Ndza khensa ku endzela nature.com. Vuhundzuluxi bya browser lebyi u byi tirhisaka byi na nseketelo wa CSS lowu nga nyawuriki. Leswaku u kuma ntokoto lowunene, hi ringanyeta ku tirhisa vuhundzuluxeri bya sweswinyana bya browser (kumbe ku tima movha wa ku fambisana eka Internet Explorer). Ku engetela kwalaho, ku tiyisisa nseketelo lowu yaka emahlweni, sayiti leyi a yi nge katsi switayele kumbe JavaScript.
Hydrogen sulfide (H2S) yina switandzhaku swotala swa physiological na pathological eka miri wa munhu. Sodium hydrosulfide (NaHS) yitirhisiwa ngopfu tani hi xitirhisiwa xa pharmacological ku kambela mbuyelo wa H2S eka swikambelo swa biological. Hambi leswi ku lahlekeriwa hi H2S kusuka eka swintshuxo swa NaHS switekaka timinete nyana ntsena, swintshuxo swa NaHS switirhisiwile tani hi ti donor compounds ta H2S eka mati yo nwa eka tidyondzo tin’wana ta swiharhi. Dyondzo leyi yi lavisisile loko mati yo nwa lawa yanga na NaHS concentration ya 30 μM lawa ya lulamisiweke eka mabodlhela ya rat/mouse yanga tshama ya tshamisekile kuringana kwalomu ka 12–24 wa tiawara, tani hileswi swiringanyetiweke hi vatsari van’wana. Lulamisa solution ya NaHS (30 μM) eka mati yo nwa kutani u hatlisa u chela eka mabodlhela ya mati ya rat/mouse. Tisampulu ti hlengeletiwile kusuka eka tip nale ndzeni ka bodlhela ra mati eka 0, 1, 2, 3, 4, 5, 6, 12 na 24 wa tiawara kuva ku pimiwa nhlayo ya sulfide hiku tirhisa ndlela ya methylene blue. Ku engetela kwalano, ti rats ta xinuna na ta xisati ti cheriwile NaHS (30 μM) kuringana mavhiki mambirhi naswona nhlayo ya serum sulfide yi pimiwe siku rin’wana na rin’wana ra vumbirhi eka vhiki rosungula xikan’we naku hela ka vhiki ravumbirhi. Nkatsakanyo wa NaHS eka xikombiso lexi kumiweke kusuka eka nhloko ya bodlhela ra mati awu tshamisekanga; yi hungutekile hi 72% na 75% endzhaku ka tiawara ta 12 na 24, hi ku landzelelana. Eka tisampulu leti kumiweke kusuka endzeni ka mabodlhela ya mati, kuhunguteka ka NaHS akuri ka nkoka eka 2 wa tiawara; hambiswiritano, yi hungutekile hi 47% na 72% endzhaku ka tiawara ta 12 na 24, hi ku landzelelana. Ku cheriwa ka NaHS aswi khumbanga serum sulfide level ya ti rats ta xinuna na ta xisati. Hiku gimeta, swintshuxo swa NaHS leswi lulamisiweke kusuka eka mati yo nwa aswifanelanga kutirhisiwa eka ku nyikela hi H2S hikuva xitolwa lexi axi tshamisekanga. Ndlela leyi ya vulawuri yita veka swifuwo eka nhlayo leyinga tshamisekangiki na leyintsongo kutlula leyi languteriweke ya NaHS.
Hydrogen sulfide (H2S) yitirhisiwile tani hi chefu kusukela hi 1700; hambiswiritano, ntirho wa yona lowu nga kotekaka tanihi molekhuli ya biosignaling ya le ndzeni wu hlamuseriwile hi Abe na Kimura hi 1996. Eka makume manharhu ya malembe lama hundzeke, mintirho yo tala ya H2S eka tisisiteme to hambana ta vanhu yi hlamuseriwile, leswi yiseke eka ku lemuka leswaku timolekhuli ta munyiki wa H2S ti nga va na matirhiselo ya tliliniki eka vutshunguri kumbe vulawuri bya mavabyi yo karhi; vona Chirino na van’wana. ku kuma nkambisiso wa sweswinyana.
Sodium hydrosulfide (NaHS) yi tirhisiwile ngopfu tani hi xitirhisiwa xa pharmacological ku kambela mbuyelo wa H2S eka tidyondzo to tala ta ndhavuko wa tisele na swiharhi5,6,7,8. Hambiswiritano, NaHS a hi munyiki lonene wa H2S hikuva yi hundzuriwa hi ku hatlisa yi va H2S/HS- eka solution, yi thyakisiwile hi ku olova hi ti polysulfides, naswona yi olova ku oxidized na volatilized4,9. Eka swikambelo swotala swa biological, NaHS yi pfanganisiwa ematini, leswi endlaka leswaku kuva na passive volatilization xikan’we naku lahlekeriwa hi H2S10,11,12, spontaneous oxidation ya H2S11,12,13, na photolysis14. Sulfide eka solution yosungula yi lahleka hiku hatlisa swinene hikokwalaho ka volatilization ya H2S11. Eka xigwitsirisi lexi pfulekeke, hafu ya vutomi (t1/2) ya H2S i kwalomu ka 5 wa timinete, naswona nhlayo ya yona yihunguteka hi kwalomu ka 13% hi minete10. Hambi leswi ku lahlekeriwa hi hydrogen sulfide kusuka eka switolwa swa NaHS switekaka timinete nyana ntsena, tidyondzo tin’wana ta swiharhi ti tirhise swintshuxo swa NaHS tani hi xihlovo xa hydrogen sulfide eka mati yo nwa kuringana 1–21 wa mavhiki, ku siva solution leyingana NaHS endzhaku ka tiawara tin’wana na tin’wana ta 12–24.15,16,17,18,19,20,21,22,23,24,25,26 Mukhuva lowu i a swi fambisani na milawu ya ndzavisiso wa sayense, tanihileswi mimpimo ya mirhi yi faneleke ku seketeriwa eka ku tirhisiwa ka yona eka tinxaka tin’wana, ngopfungopfu vanhu.27
Vulavisisi bya preclinical eka biomedicine byi kongomisa eka ku antswisa khwalithi ya nhlayiso wa muvabyi kumbe mimbuyelo ya vutshunguri. Hambiswiritano, mbuyelo wa tidyondzo to tala ta swiharhi a wu si hundzuluxeriwa eka vanhu28,29,30. Xin’wana xa swivangelo swa ku tsandzeka loku ka vuhundzuluxi i ku pfumaleka ka nyingiso eka khwalithi ya maendlelo ya tidyondzo ta swiharhi30. Hikokwalaho, xikongomelo xa ndzavisiso lowu akuri ku lavisisa loko 30 μM NaHS solutions leyi lulamisiweke eka mabodlhela ya mati ya rat/mouse yinga tshama yitshamisekile eka mati yo nwa kuringana 12–24 h, tani hileswi swinga vuriwa kumbe ku ringanyetiwa eka tidyondzo tin’wana.
Swikambelo hinkwaswo eka ndzavisiso lowu swi endliwile hi ku landza swiletelo leswi kandziyisiweke swa nhlayiso na ku tirhisiwa ka swifuwo swa le laboratori eIran31. Swiviko hinkwaswo swa swikambelo eka ndzavisiso lowu na swona swi landzele swiletelo swa ARRIVE32. Komiti ya Mahanyelo ya Nhlangano wa Sayense ya Endocrine, Yunivhesiti ya Shahid Beheshti ya Sayense ya Vutshunguri, yi pasisile maendlelo hinkwawo ya ku ringeta eka nkambisiso lowu.
Zinc acetate dihydrate (CAS: 5970-45-6) na anhydrous ferric chloride (CAS: 7705-08-0) swi xaviwile kusuka eka Biochem, Chemopahrama (Cosne-sur-Loire, France). Sodium hydrosulfide hydrate (CAS: 207683-19-0) na N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) swi xaviwile kusuka eka Sigma-Aldrich (St. Louis, MO, USA). Isoflurane yi xaviwile eka Piramal (Bethlehem, PA, U.S.A.). Hydrochloric acid (HCl) yi xaviwile eka Merck (Darmstadt, Germany).
Lulamisa solution ya NaHS (30 μM) eka mati yo nwa kutani u hatlisa u chela eka mabodlhela ya mati ya rat/mouse. Nhlayo leyi yi hlawuriwile hi ku ya hi minkandziyiso yo tala leyi tirhisaka NaHS tanihi xihlovo xa H2S; vona xiyenge lexi nge Bulo. NaHS i molecule leyi nga na mati leyi nga vaka na nhlayo yo hambana ya mati ya hydration (i.e., NaHS•xH2O); hi ku ya hi muendli, phesente ya NaHS leyi tirhisiweke eka dyondzo ya hina a yi ri 70.7% (i.e., NaHS•1.3 H2O), naswona hi tekele enhlokweni nkoka lowu eka swibalo swa hina, laha hi tirhiseke ntiko wa molekhuli wa 56.06 g/mol, ku nga ntiko wa molekhuli wa NaHS leyi nga riki na mati. Mati ya hydration (ya tlhela ya vuriwa mati ya crystallization) i ti molecule ta mati leti endlaka xivumbeko xa crystalline33. Ti hydrates tina swivumbeko swohambana hambana swa physical na thermodynamic loko ku pimanyisiwa na ti anhydrates34.
Loko unga se chela NaHS eka mati yo nwa, pima pH na mahiselo ya solvent. Hiku hatlisa chela NaHS solution eka bodlhela ra mati ya rat/mouse eka xisaka xa swifuwo. Tisampulu ti hlengeletiwile kusuka eka tip nale ndzeni ka bodlhela ra mati eka 0, 1, 2, 3, 4, 5, 6, 12, na 24 h ku pima nhlayo ya sulfide. Mipimo ya sulfide yi tekiwile hiku hatlisa endzhaku ka ku tekeriwa swikombiso swin’wana na swin’wana. Hi kumile swikombiso kusuka eka nhloko ya chubu hikuva tidyondzo tin’wana ti kombisile leswaku vukulu bya pore lebyintsongo bya chubu ra mati byi nga hunguta ku khuluka ka H2S15,19. Mhaka leyi yi vonaka yi tirha ni le ka xitshunxo lexi nga ebodlheleni. Hambiswiritano, leswi aswiri tano eka solution leyinga enhan’wini ya bodlhela ra mati, leyi ayi ri na nhlayo yale henhla ya evaporation naswona ayi autoxidizing; entiyisweni, swiharhi a swi nwa mati lawa ku sungula.
Ti rats ta xinuna na ta xisati ta Wistar titirhisiwile eka nkambisiso lowu. Ti rats ti vekiwile eka ti polypropylene cages (2–3 rats hi cage) ehansi ka swiyimo swa ntolovelo (temperature 21–26 °C, humidity 32–40%) na 12 h ya ku vonakala (7 am kuya eka 7 pm) na 12 h ya munyama (7 pm kuya eka 7 am). Ti rats ati kuma mati ya mahala ya le pompini naswona ati phameriwa standard chow (Khorak Dam Pars Company, Tehran, Iran). Ti rats ta Wistar leti ringanaka (6 wa tinhweti) (n=10, ntiko wa miri: 190–230 g) na ta xinuna (n=10, ntiko wa miri: 320–370 g) ti avanyisiwile hiku landzelelana eka mintlawa leyi tshunguriweke na NaHS (30 μM) leyi tshunguriweke (n=5 hi ntlawa). Ku kumisisa mpimo wa xikombiso, hi tirhisile endlelo ra KISS (Keep It Simple, Stupid), leri hlanganisaka ntokoto wa khale na nxopaxopo wa matimba35. Hi sungule hi endla dyondzo yo ringeta eka 3 wa ti rats naswona hi kumile mean serum total sulfide level na standard deviation (8.1 ± 0.81 μM). Kutani, hi ku tekela enhlokweni matimba ya 80% na ku teka xiyimo xa nkoka xa matlhelo mambirhi xa 5%, hi kumile mpimo wa xikombiso xo sungula (n = 5 hi ku ya hi matsalwa ya khale) lexi fambelanaka na mpimo wa mbuyelo lowu ringaniseriweke wa 2.02 na ntikelo lowu hlamuseriweke ka ha ri emahlweni lowu ringanyetiweke hi Festing ku hlayela mpimo wa xikombiso xa swiharhi swa ku ringeta35. Endzhaku ko andzisa ntikelo lowu hi SD (2.02 × 0.81), mpimo wa mbuyelo lowu voniwaka lowu vhumbhiweke (1.6 μM) a wu ri 20%, leswi amukelekaka. Leswi swi vula leswaku n = 5/ntlawa swi ringanerile ku kuma ku cinca ka 20% wa xiringaniso exikarhi ka mintlawa. Ti rats ti avanyisiwile hiku landzelelana eka mintlawa ya vulawuri na leyi tshunguriweke hi NaSH hiku tirhisa ntirho wa random wa Excel software 36 (Supplementary Fig. 1). Ku pfala mahlo swi endliwile eka xiyimo xa mbuyelo, naswona valavisisi lava endlaka mimpimo ya tikhemikhali ta vutomi a va nga swi tivi hi swiavelo swa ntlawa.
Mintlawa ya NaHS ya rimbewu hinkwaro yi tshunguriwile hi 30 μM NaHS solution leyi lulamisiweke ematini yo nwa kuringana 2 wa mavhiki; Fresh solution ayi phakeriwa endzhaku ka 24 h yin’wana na yin’wana, hi nkarhi lowu ntiko wa miri awu pimiwa. Tisampulu ta ngati ti hlengeletiwile kusuka eka tinhla ta ncila wa ti rats hinkwato ehansi ka isoflurane anesthesia siku rin’wana na rin’wana ra vumbirhi eku heleni ka mavhiki yosungula na ya vumbirhi. Tisampulu ta ngati ti centrifuged eka 3000 g kuringana 10 min, serum yi hambanisiwile kutani yi hlayisiwa eka –80°C kuva ku pimiwa endzhaku ka serum urea, creatinine (Cr), na total sulfide. Serum urea yikumiwile hi ndlela ya enzymatic urease, naswona serum creatinine yikumiwile hi ndlela ya photometric Jaffe hiku tirhisa ti kits leti xavisiwaka (Man Company, Tehran, Iran) na automatic analyzer (Selectra E, serial number 0-2124, The Netherlands). Ti intra- na interassay coefficients ta ku hambana ka urea na Cr atiri ehansi ka 2.5%.
Ndlela ya methylene blue (MB) yitirhisiwa ku pima total sulfide eka mati yo nwa na serum leyingana NaHS; MB i ndlela leyi tirhisiwaka ngopfu yo pima sulfide eka swintshuxo swa xitalo na tisampulu ta biological11,37. Ndlela ya MB yinga tirhisiwa ku ringanyeta nhlayo hinkwayo ya sulfide pool38 naku pima ti inorganic sulfides hi xivumbeko xa H2S, HS- na S2 eka xiphemu xa mati39. Eka ndlela leyi, sulphur yi khuluka tani hi zinc sulfide (ZnS) eka vukona bya zinc acetate11,38. Ku khuluka ka zinc acetate i ndlela leyi tirhisiwaka ngopfu yo hambanyisa ti sulfides kusuka eka ti chromophores tin’wana11. ZnS yitlhele yi pfanganisiwa hiku tirhisa HCl11 ehansi ka swiyimo swa acid yo tiya. Sulfide yi endla reaction na DMPD hi stoichiometric ratio ya 1:2 eka reaction leyi catalyzed hi ferric chloride (Fe3+ yi tirha tani hi oxidizing agent) ku endla dye MB, leyi voniwaka hi spectrophotometrically eka 670 nm40,41. Mpimo wa ku kumiwa ka ndlela ya MB i kwalomu ka 1 μM11.
Eka ndzavisiso lowu, 100 μL ya xikombiso xin’wana na xin’wana (solution kumbe serum) yi cheriwile eka chubu; kutani 200 μL ya zinc acetate (1% w/v eka mati ya distilled), 100 μL ya DMPD (20 mM eka 7.2 M HCl), na 133 μL ya FeCl3 (30 mM eka 1.2 M HCl) swi cheriwile. Nkatsakanyo lowu wu pfaleriwile eka 37°C emunyameni kuringana 30 min. Nkatsakanyo wu centrifuged eka 10,000 g kuringana 10 min, naswona absorbance ya supernatant yi hlayiwile eka 670 nm hiku tirhisa microplate reader (BioTek, MQX2000R2, Winooski, VT, USA). Nhlayo ya sulfide yikumiwile hiku tirhisa calibration curve ya NaHS (0–100 μM) eka ddH2O (Supplementary Fig. 2). Switolwa hinkwaswo leswi tirhisiweke eka mimpimo a swi lunghiseleriwa hi vuntshwa. Ti coefficients ta intra- na interassay ta ku hambana eka mimpimo ya sulfide akuri 2.8% na 3.4%, hiku landzelelana. Hi tlhele hi kumisisa ntsengo hinkwawo wa sulfide lowu kumiweke eka mati yo nwa lama nga na sodium thiosulfate na swikombiso swa serum hi ku tirhisa ndlela ya xikombiso lexi tiyisiweke42. Ku vuyiseriwa ka mati yo nwa lawa ya nga na sodium thiosulfate na swikombiso swa serum a ku ri 91 ± 1.1% (n = 6) na 93 ± 2.4% (n = 6), hi ku landzelelana.
Nxopaxopo wa tinhlayo wu endliwile hi ku tirhisa software ya GraphPad Prism version 8.0.2 ya Windows (GraphPad Software, San Diego, CA, USA, www.graphpad.com). Ku tirhisiwile paired t-test ku pimanisa mahiselo na pH ya mati yo nwa emahlweni na le ndzhaku ka ku engeteriwa ka NaHS. Ku lahlekeriwa hi H2S eka xitofu lexi nga na NaHS swi hlayiwile tanihi ku hunguteka ka tiphesente ku suka eka ku amukeriwa ka masungulo, naswona ku kambela loko ku lahlekeriwa ku ri ka nkoka hi tlhelo ra tinhlayo, hi endlile ANOVA ya ndlela yin’we leyi phindha-phindhiweke leyi landzeriweke hi xikambelo xa ku pimanisa ko tala xa Dunnett. Ndzilo wa miri, serum urea, serum creatinine, na total serum sulfide hiku famba ka nkarhi swi pimanyisiwile exikarhi ka ti rats leti lawuriwaka na leti tshunguriweke hi NaHS ta rimbewu rohambana hambana hiku tirhisa two-way mixed (between-within) ANOVA leyi landzeriweke hi Bonferroni post hoc test. Two-tailed P values ​​< 0.05 yi tekiwile tani hi ya nkoka hi tlhelo ra tinhlayo.
pH ya mati yo nwa ayiri 7.60 ± 0.01 loko kungase engeteriwa NaHS na 7.71 ± 0.03 endzhaku ka ku engeteriwa ka NaHS (n = 13, p = 0.0029). Mahiselo ya mati yo nwa akuri 26.5 ± 0.2 naswona ya hungutekile kuya eka 26.2 ± 0.2 endzhaku ka ku engeteriwa ka NaHS (n = 13, p = 0.0128). Lulamisa 30 μM NaHS solution eka mati yo nwa kutani u hlayisa eka bodlhela ra mati. NaHS solution ayi tshamisekangi naswona nhlayo ya yona yihunguteka hiku famba ka nkarhi. Loko ku tekiwa swikombiso kusuka enhan’wini ya bodlhela ra mati, kuhunguteka lokukulu (68.0%) swivoniwe eka awara yosungula, naswona nhlayo ya NaHS eka solution yihunguteke hi 72% na 75% endzhaku ka 12 na 24 wa tiawara, hiku landzelelana. Eka tisampulu leti kumiweke kusuka eka mabodlhela ya mati, kuhunguteka ka NaHS akuri ka nkoka kuya fika eka 2 wa tiawara, kambe endzhaku ka 12 na 24 wa tiawara aswi hungutekile hi 47% na 72%, hiku landzelelana. Data leyi yi kombisa leswaku tiphesente ta NaHS eka 30 μM solution leyi lulamisiweke eka mati yo nwa a ti hungutekile kuya kwalomu ka kotara ya nkoka wosungula endzhaku ka 24 wa tiawara, kunga langutiwi ndzhawu ya sampulu (Xifaniso 1).
Ku tshamiseka ka NaHS solution (30 μM) eka mati yo nwa eka mabodlhela ya rat/mouse. Endzhaku ka ku lulamisiwa ka solution, swikombiso swi tekiwile kusuka eka tip nale ndzeni ka bodlhela ra mati. Data yi nyikeriwile tanihi xiringaniso ± SD (n = 6/ntlawa). * na #, P < 0.05 loko ku pimanisiwa na nkarhi wa 0. Xifaniso xa bodlhela ra mati xi kombisa nhloko (leyi nga na ku pfuleka) na miri wa bodlhela. Volume ya tip yi kwalomu ka 740 μL.
Nhlayo ya NaHS eka 30 μM solution leyinga lulamisiwa akuri 30.3 ± 0.4 μM (range: 28.7–31.9 μM, n = 12). Hambiswiritano, endzhaku ka 24 h, nhlayo ya NaHS yi hungutekile kuya eka nhlayo yale hansi (xiringaniso: 3.0 ± 0.6 μM). Hilaha swi kombisiweke hakona eka Xifaniso xa 2, nhlayo ya NaHS leyi ti rats ti nga hlangana na yona a yi nga cinci hi nkarhi wa dyondzo.
Ndzilo wa miri wa ti rats ta xisati wu tlakuke swinene hiku famba ka nkarhi (kusuka eka 205.2 ± 5.2 g kuya eka 213.8 ± 7.0 g eka ntlawa wa vulawuri na kusuka eka 204.0 ± 8.6 g kuya eka 211.8 ± 7.5 g eka ntlawa lowu tshunguriweke hi NaHS); hambiswiritano, vutshunguri bya NaHS a byi nga ri na vuyelo eka ntiko wa miri (Fig. 3). Ndzilo wa miri wa ti rats ta xinuna wu tlakuke swinene hiku famba ka nkarhi (kusuka eka 338.6 ± 8.3 g kuya eka 352.4 ± 6.0 g eka ntlawa wa vulawuri na kusuka eka 352.4 ± 5.9 g kuya eka 363.2 ± 4.3 g eka ntlawa lowu tshunguriweke hi NaHS); hambiswiritano, vutshunguri bya NaHS a byi nga ri na vuyelo eka ntiko wa miri (Fig. 3).
Ku cinca ka ntiko wa miri eka ti rats ta xisati na ta xinuna endzhaku ka ku nyikiwa ka NaHS (30 μM). Data yi nyikiwile tani hi xiringaniso ± SEM naswona yi pimanisiwile hiku tirhisa tindlela timbirhi leti pfanganisiweke (exikarhi ka) nxopaxopo wa ku hambana na xikambelo xa Bonferroni post hoc. n = 5 wa rimbewu rin’wana na rin’wana eka ntlawa wun’wana na wun’wana.
Serum urea na creatine phosphate concentrations ayi ringanisiwa eka control na NaSH-treated rats eka nkambisiso hinkwawo. Ku tlula kwalaho, vutshunguri bya NaSH a byi khumbanga nhlayo ya urea ya serum na creatinechrome ( Tafula 1 ).
Baseline serum total sulfide concentrations ayi ringanisiwa exikarhi ka control na NaHS-treated ya xinuna (8.1 ± 0.5 μM vs. 9.3 ± 0.2 μM) na ya xisati (9.1 ± 1.0 μM vs. 6.1 ± 1.1 μM) rats. Ku nyikiwa ka NaHS kuringana 14 wa masiku aswinga ri na mbuyelo eka serum total sulfide levels eka ti rats ta xinuna kumbe ta xisati (Fig. 4).
Ku cinca eka serum total sulfide concentrations eka ti rats ta xinuna na ta xisati endzhaku ka ku nyikiwa ka NaHS (30 μM). Data yi nyikiwile tanihi xiringaniso ± SEM naswona yi pimanisiwile hi ku tirhisa nxopaxopo wa tindlela timbirhi lowu pfanganisiweke (endzeni-endzeni) wa ku hambana na xikambelo xa Bonferroni post hoc. Rimbewu rin’wana na rin’wana, n = 5/ntlawa.
Mahetelelo lamakulu ya ndzavisiso lowu hileswaku mati yo nwa lawa yanga na NaHS aya tshamisekangi: i kwalomu ka kotara ntsena ya nhlayo hinkwayo yosungula ya sulfide leyinga voniwaka endzhaku ka 24 wa tiawara endzhaku ka ku tekeriwa swikombiso kusuka eka nhloko nale ndzeni ka mabodlhela ya mati ya rat/mouse. Kuya emahlweni, ti rats ti langutane na ti concentrations ta NaHS letinga tshamisekangiki hikokwalaho ka ku lahlekeriwa hi H2S eka NaHS solution, naswona ku engeteriwa ka NaHS eka mati yo nwa aswi khumbanga ntiko wa miri, serum urea na creatine chromium, kumbe total serum sulfide.
Eka ndzavisiso lowu, mpimo wa ku lahlekeriwa hi H2S kusuka eka 30 μM NaHS solutions leyi lulamisiweke eka mati yo nwa ayiri kwalomu ka 3% hi awara. Eka buffered solution (100 μM sodium sulfide eka 10 mM PBS, pH 7.4), nhlayo ya sulfide yivikiwile kuva yihunguteka hi 7% hiku famba ka nkarhi eka 8 h11. Khale hi sirhelele ku nyikiwa ka NaHS endzeni ka khwiri hi ku vika leswaku mpimo wa ku lahlekeriwa hi sulfide ku suka eka 54 μM NaHS solution eka mati yo nwa a wu ri kwalomu ka 2.3% hi awara (4%/awara eka 12 h yo sungula na 1.4%/awara eka 12 h yo hetelela endzhaku ko lunghiselela)8. Mindzavisiso ya khale43 yi kumile ku lahlekeriwa hi nkarhi hinkwawo ka H2S kusuka eka swintshuxo swa NaHS, ngopfu ngopfu hikokwalaho ka volatilization na oxidation. Hambi kuri handle ko engeteriwa ka ti bubbles, sulfide eka stock solution yi lahleka hiku hatlisa hikokwalaho ka H2S volatilization11. Tidyondzo ti kombisile leswaku hi nkarhi wa endlelo ra dilution, leri tekaka kwalomu ka 30–60 wa tisekoni, kwalomu ka 5–10% wa H2S ya lahleka hikokwalaho ka evaporation6. Ku sivela ku khuluka ka H2S kusuka eka solution, valavisisi va tekile magoza yo hlaya, kukatsa naku hlanganisiwa hiku olova ka solution12, ku funengeta stock solution hi plastic film6, naku hunguta ku hlangana ka solution eka moya, tani hileswi mpimo wa ku khuluka ka H2S wu titshegeke hi air-liquid interface.13 Spontaneous oxidation ya H2S yi humelela ngopfu hikokwalaho ka ti transition metal ions, ngopfu ngopfu ferric iron, leswi nga thyaka ematini.13 Oxidation ya H2S yi endla leswaku ku va na ti polysulfides (ti athomo ta sulphur leti hlanganisiweke hi ti covalent bonds)11. Ku papalata oxidation ya yona, ti solutions letingana H2S ti lulamisiwa eka ti deoxygenated solvents44,45 kutani ti basisiwa hi argon kumbe nitrogen kuringana 20–30 min ku tiyisisa deoxygenation.11,12,37,44,45,46 Diethylenetriaminepentaacetic acid (DTPA) i metal chelator (10–4 M) leyi sivelaka HS-auoxidation eka swintshuxo swa aerobic. Loko kungari na DTPA, autooxidation rate ya HS- yiva kwalomu ka 50% eka kwalomu ka 3 h eka 25°C37,47. Kuya emahlweni, tani hileswi oxidation ya 1e-sulfide yi hlohleteriwaka hi ku vonakala ka ultraviolet, solution yifanele ku hlayisiwa eka ayisi naswona yi sirheleriwa eka ku vonakala11.
Hilaha swi kombisiweke hakona eka Xifaniso xa 5, NaHS yi hambanisiwa yi va Na+ na HS-6 loko yi khuluka ematini; ku hambana loku ku kumiwa hi pK1 ya reaction, leyi titshegeke hi mahiselo: pK1 = 3.122 + 1132/T, laha T yi sukaka eka 5 kuya eka 30°C naswona yi pimiwa hi ti degrees Kelvin (K), K = °C + 273.1548. HS- yina pK2 yale henhla (pK2 = 19), hikokwalaho eka pH < 96.49, S2- ayi vumbiwi kutani yivumbiwa hi nhlayo leyintsongo swinene. Kuhambana na sweswo, HS- yitirha tani hi base naswona yi amukela H+ kusuka eka molecule ya H2O, naswona H2O yitirha tani hi acid naswona yi hundzuriwa H2S na OH-.
Ku vumbiwa ka gasi ya H2S leyi n’okisiweke eka xitofu xa NaHS (30 μM). aq, xitofu xa mati; g, gasi, gasi; l, xihalaki. Swibalo hinkwaswo switeka leswaku pH ya mati = 7.0 naswona mahiselo ya mati = 20 °C. Yi endliwe hi BioRender.com.
Hambi leswi kungana vumbhoni bya leswaku swintshuxo swa NaHS aswi tshamisekangi, tidyondzo to hlaya ta swiharhi ti tirhise swintshuxo swa NaHS eka mati yo nwa tani hi nkatsakanyo wa munyiki wa H2S15,16,17,18,19,20,21,22,23,24,25,26 na nkarhi wo nghenelela lowu sukelaka eka 1 kuya eka 21 wa mavhiki (Table 2). Hi nkarhi wa tidyondzo leti, solution ya NaHS ayi pfuxetiwa endzhaku ka 12 h yin’wana na yin’wana, 15, 17, 18, 24, 25 h kumbe 24 h, 19, 20, 21, 22, 23 h. Mbuyelo wa hina wu kombisile leswaku ti rats ti hlangana na nhlayo ya mirhi leyi nga tshamisekangiki hikokwalaho ka ku lahlekeriwa hi H2S kusuka eka NaHS solution, naswona nhlayo ya NaHS eka mati yo nwa ya ti rats yi cinca cinca swinene eka 12 kumbe 24 h (vona Xifaniso 2). Timbirhi ta tidyondzo leti ti vikile leswaku ti levele ta H2S ematini ti tshamile ti tshamisekile eka 24 h22 kumbe leswaku i 2–3% ntsena wa ku lahlekeriwa ka H2S loku voniweke eka 12 h15, kambe a yi nyikanga datha yo seketela kumbe vuxokoxoko bya ku pima. Mindzavisiso yimbirhi yi kombisile leswaku diameter leyintsongo ya mabodlhela ya mati yinga hunguta ku khuluka ka H2S15,19. Hambiswiritano, mbuyelo wa hina wu kombisile leswaku leswi swinga hlwela ntsena ku lahlekeriwa hi H2S kusuka eka bodlhela ra mati hi 2 h kutlula 12–24 h. Tidyondzo leti hatimbirhi ti xiya leswaku hi ehleketa leswaku mpimo wa NaHS eka mati yo nwa a wu cincanga hikuva a hi vonanga ku cinca ka muhlovo ematini; hikokwalaho, oxidation ya H2S hi moya a yi nga ri ya nkoka19,20. Lexi hlamarisaka, ndlela leyi ya subjective yi kambela ku tshamiseka ka NaHS ematini ku tlula ku pima ku cinca ka nhlayo ya yona hi ku famba ka nkarhi.
Ku lahlekeriwa hi H2S eka NaHS solution swi fambelana na pH na mahiselo. Tani hilaha swi kombisiweke hakona eka dyondzo ya hina, ku n’okisiwa ka NaHS ematini swi endla leswaku ku endliwa xitofu xa alkaline50. Loko NaHS yi pfanganisiwa ematini, ku endliwa ka gasi ya H2S leyi n’okisiweke swiya hi nhlayo ya pH6. Loko pH yale hansi ya solution, xiphemu xa NaHS lexi nga kona tani hi ti molecules ta gasi ya H2S xiva xikulu naswona sulfide yotala yi lahleka kusuka eka solution ya mati11. Ku hava na yin’we ya tidyondzo leti leyi vikeke pH ya mati yo nwa lawa ya tirhisiwaka tani hi solvent ya NaHS. Kuya hi switsundzuxo swa WHO, leswi amukeriwaka hi matiko yotala, pH ya mati yo nwa yifanele kuva eka 6.5–8.551. Eka pH leyi, mpimo wa oxidation yo tisungulela ya H2S wu tlakuka kwalomu ka khume ra tiphesente13. Ku n’okisiwa ka NaHS ematini eka pH leyi swita endla leswaku kuva na nhlayo ya gasi ya H2S leyi n’okisiweke ya 1 kuya eka 22.5 μM, leswi kandziyisaka nkoka wo langutisisa pH ya mati loko kungase n’okisiwa NaHS. Ku engetela kwalaho, mpimo wa mahiselo lowu vikiweke eka dyondzo leyi nga laha henhla (18–26 °C) wu ta endla leswaku ku va na ku cinca eka nhlayo ya gasi ya H2S leyi n’okiweke eka xitofu xa kwalomu ka 10%, tanihileswi ku cinca ka mahiselo ku cincaka pK1, naswona ku cinca lokuntsongo eka pK1 ku nga va na nkucetelo lowukulu eka nhlayo ya gasi ya H2S leyi n’okiweke48. Ku engetela kwalano, nkarhi woleha wa tidyondzo tin’wana (5 wa tinhweti)22, laha eka wona ku languteriweke ku cinca cinca lokukulu ka mahiselo, na swona swi nyanyisa xiphiqo lexi.
Tidyondzo hinkwato handle ka yin’we21 ti tirhise 30 μM NaHS solution eka mati yo nwa. Ku hlamusela mpimo lowu tirhisiweke (i.e. 30 μM), vatsari van’wana va kombisile leswaku NaHS eka xiphemu xa mati yi humesa kahle nhlayo leyi fanaka ya gasi ya H2S na leswaku mpimo wa physiological wa H2S i 10 kuya eka 100 μM, hikokwalaho mpimo lowu wu le ndzeni ka mpimo wa physiological15,16. Van’wana va hlamuserile leswaku 30 μM NaHS yinga hlayisa levele ya plasma H2S endzeni ka physiological range, i.e. 5–300 μM19,20. Hi tekela enhlokweni nhlayo ya NaHS ematini ya 30 μM (pH = 7.0, T = 20 °C), leyi tirhisiweke eka tidyondzo tin’wana ku dyondza mbuyelo wa H2S. Hi nga hlayela leswaku nhlayo ya gasi ya H2S leyi n’okisiweke i 14.7 μM, leswi nga kwalomu ka 50% wa nhlayo yo sungula ya NaHS. Nhlayo leyi yi fana na ntikelo lowu hlayiweke hi vatsari van’wana ehansi ka swiyimo leswi fanaka13,48.
Eka nkambisiso wa hina, ku nyikiwa ka NaHS a ku cincanga ntiko wa miri; mbuyelo lowu wu fambisana na mbuyelo wa tidyondzo tin’wana eka timbeva ta xinuna22,23 na timbuti ta xinuna18; Hambiswiritano, tidyondzo timbirhi ti vikile leswaku NaSH yi vuyisele ku hunguteka ka ntiko wa miri eka timbuti leti tsemiweke nephrectomized24,26, kasi tidyondzo tin’wana a ti vikanga mbuyelo wa vulawuri bya NaSH eka ntiko wa miri15,16,17,19,20,21,25. Ku tlula kwalaho, eka dyondzo ya hina, ku nyikiwa ka NaSH a swi khumbanga ti levele ta serum urea na creatine chromium, leswi fambelanaka na mbuyelo wa xiviko xin’wana25.
Dyondzo yi kumile leswaku ku engeteriwa ka NaHS eka mati yo nwa kuringana 2 wa mavhiki aswi khumbanga nhlayo hinkwayo ya serum sulfide eka ti rats ta xinuna na ta xisati. Ku kumiwa loku ku fambisana na mbuyelo wa Sen et al. (16): 8 wa mavhiki ya vutshunguri hi 30 μM NaHS ematini yo nwa a swi khumbanga mpimo wa plasma sulfide eka timbambu leti lawuriwaka; hambiswiritano, va vike leswaku ku nghenelela loku ku vuyisele ku hunguteka ka mpimo wa H2S eka plasma ya timbeva leti tsemiweke nephrectomized. Li na van’wana. (22) va tlhele va vika leswaku vutshunguri bya 30 μM NaHS ematini yo nwa ku ringana tin’hweti ta 5 byi engetele ti levele ta plasma free sulfide eka timbeva leti dyuhaleke hi kwalomu ka 26%. Mindzavisiso yin’wana a yi vikanga ku cinca eka sulfide leyi rhendzelekaka endzhaku ko cheriwa NaHS eka mati yo nwa.
Tidyondzo ta nkombo ti vikile ku tirhisa Sigma NaHS15,16,19,20,21,22,23 kambe a ti nyikanga vuxokoxoko byin’wana hi mati ya hydration, naswona tidyondzo ta ntlhanu a ti boxi xihlovo xa NaHS lexi tirhisiweke eka tindlela ta tona to lunghiselela17,18,24,25,26. NaHS i molecule leyingana mati naswona mati ya yona ya hydration yanga hambana, leswi khumbhaka nhlayo ya NaHS leyi lavekaka ku lulamisa solution ya molarity leyi nyikiweke. Xikombiso, nhlayo ya NaHS eka dyondzo ya hina a yi ri NaHS•1.3 H2O. Xisweswo, nhlayo ya xiviri ya NaHS eka tidyondzo leti yi nga ha va ehansi ku tlula leyi vikiweke.
“Xana nkatsakanyo wo tano lowu hanyaka nkarhi wo koma wu nga va njhani ni vuyelo byo leha swonghasi?” Pozgay et al.21 va vutisile xivutiso lexi loko va kambisisa vuyelo bya NaHS eka colitis eka timbeva. Va tshemba leswaku tidyondzo ta nkarhi lowu taka ti ta kota ku hlamula xivutiso lexi naswona ti ringanyeta leswaku swintshuxo swa NaHS swi nga ha va na ti polysulfides leti tiyeke swinene ku engetela eka H2S na ti disulfides leti hlanganisaka mbuyelo wa NaHS21. Xin’wana lexi nga endlekaka hileswaku nhlayo yale hansi swinene ya NaHS leyi salaka eka solution na yona yingava na mbuyelo lowu pfunaka. Entiyisweni, Olson na van’wana. yi nyikile vumbhoni bya leswaku ti levele ta micromolar ta H2S engatini a hi ta physiological naswona ti fanele ku va eka nanomolar range kumbe ti nga ri kona hi ku helela13. H2S yi nga tirha hi ku tirhisa protein sulfation, ku cinca loku nga hundzukiki endzhaku ka vuhundzuluxi loku khumbaka ntirho, ku tshamiseka, na ku vekiwa ka tiphrotheyini to tala52,53,54. Entiyisweni, ehansi ka swiyimo swa physiological, kwalomu ka 10% kuya eka 25% wa ti protein totala ta xivindzi ti sulfylated53. Mindzavisiso leyimbirhi yi amukela ku onhiwa hi xihatla ka NaHS19,23 kambe hi ndlela yo hlamarisa yi vula leswaku “hi lawuleke nhlayo ya NaHS eka mati yo nwa hi ku ma siva siku na siku.”23 Nkambisiso wun’wana hi xihoxo wu vule leswaku “NaHS i munyiki wa H2S wa ntolovelo naswona yi tala ku tirhisiwa eka matirhelo ya tliliniki ku siva H2S hi yoxe.”18
Nkanelo lowu nga laha henhla wu kombisa leswaku NaHS yi lahleka kusuka eka solution hiku tirhisa volatilization, oxidation na photolysis, naswona hikokwalaho switsundzuxo swin’wana swi endliwa ku hunguta ku lahlekeriwa hi H2S kusuka eka solution. Xosungula, ku khuluka ka H2S swiya hi gas-liquid interface13 na pH ya solution11; hikokwalaho, ku hunguta ku lahlekeriwa hi evaporative, nhamu ya bodlhela ra mati yinga endliwa yiva yitsongo hilaha swinga hlamuseriwa hakona khale15,19, naswona pH ya mati yinga lulamisiwa kuya eka mpimo wale henhla lowu amukelekaka (i.e., 6.5–8.551) ku hunguta ku lahlekeriwa hi evaporative11. Xavumbirhi, oxidation yo tisungulela ya H2S yi humelela hikokwalaho ka mbuyelo wa oxygen na vukona bya ti transition metal ions eka mati yo nwa13, hikokwalaho deoxygenation ya mati yo nwa hi argon kumbe nitrogen44,45 na ku tirhisiwa ka ti metal chelators37,47 swinga hunguta oxidation ya ti sulfides. Xavunharhu, ku sivela photodecomposition ya H2S, mabodlhela ya mati yanga phutseriwa hi aluminium foil; Mukhuva lowu wu tirha na le ka switirhisiwa leswi twaka ku vonakala swo fana na streptozotocin55. Eku heteleleni, ti inorganic sulfide salts (NaHS, Na2S, na CaS) tinga nyikiwa hiku tirhisa gavage kutlula ku n’okisiwa eka mati yo nwa tani hileswi swinga vikiwa khale56,57,58; tidyondzo ti kombisile leswaku radioactive sodium sulfide leyi nyikiwaka hi gavage eka ti rats yi tswongiwa kahle naswona yi hangalasiwa eka kwalomu ka ti tissue hinkwato59. Kufikela sweswi, minkambisiso yotala yi nyikile ti inorganic sulfide salts intraperitoneally; hambiswiritano, ndlela leyi a yi tirhisiwi ngopfu eka swiyimo swa tliliniki60. Hi hala tlhelo, ndlela ya nomu i ndlela leyi tolovelekeke no tsakeriwa swinene ya vulawuri eka vanhu61. Hikokwalaho, hi ringanyeta ku kambela mbuyelo wa vaphakeri va H2S eka ti rodents hi oral gavage.
Xiphiqo hileswaku hi pimile sulfide eka aqueous solution na serum hiku tirhisa ndlela ya MB. Maendlelo yo pima sulfide ya katsa iodine titration, spectrophotometry, ndlela ya electrochemical (potentiometry, amperometry, ndlela ya coulometric na ndlela ya amperometric) na chromatography (gas chromatography na high-performance liquid chromatography), exikarhi ka swona ndlela leyi tirhisiwaka ngopfu i ndlela ya MB spectrophotometric62. Xipimelo xa ndlela ya MB yo pima H2S eka tisampulu ta biological hileswaku yi pima ti compounds hinkwato letingana sulphur naswona kungari H2S63 ya mahala hikuva yi endliwa ehansi ka swiyimo swa acid, leswi endlaka leswaku ku humesiwa sulphur kusuka eka xihlovo xa biological64. Hambiswiritano, hi ku ya hi Nhlangano wa Rihanyo ra Mani na Mani wa le Amerika, MB i ndlela leyi tolovelekeke yo pima sulfide ematini65. Hikwalaho, ku siveriwa loku a ku khumbi mimbuyelo ya hina leyikulu eka ku nga tshamiseki ka swintshuxo leswi nga na NaHS. Ku tlula kwalaho, eka dyondzo ya hina, ku vuyiseriwa ka mimpimo ya sulfide eka tisampulu ta mati na serum leti nga na NaHS a ku ri 91% na 93%, hi ku landzelelana. Mimpimo leyi yi fambisana na swiyenge leswi vikiweke khale (77–92)66, leswi kombisaka ku kongoma ka nxopaxopo loku amukelekaka42. Swi fanerile ku xiya leswaku hi tirhise timbavala ta xinuna na ta xisati hi ku landza swiletelo swa National Institutes of Health (NIH) ku papalata ku titshega ngopfu hi tidyondzo ta swiharhi swa xinuna ntsena eka tidyondzo ta le mahlweni ka tliliniki67 na ku katsa timbavala ta xinuna na ta xisati loko swi koteka68. Mhaka leyi yi kandziyisiwile hi van’wana69,70,71.
Hiku gimeta, mbuyelo wa ndzavisiso lowu wukomba leswaku swintshuxo swa NaHS leswi lulamisiweke kusuka eka mati yo nwa aswinga tirhisiwi ku humesa H2S hikokwalaho ka ku nga tshamiseki ka swona. Ndlela leyi ya vulawuri yinga veka swifuwo eka swiyimo leswinga tshamisekangiki na swale hansi kutlula leswi languteriweke swa NaHS; hikwalaho, leswi kumiweke swi nga ha va swi nga tirhi eka vanhu.
Tidathaseti leti tirhisiweke na/kumbe leti xopaxopiweke hi nkarhi wa dyondzo ya sweswi ti kumeka eka mutsari loyi a fambelanaka na yena loko a komberiwa hi ndlela leyi twalaka.
Szabo, K. Nkarhi wa ndzavisiso wa hayidirojeni sulfide (H2S): ku suka eka chefu ya mbango ku ya eka muhlanganisi wa biological. Biochemistry na Pharmacology 149, 5–19. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2017.09.010 (2018).
Abe, K. na Kimura, H. Ntirho lowu nga kotekaka wa hydrogen sulfide tanihi endogenous neuromodulator. Nyuziphepha ya Sayense ya Miehleketo, 16, 1066–1071. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. 1996 (1996).
Chirino, G., Szabo, C. na Papapetropoulos, A. Ntirho wa physiological wa hydrogen sulfide eka tisele ta swihadyani leswi mamisaka, tinyama na swirho. Ndzavisiso eka Physiology na Molecular Biology 103, 31–276. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. (2023).
Dillon, KM, Carrazzone, RJ, Matson, JB, na Kashfi, K. Xitshembiso lexi cincaka xa tisisiteme ta ku rhumela hi tisele ta nitric oxide na hydrogen sulfide: nguva leyintshwa ya vutshunguri lebyi endleriweke munhu hi xiyexe. Tikhemikhali ta swilo leswi hanyaka na Ntivo-vuxongi bya Mirhi 176, 113931. https://doi.org/10.1016/j.bcp.2020.113931 (2020).
Sun, X., na van’wana. Ku nyikiwa ka nkarhi wo leha ka munyiki wa hayidirojeni sulfide loyi a humesaka hi ku nonoka swi nga sivela ku vaviseka ka myocardial ischemia/reperfusion. Swiviko swa sayense 7, 3541. https://doi.org/10.1038/s41598-017-03941-0 (2017).
Sitdikova, GF, Fuchs, R., Kainz, W., Weiger, TM na Hermann, A. Phosphorilation ya xiteji xa BK yi lawula ku twisisa ka hayidirojeni sulfide (H2S). Mindzilakano eka Ntivo-vutomi 5, 431. https://doi.org/10.3389/fphys.2014.00431 (2014).
Sitdikova, GF, Weiger, TM na Hermann, A. Hydrogen sulfide yi tlakusa ntirho wa calcium-activated potassium (BK) channel eka tisele ta tshumba ra pituitary ra rat. Vumaki bya vumaki. Va-Pflueger lava nga ni swiphepherhele. 459, 389-397. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. (2010).
Jeddy, S., na van’wana. Hydrogen sulfide yi tlakusa mbuyelo wo sirhelela wa nitrite eka ku vaviseka ka myocardial ischemia-reperfusion eka ti rats ta type 2 ta diabetic. Okisayidi ya Nitric 124, 15–23. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. (2022).
Corvino, A., na van’wana. Mikhuva eka tikhemikhali ta munyiki wa H2S na nkucetelo wa yona eka mavabyi ya mbilu na misiha. Swidlaya-switsongwatsongwana 10, 429. https://doi.org/10.3390/antiox10030429 (2021).
DeLeon, E. R., Stoy, G. F., na Olson, K. R. (2012). Ku lahlekeriwa loku nga tirhiki ka hydrogen sulfide eka swikambelo swa biological. Nxopaxopo wa Biochemistry 421, 203–207. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. (2012).
Nagy, P., na van’wana. Swiyenge swa tikhemikhali swa mimpimo ya hydrogen sulfide eka swikombiso swa physiological. Biochimica na Xivumbeko xa swilo leswi hanyaka 1840, 876–891. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. (2014).
Kline, LL.D., na yena. Ku kumiwa ka spectrophotometric ka hydrogen sulfide eka mati ya ntumbuluko. Limnol. Oceanogr. 14, 454-458. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2. 1969.14.3.0454 (1969).
Olson, K. R. (2012) va vula leswaku. Vuleteri lebyi tirhaka eka tikhemikhali na ntivo-vutomi bya hayidirojeni sulfide. “Swidlaya-switsongwatsongwana.” Ku Endliwa ka Swikombiso swa Redox. 17, 32–44. Nkanelo wa ririmi ra Xitsonga eka swiyenge swa 1 na 2011.4401 (2012).