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Hydrogen sulfide (H2S) ili ndi zotsatira zambiri za thupi komanso matenda pa thupi la munthu. Sodium hydrosulfide (NaHS) imagwiritsidwa ntchito kwambiri ngati chida chamankhwala chowunikira zotsatira za H2S mu zoyeserera zamoyo. Ngakhale kutayika kwa H2S kuchokera ku mayankho a NaHS kumatenga mphindi zochepa chabe, mayankho a NaHS agwiritsidwa ntchito ngati mankhwala opereka H2S m'madzi akumwa m'maphunziro ena a nyama. Kafukufukuyu adafufuza ngati kumwa madzi okhala ndi kuchuluka kwa NaHS kwa 30 μM yokonzedwa m'mabotolo a makoswe/mbewa kungakhale kokhazikika kwa maola osachepera 12-24, monga momwe olemba ena adanenera. Konzani yankho la NaHS (30 μM) m'madzi akumwa ndikulithira nthawi yomweyo m'mabotolo amadzi a makoswe/mbewa. Zitsanzo zinasonkhanitsidwa kuchokera kumapeto ndi mkati mwa botolo la madzi pa maola 0, 1, 2, 3, 4, 5, 6, 12 ndi 24 kuti muyese kuchuluka kwa sulfide pogwiritsa ntchito njira ya buluu ya methylene. Kuphatikiza apo, makoswe aamuna ndi aakazi adabayidwa jakisoni wa NaHS (30 μM) kwa milungu iwiri ndipo kuchuluka kwa sulfide m'magazi kumayesedwa tsiku lililonse mkati mwa sabata yoyamba komanso kumapeto kwa sabata yachiwiri. Yankho la NaHS mu chitsanzo chomwe chidapezeka kuchokera kumapeto kwa botolo la madzi silinali lokhazikika; linachepa ndi 72% ndi 75% pambuyo pa maola 12 ndi 24, motsatana. Mu zitsanzo zomwe zidapezeka mkati mwa mabotolo amadzi, kuchepa kwa NaHS sikunali kwakukulu mkati mwa maola awiri; komabe, kunachepa ndi 47% ndi 72% pambuyo pa maola 12 ndi 24, motsatana. Jakisoni wa NaHS sunakhudze kuchuluka kwa sulfide m'magazi mwa makoswe aamuna ndi aakazi. Pomaliza, mayankho a NaHS okonzedwa kuchokera ku madzi akumwa sayenera kugwiritsidwa ntchito popereka H2S chifukwa yankholo silili lokhazikika. Njira yoperekera mankhwalawa idzapangitsa nyama kukhala ndi NaHS yosakhazikika komanso yocheperako kuposa momwe amayembekezera.
Hydrogen sulfide (H2S) yakhala ikugwiritsidwa ntchito ngati poizoni kuyambira mu 1700; komabe, ntchito yake yomwe ingakhalepo ngati molekyulu yodziwika bwino ya biosignaling inafotokozedwa ndi Abe ndi Kimura mu 1996. Kwa zaka makumi atatu zapitazi, ntchito zambiri za H2S m'machitidwe osiyanasiyana a anthu zafotokozedwa bwino, zomwe zapangitsa kuti azindikire kuti mamolekyu opereka H2S akhoza kukhala ndi ntchito zachipatala pochiza kapena kusamalira matenda ena; onani Chirino et al. kuti mumve ndemanga yaposachedwa.
Sodium hydrosulfide (NaHS) yagwiritsidwa ntchito kwambiri ngati chida chamankhwala chowunikira zotsatira za H2S m'maselo ambiri ndi maphunziro a nyama5,6,7,8. Komabe, NaHS si wopereka wabwino kwambiri wa H2S chifukwa imasinthidwa mwachangu kukhala yankho la H2S/HS- mu, imaipitsidwa mosavuta ndi polysulfides, ndipo imasungunuka mosavuta ndikusungunuka4,9. Mu zoyeserera zambiri zamoyo, NaHS imasungunuka m'madzi, zomwe zimapangitsa kuti passive volatilization ndi kutayika kwa H2S10,11,12, kusungunuka kwa H2S11,12,13, ndi photolysis14. Sulfide mu yankho loyambirira imatayika mwachangu kwambiri chifukwa cha volatilization ya H2S11. Mu chidebe chotseguka, theka la moyo (t1/2) la H2S ndi pafupifupi mphindi 5, ndipo kuchuluka kwake kumachepa ndi pafupifupi 13% pa ​​mphindi10. Ngakhale kutayika kwa hydrogen sulfide kuchokera ku mayankho a NaHS kumatenga mphindi zochepa chabe, maphunziro ena a nyama agwiritsa ntchito mayankho a NaHS ngati gwero la hydrogen sulfide m'madzi akumwa kwa milungu 1-21, m'malo mwa yankho lokhala ndi NaHS maola 12-24 aliwonse. 15,16,17,18,19,20,21,22,23,24,25,26 Mchitidwewu sugwirizana ndi mfundo za kafukufuku wa sayansi, popeza kuchuluka kwa mankhwala kuyenera kutengera kugwiritsidwa ntchito kwawo m'mitundu ina, makamaka anthu.27
Kafukufuku woyambirira wa biomedicine cholinga chake ndi kukweza ubwino wa chisamaliro cha odwala kapena zotsatira za chithandizo. Komabe, zotsatira za kafukufuku wambiri wa nyama sizinamasuliridwebe kwa anthu28,29,30. Chimodzi mwa zifukwa zomwe zapangitsa kuti kulephera kwa kumasuliraku kulephereke ndi kusayang'anira bwino njira zophunzirira za nyama30. Chifukwa chake, cholinga cha kafukufukuyu chinali kufufuza ngati mayankho a 30 μM NaHS okonzedwa m'mabotolo amadzi a makoswe/mbewa atha kukhalabe olimba m'madzi akumwa kwa maola 12-24, monga momwe zanenedwera kapena zomwe zanenedwa m'maphunziro ena.
Kuyesera konse mu kafukufukuyu kunachitika motsatira malangizo ofalitsidwa okhudza chisamaliro ndi kugwiritsa ntchito nyama za m'ma laboratories ku Iran31. Malipoti onse oyesera mu kafukufukuyu adatsatiranso malangizo a ARRIVE32. Komiti ya Makhalidwe Abwino ya Institute of Endocrine Sciences, Shahid Beheshti University of Medical Sciences, idavomereza njira zonse zoyesera mu kafukufukuyu.
Zinc acetate dihydrate (CAS: 5970-45-6) ndi anhydrous ferric chloride (CAS: 7705-08-0) zinagulidwa ku Biochem, Chemopahrama (Cosne-sur-Loire, France). Sodium hydrosulfide hydrate (CAS: 207683-19-0) ndi N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) zinagulidwa ku Sigma-Aldrich (St. Louis, MO, USA). Isoflurane inagulidwa ku Piramal (Bethlehem, PA, USA). Hydrochloric acid (HCl) inagulidwa ku Merck (Darmstadt, Germany).
Konzani yankho la NaHS (30 μM) m'madzi akumwa ndipo nthawi yomweyo muyitsanulire m'mabotolo amadzi a makoswe/mbewa. Kuchuluka kumeneku kunasankhidwa kutengera mabuku ambiri pogwiritsa ntchito NaHS ngati gwero la H2S; onani gawo la Kukambirana. NaHS ndi molekyulu yokhala ndi madzi yomwe imatha kukhala ndi madzi osiyanasiyana (monga, NaHS•xH2O); malinga ndi wopanga, kuchuluka kwa NaHS komwe kunagwiritsidwa ntchito mu kafukufuku wathu kunali 70.7% (monga, NaHS•1.3 H2O), ndipo tinaganizira izi powerengera, komwe tinagwiritsa ntchito kulemera kwa mamolekyulu a 56.06 g/mol, komwe ndi kulemera kwa mamolekyulu a NaHS yopanda madzi. Madzi osungunuka (omwe amatchedwanso madzi osungunuka) ndi mamolekyulu amadzi omwe amapanga kapangidwe ka makristalo33. Ma Hydrate ali ndi mphamvu zosiyanasiyana zakuthupi ndi za thermodynamic poyerekeza ndi ma anhydrates34.
Musanawonjezere NaHS m'madzi akumwa, yesani pH ndi kutentha kwa chosungunulira. Thirani nthawi yomweyo yankho la NaHS mu botolo la madzi la makoswe/mbewa m'khola la nyama. Zitsanzo zinasonkhanitsidwa kuchokera kumapeto ndi mkati mwa botolo la madzi pa maola 0, 1, 2, 3, 4, 5, 6, 12, ndi 24 kuti muyese kuchuluka kwa sulfide. Kuyeza kwa sulfide kunatengedwa nthawi yomweyo mutatenga zitsanzo zilizonse. Tinapeza zitsanzo kuchokera kumapeto kwa chubu chifukwa kafukufuku wina wasonyeza kuti kukula kochepa kwa chubu cha madzi kumatha kuchepetsa kuuluka kwa H2S15,19. Vutoli likuwoneka kuti likugwiranso ntchito pa yankho lomwe lili m'botolo. Komabe, sizinali choncho pa yankho lomwe lili m'khosi mwa botolo la madzi, lomwe linali ndi kuchuluka kwa kuuluka kwa madzi ndipo linkapanga oxidize yokha; kwenikweni, nyama zinayamba kumwa madzi awa.
Mu kafukufukuyu, makoswe aamuna ndi aakazi a Wistar anagwiritsidwa ntchito. Makoswewa anaikidwa m'mabokosi a polypropylene (makoswe awiri mpaka atatu pa khola lililonse) pansi pa mikhalidwe yokhazikika (kutentha 21–26 °C, chinyezi 32–40%) ndi kuwala kwa maola 12 (7 koloko m'mawa mpaka 7 koloko madzulo) ndi mdima kwa maola 12 (7 koloko madzulo mpaka 7 koloko m'mawa). Makoswewa anali ndi mwayi wopeza madzi apampopi kwaulere ndipo anadyetsedwa ndi chow wamba (Khorak Dam Pars Company, Tehran, Iran). Akazi ofanana ndi zaka (miyezi 6) (n=10, kulemera kwa thupi: 190–230 g) ndi aamuna (n=10, kulemera kwa thupi: 320–370 g) anagawidwa mwachisawawa m'magulu owongolera ndi ochiritsidwa a NaHS (30 μM) (n=5 pa gulu lililonse). Kuti tidziwe kukula kwa chitsanzo, tinagwiritsa ntchito njira ya KISS (Keep It Simple, Stupid), yomwe imaphatikiza zomwe zidachitika kale komanso kusanthula mphamvu35. Choyamba tinachita kafukufuku woyeserera pa makoswe atatu ndipo tinapeza mulingo wapakati wa sulfide mu seramu ndi kupotoka kokhazikika (8.1 ± 0.81 μM). Kenako, poganizira mphamvu ya 80% ndikutenga mulingo wofunikira wa mbali ziwiri wa 5%, tinapeza kukula koyambirira kwa chitsanzo (n = 5 kutengera zolemba zam'mbuyomu) komwe kumagwirizana ndi kukula koyenera kwa zotsatira za 2.02 ndi mtengo wofotokozedwa kale womwe unaperekedwa ndi Festing powerengera kukula kwa zitsanzo za nyama zoyesera35. Titachulukitsa mtengo uwu ndi SD (2.02 × 0.81), kukula kotsimikizika kwa zotsatira (1.6 μM) kunali 20%, zomwe ndizovomerezeka. Izi zikutanthauza kuti n = 5/gulu ndikokwanira kuzindikira kusintha kwapakati kwa 20% pakati pa magulu. Makoswe adagawidwa mwachisawawa m'magulu owongolera ndi ochiritsidwa ndi NaSH pogwiritsa ntchito ntchito yosasinthika ya pulogalamu ya Excel 36 (Chithunzi Chowonjezera 1). Kubisa kunachitika pamlingo wa zotsatira, ndipo ofufuza omwe adachita muyeso wa biochemical sanadziwe za ntchito zamagulu.
Magulu a NaHS a amuna ndi akazi onse analandira chithandizo ndi yankho la 30 μM NaHS lokonzedwa m'madzi akumwa kwa milungu iwiri; yankho latsopano linaperekedwa maola 24 aliwonse, ndipo panthawiyi kulemera kwa thupi kunayesedwa. Zitsanzo za magazi zinasonkhanitsidwa kuchokera ku nsonga za mchira wa makoswe onse pansi pa mankhwala oletsa ululu a isoflurane tsiku lililonse kumapeto kwa sabata yoyamba ndi yachiwiri. Zitsanzo za magazi zinayikidwa mu centrifuge pa 3000 g kwa mphindi 10, seramu inalekanitsidwa ndikusungidwa pa -80°C kuti iwunikenso seramu urea, creatinine (Cr), ndi total sulfide. Seramu urea inadziwika pogwiritsa ntchito njira ya enzymatic urea, ndipo seramu creatinine inadziwika pogwiritsa ntchito njira ya photometric Jaffe pogwiritsa ntchito zida zogulitsidwa (Man Company, Tehran, Iran) ndi automatic analyzer (Selectra E, serial number 0-2124, The Netherlands). Ma coefficients a intra- ndi interassay of variation a urea ndi Cr anali ochepera 2.5%.
Njira ya methylene blue (MB) imagwiritsidwa ntchito poyesa sulfide yonse m'madzi akumwa ndi seramu yokhala ndi NaHS; MB ndiyo njira yogwiritsidwa ntchito kwambiri poyesa sulfide mu mayankho ambiri ndi zitsanzo zamoyo11,37. Njira ya MB ingagwiritsidwe ntchito poyesa dziwe lonse la sulfide ndikuyezera sulfide mu mawonekedwe a H2S, HS- ndi S2 mu gawo lamadzi39. Mu njira iyi, sulfure imapangidwa ngati zinc sulfide (ZnS) pamaso pa zinc acetate11,38. Zinc acetate precipitation ndiyo njira yogwiritsidwa ntchito kwambiri polekanitsa sulfide ndi ma chromophores ena11. ZnS idasungunukanso pogwiritsa ntchito HCl11 pansi pa acidity yamphamvu. Sulfide imayanjanitsidwa ndi DMPD mu chiŵerengero cha stoichiometric cha 1:2 mu reaction yomwe imayambitsidwa ndi ferric chloride (Fe3+ imagwira ntchito ngati oxidizing agent) kuti ipange utoto wa MB, womwe umapezeka spectrophotometrically pa 670 nm40,41. Malire ozindikira njira ya MB ndi pafupifupi 1 μM11.
Mu kafukufukuyu, 100 μL ya chitsanzo chilichonse (yankho kapena seramu) inawonjezedwa mu chubu; kenako 200 μL ya zinc acetate (1% w/v m'madzi osungunuka), 100 μL ya DMPD (20 mM mu 7.2 M HCl), ndi 133 μL ya FeCl3 (30 mM mu 1.2 M HCl) zinawonjezedwa. Chosakanizacho chinayikidwa pa 37°C mumdima kwa mphindi 30. Yankho linayikidwa pa centrifuge pa 10,000 g kwa mphindi 10, ndipo kuyamwa kwa supernatant kunawerengedwa pa 670 nm pogwiritsa ntchito microplate reader (BioTek, MQX2000R2, Winooski, VT, USA). Kuchuluka kwa sulfide kunadziwika pogwiritsa ntchito calibration curve ya NaHS (0–100 μM) mu ddH2O (Chithunzi Chowonjezera 2). Mayankho onse ogwiritsidwa ntchito poyeza adakonzedwa mwatsopano. Ma coefficients a intra- ndi interassay of variation of measurements of sulfide muyeso anali 2.8% ndi 3.4%, motsatana. Tinapezanso sulfide yonse yomwe inapezeka kuchokera ku madzi akumwa okhala ndi sodium thiosulfate ndi zitsanzo za seramu pogwiritsa ntchito njira yolimba ya chitsanzo42. Kubwezeretsedwa kwa madzi akumwa okhala ndi sodium thiosulfate ndi zitsanzo za seramu kunali 91 ± 1.1% (n = 6) ndi 93 ± 2.4% (n = 6), motsatana.
Kusanthula ziwerengero kunachitika pogwiritsa ntchito pulogalamu ya GraphPad Prism version 8.0.2 ya Windows (GraphPad Software, San Diego, CA, USA, www.graphpad.com). Kuyesa kwa t komwe kunagwiritsidwa ntchito poyerekeza kutentha ndi pH ya madzi akumwa asanayambe komanso atawonjezera NaHS. Kutayika kwa H2S mu yankho lokhala ndi NaHS kunawerengedwa ngati kuchepa kwa peresenti kuchokera ku kuyamwa koyambira, ndipo kuti tiwone ngati kutayikako kunali kofunikira pa ziwerengero, tinachita ANOVA yobwerezabwereza yotsatiridwa ndi mayeso oyerekeza angapo a Dunnett. Kulemera kwa thupi, urea wa seramu, creatinine wa seramu, ndi sulfide yonse ya seramu pakapita nthawi zinayerekezeredwa pakati pa makoswe olamulira ndi ochiritsidwa ndi NaHS amitundu yosiyanasiyana pogwiritsa ntchito ANOVA yosakanikirana (pakati-mkati) yotsatiridwa ndi mayeso a Bonferroni post hoc. Ma P okhala ndi michira iwiri < 0.05 amaonedwa kuti ndi ofunika pa ziwerengero.
pH ya madzi akumwa inali 7.60 ± 0.01 NaHS isanawonjezedwe ndipo 7.71 ± 0.03 pambuyo pa NaHS yowonjezera (n = 13, p = 0.0029). Kutentha kwa madzi akumwa kunali 26.5 ± 0.2 ndipo kunachepa kufika pa 26.2 ± 0.2 pambuyo pa NaHS yowonjezera (n = 13, p = 0.0128). Konzani yankho la 30 μM NaHS m'madzi akumwa ndikusunga mu botolo la madzi. Yankho la NaHS silikhazikika ndipo kuchuluka kwake kumachepa pakapita nthawi. Mukatenga zitsanzo kuchokera pakhosi la botolo la madzi, kuchepa kwakukulu (68.0%) kunawonedwa mkati mwa ola loyamba, ndipo kuchuluka kwa NaHS mu yankho kunachepa ndi 72% ndi 75% pambuyo pa maola 12 ndi 24, motsatana. Mu zitsanzo zomwe zinapezedwa kuchokera m'mabotolo amadzi, kuchepa kwa NaHS sikunali kwakukulu mpaka maola awiri, koma pambuyo pa maola 12 ndi 24 kunachepa ndi 47% ndi 72%, motsatana. Deta iyi ikusonyeza kuti kuchuluka kwa NaHS mu yankho la 30 μM lokonzedwa m'madzi akumwa kunachepa kufika pafupifupi kotala la mtengo woyamba pambuyo pa maola 24, mosasamala kanthu za malo omwe sampuli inali (Chithunzi 1).
Kukhazikika kwa yankho la NaHS (30 μM) m'madzi akumwa m'mabotolo a makoswe/mbewa. Pambuyo pokonza yankho, zitsanzo zinatengedwa kuchokera ku nsonga ndi mkati mwa botolo la madzi. Deta imaperekedwa ngati apakati ± SD (n = 6/gulu). * ndi #, P < 0.05 poyerekeza ndi nthawi 0. Chithunzi cha botolo la madzi chikuwonetsa nsonga (yomwe ili ndi kutsegula) ndi thupi la botolo. Kuchuluka kwa nsonga ndi pafupifupi 740 μL.
Kuchuluka kwa NaHS mu yankho la 30 μM lomwe lakonzedwa kumene linali 30.3 ± 0.4 μM (kusiyana: 28.7–31.9 μM, n = 12). Komabe, patatha maola 24, kuchuluka kwa NaHS kunachepa kufika pamtengo wotsika (avereji: 3.0 ± 0.6 μM). Monga momwe zasonyezedwera pa Chithunzi 2, kuchuluka kwa NaHS komwe makoswe adakumana nako sikunali kokhazikika panthawi yophunzira.
Kulemera kwa thupi la makoswe aakazi kunawonjezeka kwambiri pakapita nthawi (kuyambira 205.2 ± 5.2 g mpaka 213.8 ± 7.0 g mu gulu lolamulira komanso kuyambira 204.0 ± 8.6 g mpaka 211.8 ± 7.5 g mu gulu lolandira chithandizo cha NaHS); komabe, chithandizo cha NaHS sichinakhudze kulemera kwa thupi (Chithunzi 3). Kulemera kwa thupi la makoswe amphongo kunawonjezeka kwambiri pakapita nthawi (kuyambira 338.6 ± 8.3 g mpaka 352.4 ± 6.0 g mu gulu lolamulira komanso kuyambira 352.4 ± 5.9 g mpaka 363.2 ± 4.3 g mu gulu lolandira chithandizo cha NaHS); komabe, chithandizo cha NaHS sichinakhudze kulemera kwa thupi (Chithunzi 3).
Kusintha kwa kulemera kwa thupi mwa makoswe aakazi ndi amphongo atalandira NaHS (30 μM). Deta imaperekedwa ngati apakati ± SEM ndipo inayerekezeredwa pogwiritsa ntchito kusanthula kwa kusiyana kwa njira ziwiri (mkati mwa pakati) ndi mayeso a Bonferroni post hoc. n = 5 ya kugonana kulikonse mu gulu lililonse.
Kuchuluka kwa urea ndi creatine phosphate m'magazi kunali kofanana pakulamulira ndi makoswe omwe adalandira NaSH mu kafukufukuyu. Kuphatikiza apo, chithandizo cha NaSH sichinakhudze kuchuluka kwa urea ndi creatinechrome m'magazi (Table 1).
Kuchuluka kwa sulfide yonse m'magazi kunali kofanana pakati pa makoswe aamuna olamulidwa ndi a NaHS (8.1 ± 0.5 μM vs. 9.3 ± 0.2 μM) ndi akazi (9.1 ± 1.0 μM vs. 6.1 ± 1.1 μM). Kupereka kwa NaHS kwa masiku 14 sikunakhudze kuchuluka kwa sulfide yonse m'magazi mwa makoswe aamuna kapena aakazi (Chithunzi 4).
Kusintha kwa kuchuluka kwa sulfide yonse m'magazi mwa makoswe aamuna ndi aakazi atatha kulandira NaHS (30 μM). Deta imaperekedwa ngati mean ± SEM ndipo inayerekezeredwa pogwiritsa ntchito kusanthula kosakanikirana (mkati-mkati) kwa kusiyana ndi mayeso a Bonferroni post hoc. Kugonana kulikonse, n = 5/gulu.
Mapeto ake a kafukufukuyu ndi akuti madzi akumwa okhala ndi NaHS ndi osakhazikika: pafupifupi kotala la kuchuluka kwa sulfide koyambirira kumatha kupezeka maola 24 mutatenga mankhwalawa kuchokera pamwamba ndi mkati mwa mabotolo amadzi a makoswe/mbewa. Kuphatikiza apo, makoswe adakumana ndi kuchuluka kwa NaHS kosakhazikika chifukwa cha kutayika kwa H2S mu yankho la NaHS, ndipo kuwonjezera kwa NaHS m'madzi akumwa sikunakhudze kulemera kwa thupi, seramu urea ndi creatine chromium, kapena seramu sulfide yonse.
Mu kafukufukuyu, kuchuluka kwa kutayika kwa H2S kuchokera ku mayankho a 30 μM NaHS okonzedwa m'madzi akumwa kunali pafupifupi 3% pa ​​ola limodzi. Mu yankho lotetezedwa (100 μM sodium sulfide mu 10 mM PBS, pH 7.4), kuchuluka kwa sulfide kunanenedwa kuti kwatsika ndi 7% pakapita nthawi kwa maola 8. Tateteza kale kuperekedwa kwa NaHS mkati mwa peritoneal pofotokoza kuti kuchuluka kwa kutayika kwa sulfide kuchokera ku yankho la 54 μM NaHS m'madzi akumwa kunali pafupifupi 2.3% pa ​​ola limodzi (4%/ola m'maola 12 oyamba ndi 1.4%/ola m'maola 12 omaliza mutakonzekera)8. Kafukufuku wakale43 adapeza kutayika kosalekeza kwa H2S kuchokera ku mayankho a NaHS, makamaka chifukwa cha kusinthasintha kwa kutentha ndi kusungunuka kwa madzi. Ngakhale popanda kuwonjezera thovu, sulfide mu yankho la stock imatayika mwachangu chifukwa cha kusinthasintha kwa kutentha ... Kafukufuku wasonyeza kuti panthawi yosungunula, yomwe imatenga pafupifupi masekondi 30-60, pafupifupi 5-10% ya H2S imatayika chifukwa cha kusungunuka6. Pofuna kupewa kusungunuka kwa H2S kuchokera ku yankho, ofufuza achita zinthu zingapo, kuphatikizapo kusakaniza pang'onopang'ono yankho12, kuphimba yankho la stock ndi pulasitiki6, ndikuchepetsa kuwonekera kwa yankho kumlengalenga, popeza kuchuluka kwa kusungunuka kwa H2S kumadalira mawonekedwe a mpweya ndi madzi.13 Kusungunuka kwa H2S kumachitika chifukwa cha ma ayoni achitsulo osinthika, makamaka chitsulo cha ferric, chomwe ndi zodetsa m'madzi.13 Kusungunuka kwa H2S kumabweretsa mapangidwe a polysulfides (maatomu a sulfure olumikizidwa ndi ma covalent bonds)11. Kuti apewe okosijeni, mayankho okhala ndi H2S amakonzedwa mu zosungunulira zopanda okosijeni44,45 kenako amatsukidwa ndi argon kapena nayitrogeni kwa mphindi 20-30 kuti atsimikizire kuti okosijeni salowa.11,12,37,44,45,46 Diethylenetriaminepentaacetic acid (DTPA) ndi metal chelator (10-4 M) yomwe imaletsa HS- autoxidation mu mayankho a aerobic. Ngati DTPA siili nayo, kuchuluka kwa okosijeni kwa HS- kuli pafupifupi 50% pa pafupifupi maola atatu pa 25°C37,47. Kuphatikiza apo, popeza okosijeni wa 1e-sulfide amathandizidwa ndi kuwala kwa ultraviolet, yankholo liyenera kusungidwa pa ayezi ndikutetezedwa ku kuwala11.
Monga momwe zasonyezedwera pa Chithunzi 5, NaHS imagawanika kukhala Na+ ndi HS-6 ikasungunuka m'madzi; kugawanika kumeneku kumatsimikiziridwa ndi pK1 ya reaction, yomwe imadalira kutentha: pK1 = 3.122 + 1132/T, komwe T imayambira pa 5 mpaka 30°C ndipo imayesedwa mu madigiri Kelvin (K), K = °C + 273.1548. HS- ili ndi pK2 yokwera (pK2 = 19), kotero pa pH < 96.49, S2- simapangidwa kapena imapangidwa pang'ono kwambiri. Mosiyana ndi zimenezi, HS- imagwira ntchito ngati maziko ndipo imalandira H+ kuchokera ku molekyulu ya H2O, ndipo H2O imagwira ntchito ngati asidi ndipo imasinthidwa kukhala H2S ndi OH-.
Kupanga mpweya wa H2S wosungunuka mu yankho la NaHS (30 µM). aq, yankho lamadzi; g, mpweya; l, madzi. Mawerengedwe onse amaganiza kuti pH ya madzi = 7.0 ndi kutentha kwa madzi = 20 °C. Yopangidwa ndi BioRender.com.
Ngakhale pali umboni wakuti mayankho a NaHS ndi osakhazikika, kafukufuku wochuluka wa nyama wagwiritsa ntchito mayankho a NaHS m'madzi akumwa ngati mankhwala operekedwa ndi H2S15,16,17,18,19,20,21,22,23,24,25,26 ndi nthawi yolowererapo kuyambira sabata 1 mpaka 21 (Table 2). Pa maphunzirowa, yankho la NaHS linasinthidwa maola 12 aliwonse, 15, 17, 18, 24, 25 kapena maola 24, 19, 20, 21, 22, 23. Zotsatira zathu zasonyeza kuti makoswe anali ndi mankhwala osakhazikika chifukwa cha kutayika kwa H2S kuchokera ku yankho la NaHS, ndipo kuchuluka kwa NaHS m'madzi akumwa a makoswe kunasinthasintha kwambiri pa maola 12 kapena 24 (onani Chithunzi 2). Kafukufuku awiri mwa awa adanena kuti kuchuluka kwa H2S m'madzi kunakhalabe kokhazikika kwa maola 24 kapena kuti kutayika kwa H2S ndi 2-3% kokha komwe kunawonedwa kwa maola 12, koma sanapereke deta yothandizira kapena tsatanetsatane wa muyeso. Kafukufuku awiri awonetsa kuti kukula kochepa kwa mabotolo amadzi kumatha kuchepetsa kuuluka kwa H2S15,19. Komabe, zotsatira zathu zawonetsa kuti izi zitha kungochedwetsa kutayika kwa H2S kuchokera ku botolo lamadzi ndi maola awiri m'malo mwa maola 12-24. Kafukufuku onsewa akuti timaganiza kuti kuchuluka kwa NaHS m'madzi akumwa sikunasinthe chifukwa sitinawone kusintha kwa mtundu m'madzi; chifukwa chake, kukhuthala kwa H2S ndi mpweya sikunali kofunikira19,20. Chodabwitsa n'chakuti, njira iyi yodziwira kukhazikika kwa NaHS m'madzi m'malo moyesa kusintha kwa kuchuluka kwake pakapita nthawi.
Kutayika kwa H2S mu yankho la NaHS kukugwirizana ndi pH ndi kutentha. Monga taonera mu kafukufuku wathu, kusungunuka kwa NaHS m'madzi kumapangitsa kuti pakhale yankho la alkaline50. Pamene NaHS isungunuka m'madzi, kupangidwa kwa mpweya wa H2S wosungunuka kumadalira pH6. pH ya yankho ikatsika, kuchuluka kwa NaHS komwe kumapezeka ngati mamolekyu a mpweya wa H2S kumawonjezeka ndipo sulfide yambiri imatayika kuchokera ku yankho lamadzi11. Palibe kafukufuku aliyense ameneyu amene adanena za pH ya madzi akumwa omwe amagwiritsidwa ntchito ngati chosungunulira cha NaHS. Malinga ndi malangizo a WHO, omwe amavomerezedwa ndi mayiko ambiri, pH ya madzi akumwa iyenera kukhala pakati pa 6.5–8.551. Mu pH iyi, kuchuluka kwa okosijeni kwa H2S kumawonjezeka pafupifupi kakhumi13. Kusungunuka kwa NaHS m'madzi mu pH iyi kudzapangitsa kuti mpweya wa H2S wosungunuka ukhale pakati pa 1 mpaka 22.5 μM, zomwe zikugogomezera kufunika koyang'anira pH ya madzi musanasungunuke NaHS. Kuphatikiza apo, kutentha komwe kwanenedwa mu kafukufukuyu (18–26 °C) kungapangitse kusintha kwa kuchuluka kwa mpweya wa H2S wosungunuka mu yankho la pafupifupi 10%, popeza kusintha kwa kutentha kumasintha pK1, ndipo kusintha pang'ono kwa pK1 kumatha kukhudza kwambiri kuchuluka kwa mpweya wa H2S wosungunuka48. Kuphatikiza apo, nthawi yayitali ya maphunziro ena (miyezi 5)22, pomwe kusintha kwakukulu kwa kutentha kumayembekezeredwa, kumawonjezeranso vutoli.
Maphunziro onse kupatulapo amodzi21 adagwiritsa ntchito yankho la 30 μM NaHS m'madzi akumwa. Pofotokoza mlingo womwe wagwiritsidwa ntchito (monga 30 μM), olemba ena adanenanso kuti NaHS mu gawo lamadzi imapanga kuchuluka komweko kwa mpweya wa H2S ndipo kuchuluka kwa thupi kwa H2S ndi 10 mpaka 100 μM, kotero mlingo uwu uli mkati mwa kuchuluka kwa thupi15,16. Ena adafotokoza kuti 30 μM NaHS imatha kusunga mulingo wa H2S m'magazi mkati mwa kuchuluka kwa thupi, mwachitsanzo 5–300 μM19,20. Tikuganizira kuchuluka kwa NaHS m'madzi a 30 μM (pH = 7.0, T = 20 °C), komwe kudagwiritsidwa ntchito m'maphunziro ena kuti tiphunzire zotsatira za H2S. Titha kuwerengera kuti kuchuluka kwa mpweya wa H2S wosungunuka ndi 14.7 μM, womwe ndi pafupifupi 50% ya kuchuluka kwa NaHS koyambirira. Mtengo uwu ndi wofanana ndi mtengo womwe unawerengedwa ndi olemba ena pansi pa mikhalidwe yomweyi13,48.
Mu kafukufuku wathu, kupereka NaHS sikunasinthe kulemera kwa thupi; zotsatirazi zikugwirizana ndi zotsatira za maphunziro ena mwa mbewa zachimuna22,23 ndi makoswe achimuna18; Komabe, maphunziro awiri adanenanso kuti NaSH idabwezeretsa kuchepa kwa kulemera kwa thupi mwa makoswe omwe adachotsedwa m'thupi24,26, pomwe maphunziro ena sananene za momwe kupereka NaSH kudakhudzira kulemera kwa thupi15,16,17,19,20,21,25. Kuphatikiza apo, mu kafukufuku wathu, kupereka NaSH sikunakhudze kuchuluka kwa urea ndi creatine chromium m'magazi, zomwe zikugwirizana ndi zotsatira za lipoti lina25.
Kafukufukuyu adapeza kuti kuwonjezera NaHS m'madzi akumwa kwa milungu iwiri sikunakhudze kuchuluka kwa sulfide m'magazi mwa makoswe aamuna ndi aakazi. Kupeza kumeneku kukugwirizana ndi zotsatira za Sen et al. (16): Masabata 8 a chithandizo ndi 30 μM NaHS m'madzi akumwa sanakhudze kuchuluka kwa sulfide m'magazi mwa makoswe olamulira; komabe, adanenanso kuti kulowererapo kumeneku kunabwezeretsa kuchepa kwa milingo ya H2S mu plasma ya mbewa zomwe sizinachiritsidwe. Li et al. (22) adanenanso kuti chithandizo ndi 30 μM NaHS m'madzi akumwa kwa miyezi 5 chinawonjezera milingo ya sulfide yopanda plasma mwa mbewa zokalamba ndi pafupifupi 26%. Kafukufuku wina sananene kusintha kwa sulfide yozungulira pambuyo powonjezera NaHS m'madzi akumwa.
Maphunziro asanu ndi awiri adanenedwa pogwiritsa ntchito Sigma NaHS15,16,19,20,21,22,23 koma sanapereke tsatanetsatane wokhudza madzi onyowa, ndipo maphunziro asanu sanatchule komwe NaHS idachokera pogwiritsa ntchito njira zawo zokonzekera17,18,24,25,26. NaHS ndi molekyulu yokhala ndi madzi onyowa ndipo kuchuluka kwa madzi onyowa kumatha kusiyana, zomwe zimakhudza kuchuluka kwa NaHS komwe kumafunikira kuti akonze yankho la molarity inayake. Mwachitsanzo, kuchuluka kwa NaHS mu kafukufuku wathu kunali NaHS•1.3 H2O. Chifukwa chake, kuchuluka kwenikweni kwa NaHS mu maphunzirowa kungakhale kotsika kuposa komwe kudanenedwa.
"Kodi mankhwala oterewa angakhale bwanji ndi zotsatira zokhalitsa chonchi?" Pozgay et al.21 anafunsa funsoli poyesa zotsatira za NaHS pa matenda a m'matumbo m'makoswe. Akuyembekeza kuti maphunziro amtsogolo adzatha kuyankha funsoli ndipo amaganiza kuti mayankho a NaHS angakhale ndi ma polysulfide okhazikika kwambiri kuwonjezera pa H2S ndi disulfides zomwe zimayambitsa zotsatira za NaHS21. Kuthekera kwina ndikuti kuchuluka kochepa kwambiri kwa NaHS komwe kutsalira mu yankho kungathandizenso. Ndipotu, Olson et al. anapereka umboni wakuti kuchuluka kwa H2S m'magazi sikuli kwa thupi ndipo kuyenera kukhala mu nanomolar range kapena kusakhalapo konse13. H2S ikhoza kugwira ntchito kudzera mu protein sulfation, kusintha komwe kumasintha pambuyo pa kumasulira komwe kumakhudza ntchito, kukhazikika, ndi malo a mapuloteni ambiri52,53,54. Ndipotu, pansi pa mikhalidwe ya thupi, pafupifupi 10% mpaka 25% ya mapuloteni ambiri a chiwindi ndi sulfylated53. Kafukufuku onse awiriwa akuvomereza kuwonongeka mwachangu kwa NaHS19,23 koma modabwitsa akunena kuti "tinayang'anira kuchuluka kwa NaHS m'madzi akumwa powasintha tsiku lililonse."23 Kafukufuku wina mwangozi adati "NaHS ndi wopereka H2S wamba ndipo amagwiritsidwa ntchito kwambiri kuchipatala kuti alowe m'malo mwa H2S yokha."18
Kukambirana pamwambapa kukuwonetsa kuti NaHS imatayika chifukwa cha yankho kudzera mu volatilization, oxidation ndi photolysis, motero malingaliro ena aperekedwa kuti achepetse kutayika kwa H2S kuchokera ku yankho. Choyamba, kutuluka kwa H2S kumadalira pa gasi-liquid interface13 ndi pH ya yankho11; chifukwa chake, kuti achepetse kutayika kwa evaporative, khosi la botolo la madzi likhoza kuchepetsedwa momwe tingathere monga tafotokozera kale15,19, ndipo pH ya madzi ikhoza kusinthidwa kukhala malire apamwamba ovomerezeka (monga, 6.5–8.551) kuti achepetse kutayika kwa evaporative11. Chachiwiri, kutuluka kwa H2S mwadzidzidzi kumachitika chifukwa cha zotsatira za mpweya ndi kukhalapo kwa ma ayoni achitsulo osinthika m'madzi akumwa13, kotero kuchotsa okosijeni m'madzi akumwa ndi argon kapena nitrogen44,45 ndi kugwiritsa ntchito zitsulo zotchedwa chelators37,47 kungachepetse kutuluka kwa sulfides. Chachitatu, kuti tipewe kuwonongeka kwa H2S, mabotolo amadzi akhoza kukulungidwa ndi zojambulazo za aluminiyamu; Mchitidwewu umagwiranso ntchito pazinthu zomwe zimakhudzidwa ndi kuwala monga streptozotocin55. Pomaliza, mchere wa sulfide wosapangidwa kuchokera ku zinthu zachilengedwe (NaHS, Na2S, ndi CaS) ukhoza kuperekedwa kudzera mu gavage m'malo mosungunuka m'madzi akumwa monga momwe zidanenedwera kale56,57,58; kafukufuku wasonyeza kuti sodium sulfide yochokera ku radioactive yomwe imaperekedwa kudzera mu gavage kwa makoswe imayamwa bwino ndikugawidwa pafupifupi minofu yonse59. Mpaka pano, maphunziro ambiri apereka mchere wa sulfide wosapangidwa kuchokera ku zinthu zachilengedwe mkati mwa chiberekero; komabe, njira iyi sigwiritsidwa ntchito kawirikawiri m'malo azachipatala60. Kumbali inayi, njira yomwa ndiyo njira yodziwika bwino komanso yokondedwa yoperekera mankhwala mwa anthu61. Chifukwa chake, tikukulimbikitsani kuwunika momwe opereka H2S amakhudzira makoswe pogwiritsa ntchito gavage yomwa.
Choletsa ndichakuti tinayesa sulfide mu yankho lamadzi ndi seramu pogwiritsa ntchito njira ya MB. Njira zoyezera sulfide zikuphatikizapo ayodini titration, spectrophotometry, njira ya electrochemical (potentiometry, amperometry, njira ya coulometric ndi njira ya amperometric) ndi chromatography (gas chromatography ndi high-performance liquid chromatography), pakati pa njira yomwe imagwiritsidwa ntchito kwambiri ndi njira ya MB spectrophotometric62. Choletsa cha njira ya MB poyezera H2S mu zitsanzo zamoyo ndikuti imayesa mankhwala onse okhala ndi sulfure osati H2S63 yaulere chifukwa imachitidwa pansi pa acidity, zomwe zimapangitsa kuti sulfure ichotsedwe kuchokera ku gwero lachilengedwe64. Komabe, malinga ndi American Public Health Association, MB ndiye njira yodziwika bwino yoyezera sulfide m'madzi65. Chifukwa chake, choletsa ichi sichikhudza zotsatira zathu zazikulu pa kusakhazikika kwa mayankho okhala ndi NaHS. Kuphatikiza apo, mu kafukufuku wathu, kubwezeretsa kwa muyeso wa sulfide mu zitsanzo zamadzi ndi seramu zokhala ndi NaHS kunali 91% ndi 93%, motsatana. Makhalidwe amenewa akugwirizana ndi ma ranges omwe adanenedwa kale (77–92)66, zomwe zikusonyeza kulondola kovomerezeka kwa kusanthula42. Ndikofunikira kudziwa kuti tidagwiritsa ntchito makoswe aamuna ndi aakazi motsatira malangizo a National Institutes of Health (NIH) kuti tipewe kudalira kwambiri maphunziro a nyama za amuna okha m'maphunziro asanayambe kuchipatala67 komanso kuphatikiza makoswe aamuna ndi aakazi nthawi iliyonse yomwe zingatheke68. Mfundo iyi yagogomezeredwa ndi ena69,70,71.
Pomaliza, zotsatira za kafukufukuyu zikusonyeza kuti njira za NaHS zokonzedwa kuchokera ku madzi akumwa sizingagwiritsidwe ntchito popanga H2S chifukwa cha kusakhazikika kwawo. Njira imeneyi yoperekera mankhwala ingapangitse nyama kukhala ndi milingo yotsika ya NaHS kuposa momwe amayembekezera; chifukwa chake, zomwe zapezekazi sizingagwire ntchito kwa anthu.
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