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Hydrogen sulfide (H2S) alina ebikosa omubiri n’obulwadde ebingi ku mubiri gw’omuntu. Sodium hydrosulfide (NaHS) ekozesebwa nnyo ng’ekintu eky’eddagala okwekenneenya ebiva mu H2S mu kugezesa ebiramu. Wadde ng’okufiirwa H2S okuva mu biwujjo bya NaHS kitwala eddakiika ntono zokka, ebisengejja bya NaHS bibadde bikozesebwa ng’ebirungo ebigaba H2S mu mazzi ag’okunywa mu kunoonyereza okumu ku bisolo. Okunoonyereza kuno kwanoonyereza oba amazzi ag’okunywa nga galina NaHS concentration ya 30 μM agategekeddwa mu bucupa bw’envunza/ebibe gasobola okusigala nga tegakyukakyuka okumala waakiri essaawa 12–24, nga bwe kyateesebwa abawandiisi abamu. Tegeka ekisengejjero kya NaHS (30 μM) mu mazzi ag’okunywa era amangu ago oyiwe mu bidomola by’amazzi g’envunza/ebibe. Sampuli zaakuŋŋaanyizibwa okuva ku nsonga ne munda mu ccupa y’amazzi ku ssaawa 0, 1, 2, 3, 4, 5, 6, 12 ne 24 okupima ebirimu salufayidi nga tukozesa enkola ya bbululu wa methylene. Okugatta ku ekyo, envunza ensajja n’enkazi zaafukibwako NaHS (30 μM) okumala wiiki bbiri era ebipimo bya serum sulfide byapimibwa buli luvannyuma lwa nnaku bbiri mu wiiki esooka ne ku nkomerero ya wiiki eyookubiri. Ekisengejjo kya NaHS mu sampuli eyafunibwa okuva ku nsonga y’eccupa y’amazzi tekyali kinywevu; kyakendeera ebitundu 72% ne 75% oluvannyuma lw’essaawa 12 ne 24. Mu sampuli ezafunibwa okuva munda mu bidomola by’amazzi, okukendeera kwa NaHS tekwali kwa maanyi mu ssaawa 2; wabula kyakendeera ebitundu 47% ne 72% oluvannyuma lw’essaawa 12 ne 24. Empiso ya NaHS teyakosa mutindo gwa serum sulfide mu nvunza ensajja n’enkazi. Mu kumaliriza, ebizigo bya NaHS ebitegekebwa okuva mu mazzi ag’okunywa tebirina kukozesebwa mu kuwaayo H2S kubanga eddagala lino tebinywevu. Ekkubo lino ery’okubigaba lijja kussa ebisolo ku bungi bwa NaHS obutali bwa bulijjo era obutono okusinga bwe busuubirwa.
Hydrogen sulfide (H2S) ekozesebwa ng’obutwa okuva mu mwaka gwa 1700; wabula, omulimu gwayo ogusoboka nga molekyu y’obubonero bw’obulamu obw’omunda gwannyonnyolwa Abe ne Kimura mu 1996. Mu myaka amakumi asatu egiyise, emirimu mingi egya H2S mu nkola z’abantu ez’enjawulo gibadde ginnyonnyoddwa, ekiviiriddeko okukimanya nti molekyu ezigaba H2S ziyinza okuba n’okukozesebwa mu bujjanjabi mu kujjanjaba oba okuddukanya endwadde ezimu; laba Chirino ne banne. okusobola okwekenneenya gye buvuddeko.
Sodium hydrosulfide (NaHS) ebadde ekozesebwa nnyo ng’ekintu eky’eddagala okwekenneenya ebiva mu H2S mu kunoonyereza kungi okw’okukuza obutoffaali n’ebisolo5,6,7,8. Naye NaHS si mugabi wa H2S mulungi kubanga ekyusibwa mangu n’efuuka H2S/HS- mu solution, kyangu okufuuka ekicaafu ne polysulfides, era kyangu okufuuka oxidized ne volatilized4,9. Mu kugezesa kungi okw’ebiramu, NaHS esaanuuka mu mazzi, ekivaamu okuwuuma okutali kwa maanyi n’okufiirwa H2S10,11,12, okufuuka okw’okwekolako okwa H2S11,12,13, n’okusengejja ekitangaala14. Sulfide mu solution eyasooka ebulwa mangu nnyo olw’okufuumuuka kwa H2S11. Mu kibya ekiggule, ekitundu ky’obulamu (t1/2) ekya H2S kiba ddakiika nga 5, era obuzito bwayo bukendeera ebitundu nga 13% buli ddakiika10. Newankubadde okufiirwa kwa hydrogen sulfide okuva mu NaHS solutions kutwala eddakiika ntono zokka, okunoonyereza okumu ku bisolo kukozesezza NaHS solutions nga ensibuko ya hydrogen sulfide mu mazzi ag’okunywa okumala wiiki 1–21, nga zikyusa solution erimu NaHS buli ssaawa 12–24.15,16,17,18,19,20,21,22,23,24,25,26 Enkola eno eri tekikwatagana na misingi gya kunoonyereza kwa ssaayansi, okuva emiwendo gy’eddagala bwe girina okusinziira ku nkozesa yaago mu bika ebirala naddala abantu.27
Okunoonyereza okusooka mu bujjanjabi bw’ebiramu kugenderera okutumbula omutindo gw’okulabirira omulwadde oba ebiva mu bujjanjabi. Wabula ebivudde mu kunoonyereza okusinga ku bisolo tebinnaba kuvvuunulwa mu bantu28,29,30. Emu ku nsonga lwaki okuvvuunula kuno kulemererwa kwe butafaayo ku mutindo gw’enkola y’okunoonyereza ku bisolo30. Nolwekyo, ekigendererwa ky’okunoonyereza kuno kyali kya kunoonyereza oba 30 μM NaHS solutions ezitegekeddwa mu bucupa bw’amazzi g’envunza/ebibe zisobola okusigala nga zinywevu mu mazzi ag’okunywa okumala 12–24 h, nga bwe kyagambibwa oba bwe kyateesebwa mu kunoonyereza okumu.
Okugezesa kwonna mu kunoonyereza kuno kwakolebwa nga kugoberera ebiragiro ebyafulumiziddwa ku ndabirira n’okukozesa ebisolo bya laboratory mu Iran31. Lipoota zonna ez’okugezesa mu kunoonyereza kuno nazo zagoberera enkola ya ARRIVE32. Akakiiko akakwasisa empisa mu Institute of Endocrine Sciences, Shahid Beheshti University of Medical Sciences, kakkiriza enkola zonna ez’okugezesa mu kunoonyereza kuno.
Zinc acetate dihydrate (CAS: 5970-45-6) ne ferric chloride atalina mazzi (CAS: 7705-08-0) byagulibwa okuva mu Biochem, Chemopahrama (Cosne-sur-Loire, Bufalansa). Sodium hydrosulfide hydrate (CAS: 207683-19-0) ne N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) byagulibwa okuva mu Sigma-Aldrich (St. Louis, MO, Amerika). Isoflurane yagulibwa okuva mu Piramal (Bethlehem, PA, USA). Asidi wa hydrochloric (HCl) yagulibwa okuva mu kkampuni ya Merck (Darmstadt, Germany).
Tegeka ekisengejjero kya NaHS (30 μM) mu mazzi ag’okunywa era amangu ago oyiwe mu bidomola by’amazzi g’envunza/ebibe. Ekisengejjo kino kyalondebwa okusinziira ku bitabo bingi ebifulumiziddwa nga tukozesa NaHS ng’ensibuko ya H2S; laba ekitundu eky’okukubaganya ebirowoozo. NaHS molekyu erimu amazzi esobola okubeeramu amazzi ag’amazzi ag’enjawulo (kwe kugamba, NaHS•xH2O); okusinziira ku omukozi, ebitundu bya NaHS ebyakozesebwa mu kunoonyereza kwaffe byali 70.7% (kwe kugamba, NaHS•1.3 H2O), era omuwendo guno twagutwala mu nkola mu kubala kwaffe, gye twakozesa obuzito bwa molekyu obwa 56.06 g/mol, nga buno bwe buzito bwa molekyu bwa NaHS etaliimu mazzi. Amazzi g’amazzi (agayitibwa amazzi g’obutafaali) ge molekyo z’amazzi ezikola ensengekera ya kirisitaalo33. Hydrates zirina eby’enjawulo eby’omubiri n’eby’ebbugumu bw’ogeraageranya ne anhydrates34.
Nga tonnaba kuteeka NaHS mu mazzi g’okunywa, pima pH n’ebbugumu ly’ekizimbulukusa. Amangu ago yiwa eddagala lya NaHS mu ccupa y’amazzi g’envunza/ebibe mu kiyumba ky’ebisolo. Sampuli zaakuŋŋaanyizibwa okuva ku ntikko ne munda mu ccupa y’amazzi ku ssaawa 0, 1, 2, 3, 4, 5, 6, 12, ne 24 okupima ekirungo kya sulfide. Ebipimo bya sulfide byakolebwa amangu ddala nga buli luvannyuma lw’okutwala sampuli. Twafuna sampuli okuva ku ntikko ya ttanka kubanga okunoonyereza okumu kulaga nti obutuli obutono obwa ttanka y’amazzi busobola okukendeeza ku kufuumuuka kwa H2S15,19. Ensonga eno erabika ekwata ne ku solution eri mu ccupa. Wabula kino tekyali bwe kityo ku solution mu bulago bw’eccupa y’amazzi, eyalina omutindo gw’okufuumuuka ogw’amaanyi era nga gukola autoxidizing; mu butuufu, ebisolo bye byasooka okunywa amazzi gano.
Envunza ensajja n’enkazi eza Wistar ze zaakozesebwa mu kunoonyereza kuno. Envunza zaateekebwa mu biyumba bya polypropylene (envunza 2–3 buli kiyumba) mu mbeera eza bulijjo (ebbugumu 21–26 °C, obunnyogovu 32–40%) nga zirina ekitangaala essaawa 12 (7 ez’oku makya okutuuka ku ssaawa 7 ez’emisana) n’essaawa 12 ez’ekizikiza (7 ez’emisana okutuuka ku ssaawa 7 ez’oku makya). Envunza zaali zifuna amazzi ga ttaapu ku bwereere era nga ziriisibwa chow eya bulijjo (Khorak Dam Pars Company, Tehran, Iran). Envunza za Wistar ezikwatagana (emyezi 6) (n=10, obuzito bw’omubiri: 190–230 g) n’ensajja (n=10, obuzito bw’omubiri: 320–370 g) zaagabanyizibwamu mu ngeri ey’ekifuulannenge mu bibinja ebifuga ne NaHS (30 μM) ebyajjanjabwa (n=5 buli kibinja). Okuzuula obunene bwa sampuli, twakozesa enkola ya KISS (Keep It Simple, Stupid), egatta obumanyirivu obw’emabega n’okwekenneenya amaanyi35. Twasooka kukola okunoonyereza okw’okugezesa ku nvunza 3 ne tuzuula omutindo gwa serum total sulfide level ne standard deviation (8.1 ± 0.81 μM). Awo, nga tulowooza ku maanyi ga 80% era nga tufudde nti omutindo gw’amakulu ga 5% ogw’enjuyi bbiri, twasalawo obunene bwa sampuli obusookerwako (n = 5 okusinziira ku biwandiiko eby’emabega) obwali bukwatagana n’obunene bw’ekikolwa obutuufu obwa 2.02 n’omuwendo ogwategekebwa edda ogwateesebwako Festing olw’okubalirira obunene bwa sampuli y’ebisolo eby’okugezesa35. Oluvannyuma lw’okukubisaamu omuwendo guno ne SD (2.02 × 0.81), obunene bw’ekikolwa ekizuuliddwa (1.6 μM) bwali 20%, ekikkirizibwa. Kino kitegeeza nti n = 5/ekibinja kimala okuzuula enkyukakyuka ya 20% wakati w’ebibinja. Envunza zaawulwamu mu ngeri ey’ekifuulannenge mu bibinja ebifuga n’ebijjanjabiddwa NaSH nga bakozesa omulimu ogw’ekifuulannenge ogwa Excel software 36 (Supplementary Fig. 1). Okuziba amaaso kwakolebwa ku ddaala ly’ebivaamu, era abanoonyereza abaali bakola ebipimo by’ebiramu tebaamanyi ku mirimu gya kibinja.
Ebibinja bya NaHS eby’ekikula kyombi byajjanjabwa n’omubisi gwa NaHS ogwa 30 μM ogwategekebwa mu mazzi ag’okunywa okumala wiiki 2; Fresh solution was supplied every 24 h, mu kiseera ekyo obuzito bw’omubiri bwapimibwa. Sampuli z’omusaayi zaakuŋŋaanyizibwa okuva ku nsonga z’omukira gw’envunza zonna ezaali zisumululwa eddagala lya isoflurane buli luvannyuma lwa nnaku bbiri ku nkomerero ya wiiki esooka n’eyookubiri. Sampuli z’omusaayi zasengejebwa ku 3000 g okumala edakiika 10, serum yayawulwamu n’eterekebwa ku –80°C okupima oluvannyuma kwa serum urea, creatinine (Cr), ne total sulfide. Serum urea yazuulibwa mu nkola ya enzymatic urease, ate serum creatinine yazuulibwa mu nkola ya photometric Jaffe nga tukozesa ebikozesebwa ebitundibwa (Man Company, Tehran, Iran) n’ekyuma ekikebera otomatiki (Selectra E, serial number 0-2124, The Netherlands). Emiwendo gy’enkyukakyuka mu kukebera mu n’okukebera wakati ku urea ne Cr gyali wansi wa 2.5%.
Enkola ya methylene blue (MB) ekozesebwa okupima total sulfide mu mazzi ag’okunywa ne serum erimu NaHS; MB y’enkola esinga okukozesebwa okupima salufayidi mu biwujjo ebinene ne sampuli z’ebiramu11,37. Enkola ya MB esobola okukozesebwa okubalirira ekidiba kya salufayidi kyonna38 n’okupima salufayidi ezitali za kiramu mu ngeri ya H2S, HS- ne S2 mu kitundu eky’amazzi39. Mu nkola eno, salufa atonnya nga zinki salufaayidi (ZnS) nga waliwo zinki acetate11,38. Okutonnya kwa zinc acetate y’enkola esinga okukozesebwa okwawula sulfides okuva mu chromophores endala11. ZnS yaddamu okusaanuusibwa nga tukozesa HCl11 mu mbeera za asidi ow’amaanyi. Salfayidi akwatagana ne DMPD mu mugerageranyo gwa stoichiometric ogwa 1:2 mu nsengekera etabulwa ferric chloride (Fe3+ ekola nga ekirungo ekifuula oxidizing agent) okukola langi MB, ezuulibwa mu ngeri ya spectrophotometric ku 670 nm40,41. Ekkomo ly’okuzuula ery’enkola ya MB liri nga 1 μM11.
Mu kunoonyereza kuno, 100 μL za buli sampuli (solution oba serum) zaateekebwa mu ttanka; olwo 200 μL za zinc acetate (1% w/v mu mazzi agafumbiddwa), 100 μL za DMPD (20 mM mu 7.2 M HCl), ne 133 μL za FeCl3 (30 mM mu 1.2 M HCl) ne ziteekebwamu. Omutabula gwafumbirwa ku 37°C mu nzikiza okumala edakiika 30. Ekisengejjero kyafukibwa ku 10,000 g okumala edakiika 10, era okunyiga kw’amazzi agasukkulumye ku mazzi ne kusomebwa ku 670 nm nga tukozesa microplate reader (BioTek, MQX2000R2, Winooski, VT, USA). Ebirungo bya sulfide byazuulibwa nga tukozesa enkola ya calibration curve ya NaHS (0–100 μM) mu ddH2O (Supplementary Fig. 2). Ebisengejjero byonna ebyakozesebwa mu kupima byali bipya okutegekebwa. Emiwendo gy’enkyukakyuka mu kukebera mu n’okugezesebwa wakati w’ebipimo bya salufayidi gyali 2.8% ne 3.4%, okusinziira ku. Twazuula n’omuwendo gwa sulfide gwonna ogwazuulibwa okuva mu sampuli z’amazzi ag’okunywa ne serum ezirimu sodium thiosulfate nga tukozesa enkola ya sampuli ezinywevu42. Okuwona kwa sampuli z’amazzi ag’okunywa ne serum ezirimu sodium thiosulfate zaali 91 ± 1.1% (n = 6) ne 93 ± 2.4% (n = 6), okusinziira ku.
Okwekenenya ebibalo kwakolebwa nga tukozesa pulogulaamu ya GraphPad Prism version 8.0.2 for Windows (GraphPad Software, San Diego, CA, USA, www.graphpad.com). Ekigezo kya t-test ekya paired t-test kyakozesebwa okugeraageranya ebbugumu ne pH y’amazzi ag’okunywa nga tebannaba kwongerako NaHS n’oluvannyuma lw’okugattibwa. Okufiirwa kwa H2S mu NaHS-containing solution kwabalirirwa nga okukendeera kwa bitundu ku kikumi okuva ku baseline uptake, era okwekenneenya oba okufiirwa kwali kwa maanyi mu bibalo, twakola one-way repeated-measures ANOVA nga tugobererwa Dunnett’s multiple comparison test. Obuzito bw’omubiri, serum urea, serum creatinine, ne total serum sulfide okumala ekiseera byageraageranyizibwa wakati w’envunza ezifuga n’ezijjanjabiddwa NaHS ez’ekikula eky’enjawulo nga bakozesa enkola ya two-way mixed (between-within) ANOVA nga egobererwa okugezesebwa kwa Bonferroni post hoc. Emiwendo gya P egy’emikira ebiri < 0.05 gyatwalibwa nga egy’omugaso mu bibalo.
pH y’amazzi ag’okunywa yali 7.60 ± 0.01 nga tebannaba kwongerako NaHS ne 7.71 ± 0.03 oluvannyuma lw’okugattako NaHS (n = 13, p = 0.0029). Ebbugumu ly’amazzi ag’okunywa lyali 26.5 ± 0.2 era lyakendeera okutuuka ku 26.2 ± 0.2 oluvannyuma lw’okugattako NaHS (n = 13, p = 0.0128). Tegeka 30 μM NaHS solution mu mazzi ag’okunywa era ogitereke mu ccupa y’amazzi. NaHS solution tenywevu era ekirungo kyayo kikendeera okumala ekiseera. Nga batwala sampuli okuva mu bulago bw’eccupa y’amazzi, okukendeera okw’amaanyi (68.0%) kwalabiddwa mu ssaawa esooka, era NaHS mu solution yakendeera ebitundu 72% ne 75% oluvannyuma lw’essaawa 12 ne 24. Mu sampuli ezafunibwa okuva mu bidomola by’amazzi, okukendeera kwa NaHS tekwali kwa maanyi okutuuka ku ssaawa 2, naye oluvannyuma lw’essaawa 12 ne 24 kwali kukendedde ebitundu 47% ne 72%, okusinziira ku. Ebiwandiiko bino biraga nti ebitundu bya NaHS mu 30 μM solution eyategekebwa mu mazzi ag’okunywa byali bikendedde okutuuka ku nga kimu kyakuna eky’omuwendo ogwasooka oluvannyuma lw’essaawa 24, awatali kulowooza ku kifo kya sampuli (Figure 1).
Okutebenkera kw’ekisengejjero kya NaHS (30 μM) mu mazzi ag’okunywa mu bidomola by’envunza/ebibe. Oluvannyuma lw’okuteekateeka ekisengejjero, sampuli zaggyibwa ku ntikko ne munda mu ccupa y’amazzi. Ebiwandiiko biragiddwa nga kigero ± SD (n = 6/ekibinja). * ne #, P < 0.05 bw’ogeraageranya n’obudde 0. Ekifaananyi ky’eccupa y’amazzi kiraga ensonga (erimu ekiggule) n’omubiri gw’eccupa. Voliyumu y’ensonga eri nga 740 μL.
Ekirungo kya NaHS mu 30 μM solution eyaakategekebwa kyali 30.3 ± 0.4 μM (range: 28.7–31.9 μM, n = 12). Wabula oluvannyuma lw’essaawa 24, ekirungo kya NaHS kyakendeera okutuuka ku muwendo ogwa wansi (kitegeeza: 3.0 ± 0.6 μM). Nga bwe kiragibwa mu kifaananyi 2, ebirungo bya NaHS envunza bye byakwatibwa tebyali bya bulijjo mu kiseera ky’okunoonyereza.
Obuzito bw’omubiri gw’envunza enkazi bweyongera nnyo okumala ekiseera (okuva ku 205.2 ± 5.2 g okutuuka ku 213.8 ± 7.0 g mu kibinja ekifuga ate okuva ku 204.0 ± 8.6 g okutuuka ku 211.8 ± 7.5 g mu kibinja ekyajjanjabwa NaHS); wabula, obujjanjabi bwa NaHS tebwalina kye bwakola ku buzito bw’omubiri (Fig. 3). Obuzito bw’omubiri gw’envunza ensajja bweyongera nnyo okumala ekiseera (okuva ku 338.6 ± 8.3 g okutuuka ku 352.4 ± 6.0 g mu kibinja ekifuga n’okuva ku 352.4 ± 5.9 g okutuuka ku 363.2 ± 4.3 g mu kibinja ekyajjanjabwa NaHS); wabula, obujjanjabi bwa NaHS tebwalina kye bwakola ku buzito bw’omubiri (Fig. 3).
Enkyukakyuka mu buzito bw’omubiri mu nvunza enkazi n’ensajja oluvannyuma lw’okuweebwa NaHS (30 μM). Ebiwandiiko biragiddwa nga kigero ± SEM era ne bigeraageranyizibwa nga tukozesa okwekenneenya okw’enjawulo okutabuddwamu (mu-wakati) okw’engeri bbiri n’okugezesebwa kwa Bonferroni post hoc. n = 5 eza buli kikula mu buli kibinja.
Ebirungo bya serum urea ne creatine phosphate byageraageranyizibwa mu nvunza ezifuga n’ezijjanjabiddwa NaSH mu kunoonyereza kwonna. Ekirala, obujjanjabi bwa NaSH tebwakosa bungi bwa serum urea ne creatinechrome (Table 1).
Baseline serum total sulfide concentrations zaali zigeraageranyizibwa wakati w’envunza ensajja (8.1 ± 0.5 μM vs. 9.3 ± 0.2 μM) n’enkazi (9.1 ± 1.0 μM vs. 6.1 ± 1.1 μM) control ne NaHS-treated. Okugaba NaHS okumala ennaku 14 tekwalina kye kwakola ku miwendo gya serum total sulfide mu nvunza ensajja oba enkazi (Fig. 4).
Enkyukakyuka mu serum total sulfide concentrations mu nvunza ensajja n’enkazi oluvannyuma lw’okuweebwa NaHS (30 μM). Ebiwandiiko biragiddwa nga kigero ± SEM era ne bigeraageranyizibwa nga tukozesa okwekenneenya okw’enjawulo okutabuddwamu (munda-munda) okw’engeri bbiri n’okugezesebwa kwa Bonferroni post hoc. Buli kikula, n = 5/ekibinja.
Ekikulu ekimaliriziddwa mu kunoonyereza kuno kiri nti amazzi ag’okunywa agalimu NaHS teganywevu: nga kimu kyakuna kyokka ku muwendo gwonna ogwa sulfide mu kusooka bye bisobola okuzuulibwa oluvannyuma lw’essaawa 24 oluvannyuma lw’okutwala sampuli okuva ku ntikko ne munda mu bidomola by’amazzi g’envunza/ebibe. Ekirala, envunza zaali zifunye NaHS ezitali nnywevu olw’okufiirwa H2S mu NaHS solution, era okugatta NaHS mu mazzi ag’okunywa tekyakosa buzito bwa mubiri, serum urea ne creatine chromium, oba total serum sulfide.
Mu kunoonyereza kuno, omuwendo gw’okufiirwa H2S okuva mu 30 μM NaHS solutions ezitegekeddwa mu mazzi ag’okunywa gwali nga 3% buli ssaawa. Mu solution eya buffered (100 μM sodium sulfide mu 10 mM PBS, pH 7.4), ekirungo kya sulfide kyategeezebwa nti kikendeera ebitundu 7% okumala ekiseera mu 8 h11. Emabegako twawolereza okugaba NaHS mu lubuto nga tutegeeza nti omuwendo gw’okufiirwa sulfide okuva mu 54 μM NaHS solution mu mazzi ag’okunywa gwali nga 2.3% buli ssaawa (4%/essaawa mu ssaawa 12 ezisooka ne 1.4%/essaawa mu ssaawa 12 ezisembayo oluvannyuma lw’okuteekateeka)8. Okunoonyereza okwasooka43 kwazuula okufiirwa kwa H2S buli kiseera okuva mu biwujjo bya NaHS, okusinga olw’okuwunya n’okufuumuuka. Ne bwe waba tewali kugatta biwujjo, sulfide mu stock solution ebula mangu olw’okuwunya kwa H2S11. Okunoonyereza kulaga nti mu nkola y’okufuumuula, etwala sekondi nga 30–60, ebitundu nga 5–10% ebya H2S bibula olw’okufuumuuka6. Okuziyiza okufuumuuka kwa H2S okuva mu kisoolo, abanoonyereza bakoze ebikolwa ebiwerako, omuli okutabula mpola ekisengejjero12, okubikka ekisengejjero kya sitokisi ne firimu ya pulasitiika6, n’okukendeeza ku kufuumuuka kw’ekisengejjero mu mpewo, okuva omutindo gw’okufuumuuka kwa H2S bwe gusinziira ku nkolagana y’empewo n’amazzi.13 Okwokya okw’okwekolako okwa H2S kubaawo okusinga olw’ayoni z’ebyuma ezikyusa naddala ferric ekyuma, nga bino biba bucaafu mu mazzi.13 Okisayidi ya H2S kivaamu okutondebwa kwa polysulfides (atomu za salufa eziyungiddwa ku covalent bonds)11. Okwewala okufuuka omukka gwayo, ebisengejjero ebirimu H2S bitegekebwa mu bizimbulukusa ebitaliimu okisigyeni44,45 oluvannyuma ne birongoosebwa ne argon oba nayitrojeni okumala edakiika 20–30 okukakasa nti omukka gwayo gufuuse omukka.11,12,37,44,45,46 Diethylenetriaminepentaacetic acid (DTPA) ye metal chelator (10–4 M) eziyiza HS-auoxidation mu ebizigo ebiyitibwa aerobic solutions. Mu butabeerawo DTPA, omuwendo gwa autooxidation ogwa HS- guli nga 50% mu ssaawa nga 3 ku 25°C37,47. Ekirala, okuva okuwunyiriza kwa 1e-sulfide bwe kutabulwa ekitangaala kya ultraviolet, ekisengejjero kisaana okuterekebwa ku ice ne kikuumibwa okuva ku kitangaala11.
Nga bwe kiragibwa mu kifaananyi 5, NaHS eyawukana mu Na+ ne HS-6 nga esaanuuse mu mazzi; okukutula kuno kusalibwawo pK1 y’ensengekera, nga eno yeesigamye ku bbugumu: pK1 = 3.122 + 1132/T, nga T eva ku 5 okutuuka ku 30°C era epimibwa mu diguli Kelvin (K), K = °C + 273.1548. HS- erina pK2 enkulu (pK2 = 19), kale ku pH < 96.49, S2- tetondebwa oba ekolebwa mu bungi obutono ennyo. Okwawukana ku ekyo, HS- ekola nga base era n’ekkiriza H+ okuva mu molekyo ya H2O, ate H2O ekola nga asidi n’akyusibwa okufuuka H2S ne OH-.
Okutondebwa kw’omukka gwa H2S ogusaanuuse mu kisoolo kya NaHS (30 μM). aq, ekisengejjero eky’amazzi; g, ggaasi; l, amazzi. Okubala kwonna kuteebereza nti pH y’amazzi = 7.0 ate ebbugumu ly’amazzi = 20 °C. Yatondebwawo ne BioRender.com.
Wadde nga waliwo obujulizi nti ebisengejja bya NaHS tebinywevu, okunoonyereza okuwerako ku bisolo kukozesezza ebisengejja bya NaHS mu mazzi ag’okunywa ng’ekirungo ekigaba H2S15,16,17,18,19,20,21,22,23,24,25,26 nga ebiseera by’okuyingira mu nsonga biva ku wiiki 1 okutuuka ku 21 (Table 2). Mu kunoonyereza kuno, ekisengejjero kya NaHS kyazzibwa buggya buli ssaawa 12, 15, 17, 18, 24, 25 oba 24 h, 19, 20, 21, 22, 23 h. Ebivudde mu kunoonyereza kwaffe biraga nti envunza zaali zifunye eddagala eritali ntebenkevu olw’okufiirwa H2S okuva mu NaHS solution, era NaHS ebiri mu mazzi g’envunza ag’okunywa byakyukakyuka nnyo mu ssaawa 12 oba 24 (laba ekifaananyi 2). Okunoonyereza kubiri ku kuno kwalaga nti emiwendo gya H2S mu mazzi gyasigala nga gitebenkedde okumala essaawa 24 h22 oba nti okufiirwa kwa H2S ebitundu 2–3% byokka kwe kwalabibwa okumala essaawa 12 h15, naye tekwawa biwandiiko biwagira oba ebikwata ku kupima. Okunoonyereza kubiri kulaga nti obuwanvu obutono obw’amacupa g’amazzi busobola okukendeeza ku kufuumuuka kwa H2S15,19. Wabula, ebivudde mu kunoonyereza kwaffe biraga nti kino kiyinza okulwawo okufiirwa H2S okuva mu ccupa y’amazzi 2 h zokka okusinga 12–24 h. Okunoonyereza kwombi kulaga nti tuteebereza nti omutindo gwa NaHS mu mazzi ag’okunywa tegwakyuka kubanga tetwalaba nkyukakyuka ya langi mu mazzi; n’olwekyo, oxidation ya H2S mu mpewo teyali ya maanyi19,20. Ekyewuunyisa, enkola eno ey’omutwe yeetegereza obutebenkevu bwa NaHS mu mazzi okusinga okupima enkyukakyuka mu bungi bwayo mu kiseera.
Okufiirwa kwa H2S mu kisoolo kya NaHS kikwatagana ne pH n’ebbugumu. Nga bwe kyalagulwa mu kunoonyereza kwaffe, okusaanuusa NaHS mu mazzi kivaamu okutondebwawo kw’ekisengejjero kya alkaline50. NaHS bw’esaanuuka mu mazzi, okutondebwa kw’omukka gwa H2S ogusaanuuse kisinziira ku muwendo gwa pH6. pH y’ekisengejjero gy’ekoma okuba wansi, ekitundu kya NaHS gye kikoma okuba ekinene nga molekyo za ggaasi ya H2S era salufaayidi gy’akoma okubula okuva mu kisoolo eky’amazzi11. Tewali n’emu ku kunoonyereza kuno yalaga pH y’amazzi ag’okunywa agakozesebwa ng’ekizimbulukusa ku NaHS. Okusinziira ku biteeso by’ekitongole ky’ebyobulamu eky’ensi yonna, ebikkirizibwa amawanga agasinga obungi, pH y’amazzi ag’okunywa erina okuba wakati wa 6.5–8.551. Mu pH eno, omutindo gw’okufuuka okw’okwekolako okwa H2S gweyongera emirundi nga kkumi13. Okusaanuusa NaHS mu mazzi mu pH eno kijja kuvaamu omukka gwa H2S ogutabuddwa okuva ku 1 okutuuka ku 22.5 μM, ekiggumiza obukulu bw’okulondoola pH y’amazzi nga tonnasaanuusa NaHS. Okugatta ku ekyo, ebbugumu eryategeezeddwa mu kunoonyereza waggulu (18–26 °C) lyandivuddemu enkyukakyuka mu bungi bwa ggaasi wa H2S asaanuuse mu kisoolo nga 10%, okuva enkyukakyuka mu bbugumu bwe zikyusa pK1, era enkyukakyuka entonotono mu pK1 ziyinza okuba n’akakwate akakulu ku bungi bwa ggaasi wa H2S asaanuuse48. Okugatta ku ekyo, ebbanga eddene ery’okunoonyereza okumu (emyezi 5)22, nga mu kiseera kino enkyukakyuka ennene ey’ebbugumu esuubirwa, nakyo kyongera ku kizibu kino.
Okunoonyereza kwonna okuggyako one21 kwakozesa 30 μM NaHS solution mu mazzi ag’okunywa. Okunnyonnyola ddoozi ekozesebwa (kwe kugamba 30 μM), abawandiisi abamu baalaga nti NaHS mu mutendera gw’amazzi ekola ekirungo kye kimu ddala ekya ggaasi ya H2S era nti ekitundu ky’omubiri ekya H2S kiri 10 ku 100 μM, kale ddoozi eno eri mu bbanga ly’omubiri15,16. Abalala bannyonnyola nti 30 μM NaHS esobola okukuuma omutindo gwa plasma H2S mu bbanga ly’omubiri, kwe kugamba 5–300 μM19,20. Tulowooza ku bungi bwa NaHS mu mazzi aga 30 μM (pH = 7.0, T = 20 °C), ekyakozesebwa mu kunoonyereza okumu okunoonyereza ku bikolwa bya H2S. Tusobola okubala nti obuzito bwa ggaasi wa H2S asaanuuse buli 14.7 μM, nga buno buba nga 50% ku bungi bwa NaHS obwasooka. Omuwendo guno gufaananako n’omuwendo ogwabalirirwa abawandiisi abalala mu mbeera ze zimu13,48.
Mu kunoonyereza kwaffe, okugaba NaHS tekyakyusa buzito bwa mubiri; ekivuddemu kino kikwatagana n’ebyava mu kunoonyereza okulala mu mbeba ensajja22,23 n’envunza ensajja18; Wabula okunoonyereza kubiri kwalaga nti NaSH yazzaawo obuzito bw’omubiri obwali bukendedde mu nvunza ezasaliddwako ekibumba24,26, so ng’ate okunoonyereza okulala tekwalaga ngeri NaSH gye yakwata ku buzito bw’omubiri15,16,17,19,20,21,25. Ekirala, mu kunoonyereza kwaffe, okugaba NaSH tekyakosa miwendo gya serum urea ne creatine chromium, ekikwatagana n’ebyava mu lipoota endala25.
Okunoonyereza kuno kwazuula nti okugatta NaHS mu mazzi ag’okunywa okumala wiiki 2 tekyakosa bungi bwa serum sulfide bwonna mu nvunza ensajja n’enkazi. Kino kye bazudde kikwatagana n’ebyava mu kunoonyereza kwa Sen et al. (16): Wiiki 8 ez’okujjanjabwa ne 30 μM NaHS mu mazzi ag’okunywa tezaakosa miwendo gya plasma sulfide mu nvunza ezifuga; wabula, baategeeza nti okuyingira mu nsonga kuno kwazzaawo emiwendo gya H2S egyakendeera mu plasma y’ebibe ebyasalibwako ekibumba. Li ne banne. (22) era baategeeza nti okujjanjaba 30 μM NaHS mu mazzi ag’okunywa okumala emyezi 5 kyayongera ku plasma free sulfide levels mu mbeba ezikaddiye ebitundu nga 26%. Okunoonyereza okulala tekulaze nkyukakyuka mu sulfide etambula oluvannyuma lw’okugatta NaHS mu mazzi ag’okunywa.
Okunoonyereza musanvu kwalaga nti bakozesa Sigma NaHS15,16,19,20,21,22,23 naye tekwawa bisingawo ku mazzi ga hydration, ate okunoonyereza kutaano tekwayogera ku nsibuko ya NaHS ekozesebwa mu nkola zaabwe ez’okuteekateeka17,18,24,25,26. NaHS molekyu erimu amazzi era amazzi gaayo ag’amazzi gayinza okwawukana, ekikosa obungi bwa NaHS obwetaagisa okuteekateeka ekisengejjero kya molarity eweereddwa. Okugeza, ebirimu NaHS mu kunoonyereza kwaffe byali NaHS•1.3 H2O. Bwe kityo, ebirungo bya NaHS ebituufu mu kunoonyereza kuno biyinza okuba wansi okusinga ebyo ebyaloopebwa.
“Ekirungo ekiwangaala ekitono bwe kiti kiyinza kitya okuba n’ekikolwa ekiwangaala bwe kityo?” Pozgay et al.21 baabuuza ekibuuzo kino nga beetegereza ebikolwa bya NaHS ku bulwadde bw’olubuto mu mbeba. Basuubira nti okunoonyereza mu biseera eby’omu maaso kujja kusobola okuddamu ekibuuzo kino era ne bateebereza nti ebisengejja bya NaHS biyinza okubaamu polysulfides ezisingako obutebenkevu nga kwotadde ne H2S ne disulfides ezitabaganya ekikolwa kya NaHS21. Ekirala ekiyinza okubaawo kiri nti ebirungo ebitono ennyo ebya NaHS ebisigadde mu solution nabyo biyinza okuba n’ekikolwa eky’omugaso. Mu butuufu, Olson et al. yawa obujulizi nti emiwendo gya micromolar egya H2S mu musaayi si gya mubiri era girina okuba mu nanomolar range oba nga tegiriiwo yonna13. H2S eyinza okukola okuyita mu protein sulfation, enkyukakyuka ekyukakyuka oluvannyuma lw’okuvvuunula ekosa enkola, okutebenkera, n’okubeera mu kifo kya puloteyina nnyingi52,53,54. Mu butuufu, mu mbeera z’omubiri, ebitundu nga 10% ku 25% ku puloteyina nnyingi ez’ekibumba zirimu sulfylated53. Okunoonyereza kwombi kukkiriza okusaanawo okw’amangu kwa NaHS19,23 naye ekyewuunyisa kugamba nti “twafuga obungi bwa NaHS mu mazzi ag’okunywa nga tugakyusa buli lunaku.”23 Okunoonyereza okumu kwagamba mu butanwa nti “NaHS is a standard H2S donor and is commonly used in clinical practice to replace H2S itself.”18
Okukubaganya ebirowoozo waggulu kulaga nti NaHS ebula okuva mu solution okuyita mu volatilization, oxidation ne photolysis, era n’olwekyo ebimu ku biteeso bikolebwa okukendeeza ku kufiirwa kwa H2S okuva mu solution. Ekisooka, okufuumuuka kwa H2S kwesigamye ku nsengekera ya ggaasi n’amazzi13 ne pH y’ekisengejjero11; n’olwekyo, okukendeeza ku kufiirwa kw’okufuumuuka, ensingo y’eccupa y’amazzi esobola okufuulibwa entono nga bwe kyayogerwako emabegako15,19, era pH y’amazzi esobola okutereezebwa okutuuka ku kkomo erya waggulu erikkirizibwa (kwe kugamba, 6.5–8.551) okukendeeza ku kufiirwa kw’okufuumuuka11. Ekyokubiri, okufuuka okw’okwekolako okwa H2S kubaawo olw’ebikolwa bya okisigyeni n’okubeerawo kwa ayoni z’ebyuma ebikyukakyuka mu mazzi ag’okunywa13, kale okuggya okisigyeni mu mazzi ag’okunywa ne argon oba nayitrojeni44,45 n’okukozesa ebyuma ebiyitibwa chelators37,47 bisobola okukendeeza ku oxidation ya sulfides. Ekyokusatu, okuziyiza okuvunda kw’ekitangaala kwa H2S, obucupa bw’amazzi busobola okuzingibwa n’ekipande kya aluminiyamu; Enkola eno era ekwata ku bintu ebikwata ekitangaala nga streptozotocin55. N’ekisembayo, eminnyo gya salufayidi egy’obutonde (NaHS, Na2S, ne CaS) giyinza okuweebwa nga giyita mu gavage okusinga okusaanuusibwa mu mazzi ag’okunywa nga bwe kyaloopebwa emabegako56,57,58; okunoonyereza kulaga nti radioactive sodium sulfide ewereddwa okuyita mu gavage eri envunza ayingizibwa bulungi era egabibwa kumpi mu bitundu byonna59. N’okutuusa kati, okunoonyereza okusinga kubadde kugaba eminnyo gya sulfide egitali gya kiramu mu lubuto; wabula, ekkubo lino teritera kukozesebwa mu mbeera z‟obujjanjabi60. Ku luuyi olulala, ekkubo ery’omu kamwa lye kkubo erisinga okukozesebwa era erisinga okwettanirwa okuweebwa mu bantu61. N’olwekyo, tusaba okwekenneenya ebiva mu bagaba H2S mu biwuka nga bakozesa eddagala eriyitibwa oral gavage.
Ekikoma kiri nti twapima sulfide mu solution y’amazzi ne serum nga tukozesa enkola ya MB. Enkola z’okupima salufayidi mulimu okupima ayodini, okupima ekitangaala, enkola y’amasannyalaze (potentiometry, amperometry, enkola ya coulometric n’enkola ya amperometric) ne chromatography (gas chromatography ne high-performance liquid chromatography), nga mu bino enkola esinga okukozesebwa ye MB spectrophotometric method62. Ekikoma ku nkola ya MB ey’okupima H2S mu sampuli z’ebiramu kwe kuba nti epima ebirungo byonna ebirimu salufa so si H2S63 ow’eddembe kubanga ekolebwa mu mbeera ya asidi, ekivaamu okuggya ekibiriiti okuva mu nsibuko y’ebiramu64. Wabula okusinziira ku kibiina ky’ebyobulamu mu Amerika, MB y’enkola eya bulijjo ey’okupima sulfide mu mazzi65. N’olwekyo, okukoma kuno tekukosa bivuddemu byaffe ebikulu ku butabeera mu ntebenkevu bw’ebisengejja ebirimu NaHS. Ekirala, mu kunoonyereza kwaffe, okuzzaawo ebipimo bya sulfide mu sampuli z’amazzi ne serum ezirimu NaHS kwali 91% ne 93%, okusinziira ku. Emiwendo gino gikwatagana n’obuwanvu obwaloopebwa emabegako (77–92)66, ekiraga obutuufu obw’okwekenneenya obukkirizibwa42. Kinajjukirwa nti twakozesa envunza ensajja n’enkazi okusinziira ku ndagiriro z’ekitongole ky’ebyobulamu ekya National Institutes of Health (NIH) okwewala okwesigama ennyo ku kunoonyereza ku bisolo ebisajja byokka mu kunoonyereza okusooka67 n’okussaamu envunza ensajja n’enkazi buli lwe kiba kisoboka68. Ensonga eno ebadde eggumiza abalala69,70,71.
Mu kumaliriza, ebivudde mu kunoonyereza kuno biraga nti ebisengejja bya NaHS ebitegekebwa okuva mu mazzi ag’okunywa tebisobola kukozesebwa kukola H2S olw’obutabeera mu ntebenkevu. Ekkubo lino ery’okugaba lyandireese ebisolo ku miwendo gya NaHS egitanywevu era egya wansi okusinga bwe gyali gisuubirwa; n’olwekyo, ebizuuliddwa biyinza obutakwata ku bantu.
Datasets ezikozesebwa ne/oba ezeekenneenyezebwa mu kiseera ky’okunoonyereza okuliwo kati zifunibwa okuva eri omuwandiisi akwatagana nga osabye mu ngeri entuufu.
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Dillon, KM, Carrazzone, RJ, Matson, JB, ne Kashfi, K. Okusuubiza okukulaakulana okw’enkola z’okutuusa obutoffaali ku nitric oxide ne hydrogen sulfide: omulembe omupya ogw’eddagala erikwata ku muntu. Ebiramu n’eddagala 176, 113931. https://doi.org/10.1016/j.bcp.2020.113931 (2020).
Sun, X., n’abalala. Okugaba okumala ebbanga eddene eky’omugabi wa hydrogen sulfide afuluma mpola kiyinza okuziyiza obuvune mu myocardial ischemia/reperfusion. Lipoota za ssaayansi 7, 3541. https://doi.org/10.1038/s41598-017-03941-0 (2017).
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Sitdikova, GF, Weiger, TM ne Hermann, A. Hydrogen sulfide ayongera ku mikutu gya calcium-activated potassium (BK) mu butoffaali bw’ekizimba ky’envunza. Archit. Ebiwuka ebiyitibwa Pfluegers. 459, 389-397. 10.1007/s00424-009-0737-0 (2010) Enkola y’okukuuma obutonde bw’ensi mu bitundu by’eggwanga eby’enjawulo.
Jeddy, S., n’abalala. Hydrogen sulfide ayongera ku buzibu bw’obukuumi bwa nitrite okuva ku buvune obuva ku myocardial ischemia-reperfusion mu nvunza ezirina ssukaali ow’ekika eky’okubiri. Okisayidi ya Nitric 124, 15-23. 10.1016/j.niox.2022.04.004 (2022) Enkola y’okukuuma obutonde bw’ensi mu bitundu by’eggwanga eby’enjawulo.
Corvino, A., n’abalala. Emitendera mu kemiko y’abagaba H2S n’enkosa yaayo ku ndwadde z’emisuwa. Ebirungo ebiziyiza obuwuka obuleeta endwadde 10, 429. https://doi.org/10.3390/antiox10030429 (2021).
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Nagy, P., n’abalala. Ebintu ebikwata ku kemiko eby’okupima haidrojeni salufayidi mu sampuli z’omubiri. Biochimica n’Ebyobulamu Acta 1840, 876-891. 10.1016/j.bbagen.2013.05.037 (2014) Enkola y’okukuuma obutonde bw’ensi mu bitundu by’eggwanga eby’enjawulo.
Kline, LL.D., era nga ye muwandiisi w’ebitabo. Okuzuula kwa spectrophotometric kwa hydrogen sulfide mu mazzi ag’obutonde. Limnol. Oceanogr. 14, 454-458. 1969.14.3.0454 (1969) Enkola y’okukuuma obutonde bw’ensi mu bitundu by’eggwanga eby’enjawulo.
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