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Sulfide ya Hydrogen (H2S) igira ingaruka nyinshi ku mubiri w'umuntu ku miterere y'umubiri n'indwara. Sodium hydrosulfide (NaHS) ikoreshwa cyane nk'igikoresho cy'imiti mu gusuzuma ingaruka za H2S mu bushakashatsi bw'ibinyabuzima. Nubwo gutakaza H2S bivuye ku bisubizo bya NaHS bifata iminota mike gusa, ibisubizo bya NaHS byakoreshejwe nk'ibice by'inyongera bya H2S mu mazi yo kunywa muri bimwe mu bushakashatsi bw'inyamaswa. Ubu bushakashatsi bwagaragaje niba kunywa amazi afite 30 μM ya NaHS yateguwe mu macupa y'imbeba/imbeba bishobora kuguma mu buryo buhamye nibura amasaha 12-24, nk'uko byagaragajwe n'abanditsi bamwe. Tegura umuti wa NaHS (30 μM) mu mazi yo kunywa hanyuma uhite uwusuka mu macupa y'amazi y'imbeba/imbeba. Ingero zakusanyijwe ku mutwe no imbere mu icupa ry'amazi ku masaha 0, 1, 2, 3, 4, 5, 6, 12 na 24 kugira ngo upime ingano ya sulfide hakoreshejwe uburyo bwa methylene blue. Byongeye kandi, imbeba z'ingabo n'iz'ingore zatewe NaHS (30 μM) mu gihe cy'ibyumweru bibiri kandi igipimo cya sulfide mu maraso cyapimwe buri munsi mu cyumweru cya mbere no mu mpera z'icyumweru cya kabiri. Umuti wa NaHS mu gipimo cyavuye ku isonga ry'icupa ry'amazi wari uhindagurika; wagabanutseho 72% na 75% nyuma y'amasaha 12 na 24, uko bikurikirana. Mu bipimo byafashwe mu macupa y'amazi, igabanuka rya NaHS ntiryari rigaragara mu masaha 2; icyakora, ryagabanutseho 47% na 72% nyuma y'amasaha 12 na 24, uko bikurikirana. Urushinge rwa NaHS ntirwagize ingaruka ku gipimo cya sulfide mu maraso y'imbeba z'ingabo n'iz'ingore. Muri make, umuti wa NaHS wakozwe mu mazi yo kunywa ntugomba gukoreshwa mu gutanga H2S kuko umuti uhindagurika. Ubu buryo bwo gutanga umuti buzashyira amatungo ku rugero rudasanzwe kandi ruciriritse ugereranije n'uko byari byitezwe rwa NaHS.
Hydrogen sulfide (H2S) yakoreshejwe nk'uburozi kuva mu 1700; icyakora, uruhare rwayo rushoboka nk'ikintu cy'umwimerere gitanga ibimenyetso bya biosignaling rwasobanuwe na Abe na Kimura mu 1996. Mu myaka mirongo itatu ishize, imikorere myinshi ya H2S mu buryo butandukanye bw'abantu yarasobanuwe, bituma hamenyekana ko molekile zitanga H2S zishobora kugira akamaro mu buvuzi mu kuvura cyangwa mu kuvura indwara zimwe na zimwe; reba Chirino et al. kugira ngo ubone isuzuma riheruka.
Sodium hydrosulfide (NaHS) yakoreshejwe cyane nk'igikoresho cy'imiti mu gusuzuma ingaruka za H2S mu miterere myinshi y'uturemangingo n'ubushakashatsi ku nyamaswa5,6,7,8. Ariko, NaHS si umutanga H2S mwiza kuko ihinduka vuba ikaba umuti wa H2S/HS- muri, ihumanywa byoroshye na polysulfide, kandi irashwanyagurika byoroshye no guhinduka4,9. Mu bushakashatsi bwinshi bw'ibinyabuzima, NaHS ishongeshwa mu mazi, bigatuma habaho guhinduka kw'ibintu mu buryo butunguranye no kubura H2S10,11,12, oxidation yihuta ya H2S11,12,13, na photolysis14. Sulfide mu muti w'umwimerere itakaza vuba cyane bitewe no guhinduka kw'ibintu mu buryo butunguranye bya H2S11. Mu gikoresho gifunguye, igice cy'ubuzima (t1/2) cya H2S ni iminota 5, kandi ubwinshi bwayo bugabanukaho hafi 13% ku munota10. Nubwo kubura sulfide ya hydrogen kuva mu bisubizo bya NaHS bifata iminota mike gusa, ubushakashatsi bumwe na bumwe ku nyamaswa bwakoresheje ibisubizo bya NaHS nk'isoko ya sulfide ya hydrogen mu mazi yo kunywa mu gihe cy'ibyumweru 1-21, bisimbura umuti urimo NaHS buri masaha 12-24. 15,16,17,18,19,20,21,22,23,24,25,26 Iyi mikorere ntabwo ijyanye n'amahame y'ubushakashatsi bwa siyansi, kuko ingano y'imiti igomba gushingira ku ikoreshwa ryayo mu zindi nyamaswa, cyane cyane abantu.27
Ubushakashatsi bwakozwe mbere yo kuvura indwara mu buvuzi bushingiye ku binyabuzima bugamije kunoza ireme ry’ubuvuzi bw’abarwayi cyangwa umusaruro w’ubuvuzi. Ariko, ibyavuye mu bushakashatsi bwinshi ku nyamaswa ntibiramenyekana ku bantu28,29,30. Imwe mu mpamvu zatumye ubu buryo bwo guhindura ibintu bunanirwa ni ukudashyira mu bikorwa ireme ry’uburyo bwo gusuzuma inyamaswa30. Kubwibyo, intego y’ubu bushakashatsi kwari ukureba niba ibisubizo bya 30 μM NaHS byateguwe mu macupa y’amazi y’imbeba/imbeba bishobora kuguma mu mazi meza mu gihe cy’amasaha 12-24, nk’uko bivugwa cyangwa byavuzwe muri bimwe mu bushakashatsi.
Igerageza ryose muri ubu bushakashatsi ryakozwe hakurikijwe amabwiriza yasohotse mu kwita no gukoresha inyamaswa zo muri laboratwari muri Irani31. Raporo zose z'igerageza muri ubu bushakashatsi zakurikije amabwiriza ya ARRIVE32. Komite Ishinzwe Imyitwarire mu Kigo cy'Ubumenyi bw'Imisemburo, Kaminuza ya Shahid Beheshti y'Ubumenyi bw'Ubuvuzi, yemeje inzira zose z'igerageza muri ubu bushakashatsi.
Zinc acetate dihydrate (CAS: 5970-45-6) na anhydrous ferric chloride (CAS: 7705-08-0) byaguzwe muri Biochem, Chemopahrama (Cosne-sur-Loire, mu Bufaransa). Sodium hydrosulfide hydrate (CAS: 207683-19-0) na N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) byaguzwe muri Sigma-Aldrich (St. Louis, MO, muri Amerika). Isoflurane yaguzwe muri Piramal (Bethlehem, PA, muri Amerika). Aside Hydrochloric (HCl) yaguzwe muri Merck (Darmstadt, mu Budage).
Tegura umuti wa NaHS (30 μM) mu mazi yo kunywa hanyuma uhite uwusuka mu macupa y'amazi y'imbeba/imbeba. Iyi ngano yatoranijwe hashingiwe ku bitabo byinshi byakoreshejwe na NaHS nk'isoko ya H2S; reba igice cy'Ibiganiro. NaHS ni molekile ifite amazi ashobora kuba arimo amazi atandukanye (ni ukuvuga NaHS • xH2O); nk'uko uwakoze ubushakashatsi yabitangaje, igipimo cya NaHS cyakoreshejwe mu bushakashatsi bwacu cyari 70.7% (ni ukuvuga NaHS • 1.3 H2O), kandi twafashe iki giciro mu mibare yacu, aho twakoresheje uburemere bwa molekile ya 56.06 g / mol, ari nabwo buremere bwa molekile ya NaHS idakora amazi. Amazi yo kunywa (nanone yitwa amazi yo kristalo) ni molekile z'amazi zigize imiterere ya kristalo33. Hydrate zifite imiterere itandukanye ya fiziki na thermodynamic ugereranije na anhydrates34.
Mbere yo kongeramo NaHS mu mazi yo kunywa, pima pH n'ubushyuhe bw'umushongi. Suka umuti wa NaHS ako kanya mu icupa ry'amazi ry'imbeba/imbeba riri mu kazu k'inyamaswa. Ingero zakusanyijwe uhereye ku mutwe no imbere mu icupa ry'amazi ku masaha 0, 1, 2, 3, 4, 5, 6, 12, na 24 kugira ngo upime ingano ya sulfide. Ibipimo bya sulfide byafashwe ako kanya nyuma ya buri ngero. Twabonye ingero uhereye ku mutwe w'umuyoboro kuko ubushakashatsi bumwe bwagaragaje ko ingano nto y'umuyoboro w'amazi ishobora kugabanya H2S evaporation15,19. Iki kibazo gisa nkaho kireba umuti uri mu icupa. Ariko, si ko byagenze ku muti wari mu ijosi ry'icupa ry'amazi, wari ufite igipimo cyo hejuru cy'evaporation kandi warahitaga uhinduka; mu by'ukuri, inyamaswa zabanje kunywa aya mazi.
Imbeba z’ingabo n’ingore za Wistar zakoreshejwe muri ubwo bushakashatsi. Imbeba zashyizwe mu tuzu twa polypropylene (imbeba 2-3 kuri buri kazu) mu buryo busanzwe (ubushyuhe bwa 21-26 °C, ubushyuhe bwa 32-40%) hamwe n’urumuri rw’amasaha 12 (7h00 kugeza 7h00) n’umwijima w’amasaha 12 (7h00 kugeza 7h00). Imbeba zari zifite amazi yo kuvogera ku buntu kandi zagaburirwaga chow isanzwe (Khorak Dam Pars Company, Tehran, Irani). Imbeba z’ingore zihuye n’imyaka (amezi 6) (n=10, uburemere bw’umubiri: 190–230 g) n’iz’ingabo (n=10, uburemere bw’umubiri: 320–370 g) zagabanyijwemo amatsinda yo kugenzura no kuvurwa na NaHS (30 μM) (n=5 kuri buri tsinda). Kugira ngo tumenye ingano y’icyitegererezo, twakoresheje uburyo bwa KISS (Keep It Simple, Stupid), buhuza ubunararibonye bwabanje n’isesengura ry’imbaraga35. Twabanje gukora inyigo y’igerageza ku mbeba 3 maze tumenya urwego rw’ikigereranyo cya sulfide mu maraso n’itandukaniro risanzwe (8.1 ± 0.81 μM). Hanyuma, dutekereje ku mbaraga za 80% kandi dufata urwego rw’uburemere bwa 5% ku mpande zombi, twasanze ingano y’icyitegererezo cy’ibanze (n = 5 dushingiye ku nyandiko zabanje) ihuye n’ingano y’ingaruka isanzwe ya 2.02 hamwe n’agaciro kagenwe mbere na Festing ko kubara ingano y’icyitegererezo cy’inyamaswa zakorewe igerageza35. Nyuma yo gukuba iki gipimo na SD (2.02 × 0.81), ingano y’ingaruka yahanuwe (1.6 μM) yari 20%, ibyo bikaba byemewe. Ibi bivuze ko n = 5/itsinda ihagije kugira ngo hamenyekane impinduka ingana na 20% hagati y’amatsinda. Imbeba zagabanyijwemo mu buryo butunguranye amatsinda yagenzuwe n’ayavuwe na NaSH hakoreshejwe imikorere idasanzwe ya porogaramu ya Excel 36 (Ishusho y’inyongera ya 1). Gupima uruhu rw’inyuma byakozwe ku rwego rw’umusaruro, kandi abashakashatsi bakoze ibipimo bya biochemical ntibari bazi inshingano z’amatsinda.
Amatsinda ya NaHS y’ibitsina byombi yavuwe n’umuti wa NaHS wa 30 μM wateguriwe mu mazi yo kunywa mu gihe cy’ibyumweru 2; Umuti mushya watangwaga buri masaha 24, muri icyo gihe hapimwaga uburemere bw’umubiri. Ingero z’amaraso zakusanyijwe mu mpera z’imbeba zose hakoreshejwe ikinyabutabire cya isoflurane buri munsi ukurikiyeho mu mpera z’icyumweru cya mbere n’icya kabiri. Ingero z’amaraso zashyizwe muri centrifuge kuri 3000 g mu gihe cy’iminota 10, amaraso yatandukanyijwe maze abikwa kuri -80°C kugira ngo hakorwe urea mu maraso, creatinine (Cr), na sulfide yose. Urea mu maraso yagenzuwe hakoreshejwe uburyo bwa enzymatic urea, naho creatinine mu maraso yagenzuwe hakoreshejwe uburyo bwa photometric Jaffe hakoreshejwe ibikoresho bigurishwa (Man Company, Tehran, Irani) na automatic analyzer (Selectra E, serial number 0-2124, The Netherlands). Ingano y’impinduka za urea na Cr mu maraso yari munsi ya 2.5%.
Uburyo bwa methylene blue (MB) bukoreshwa mu gupima sulfide yose mu mazi yo kunywa no mu maraso irimo NaHS; MB ni bwo buryo bukoreshwa cyane mu gupima sulfide mu bisubizo byinshi n'ibipimo by'ibinyabuzima11,37. Uburyo bwa MB bushobora gukoreshwa mu gupima pool ya sulfide yose38 no gupima sulfide inorganic mu buryo bwa H2S, HS- na S2 mu gice cy'amazi39. Muri ubu buryo, sulfure ihinduka zinc sulfide (ZnS) mu gihe hari zinc acetate11,38. Zinc acetate precipitation ni bwo buryo bukoreshwa cyane mu gutandukanya sulfide n'izindi chromophores11. ZnS yongeye gushonga hakoreshejwe HCl11 mu bihe bya aside nyinshi. Sulfide ikorana na DMPD mu gipimo cya stoichiometric cya 1:2 mu buryo bwakoreshejwe na ferric chloride (Fe3+ ikora nk'ikintu gitanga oxidizing) kugira ngo ikore irangi MB, riboneka muri spectrophotometrically kuri 670 nm40,41. Umupaka ntarengwa wo kumenya uburyo bwa MB ni hafi 1 μM11.
Muri ubu bushakashatsi, 100 μL ya buri gipimo (umuti cyangwa serumu) byongewe mu muyoboro; hanyuma hongewemo 200 μL ya zinc acetate (1% w/v mu mazi yaciwe), 100 μL ya DMPD (20 mM muri 7.2 M HCl), na 133 μL ya FeCl3 (30 mM muri 1.2 M HCl). Uruvange rwashyizwe mu bushyuhe bwa 37°C mu mwijima mu gihe cy'iminota 30. Umuti washyizwe muri centrifuge kuri 10.000 g mu gihe cy'iminota 10, maze ubushobozi bwo kwinjiza amazi bwasomwe kuri 670 nm hakoreshejwe microplate reader (BioTek, MQX2000R2, Winooski, VT, USA). Ingano ya sulfide yagenwe hakoreshejwe calibration curve ya NaHS (0–100 μM) muri ddH2O (Ishusho y'inyongera ya 2). Ibisubizo byose byakoreshejwe mu gupima byateguwe vuba. Ingano y’impinduka mu bipimo bya sulfide hagati na hagati yari 2.8% na 3.4%, uko bikurikirana. Twanamenye sulfide yose yavuye mu mazi yo kunywa arimo sodium thiosulfate n’ibipimo bya serum dukoresheje uburyo bwo gupima bwashyizwemo imbaraga42. Ingano y’amazi yo kunywa arimo sodium thiosulfate n’ibipimo bya serum yari 91 ± 1.1% (n = 6) na 93 ± 2.4% (n = 6), uko bikurikirana.
Isesengura ry’imibare ryakozwe hakoreshejwe porogaramu ya GraphPad Prism verisiyo ya 8.0.2 kuri Windows (GraphPad Software, San Diego, CA, USA, www.graphpad.com). Hakoreshejwe ikizamini cya t-test kugira ngo hagereranywe ubushyuhe na pH y’amazi yo kunywa mbere na nyuma yo kongeramo NaHS. Igihombo cya H2S mu gisubizo kirimo NaHS cyabazwe nk'igabanuka ry'ijanisha ugereranije n’ikoreshwa ry’ibanze, kandi kugira ngo harebwe niba igihombo cyari gifite agaciro gakomeye mu mibare, twakoze ANOVA isubiramo mu buryo bumwe hagakurikiraho ikizamini cya Dunnett cyo kugereranya inshuro nyinshi. Uburemere bw'umubiri, urea mu maraso, creatinine mu maraso, na sulfide yose mu maraso uko igihe cyagiye gihita byagereranyijwe hagati y’imbeba zafashwe na NaHS z’ibitsina bitandukanye hakoreshejwe ANOVA ivanze (hagati-hamwe) hagakurikiraho ikizamini cya Bonferroni nyuma ya hoc. Agaciro ka P gafite imirizo ibiri < 0.05 kafashwe nk'akafite agaciro gakomeye mu mibare.
pH y'amazi yo kunywa yari 7.60 ± 0.01 mbere yo kongeramo NaHS na 7.71 ± 0.03 nyuma yo kongeramo NaHS (n = 13, p = 0.0029). Ubushyuhe bw'amazi yo kunywa bwari 26.5 ± 0.2 kandi bwaragabanutse bugera kuri 26.2 ± 0.2 nyuma yo kongeramo NaHS (n = 13, p = 0.0128). Tegura umuti wa NaHS wa 30 μM mu mazi yo kunywa hanyuma uwubike mu icupa ry'amazi. Umuti wa NaHS ntuhindagurika kandi ubwinshi bwawo buragabanuka uko igihe kigenda gihita. Iyo hafatwaga ingero mu ijosi ry'icupa ry'amazi, hagaragaye igabanuka rikomeye (68.0%) mu isaha ya mbere, kandi ingano ya NaHS mu muti yagabanutseho 72% na 75% nyuma y'amasaha 12 na 24. Mu bipimo byabonetse mu macupa y'amazi, igabanuka rya NaHS ntiryari rinini kugeza ku masaha 2, ariko nyuma y'amasaha 12 na 24 ryagabanutseho 47% na 72%, uko bikurikirana. Aya makuru agaragaza ko ijanisha rya NaHS mu gisubizo cya 30 μM cyateguriwe mu mazi yo kunywa ryagabanutse rigera kuri kimwe cya kane cy'agaciro ka mbere nyuma y'amasaha 24, hatitawe ku hantu ho gufatira ingero (Ishusho ya 1).
Ubushobozi bwo gupima NaHS (30 μM) mu mazi yo kunywa mu macupa y'imbeba/imbeba. Nyuma yo gutegura umuti, ingero zafashwe ku mutwe no imbere mu icupa ry'amazi. Amakuru agaragazwa nk'ikigereranyo ± SD (n = 6/itsinda). * na #, P < 0.05 ugereranije n'igihe 0. Ifoto y'icupa ry'amazi igaragaza umutwe (ufite umwanya ufunguye) n'umubiri w'icupa. Ingano y'umutwe ni hafi 740 μL.
Igipimo cya NaHS mu gisubizo cya 30 μM cyateguwe vuba cyari 30.3 ± 0.4 μM (urugero: 28.7–31.9 μM, n = 12). Ariko, nyuma y'amasaha 24, igipimo cya NaHS cyaragabanutse kigera ku gipimo gito (impuzandengo: 3.0 ± 0.6 μM). Nkuko bigaragara ku Ishusho ya 2, igipimo cya NaHS imbeba zahuye nacyo nticyari gihoraho mu gihe cy'ubushakashatsi.
Uburemere bw'umubiri w'imbeba z'ingore bwariyongereye cyane uko igihe cyagiye gihita (kuva kuri 205.2 ± 5.2 g kugeza kuri 213.8 ± 7.0 g mu itsinda ryagenzuwe no kuva kuri 204.0 ± 8.6 g kugeza kuri 211.8 ± 7.5 g mu itsinda ryavuwe na NaHS); icyakora, ubuvuzi bwa NaHS nta ngaruka bwagize ku buremere bw'umubiri (Ishusho ya 3). Uburemere bw'umubiri bw'imbeba z'ingore bwariyongereye cyane uko igihe cyagiye gihita (kuva kuri 338.6 ± 8.3 g kugeza kuri 352.4 ± 6.0 g mu itsinda ryagenzuwe no kuva kuri 352.4 ± 5.9 g kugeza kuri 363.2 ± 4.3 g mu itsinda ryavuwe na NaHS); icyakora, ubuvuzi bwa NaHS nta ngaruka bwagize ku buremere bw'umubiri (Ishusho ya 3).
Impinduka mu buremere bw'umubiri mu mbeba z'ingore n'iz'ingabo nyuma yo guhabwa NaHS (30 μM). Amakuru agaragazwa nk'ikigereranyo cya ± SEM kandi yagereranyijwe hakoreshejwe isesengura ry'uburyo bubiri (muri hagati) bw'impinduka hifashishijwe ikizamini cya Bonferroni nyuma ya hoc. n = 5 bya buri gitsina muri buri tsinda.
Igipimo cya urea mu maraso na fosfeti ya creatine mu maraso cyanganaga mu kugenzura no mu mbeba zavuwe na NaSH mu bushakashatsi bwose. Byongeye kandi, uburyo bwo kuvura NaSH ntibwagize ingaruka ku gipimo cya urea mu maraso na creatinechrome (Imbonerahamwe ya 1).
Igipimo cy’ibanze cya sulfide mu maraso cyagereranywaga hagati y’imbeba z’abagabo zavuwe na NaHS (8.1 ± 0.5 μM ugereranyije na 9.3 ± 0.2 μM) n’imbeba z’ingore (9.1 ± 1.0 μM ugereranyije na 6.1 ± 1.1 μM). Gutanga NaHS mu minsi 14 nta ngaruka byagize ku gipimo cy’sulfide mu maraso mu mbeba z’abagabo cyangwa iz’ingore (Ishusho ya 4).
Impinduka mu gipimo cya sulfide yose mu maraso mu mbeba z'ingabo n'iz'ingore nyuma yo guhabwa NaHS (30 μM). Amakuru agaragazwa nk'ikigereranyo cya ± SEM kandi yagereranyijwe hakoreshejwe isesengura ry'uburyo bubiri (muri-muri-muri) ry'impinduka hakoreshejwe ikizamini cya Bonferroni nyuma ya hoc. Buri gitsina, n = 5/itsinda.
Umwanzuro nyamukuru w'ubu bushakashatsi ni uko amazi yo kunywa arimo NaHS adahindagurika: kimwe cya kane cy'ingano ya sulfide ya mbere ni cyo gishobora kugaragara nyuma y'amasaha 24 nyuma yo gupimwa uhereye ku mutwe no imbere mu macupa y'amazi y'imbeba/imbeba. Byongeye kandi, imbeba zahuye n'ingano idahindagurika ya NaHS bitewe no gutakaza H2S mu gisubizo cya NaHS, kandi kongeramo NaHS mu mazi yo kunywa ntibyagize ingaruka ku buremere bw'umubiri, urea mu maraso na chromium ya creatine, cyangwa sulfide yose mu maraso.
Muri ubu bushakashatsi, igipimo cy'igihombo cya H2S cyavuye kuri 30 μM NaHS cyateguriwe mu mazi yo kunywa cyari hafi 3% ku isaha. Mu gisubizo cyabugenewe (100 μM sodium sulfide muri 10 mM PBS, pH 7.4), igipimo cya sulfide cyavuzwe ko cyagabanutseho 7% mu gihe kirekire mu masaha 8 na 11. Twari twarirengagije itangwa rya NaHS mu nda y'umura dutangaza ko igipimo cy'igihombo cya sulfide cyavuye kuri 54 μM NaHS mu mazi yo kunywa cyari hafi 2.3% ku isaha (4%/isaha mu masaha 12 ya mbere na 1.4%/isaha mu masaha 12 ya nyuma nyuma yo gutegura)8. Ubushakashatsi bwa mbere43 bwagaragaje igihombo gihoraho cya H2S cyaturutse kuri NaHS, ahanini bitewe no guhindagurika no gushonga. Nubwo hatabayeho kongeramo utubumbe, sulfide mu gisubizo cyabugenewe iratakaza vuba bitewe no guhindagurika kwa H2S11. Ubushakashatsi bwagaragaje ko mu gihe cyo gushonga, bifata amasegonda agera kuri 30-60, hafi 5-10% bya H2S bibura bitewe no guhumeka6. Kugira ngo hirindwe guhumeka kwa H2S mu gisubizo, abashakashatsi bafashe ingamba zitandukanye, harimo no gukangura buhoro buhoro umuti12, gupfuka umuti wa pulasitiki6, no kugabanya uburyo umuti ugera mu mwuka, kubera ko igipimo cyo guhumeka kwa H2S giterwa n'aho umwuka n'amazi bihurira.13 Gushonga kwa H2S mu buryo butunguranye bibaho ahanini bitewe na iyoni z'icyuma zihinduka, cyane cyane icyuma cya feri, ari byo bihumanya mu mazi.13 Gushonga kwa H2S bivamo ikorwa rya polysulfide (atome za sulfure zihujwe n'imigozi ya covalent)11. Kugira ngo hirindwe ogisijeni yayo, imiti irimo H2S itegurwa mu bintu bidafite ogisijeni44,45 hanyuma igasukurwa na argon cyangwa azote mu gihe cy'iminota 20-30 kugira ngo habeho ogisijeni.11,12,37,44,45,46 Diethylenetriamine aside pentaacetic (DTPA) ni metal chelator (10-4 M) ibuza ogisijeni ya HS mu bisubizo bya aerobic. Iyo DTPA idafite, igipimo cya ogisijeni ya HS- ni hafi 50% mu gihe kingana n'amasaha 3 kuri 25°C37,47. Byongeye kandi, kubera ko ogisijeni ya 1e-sulfide iterwa imbaraga n'urumuri rwa ultraviolet, umuti ugomba kubikwa ku rubura kandi ukingirwa urumuri11.
Nkuko bigaragara ku Ishusho ya 5, NaHS yitandukanya na Na+ na HS-6 iyo ishongeshejwe mu mazi; uku kwitandukanya kugaragazwa na pK1 y'uburyo ibintu bigenda, bitewe n'ubushyuhe: pK1 = 3.122 + 1132/T, aho T iri hagati ya 5 na 30°C kandi ipimwa muri dogere Kelvin (K), K = °C + 273.1548. HS- ifite pK2 iri hejuru (pK2 = 19), bityo kuri pH < 96.49, S2- ntabwo ibaho cyangwa ngo ikorerwe mu rugero ruto cyane. Mu buryo bunyuranye, HS- ikora nk'ishingiro kandi yakira H+ ivuye muri molekyuli ya H2O, naho H2O ikora nk'aside igahinduka H2S na OH-.
Ikorwa rya gaze ya H2S yashongeshejwe mu gisubizo cya NaHS (30 µM). aq, igisubizo cy'amazi; g, gaze; l, amazi. Imibare yose ifata ko pH y'amazi = 7.0 n'ubushyuhe bw'amazi = 20 °C. Byakozwe na BioRender.com.
Nubwo hari ibimenyetso bigaragaza ko ibisubizo bya NaHS bidahamye, ubushakashatsi bwinshi ku nyamaswa bwakoresheje ibisubizo bya NaHS mu mazi yo kunywa nk'umuterankunga wa H2S15,16,17,18,19,20,21,22,23,24,25,26 hamwe n'igihe cyo kuvura kiri hagati y'icyumweru 1 na 21 (Imbonerahamwe ya 2). Muri ubu bushakashatsi, igisubizo cya NaHS cyavugururwaga buri masaha 12, 15, 17, 18, 24, 25 cyangwa amasaha 24, 19, 20, 21, 22, 23. Ibisubizo byacu byagaragaje ko imbeba zahuye n'ibipimo bidahamye by'imiti bitewe no gutakaza H2S bivuye ku gisubizo cya NaHS, kandi ingano ya NaHS mu mazi yo kunywa y'imbeba yarahindutse cyane mu masaha 12 cyangwa 24 (reba Ishusho ya 2). Ubushakashatsi bubiri muri ubu bushakashatsi bwagaragaje ko urugero rwa H2S mu mazi rwagumye ruhamye mu masaha 24 n'abiri cyangwa ko igihombo cya H2S cya 2-3% gusa ari cyo cyagaragaye mu masaha 12 n'abiri n'amanota, ariko ntibwatanze amakuru afatika cyangwa ibisobanuro birambuye ku bipimo. Ubushakashatsi bubiri bwagaragaje ko uburebure buto bw'amacupa y'amazi bushobora kugabanya imyuka ya H2S15,19. Ariko, ibyavuye mu bushakashatsi bwacu byagaragaje ko ibi bishobora gutinza igihombo cya H2S mu icupa ry'amazi mu masaha 2 aho gutinza amasaha 12-24. Ubushakashatsi bwombi bugaragaza ko dutekereza ko urugero rwa NaHS mu mazi yo kunywa rutahindutse kuko tutabonye impinduka mu ibara mu mazi; bityo, ogisijeni ya H2S n'umwuka ntabwo yari ifatika19,20. Igitangaje ni uko ubu buryo busuzuma ubuziranenge bwa NaHS mu mazi aho gupima impinduka mu bwinshi bwayo uko igihe kigenda gihita.
Gutakaza H2S mu gisubizo cya NaHS bifitanye isano na pH n'ubushyuhe. Nkuko byagaragajwe mu bushakashatsi bwacu, gushonga NaHS mu mazi bituma habaho umuti wa alkaline50. Iyo NaHS ishongeshejwe mu mazi, imiterere ya gaze ya H2S ishongeshejwe iterwa n'agaciro ka pH6. Uko pH y'igisubizo iba nto, ni ko igipimo cya NaHS kigaragara nk'uduce twa gaze ya H2S kiba kinini kandi sulfide nyinshi ikabura mu gisubizo cy'amazi11. Nta na kimwe muri ubu bushakashatsi cyagaragaje pH y'amazi yo kunywa akoreshwa nk'umushongi wa NaHS. Dukurikije inama za OMS, zemejwe n'ibihugu byinshi, pH y'amazi yo kunywa igomba kuba iri hagati ya 6.5–8.551. Muri uru rugero rwa pH, igipimo cya oxidation ya H2S cyiyongera inshuro zigera ku icumi13. Gushonga NaHS mu mazi muri uru rugero rwa pH bizatuma habaho igipimo cya gaze ya H2S ishongeshejwe kiri hagati ya 1 na 22.5 μM, ibi bikaba bishimangira akamaro ko kugenzura pH y'amazi mbere yo gushonga NaHS. Byongeye kandi, urugero rw'ubushyuhe rwavuzwe mu bushakashatsi bwavuzwe haruguru (18–26 °C) rwatuma habaho impinduka mu rugero rwa gaze ya H2S yashongeshejwe mu gisubizo cya hafi 10%, kubera ko impinduka z'ubushyuhe zihindura pK1, kandi impinduka nto muri pK1 zishobora kugira ingaruka zikomeye ku rugero rwa gaze ya H2S yashongeshejwe48. Byongeye kandi, igihe kirekire cy'ubushakashatsi bumwe na bumwe (amezi 5)22, aho ubushyuhe butegerejwe guhindagurika cyane, nabyo byongera iki kibazo.
Ubushakashatsi bwose uretse bumwe21 bwakoresheje umuti wa 30 μM NaHS mu mazi yo kunywa. Kugira ngo basobanure urugero rwakoreshejwe (ni ukuvuga 30 μM), abanditsi bamwe bavuze ko NaHS mu gice cy'amazi itanga ingano imwe ya gaze ya H2S kandi ko urugero rwa H2S mu mikorobe 10 kugeza 100, bityo iki gipimo kiri mu rugero rwa physiologiya15,16. Abandi basobanuye ko 30 μM NaHS ishobora kugumana urugero rwa H2S mu mikorobe physiologiya, ni ukuvuga 5–300 μM19,20. Dusuzuma urugero rwa NaHS mu mazi rwa 30 μM (pH = 7.0, T = 20 °C), rwakoreshejwe mu bushakashatsi bumwe na bumwe kugira ngo twige ingaruka za H2S. Dushobora kubara ko ingano ya gaze ya H2S yashongeshejwe ari 14.7 μM, bingana na 50% by'ingano ya NaHS ya mbere. Iki gipimo gisa n'agaciro kabazwe n'abandi banditsi muri ubwo buryo13,48.
Mu bushakashatsi bwacu, gutanga NaHS ntabwo byahinduye ibiro by'umubiri; ibi bihuye n'ibyavuye mu bundi bushakashatsi ku mbeba z'abagabo22,23 n'imbeba z'abagabo18; Icyakora, ubushakashatsi bubiri bwagaragaje ko NaSH yagaruye ibiro by'umubiri byagabanutse ku mbeba zangiritse24,26, mu gihe ubundi bushakashatsi butagaragaje ingaruka zo gutanga NaSH ku buremere bw'umubiri15,16,17,19,20,21,25. Byongeye kandi, mu bushakashatsi bwacu, gutanga NaSH ntacyo byahinduye ku rugero rwa urea na creatine chromium mu maraso, ibi bikaba bihuye n'ibyavuye mu bundi bushakashatsi25.
Ubushakashatsi bwagaragaje ko kongera NaHS mu mazi yo kunywa mu byumweru 2 bitagize ingaruka ku gipimo cya sulfide mu maraso mu mbeba z'abagabo n'ingore. Ibi bihuye n'ibyavuye muri Sen et al. (16): ibyumweru 8 byo kuvurwa hakoreshejwe 30 μM NaHS mu mazi yo kunywa ntacyo byagize ku gipimo cya sulfide mu maraso mu mbeba zigenzura; icyakora, bavuze ko ubu buryo bwo kuvura bwasubije urugero rwa H2S mu maraso rw'imbeba zangiritse. Li et al. (22) nabo bavuze ko kuvurwa hakoreshejwe 30 μM NaHS mu mazi yo kunywa mu gihe cy'amezi 5 byongera urugero rwa sulfide idafite plasma mu mbeba zishaje ku kigero cya 26%. Ubundi bushakashatsi ntibwagaragaje impinduka mu maraso nyuma yo kongeramo NaHS mu mazi yo kunywa.
Ubushakashatsi burindwi bwatanzwe hakoreshejwe Sigma NaHS15,16,19,20,21,22,23 ariko ntibwatanze ibisobanuro birambuye ku mazi yo gutonyanga, kandi ubushakashatsi butanu ntibwavuze aho NaHS ikomoka bwakoreshejwe mu buryo bwo gutegura 17,18,24,25,26. NaHS ni molekile ifite amazi menshi kandi amazi yayo ashobora gutonyanga, ibyo bikaba bigira ingaruka ku rugero rwa NaHS ikenewe kugira ngo hategurwe umuti wa molarite runaka. Urugero, ingano ya NaHS mu bushakashatsi bwacu yari NaHS •1.3 H2O. Bityo, ingano nyayo ya NaHS muri ubu bushakashatsi ishobora kuba iri hasi ugereranyije n’iyatanzwe.
“Ni gute ikintu nk’iki kimara igihe gito gishobora kugira ingaruka zirambye?” Pozgay n’abandi.21 babajije iki kibazo ubwo basuzumaga ingaruka za NaHS ku ndwara ya colitis mu mbeba. Bizeye ko ubushakashatsi bw’ejo hazaza buzashobora gusubiza iki kibazo kandi bakeka ko ibisubizo bya NaHS bishobora kuba birimo polysulfides zihamye kurushaho uretse H2S na disulfides zihuza ingaruka za NaHS21. Ikindi gishoboka ni uko ingano nto cyane ya NaHS isigaye mu gisubizo ishobora no kugira ingaruka nziza. Mu by’ukuri, Olson n’abandi batanze ibimenyetso byerekana ko urugero rwa micromolar rwa H2S mu maraso atari urw’umubiri kandi ko rugomba kuba ruri mu rugero rwa nanomolar cyangwa rutabaho rwose13. H2S ishobora gukora binyuze muri poroteyine sulfation, impinduka ishobora guhinduka nyuma yo guhindura imikorere, ituze, n’aho poroteyine nyinshi ziherereye52,53,54. Mu by’ukuri, mu bihe by’umubiri, hafi 10% kugeza 25% bya poroteyine nyinshi z’umwijima ziba za sulfylated53. Ubushakashatsi bwombi bwemeza ko NaHS19,23 yangiritse vuba ariko butangaje buvuga ko “twagenzuye ubwinshi bwa NaHS mu mazi yo kunywa tuyasimbuza buri munsi.”23 Ubushakashatsi bumwe bwavuze mu buryo butunguranye ko “NaHS ari umutanga H2S usanzwe kandi ukoreshwa cyane mu buvuzi kugira ngo usimbuze H2S ubwayo.”18
Ibiganiro byavuzwe haruguru bigaragaza ko NaHS ibura bitewe n’umuti uva mu kirere binyuze mu guhindagurika kw’ikirere, ogisijeni na fotolisiyo, bityo hari ibitekerezo bimwe na bimwe bitangwa kugira ngo bigabanye gutakaza H2S bivuye mu muti. Ubwa mbere, gushonga kwa H2S biterwa n’aho gazi n’amazi bihurira13 na pH y’umuti11; bityo, kugira ngo bigabanye gutakaza kwa evaporative, ijosi ry’icupa ry’amazi rishobora gukorwa rito uko bishoboka kose nkuko byasobanuwe mbere15,19, kandi pH y’amazi ishobora guhindurwa ku rugero rwo hejuru rwemewe (ni ukuvuga 6.5–8.551) kugira ngo bigabanye gutakaza kwa evaporative11. Icya kabiri, ogisijeni ya H2S ibaho bitewe n’ingaruka za ogisijeni n’uko hariho iyoni z’icyuma zihinduka mu mazi yo kunywa13, bityo gusohora ogisijeni mu mazi yo kunywa hakoreshejwe argon cyangwa azote44,45 no gukoresha ibyuma bigabanya ogisijeni37,47 bishobora kugabanya ogisijeni ya sulfide. Icya gatatu, kugira ngo hirindwe ko H2S ibora, amacupa y’amazi ashobora gupfunyikwa n’urupapuro rwa aluminiyumu; Iyi gahunda ikoreshwa no ku bikoresho bishobora kwanduza urumuri nka streptozotocin55. Amaherezo, umunyu wa sulfide inorganic (NaHS, Na2S, na CaS) ushobora gutangwa binyuze muri gavage aho gushongeshwa mu mazi yo kunywa nk'uko byatangajwe mbere56,57,58; ubushakashatsi bwagaragaje ko sodium sulfide itangwa binyuze muri gavage ku mbeba yinjizwa neza kandi igakwirakwizwa hafi ya zose59. Kugeza ubu, ubushakashatsi bwinshi bwatanze umunyu wa sulfide inorganic mu nda y'umura; icyakora, ubu buryo ntibukunze gukoreshwa mu buvuzi60. Ku rundi ruhande, inzira yo kunywa ni yo nzira ikunze gukoreshwa kandi ikunzwe cyane mu bantu61. Kubwibyo, turasaba gusuzuma ingaruka z'abatanga H2S ku mbeba hakoreshejwe gavage yo mu kanwa.
Imbogamizi ni uko twapimye sulfide mu gisubizo cy’amazi na serum dukoresheje uburyo bwa MB. Uburyo bwo gupima sulfide burimo titration ya iodine, spectrophotometry, uburyo bwa electrochemical (potentiometry, amperometry, uburyo bwa coulometric na amperometric) na chromatography (gas chromatography na high-performance liquid chromatography), muri zo uburyo bukoreshwa cyane ni uburyo bwa MB spectrophotometric62. Imbogamizi ku buryo bwa MB bwo gupima H2S mu bipimo bya biological ni uko bupima ibintu byose birimo sulfur kandi ntabwo ari H2S63 yigenga kuko ikorwa mu bihe bya aside, bigatuma isulfure ikurwa mu isoko ya biological64. Ariko, nk'uko bivugwa n'Ishyirahamwe ry'Ubuzima rya Amerika, MB ni bwo buryo busanzwe bwo gupima sulfide mu mazi65. Kubwibyo, iyi mbogamizi ntigira ingaruka ku bisubizo byacu by'ingenzi ku kudahinduka kw'ibisubizo birimo NaHS. Byongeye kandi, mu bushakashatsi bwacu, kugaruza ibipimo bya sulfide mu bipimo by'amazi na serum birimo NaHS byari 91% na 93%, uko bikurikirana. Izi ndangagaciro zijyanye n’ibipimo byavuzwe mbere (77–92)66, bigaragaza uburyo bwo gusesengura bukwiye42. Ni ngombwa kumenya ko twakoresheje imbeba z’ingabo n’iz’ingore dukurikije amabwiriza y’Ikigo cy’Igihugu cy’Ubuzima (NIH) kugira ngo twirinde kwishingikiriza cyane ku bushakashatsi ku nyamaswa z’ingabo gusa mu bushakashatsi bwakozwe mbere yo kwa muganga67 no gushyiramo imbeba z’ingabo n’iz’ingore igihe cyose bishoboka68. Iyi ngingo yashimangiwe n’abandi69,70,71.
Mu gusoza, ibyavuye muri ubu bushakashatsi bigaragaza ko ibisubizo bya NaHS byakozwe mu mazi yo kunywa bidashobora gukoreshwa mu gukora H2S bitewe n’uko zidahindagurika. Ubu buryo bwo gutanga imiti bwashyira inyamaswa mu kaga gakomeye kandi kari munsi y’urugero rwa NaHS rwari rwitezwe; bityo, ibyavuye muri ubu bushakashatsi bishobora kudakoreshwa ku bantu.
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