zahiri da na thermodynamic daban-daban idan aka kwatanta da anhydrates34.
Kafin a ƙara NaHS a cikin ruwan sha, a auna pH da zafin ruwan. Nan da nan a zuba maganin NaHS a cikin kwalbar ruwan bera/linzami a cikin kejin dabbobi. An tattara samfura daga ƙarshen kwalbar da kuma daga cikin kwalbar ruwan a awanni 0, 1, 2, 3, 4, 5, 6, 12, da 24 don auna yawan sinadarin sulfide. An ɗauki ma'aunin sulfide nan da nan bayan kowace samfurin. Mun sami samfura daga ƙarshen bututun saboda wasu bincike sun nuna cewa ƙaramin girman ramin bututun ruwa zai iya rage ƙafewar H2S15,19. Wannan batu ya shafi maganin da ke cikin kwalbar. Duk da haka, ba haka lamarin yake ba ga maganin da ke wuyan kwalbar ruwa, wanda ke da ƙimar ƙafewar ruwa mafi girma kuma yana sa iska ta yi ta aiki; a zahiri, dabbobin sun fara shan wannan ruwan.
An yi amfani da berayen Wistar maza da mata a cikin binciken. An ajiye berayen a cikin kejin polypropylene (bera 2-3 a kowace keji) a ƙarƙashin yanayi na yau da kullun (zafin jiki 21-26 °C, zafi 32-40%) tare da haske na awanni 12 (7 na safe zuwa 7 na yamma) da kuma duhu na awanni 12 (7 na yamma zuwa 7 na safe). Beran suna da damar samun ruwan famfo kyauta kuma an ciyar da su da abincin da aka saba (Khorak Dam Pars Company, Tehran, Iran). Mace (n=10, nauyin jiki: 190–230 g) da namiji (n=10, nauyin jiki: 320–370 g) An raba berayen Wistar bazuwar zuwa ƙungiyoyin sarrafawa da NaHS (30 μM) waɗanda aka yi wa magani (n=5 a kowace rukuni). Don tantance girman samfurin, mun yi amfani da hanyar KISS (Keep It Simple, Stupid), wacce ta haɗa gogewa ta baya da nazarin ƙarfi35. Da farko mun gudanar da wani bincike na gwaji kan beraye 3 kuma muka tantance matsakaicin matakin sulfide na jini da karkacewar da aka saba (8.1 ± 0.81 μM). Sannan, idan aka yi la'akari da ƙarfin kashi 80% kuma muka ɗauka cewa matakin mahimmanci na kashi 5% mai gefe biyu ne, mun ƙayyade girman samfurin farko (n = 5 bisa ga wallafe-wallafen da suka gabata) wanda ya yi daidai da girman tasirin da aka daidaita na 2.02 tare da ƙimar da aka riga aka ƙayyade da Festing ya ba da shawara don ƙididdige girman samfurin dabbobin gwaji35. Bayan ninka wannan ƙimar da SD (2.02 × 0.81), girman tasirin da aka annabta (1.6 μM) ya kasance 20%, wanda abin karɓa ne. Wannan yana nufin cewa n = 5/rukuni ya isa ya gano matsakaicin canji na kashi 20% tsakanin ƙungiyoyi. An raba beraye bazuwar zuwa ƙungiyoyin sarrafawa da kuma ƙungiyoyin da aka yi wa magani da NaSH ta amfani da aikin bazuwar na software na Excel 36 (Ƙarin Hoto na 1). An yi makanta a matakin sakamako, kuma masu binciken da ke yin ma'aunin sinadarai ba su san ayyukan rukuni ba.
An yi wa ƙungiyoyin NaHS na jinsi biyu magani da maganin NaHS 30 μM da aka shirya a cikin ruwan sha na tsawon makonni 2; An ba da sabon maganin a kowace sa'o'i 24, a lokacin da ake auna nauyin jiki. An tattara samfuran jini daga ƙarshen wutsiyar dukkan beraye a ƙarƙashin maganin isoflurane kowace rana a ƙarshen makonni na farko da na biyu. An sanya samfuran jini a centrifuge a 3000 g na minti 10, an raba serum ɗin kuma an adana shi a -80°C don auna urea na jini, creatinine (Cr), da jimlar sulfide. An ƙayyade urea na jini ta hanyar hanyar enzymatic urease, kuma an ƙayyade creatinine na jini ta hanyar hanyar photometric Jaffe ta amfani da kayan aiki da ake samu a kasuwa (Man Company, Tehran, Iran) da kuma na'urar nazarin atomatik (Selectra E, lambar serial 0-2124, Netherlands). Ma'aunin bambancin urea da Cr a cikin jini da Cr sun kasance ƙasa da 2.5%.
Ana amfani da hanyar methylene blue (MB) don auna jimlar sulfide a cikin ruwan sha da kuma serum ɗin da ke ɗauke da NaHS; MB ita ce hanyar da aka fi amfani da ita don auna sulfide a cikin mafita masu yawa da samfuran halittu11,37. Ana iya amfani da hanyar MB don kimanta jimlar sulfide pool38 da kuma auna sulfide marasa tsari a cikin nau'in H2S, HS- da S2 a cikin matakin ruwa39. A cikin wannan hanyar, ana haƙa sulfur a matsayin zinc sulfide (ZnS) a gaban zinc acetate11,38. Ruwan zinc acetate shine hanyar da aka fi amfani da ita don raba sulfide daga wasu chromophores11. An sake wargaza ZnS ta amfani da HCl11 a ƙarƙashin yanayin acidic mai ƙarfi. Sulfide yana amsawa da DMPD a cikin rabon stoichiometric na 1:2 a cikin amsawar da ferric chloride ya haɓaka (Fe3+ yana aiki azaman wakili mai hana iska) don samar da rini MB, wanda aka gano ta hanyar spectrophotometric a 670 nm40,41. Iyakar gano hanyar MB shine kimanin 1 μM11.
A cikin wannan binciken, an ƙara 100 μL na kowane samfurin (maganin ko magani) a cikin bututu; sannan aka ƙara 200 μL na zinc acetate (1% w/v a cikin ruwan da aka tace), 100 μL na DMPD (20 mM a cikin 7.2 M HCl), da 133 μL na FeCl3 (30 mM a cikin 1.2 M HCl). An saka cakuda a 37°C a cikin duhu na tsawon minti 30. An sanya centrifuge a 10,000 g na tsawon minti 10, kuma an karanta shan ruwan a 670 nm ta amfani da na'urar karanta microplate (BioTek, MQX2000R2, Winooski, VT, Amurka). An ƙayyade yawan sulfide ta amfani da lanƙwasa na daidaitawa na NaHS (0–100 μM) a cikin ddH2O (Hoto na Ƙari na 2). Duk mafita da aka yi amfani da su don ma'aunin an shirya su sabo. Ma'aunin bambancin da ke tsakanin gwaje-gwajen da kuma gwaje-gwajen da aka yi amfani da su wajen auna sulfide ya kasance kashi 2.8% da 3.4%, bi da bi. Mun kuma tantance jimillar sulfide da aka samu daga ruwan sha da kuma samfuran jini da ke dauke da sodium thiosulfate ta amfani da hanyar samfurin da aka ƙarfafa42. An gano ruwan sha da samfuran jini da ke dauke da sodium thiosulfate sun kasance kashi 91 ± 1.1% (n = 6) da kuma kashi 93 ± 2.4% (n = 6), bi da bi.
An gudanar da nazarin ƙididdiga ta amfani da software na GraphPad Prism sigar 8.0.2 don Windows (GraphPad Software, San Diego, CA, Amurka, www.graphpad.com). An yi amfani da gwajin t-haɗi don kwatanta zafin jiki da pH na ruwan sha kafin da kuma bayan ƙara NaHS. An ƙididdige asarar H2S a cikin maganin da ke ɗauke da NaHS a matsayin raguwar kashi daga shan ruwa na farko, kuma don tantance ko asarar tana da mahimmanci a kididdiga, mun yi ANOVA mai maimaitawa ta hanya ɗaya sannan gwajin kwatantawa da yawa na Dunnett. An kwatanta nauyin jiki, urea na jini, creatinine na jini, da jimillar sulfide na jini a tsawon lokaci tsakanin berayen da aka yi wa magani da NaHS na jinsi daban-daban ta amfani da ANOVA mai gauraya (tsakanin-ciki) sannan gwajin Bonferroni bayan hoc. An ɗauki ƙimar P mai wutsiya biyu < 0.05 a matsayin mahimmanci a kididdiga.
pH na ruwan sha shine 7.60 ± 0.01 kafin ƙara NaHS da 7.71 ± 0.03 bayan ƙara NaHS (n = 13, p = 0.0029). Zafin ruwan sha shine 26.5 ± 0.2 kuma ya ragu zuwa 26.2 ± 0.2 bayan ƙara NaHS (n = 13, p = 0.0128). Shirya maganin NaHS 30 μM a cikin ruwan sha kuma adana shi a cikin kwalban ruwa. Maganin NaHS ba shi da ƙarfi kuma yawansa yana raguwa akan lokaci. Lokacin da aka ɗauki samfurin daga wuyan kwalban ruwa, an lura da raguwa mai yawa (68.0%) a cikin awa ɗaya na farko, kuma abun ciki na NaHS a cikin maganin ya ragu da 72% da 75% bayan awanni 12 da 24, bi da bi. A cikin samfuran da aka samo daga kwalaben ruwa, raguwar NaHS ba ta da mahimmanci har zuwa awanni 2, amma bayan awanni 12 da 24 ya ragu da 47% da 72%, bi da bi. Waɗannan bayanai sun nuna cewa kaso na NaHS a cikin maganin 30 μM da aka shirya a cikin ruwan sha ya ragu zuwa kusan kashi ɗaya cikin huɗu na ƙimar farko bayan awanni 24, ba tare da la'akari da wurin da aka ɗauki samfurin ba (Hoto na 1).
Kwanciyar ruwan NaHS (30 μM) a cikin ruwan sha a cikin kwalaben bera/linzami. Bayan shirya maganin, an ɗauki samfura daga ƙarshen kwalbar da kuma cikin kwalbar. An gabatar da bayanai a matsayin matsakaicin ± SD (n = 6/rukuni). * da #, P < 0.05 idan aka kwatanta da lokaci 0. Hoton kwalbar ruwan yana nuna ƙarshen (tare da buɗewa) da jikin kwalbar. Girman ƙarshen shine kimanin 740 μL.
Yawan NaHS a cikin maganin 30 μM da aka shirya kwanan nan shine 30.3 ± 0.4 μM (tsawo: 28.7–31.9 μM, n = 12). Duk da haka, bayan awanni 24, yawan NaHS ya ragu zuwa ƙarancin ƙima (matsakaicin: 3.0 ± 0.6 μM). Kamar yadda aka nuna a Hoto na 2, yawan NaHS da berayen suka fallasa ba su da tabbas a lokacin binciken.
Nauyin jikin beraye mata ya ƙaru sosai a tsawon lokaci (daga 205.2 ± 5.2 g zuwa 213.8 ± 7.0 g a cikin ƙungiyar kulawa da kuma daga 204.0 ± 8.6 g zuwa 211.8 ± 7.5 g a cikin ƙungiyar da NaHS ya yi wa magani); duk da haka, maganin NaHS bai yi wani tasiri ga nauyin jiki ba (Hoto na 3). Nauyin jikin beraye maza ya ƙaru sosai a tsawon lokaci (daga 338.6 ± 8.3 g zuwa 352.4 ± 6.0 g a cikin ƙungiyar kulawa da kuma daga 352.4 ± 5.9 g zuwa 363.2 ± 4.3 g a cikin ƙungiyar da NaHS ya yi wa magani); duk da haka, maganin NaHS bai yi wani tasiri ga nauyin jiki ba (Hoto na 3).
Canje-canje a cikin nauyin jiki a cikin beraye mata da maza bayan shan NaHS (30 μM). An gabatar da bayanai a matsayin matsakaicin ± SEM kuma an kwatanta su ta amfani da nazarin bambance-bambancen da aka haɗa ta hanyoyi biyu (cikin-tsakanin) tare da gwajin Bonferroni bayan hoc. n = 5 na kowace jinsi a cikin kowace ƙungiya.
Yawan urea da creatine phosphate a cikin jini sun yi daidai a cikin sarrafawa da kuma beraye da aka yi wa magani da NaSH a duk tsawon binciken. Bugu da ƙari, maganin NaSH bai shafi yawan urea da creatinechrome a cikin jini ba (Tebur 1).
Jimlar yawan sinadarin sulfide a cikin jini ya kasance iri ɗaya tsakanin berayen da aka yi wa magani da NaHS (8.1 ± 0.5 μM vs. 9.3 ± 0.2 μM) da na mace (9.1 ± 1.0 μM vs. 6.1 ± 1.1 μM). Shan NaHS na tsawon kwanaki 14 bai yi wani tasiri ga jimillar matakan sulfide a cikin jini ba a cikin berayen maza ko mata (Hoto na 4).
Canje-canje a cikin jimlar yawan sulfide a cikin jini a cikin beraye maza da mata bayan shan NaHS (30 μM). An gabatar da bayanai a matsayin matsakaicin ± SEM kuma an kwatanta su ta amfani da nazarin bambance-bambancen da aka haɗa ta hanyoyi biyu (cikin-ciki) tare da gwajin Bonferroni bayan hoc. Kowane jinsi, n = 5/rukuni.
Babban sakamakon wannan binciken shi ne cewa ruwan sha da ke ɗauke da NaHS ba shi da ƙarfi: kusan kashi ɗaya bisa huɗu na jimillar sinadarin sulfide na farko ne kawai za a iya gano sa'o'i 24 bayan an ɗauki samfurin daga ƙarshen kwalban ruwan bera/linzami. Bugu da ƙari, beraye sun fuskanci yawan sinadarin NaHS mara ƙarfi saboda asarar H2S a cikin ruwan NaHS, kuma ƙara NaHS a cikin ruwan sha bai shafi nauyin jiki ba, urea na jini da creatine chromium, ko kuma jimillar sinadarin sulfide a cikin jini ba.
A cikin wannan binciken, ƙimar asarar H2S daga maganin NaHS 30 μM da aka shirya a cikin ruwan sha ya kai kusan kashi 3% a kowace awa. A cikin maganin da aka buffered (100 μM sodium sulfide a cikin 10 mM PBS, pH 7.4), an ruwaito cewa yawan sulfide ya ragu da kashi 7% a tsawon lokaci a cikin awanni 8. Mun riga mun kare gudanar da NaHS a cikin mahaifa ta hanyar bayar da rahoton cewa ƙimar asarar sulfide daga maganin NaHS 54 μM a cikin ruwan sha ya kasance kusan kashi 2.3% a kowace awa (4%/awa a cikin awanni 12 na farko da 1.4%/awa a cikin awanni 12 na ƙarshe bayan shiri)8. Nazarin da aka yi a baya43 ya gano asarar H2S akai-akai daga maganin NaHS, galibi saboda volatilization da oxidation. Ko da ba tare da ƙara kumfa ba, sulfide a cikin maganin yana ɓacewa da sauri saboda volatilization H2S11. Bincike ya nuna cewa a lokacin tsarin narkewar, wanda ke ɗaukar kimanin daƙiƙa 30-60, kusan kashi 5-10% na H2S yana ɓacewa saboda ƙafewa6. Don hana ƙafewa H2S daga maganin, masu bincike sun ɗauki matakai da dama, ciki har da jujjuyawar maganin a hankali12, rufe maganin da filastik6, da rage fallasa maganin ga iska, tunda ƙimar ƙafewa H2S ya dogara da haɗin iska da ruwa.13 Iskar shaka ta H2S ba zato ba tsammani tana faruwa ne galibi saboda ions na ƙarfe masu canzawa, musamman ƙarfe mai kama da ƙarfe, waɗanda ƙazanta ne a cikin ruwa.13 Iskar shaka ta H2S tana haifar da samuwar polysulfides (atom ɗin sulfur da aka haɗa ta hanyar haɗin covalent)11. Domin guje wa iskar shaka, ana shirya mafita masu ɗauke da H2S a cikin abubuwan narkewar da aka cire daga iskar shaka44,45 sannan a tsaftace su da argon ko nitrogen na tsawon mintuna 20-30 don tabbatar da rage iskar shaka.11,12,37,44,45,46 Diethylenetriaminepentaacetic acid (DTPA) wani abu ne mai ƙarfe (10-4 M) wanda ke hana HS- autooxidation a cikin mafita na aerobic. Idan babu DTPA, ƙimar autooxidation na HS- yana kusan 50% a cikin kimanin awanni 3 a 25°C37,47. Bugu da ƙari, tunda iskar shaka ta 1e-sulfide tana da tasirin hasken ultraviolet, ya kamata a adana maganin a kan kankara kuma a kare shi daga haske11.
Kamar yadda aka nuna a Hoto na 5, NaHS yana rabuwa zuwa Na+ da HS-6 lokacin da aka narkar da shi a cikin ruwa; wannan rabuwar ana ƙaddara ta hanyar pK1 na amsawar, wanda ya dogara da zafin jiki: pK1 = 3.122 + 1132/T, inda T ke tsakanin 5 zuwa 30°C kuma ana auna shi a digiri Kelvin (K), K = °C + 273.1548. HS- yana da babban pK2 (pK2 = 19), don haka a pH < 96.49, S2- ba a samar da shi ko kuma an samar da shi a ƙananan adadi. Sabanin haka, HS- yana aiki a matsayin tushe kuma yana karɓar H+ daga ƙwayar H2O, kuma H2O yana aiki a matsayin acid kuma ana canza shi zuwa H2S da OH-.
Samuwar iskar gas ta H2S da aka narkar a cikin ruwan NaHS (30 µM). aq, ruwan magani; g, iskar gas; l, ruwa. Duk lissafin yana ɗauka cewa pH na ruwa = 7.0 da zafin ruwa = 20 °C. An ƙirƙira shi da BioRender.com.
Duk da shaidar da ke nuna cewa maganin NaHS ba shi da ƙarfi, binciken dabbobi da dama sun yi amfani da maganin NaHS a cikin ruwan sha a matsayin mahaɗin mai ba da gudummawa na H2S15,16,17,18,19,20,21,22,23,24,25,26 tare da tsawon lokacin shiga tsakani daga makonni 1 zuwa 21 (Tebur 2). A lokacin waɗannan nazarin, an sabunta maganin NaHS a kowace sa'o'i 12, 15, 17, 18, 24, 25 ko sa'o'i 24, 19, 20, 21, 22, 23. Sakamakonmu ya nuna cewa beraye sun fuskanci yawan magunguna marasa ƙarfi saboda asarar H2S daga maganin NaHS, kuma abubuwan da ke cikin ruwan sha na beraye sun canza sosai a cikin sa'o'i 12 ko 24 (duba Hoto na 2). Biyu daga cikin waɗannan nazarin sun ba da rahoton cewa matakan H2S a cikin ruwa sun kasance daidai a cikin awanni 24 ko kuma cewa asarar H2S kashi 2-3% ne kawai aka lura a cikin awanni 12, amma ba su bayar da bayanai masu goyan baya ko cikakkun bayanai na aunawa ba. Nazari biyu sun nuna cewa ƙaramin diamita na kwalaben ruwa na iya rage ƙafewar H2S15,19. Duk da haka, sakamakonmu ya nuna cewa wannan na iya jinkirta asarar H2S daga kwalban ruwa da awanni 2 maimakon awanni 12-24. Duk waɗannan binciken sun lura cewa mun ɗauka cewa matakin NaHS a cikin ruwan sha bai canza ba saboda ba mu lura da canjin launi a cikin ruwan ba; saboda haka, iskar shakar H2S ta iska ba ta da mahimmanci19,20. Abin mamaki, wannan hanyar tunani tana tantance daidaiton NaHS a cikin ruwa maimakon auna canjin yawansa akan lokaci.
Asarar H2S a cikin maganin NaHS yana da alaƙa da pH da zafin jiki. Kamar yadda aka lura a cikin bincikenmu, narkar da NaHS a cikin ruwa yana haifar da samuwar maganin alkaline50. Lokacin da NaHS ya narke a cikin ruwa, samuwar iskar H2S da aka narkar ya dogara da ƙimar pH6. Ƙananan pH na maganin, mafi girman adadin NaHS da ke kasancewa a matsayin ƙwayoyin iskar H2S kuma ana rasa ƙarin sulfide daga maganin ruwa11. Babu ɗayan waɗannan nazarin da ya ba da rahoton pH na ruwan sha da aka yi amfani da shi azaman mai narkewa ga NaHS. A cewar shawarwarin WHO, waɗanda yawancin ƙasashe suka amince da su, pH na ruwan sha ya kamata ya kasance a cikin kewayon 6.5–8.551. A cikin wannan kewayon pH, ƙimar iskar shaye-shaye ta H2S ta tashi kusan ninki goma13. Narkar da NaHS a cikin ruwa a cikin wannan kewayon pH zai haifar da yawan iskar H2S da aka narkar daga 1 zuwa 22.5 μM, wanda ke jaddada mahimmancin sa ido kan pH na ruwan kafin narkar da NaHS. Bugu da ƙari, yanayin zafin da aka ruwaito a cikin binciken da ke sama (18–26 °C) zai haifar da canji a cikin yawan iskar gas ta H2S da aka narkar a cikin maganin kusan 10%, tunda canje-canjen zafin jiki suna canza pK1, kuma ƙananan canje-canje a cikin pK1 na iya yin tasiri mai mahimmanci akan yawan iskar gas ta H2S da aka narkar48. Bugu da ƙari, tsawon lokacin wasu nazarin (watanni 5)22, wanda a lokacin ake tsammanin babban bambancin zafin jiki, shi ma yana ƙara ta'azzara wannan matsalar.
Duk wani bincike banda one21 ya yi amfani da maganin NaHS 30 μM a cikin ruwan sha. Don bayyana adadin da aka yi amfani da shi (watau 30 μM), wasu marubuta sun nuna cewa NaHS a cikin matakin ruwa yana samar da daidai adadin iskar H2S kuma cewa kewayon ilimin halittar H2S shine 10 zuwa 100 μM, don haka wannan adadin yana cikin kewayon ilimin halittar jiki15,16. Wasu sun bayyana cewa 30 μM NaHS na iya kiyaye matakin H2S na plasma a cikin kewayon ilimin halittar jiki, watau 5-300 μM19,20. Mun yi la'akari da yawan NaHS a cikin ruwa na 30 μM (pH = 7.0, T = 20 °C), wanda aka yi amfani da shi a wasu nazarin don nazarin tasirin H2S. Za mu iya ƙididdige cewa yawan iskar H2S da aka narkar shine 14.7 μM, wanda shine kusan 50% na yawan NaHS na farko. Wannan ƙimar tana kama da ƙimar da wasu marubuta suka ƙididdige a ƙarƙashin yanayi iri ɗaya13,48.
A cikin bincikenmu, gudanar da NaHS bai canza nauyin jiki ba; wannan sakamakon ya yi daidai da sakamakon wasu bincike a kan beraye maza22,23 da beraye maza18; Duk da haka, bincike biyu sun ba da rahoton cewa NaSH ya dawo da raguwar nauyin jiki a cikin beraye da aka yi wa tiyata24,26, yayin da wasu bincike ba su bayar da rahoton tasirin gudanar da NaSH akan nauyin jiki15,16,17,19,20,21,25 ba. Bugu da ƙari, a cikin bincikenmu, gudanar da NaSH bai shafi matakan urea da creatine chromium na jini ba, wanda ya yi daidai da sakamakon wani rahoto25.
Binciken ya gano cewa ƙara NaHS a cikin ruwan sha na tsawon makonni 2 bai shafi jimillar yawan sinadarin sulfide a cikin beraye maza da mata ba. Wannan binciken ya yi daidai da sakamakon Sen et al. (16): makonni 8 na magani da 30 μM NaHS a cikin ruwan sha bai shafi matakan sulfide na plasma a cikin beraye masu sarrafawa ba; duk da haka, sun ba da rahoton cewa wannan shiga tsakani ya dawo da raguwar matakan H2S a cikin jini na beraye masu nephrectomized. Li et al. (22) sun kuma ba da rahoton cewa magani da 30 μM NaHS a cikin ruwan sha na tsawon watanni 5 ya ƙara matakan sulfide marasa plasma a cikin beraye tsofaffi da kusan 26%. Sauran nazarin ba su bayar da rahoton canje-canje a cikin sulfide mai yawo ba bayan ƙara NaHS a cikin ruwan sha.
Nazarce-nazarce bakwai sun ruwaito ta amfani da Sigma NaHS15,16,19,20,21,22,23 amma ba su bayar da ƙarin bayani game da ruwan da ke ɗauke da sinadarin ba, kuma nazarce-nazarce guda biyar ba su ambaci tushen NaHS da ake amfani da shi a hanyoyin shirya su ba17,18,24,25,26. NaHS kwayar halitta ce mai ɗauke da sinadarin kuma ruwan da ke ɗauke da sinadarin na iya bambanta, wanda ke shafar adadin NaHS da ake buƙata don shirya maganin wani sinadarin da aka bayar. Misali, abubuwan da ke ɗauke da sinadarin NaHS a cikin bincikenmu sune NaHS•1.3 H2O. Don haka, ainihin yawan NaHS a cikin waɗannan nazarce-nazarcen na iya zama ƙasa da waɗanda aka ruwaito.
"Ta yaya irin wannan sinadari mai ɗan gajeren lokaci zai iya yin tasiri mai ɗorewa?" Pozgay da abokan aikinsa 21 sun yi wannan tambayar lokacin da suke kimanta tasirin NaHS akan colitis a cikin beraye. Suna fatan cewa nazarin da za a yi nan gaba zai iya amsa wannan tambayar kuma ya yi hasashen cewa mafita na NaHS na iya ƙunsar ƙarin polysulfides masu karko ban da H2S da disulfides waɗanda ke shiga tsakani kan tasirin NaHS21. Wata yuwuwar kuma ita ce ƙarancin yawan NaHS da ke cikin maganin na iya samun tasiri mai amfani. A zahiri, Olson da abokan aikinsa sun ba da shaida cewa matakan micromolar na H2S a cikin jini ba na jiki ba ne kuma ya kamata su kasance a cikin kewayon nanomolar ko kuma ba su nan gaba ɗaya13. H2S na iya aiki ta hanyar furotin sulfation, wani canji mai canzawa bayan fassara wanda ke shafar aiki, kwanciyar hankali, da kuma gano sunadaran da yawa52,53,54. A gaskiya ma, a ƙarƙashin yanayin ilimin halittar jiki, kusan kashi 10% zuwa 25% na furotin hanta da yawa suna sulfylated53. Duk binciken biyu sun amince da lalata NaHS19,23 cikin sauri amma abin mamaki sun bayyana cewa "mun sarrafa yawan NaHS a cikin ruwan sha ta hanyar maye gurbinsa kowace rana."23 Wani bincike ya bayyana ba zato ba tsammani cewa "NaHS misali ne na bayar da gudummawar H2S kuma ana amfani da shi a aikace a asibiti don maye gurbin H2S da kansa."18
Tattaunawar da ke sama ta nuna cewa NaHS yana ɓacewa daga ruwan ta hanyar volatilization, oxidation da photolysis, don haka an ba da wasu shawarwari don rage asarar H2S daga ruwan. Na farko, fitar da H2S ya dogara ne akan haɗin gas-ruwa13 da pH na ruwan11; saboda haka, don rage asarar tururin, ana iya sanya wuyan kwalbar ruwa ƙanƙanta kamar yadda aka bayyana a baya15,19, kuma ana iya daidaita pH na ruwan zuwa iyakar sama mai karɓuwa (watau, 6.5–8.551) don rage asarar tururin11. Na biyu, iskar shaka ta H2S ta bazata tana faruwa ne saboda tasirin iskar oxygen da kasancewar ions na ƙarfe masu canzawa a cikin ruwan sha13, don haka cire iskar shaka ta hanyar amfani da argon ko nitrogen44,45 da amfani da chelators na ƙarfe37,47 na iya rage iskar shaka ta sulfide. Na uku, don hana rugujewar H2S, ana iya naɗe kwalaben ruwa da foil na aluminum; Wannan aikin ya shafi kayan da ke da sauƙin haske kamar streptozotocin55. A ƙarshe, ana iya ba da gishirin sulfide marasa amfani (NaHS, Na2S, da CaS) ta hanyar gavage maimakon narkewa a cikin ruwan sha kamar yadda aka ruwaito a baya56,57,58; bincike ya nuna cewa sodium sulfide mai rediyoaktif da aka bayar ta hanyar gavage ga beraye yana sha sosai kuma yana rarrabawa ga kusan dukkan kyallen takarda59. Zuwa yanzu, yawancin bincike sun ba da gishirin sulfide marasa amfani a cikin fitsari; duk da haka, ba kasafai ake amfani da wannan hanyar a wuraren asibiti60 ba. A gefe guda kuma, hanyar baki ita ce hanyar da aka fi amfani da ita kuma aka fi so a cikin mutane61. Saboda haka, muna ba da shawarar kimanta tasirin masu ba da gudummawar H2S a cikin beraye ta hanyar gavage ta baki.
Iyakancewa shine mun auna sulfide a cikin ruwan da aka yi amfani da shi da kuma maganin ta hanyar amfani da hanyar MB. Hanyoyin auna sulfide sun haɗa da titration na iodine, spectrophotometry, hanyar electrochemical (potentiometry, amperometry, hanyar coulometric da hanyar amperometric) da chromatography (gas chromatography da high-performance liquid chromatography), daga cikinsu hanyar da aka fi amfani da ita ita ce hanyar MB spectrophotometric62. Iyakance hanyar MB don auna H2S a cikin samfuran halittu shine cewa tana auna duk mahaɗan da ke ɗauke da sulfur kuma ba kyauta H2S63 ba saboda ana yin ta a ƙarƙashin yanayin acidic, wanda ke haifar da cire sulfur daga tushen halittu64. Duk da haka, a cewar Ƙungiyar Lafiyar Jama'a ta Amurka, MB ita ce hanyar da aka saba amfani da ita don auna sulfide a cikin ruwa65. Saboda haka, wannan iyakancewa ba ta shafi manyan sakamakonmu kan rashin daidaiton mafita da ke ɗauke da NaHS ba. Bugu da ƙari, a cikin bincikenmu, dawo da ma'aunin sulfide a cikin samfuran ruwa da na jini da ke ɗauke da NaHS shine 91% da 93%, bi da bi. Waɗannan ƙimar sun yi daidai da jeri da aka ruwaito a baya (77–92)66, wanda ke nuna daidaiton nazari mai karɓuwa42. Ya kamata a lura cewa mun yi amfani da beraye maza da mata bisa ga jagororin Cibiyar Lafiya ta Ƙasa (NIH) don guje wa dogaro da nazarin dabbobi na maza kawai a cikin nazarin asibiti67 da kuma haɗa beraye maza da mata duk lokacin da zai yiwu68. Wasu sun jaddada wannan batu69,70,71.
A ƙarshe, sakamakon wannan binciken ya nuna cewa ba za a iya amfani da maganin NaHS da aka shirya daga ruwan sha don samar da H2S ba saboda rashin kwanciyar hankali. Wannan hanyar da ake amfani da ita wajen ba da magani za ta fallasa dabbobi ga matakan NaHS marasa ƙarfi da ƙasa da yadda ake tsammani; saboda haka, binciken ba zai iya amfani da mutane ba.
Bayanan da aka yi amfani da su da/ko aka yi nazari a kansu a lokacin binciken na yanzu suna samuwa daga marubucin da ya dace bisa ga buƙata mai ma'ana.
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