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He nui nā hopena physiological a me nā pathological o ka hydrogen sulfide (H2S) i ke kino o ke kanaka. Hoʻohana nui ʻia ka sodium hydrosulfide (NaHS) ma ke ʻano he mea hana pharmacological e loiloi i nā hopena o H2S i nā hoʻokolohua olaola. ʻOiai ʻo ka nalowale ʻana o H2S mai nā hopena NaHS he mau minuke wale nō, ua hoʻohana ʻia nā hopena NaHS ma ke ʻano he mau mea hāʻawi no H2S i ka wai inu i kekahi mau haʻawina holoholona. Ua noiʻi kēia haʻawina inā hiki i ka wai inu me ka nui o NaHS o 30 μM i hoʻomākaukau ʻia i loko o nā ʻōmole ʻiole/ʻiole ke noho paʻa no ka liʻiliʻi he 12-24 mau hola, e like me ka manaʻo o kekahi mau mea kākau. E hoʻomākaukau i kahi hopena o NaHS (30 μM) i loko o ka wai inu a ninini koke i loko o nā ʻōmole wai ʻiole/ʻiole. Ua hōʻiliʻili ʻia nā laʻana mai ka piko a me loko o ka ʻōmole wai ma 0, 1, 2, 3, 4, 5, 6, 12 a me 24 mau hola e ana i ka ʻike sulfide me ka hoʻohana ʻana i ke ʻano methylene blue. Eia kekahi, ua hoʻokomo ʻia nā ʻiole kāne a me nā wahine me NaHS (30 μM) no ʻelua pule a ua ana ʻia nā ʻano serum sulfide i kēlā me kēia lā ʻē aʻe i ka pule mua a ma ka hopena o ka lua o ka pule. ʻAʻole paʻa ka hopena NaHS i loko o ka hāpana i loaʻa mai ka piko o ka ʻōmole wai; ua emi iho ia ma 72% a me 75% ma hope o 12 a me 24 mau hola, kēlā me kēia. I loko o nā hāpana i loaʻa mai loko mai o nā ʻōmole wai, ʻaʻole koʻikoʻi ka emi ʻana o NaHS i loko o 2 mau hola; akā naʻe, ua emi iho ia ma 47% a me 72% ma hope o 12 a me 24 mau hola, kēlā me kēia. ʻAʻole i hoʻopilikia ka inikini NaHS i ka pae serum sulfide o nā ʻiole kāne a me nā wahine. I ka hopena, ʻaʻole pono e hoʻohana ʻia nā hopena NaHS i hoʻomākaukau ʻia mai ka wai inu no ka hāʻawi ʻana i ka H2S no ka mea he paʻa ʻole ka hopena. E hōʻike kēia ala o ka lawelawe ʻana i nā holoholona i nā nui NaHS i kūlike ʻole a liʻiliʻi ma mua o ka mea i manaʻo ʻia.
Ua hoʻohana ʻia ka hydrogen sulfide (H2S) ma ke ʻano he toxin mai ka makahiki 1700; akā naʻe, ua wehewehe ʻia kona kūlana hiki ke lilo i mole biosignaling endogenous e Abe lāua ʻo Kimura i ka makahiki 1996. I loko o nā makahiki he kanakolu i hala iho nei, ua wehewehe ʻia nā hana he nui o H2S i nā ʻōnaehana kanaka like ʻole, e alakaʻi ana i ka ʻike ʻana he hiki i nā mole hāʻawi H2S ke loaʻa nā noi lapaʻau i ka mālama ʻana a i ʻole ka hoʻokele ʻana i kekahi mau maʻi; e ʻike iā Chirino et al. no kahi loiloi hou.
Ua hoʻohana nui ʻia ʻo Sodium hydrosulfide (NaHS) ma ke ʻano he mea hana pharmacological e loiloi i nā hopena o H2S i nā haʻawina moʻomeheu cell a me nā holoholona he nui5,6,7,8. Eia nō naʻe, ʻaʻole ʻo NaHS kahi mea hāʻawi H2S kūpono no ka mea ua hoʻololi koke ʻia i H2S/HS- i loko o ka hopena, ua haumia maʻalahi me nā polysulfides, a ua maʻalahi ka oxidized a me ka volatilized4,9. I loko o nā hoʻokolohua olaola he nui, ua hoʻoheheʻe ʻia ʻo NaHS i loko o ka wai, e hopena ana i ka volatilization passive a me ka nalowale o H2S10,11,12, ka oxidation spontaneous o H2S11,12,13, a me ka photolysis14. Ua nalowale koke ka sulfide i loko o ka hopena mua ma muli o ka volatilization o H2S11. I loko o kahi pahu hāmama, ʻo ka hapalua ola (t1/2) o H2S ma kahi o 5 mau minuke, a ua emi kona ʻano ma kahi o 13% i kēlā me kēia minuke10. ʻOiai ʻo ka nalowale ʻana o ka hydrogen sulfide mai nā hopena NaHS he mau minuke wale nō ia, ua hoʻohana kekahi mau haʻawina holoholona i nā hopena NaHS ma ke ʻano he kumu o ka hydrogen sulfide i ka wai inu no 1-21 mau pule, e pani ana i ka hopena i loaʻa iā NaHS i kēlā me kēia 12-24 mau hola.15,16,17,18,19,20,21,22,23,24,25,26 ʻAʻole kūlike kēia hana me nā kumumanaʻo o ka noiʻi ʻepekema, ʻoiai pono e hoʻokumu ʻia nā dosages lāʻau ma kā lākou hoʻohana ʻana i nā ʻano ʻē aʻe, ʻoi aku hoʻi nā kānaka.27
ʻO ka noiʻi preclinical i ka biomedicine ka pahuhopu e hoʻomaikaʻi i ka maikaʻi o ka mālama ʻana i ka mea maʻi a i ʻole nā ​​hopena lapaʻau. Eia nō naʻe, ʻaʻole i unuhi ʻia nā hopena o ka hapa nui o nā haʻawina holoholona i nā kānaka28,29,30. ʻO kekahi o nā kumu no kēia hāʻule ʻana o ka unuhi ʻana, ʻo ia ka nele o ka nānā ʻana i ka maikaʻi o ke ʻano hana o nā haʻawina holoholona30. No laila, ʻo ka pahuhopu o kēia haʻawina, ʻo ia ke noiʻi inā hiki i nā hopena 30 μM NaHS i hoʻomākaukau ʻia i loko o nā ʻōmole wai ʻiole/ʻiole ke noho paʻa i ka wai inu no 12-24 hola, e like me ka ʻōlelo a i ʻole i manaʻo ʻia i loko o kekahi mau haʻawina.
Ua mālama ʻia nā hoʻokolohua āpau ma kēia haʻawina e like me nā alakaʻi i paʻi ʻia no ka mālama ʻana a me ka hoʻohana ʻana i nā holoholona hoʻokolohua ma Iran31. Ua hahai pū nā hōʻike hoʻokolohua āpau ma kēia haʻawina i nā alakaʻi ARRIVE32. Ua ʻāpono ke Kōmike Ethics o ka Institute of Endocrine Sciences, Shahid Beheshti University of Medical Sciences, i nā kaʻina hana hoʻokolohua āpau ma kēia haʻawina.
Ua kūʻai ʻia ʻo Zinc acetate dihydrate (CAS: 5970-45-6) a me ka ferric chloride anhydrous (CAS: 7705-08-0) mai Biochem, Chemopahrama (Cosne-sur-Loire, Palani). Ua kūʻai ʻia ʻo Sodium hydrosulfide hydrate (CAS: 207683-19-0) a me N,N-dimethyl-p-phenylenediamine (DMPD) (CAS: 535-47-0) mai Sigma-Aldrich (St. Louis, MO, USA). Ua kūʻai ʻia ʻo Isoflurane mai Piramal (Bethlehem, PA, USA). Ua kūʻai ʻia ka waikawa Hydrochloric (HCl) mai Merck (Darmstadt, Kelemānia).
E hoʻomākaukau i kahi hopena o NaHS (30 μM) i loko o ka wai inu a ninini koke i loko o nā ʻōmole wai ʻiole/ʻiole. Ua koho ʻia kēia ʻano hui ʻana ma muli o nā puke pai he nui e hoʻohana ana iā NaHS ma ke ʻano he kumu o H2S; e ʻike i ka ʻāpana Kūkākūkā. ʻO NaHS kahi mole hydrated hiki ke loaʻa nā ʻano wai like ʻole o ka hydration (ʻo ia hoʻi, NaHS•xH2O); e like me ka mea hana, ʻo ka pakeneka o NaHS i hoʻohana ʻia i kā mākou noiʻi he 70.7% (ʻo ia hoʻi, NaHS•1.3 H2O), a ua noʻonoʻo mākou i kēia waiwai i kā mākou helu ʻana, kahi a mākou i hoʻohana ai i ke kaumaha molekala o 56.06 g/mol, ʻo ia ke kaumaha molekala o NaHS anhydrous. ʻO ka wai o ka hydration (i kapa ʻia hoʻi he wai o ka crystallization) nā molekala wai e hana ana i ka ʻano crystalline33. He ʻokoʻa nā waiwai kino a me ka thermodynamic o nā hydrates i hoʻohālikelike ʻia me nā anhydrates34.
Ma mua o ka hoʻohui ʻana iā NaHS i ka wai inu, e ana i ka pH a me ka mahana o ka mea hoʻoheheʻe. E ninini koke i ka hopena NaHS i loko o ka ʻōmole wai ʻiole/ʻiole i loko o ka pā holoholona. Ua hōʻiliʻili ʻia nā laʻana mai ka piko a mai loko mai o ka ʻōmole wai ma 0, 1, 2, 3, 4, 5, 6, 12, a me 24 hola e ana i ka nui o ka sulfide. Ua lawe koke ʻia nā ana sulfide ma hope o kēlā me kēia laʻana. Ua loaʻa iā mākou nā laʻana mai ka piko o ka ʻōmole no ka mea ua hōʻike kekahi mau haʻawina e hiki i ka liʻiliʻi o ka lua o ka ʻōmole wai ke hōʻemi i ka mahu ʻana o H2S15,19. Me he mea lā e pili ana kēia pilikia i ka hopena i loko o ka ʻōmole. Eia naʻe, ʻaʻole kēia ka hihia no ka hopena ma ka ʻāʻī o ka ʻōmole wai, kahi i ʻoi aku ka nui o ka mahu ʻana a ua autooxidizing; ʻo ka ʻoiaʻiʻo, ua inu mua nā holoholona i kēia wai.
Ua hoʻohana ʻia nā ʻiole Wistar kāne a me wahine i loko o ke aʻo ʻana. Ua hoʻokomo ʻia nā ʻiole i loko o nā pā polypropylene (2-3 ʻiole no kēlā me kēia pā) ma lalo o nā kūlana maʻamau (mahana 21-26 °C, haʻahaʻa 32-40%) me 12 hola o ke kukui (7 am a 7 pm) a me 12 hola o ka pouli (7 pm a 7 am). Loaʻa i nā ʻiole ke komo manuahi i ka wai piula a ua hānai ʻia me ka chow maʻamau (Khorak Dam Pars Company, Tehran, Iran). Ua hoʻokaʻawale ʻia nā ʻiole Wistar kāne (6 mahina) (n = 10, kaumaha kino: 190-230 g) a me nā ʻiole kāne (n = 10, kaumaha kino: 320-370 g) i nā hui i mālama ʻia me ka mana a me NaHS (30 μM) (n = 5 no kēlā me kēia hui). No ka hoʻoholo ʻana i ka nui o ka laʻana, ua hoʻohana mākou i ke ʻano KISS (Keep It Simple, Stupid), kahi e hoʻohui ai i ka ʻike ma mua a me ka loiloi mana35. Ua hana mua mākou i kahi noiʻi hoʻāʻo ma luna o 3 mau ʻiole a ua hoʻoholo i ka awelika o ka pae sulfide serum holoʻokoʻa a me ka ʻokoʻa maʻamau (8.1 ± 0.81 μM). A laila, e noʻonoʻo ana i ka mana 80% a me ka manaʻo ʻana i kahi pae koʻikoʻi ʻelua ʻaoʻao 5%, ua hoʻoholo mākou i kahi nui o ka laʻana mua (n = 5 ma muli o nā palapala mua) i kūlike i ka nui o ka hopena maʻamau o 2.02 me ka waiwai i wehewehe mua ʻia e Festing no ka helu ʻana i ka nui o ka laʻana o nā holoholona hoʻokolohua35. Ma hope o ka hoʻonui ʻana i kēia waiwai e ka SD (2.02 × 0.81), ʻo ka nui o ka hopena i ʻike ʻia (1.6 μM) he 20%, kahi mea i ʻae ʻia. ʻO ke ʻano kēia ua lawa ka n = 5/hui e ʻike i kahi loli awelika 20% ma waena o nā hui. Ua māhele ʻia nā ʻiole i loko o nā hui kaohi a me nā hui i mālama ʻia me NaSH me ka hoʻohana ʻana i ka hana maʻamau o ka polokalamu Excel 36 (Kiʻi Hoʻohui 1). Ua hana ʻia ka Blinding ma ka pae hopena, a ʻaʻole ʻike nā mea noiʻi e hana ana i nā ana biochemical i nā hoʻokaʻawale hui.
Ua mālama ʻia nā hui NaHS o nā kāne ʻelua me ka hopena 30 μM NaHS i hoʻomākaukau ʻia i loko o ka wai inu no 2 mau pule; Ua hoʻolako ʻia ka hopena hou i kēlā me kēia 24 hola, i ka manawa i ana ʻia ai ke kaumaha o ke kino. Ua hōʻiliʻili ʻia nā laʻana koko mai nā wēlau huelo o nā ʻiole āpau ma lalo o ka anesthesia isoflurane i kēlā me kēia lā ʻē aʻe i ka hopena o nā pule mua a me ka lua. Ua centrifuged ʻia nā laʻana koko ma 3000 g no 10 min, ua hoʻokaʻawale ʻia ka serum a mālama ʻia ma -80°C no ke ana ʻana o ka serum urea, creatinine (Cr), a me ka sulfide holoʻokoʻa. Ua hoʻoholo ʻia ka serum urea e ke ʻano urease enzymatic, a ua hoʻoholo ʻia ka serum creatinine e ke ʻano Jaffe photometric me ka hoʻohana ʻana i nā kits i loaʻa ma ke kālepa (Man Company, Tehran, Iran) a me kahi mea kālailai aunoa (Selectra E, helu serial 0-2124, Netherlands). ʻO nā coefficients intra- a me interassay o ka loli no ka urea a me Cr he emi ma mua o 2.5%.
Hoʻohana ʻia ke ʻano methylene blue (MB) e ana i ka sulfide holoʻokoʻa i loko o ka wai inu a me ke serum e loaʻa ana iā NaHS; ʻO MB ke ʻano i hoʻohana pinepine ʻia no ke ana ʻana i ka sulfide i loko o nā hopena nui a me nā laʻana biological11,37. Hiki ke hoʻohana ʻia ke ʻano MB e kuhi i ka waihona sulfide holoʻokoʻa38 a ana i nā sulfides inorganic ma ke ʻano o H2S, HS- a me S2 i ka pae wai39. Ma kēia ʻano, hoʻoheheʻe ʻia ka sulfur ma ke ʻano he zinc sulfide (ZnS) i ke alo o ka zinc acetate11,38. ʻO ka zinc acetate precipitation ke ʻano i hoʻohana nui ʻia no ka hoʻokaʻawale ʻana i nā sulfides mai nā chromophores ʻē aʻe11. Ua hoʻoheheʻe hou ʻia ʻo ZnS me ka hoʻohana ʻana iā HCl11 ma lalo o nā kūlana waikawa ikaika. Hoʻopili ka sulfide me DMPD ma kahi ratio stoichiometric o 1:2 i loko o kahi hopena i hoʻoulu ʻia e ka ferric chloride (hana ʻo Fe3+ ma ke ʻano he oxidizing agent) e hana i ka dye MB, kahi i ʻike ʻia spectrophotometrically ma 670 nm40,41. ʻO ka palena ʻike o ke ʻano MB ma kahi o 1 μM11.
Ma kēia haʻawina, ua hoʻohui ʻia he 100 μL o kēlā me kēia laʻana (waihoʻonā a i ʻole serum) i loko o kahi ʻōmole; a laila ua hoʻohui ʻia he 200 μL o ka zinc acetate (1% w/v i loko o ka wai i hoʻoheheʻe ʻia), 100 μL o DMPD (20 mM i loko o 7.2 M HCl), a me 133 μL o FeCl3 (30 mM i loko o 1.2 M HCl). Ua hoʻomoʻa ʻia ka hui ʻana ma 37°C i ka pouli no 30 min. Ua centrifuged ʻia ka waihoʻonā ma 10,000 g no 10 min, a ua heluhelu ʻia ka absorbance o ka supernatant ma 670 nm me ka hoʻohana ʻana i kahi mea heluhelu microplate (BioTek, MQX2000R2, Winooski, VT, USA). Ua hoʻoholo ʻia nā ʻano sulfide me ka hoʻohana ʻana i kahi piʻo calibration o NaHS (0–100 μM) ma ddH2O (Kiʻi Hoʻohui 2). Ua hoʻomākaukau hou ʻia nā waihoʻonā a pau i hoʻohana ʻia no nā ana. ʻO nā coefficients intra- a me interassay o ka loli no nā ana sulfide he 2.8% a me 3.4%, kēlā me kēia. Ua hoʻoholo pū mākou i ka huina sulfide i loaʻa mai ka wai inu sodium thiosulfate a me nā laʻana serum me ka hoʻohana ʻana i ke ʻano laʻana i hoʻoikaika ʻia42. ʻO nā hoʻōla no ka wai inu sodium thiosulfate a me nā laʻana serum he 91 ± 1.1% (n = 6) a me 93 ± 2.4% (n = 6), kēlā me kēia.
Ua hana ʻia ka loiloi helu me ka hoʻohana ʻana i ka polokalamu GraphPad Prism mana 8.0.2 no Windows (GraphPad Software, San Diego, CA, USA, www.graphpad.com). Ua hoʻohana ʻia kahi hoʻāʻo t i hui pū ʻia e hoʻohālikelike i ka mahana a me ka pH o ka wai inu ma mua a ma hope o ka hoʻohui ʻana o NaHS. Ua helu ʻia ka nalowale o H2S i loko o ka hopena i loaʻa iā NaHS ma ke ʻano he pakeneka emi mai ka lawe ʻana o ka baseline, a no ka loiloi inā he koʻikoʻi ka nalowale, ua hana mākou i kahi ANOVA hana hou ʻia hoʻokahi ala i ukali ʻia e ka hoʻāʻo hoʻohālikelike lehulehu a Dunnett. Ua hoʻohālikelike ʻia ke kaumaha o ke kino, ka urea serum, ka creatinine serum, a me ka serum sulfide holoʻokoʻa i ka hala ʻana o ka manawa ma waena o nā ʻiole kaohi a me nā ʻiole i mālama ʻia me NaHS o nā kāne like ʻole me ka hoʻohana ʻana i kahi ANOVA hui ʻelua ala (ma waena o loko) i ukali ʻia e kahi hoʻāʻo Bonferroni post hoc. Ua manaʻo ʻia he koʻikoʻi ka helu ʻana o nā waiwai P ʻelua-huelo < 0.05.
ʻO ka pH o ka wai inu he 7.60 ± 0.01 ma mua o ka hoʻohui ʻana o NaHS a me 7.71 ± 0.03 ma hope o ka hoʻohui ʻana o NaHS (n = 13, p = 0.0029). ʻO ka mahana o ka wai inu he 26.5 ± 0.2 a ua emi iho i 26.2 ± 0.2 ma hope o ka hoʻohui ʻana o NaHS (n = 13, p = 0.0128). E hoʻomākaukau i ka 30 μM NaHS solution i loko o ka wai inu a mālama iā ia i loko o kahi ʻōmole wai. He paʻa ʻole ka solution NaHS a emi iho kona nui i ka hala ʻana o ka manawa. I ka lawe ʻana i ka laʻana mai ka ʻāʻī o ka ʻōmole wai, ua ʻike ʻia kahi emi nui (68.0%) i loko o ka hola mua, a ua emi ka nui o NaHS i loko o ka solution ma 72% a me 75% ma hope o 12 a me 24 mau hola. I nā laʻana i loaʻa mai nā ʻōmole wai, ʻaʻole koʻikoʻi ka emi ʻana o NaHS a hiki i 2 mau hola, akā ma hope o 12 a me 24 mau hola ua emi iho ma 47% a me 72%, kēlā me kēia. Hōʻike kēia ʻikepili ua emi ka pakeneka o NaHS i loko o kahi hopena 30 μM i hoʻomākaukau ʻia i loko o ka wai inu i kahi hapaha o ka waiwai mua ma hope o 24 mau hola, me ka nānā ʻole i ka wahi laʻana (Kiʻi 1).
Ke kūpaʻa o ka hopena NaHS (30 μM) i loko o ka wai inu i loko o nā ʻōmole ʻiole/ʻiole. Ma hope o ka hoʻomākaukau ʻana i ka hopena, ua lawe ʻia nā laʻana mai ka piko a me loko o ka ʻōmole wai. Hōʻike ʻia nā ʻikepili ma ke ʻano he awelika ± SD (n = 6/hui). * a me #, P < 0.05 i hoʻohālikelike ʻia me ka manawa 0. Hōʻike ke kiʻi o ka ʻōmole wai i ka piko (me ka wehe ʻana) a me ke kino o ka ʻōmole. ʻO ka nui o ka piko ma kahi o 740 μL.
ʻO ka nui o NaHS i loko o ka hopena 30 μM i hoʻomākaukau hou ʻia he 30.3 ± 0.4 μM (pae: 28.7–31.9 μM, n = 12). Eia naʻe, ma hope o 24 hola, ua emi ka nui o NaHS i kahi waiwai haʻahaʻa (awelika: 3.0 ± 0.6 μM). E like me ka mea i hōʻike ʻia ma ke Kiʻi 2, ʻaʻole mau ka nui o NaHS i hōʻike ʻia ai nā ʻiole i ka wā o ke aʻo ʻana.
Ua piʻi nui ke kaumaha o ke kino o nā ʻiole wahine i ka hala ʻana o ka manawa (mai 205.2 ± 5.2 g a i 213.8 ± 7.0 g i ka hui kaohi a mai 204.0 ± 8.6 g a i 211.8 ± 7.5 g i ka hui i mālama ʻia me NaHS); akā naʻe, ʻaʻohe hopena o ka lāʻau NaHS i ke kaumaha o ke kino (Kiʻi 3). Ua piʻi nui ke kaumaha o ke kino o nā ʻiole kāne i ka hala ʻana o ka manawa (mai 338.6 ± 8.3 g a i 352.4 ± 6.0 g i ka hui kaohi a mai 352.4 ± 5.9 g a i 363.2 ± 4.3 g i ka hui i mālama ʻia me NaHS); akā naʻe, ʻaʻohe hopena o ka lāʻau NaHS i ke kaumaha o ke kino (Kiʻi 3).
Nā loli i ke kaumaha o ke kino i nā ʻiole wahine a me nā ʻiole kāne ma hope o ka lawelawe ʻana o NaHS (30 μM). Hōʻike ʻia nā ʻikepili ma ke ʻano he awelika ± SEM a ua hoʻohālikelike ʻia me ka hoʻohana ʻana i ka loiloi ʻelua ala i hui pū ʻia (i loko o waena) o ka variance me ka hoʻāʻo Bonferroni post hoc. n = 5 o kēlā me kēia kāne ma kēlā me kēia hui.
Ua like ka nui o ka urea serum a me ka creatine phosphate i nā ʻiole kaohi a me nā ʻiole i mālama ʻia me NaSH ma o ke aʻo ʻana. Eia kekahi, ʻaʻole i hoʻopilikia ka lāʻau NaSH i ka nui o ka urea serum a me ka creatinechrome (Papa 1).
Ua like nā pae sulfide serum kumu ma waena o nā ʻiole kāne (8.1 ± 0.5 μM vs. 9.3 ± 0.2 μM) a me nā ʻiole wahine (9.1 ± 1.0 μM vs. 6.1 ± 1.1 μM). ʻAʻohe hopena o ka hoʻokele ʻana o NaHS no 14 mau lā i nā pae sulfide serum i nā ʻiole kāne a wahine paha (Kiʻi 4).
Nā loli i nā nui o ka sulfide serum holoʻokoʻa i nā ʻiole kāne a me nā wahine ma hope o ka lawelawe ʻana o NaHS (30 μM). Hōʻike ʻia nā ʻikepili ma ke ʻano he awelika ± SEM a ua hoʻohālikelike ʻia me ka hoʻohana ʻana i kahi loiloi ʻelua ala i hui pū ʻia (i loko o loko) o ka variance me ka hoʻāʻo post hoc Bonferroni. Kēlā me kēia kāne, n = 5/hui.
ʻO ka hopena nui o kēia noiʻi, ʻo ia ka wai inu i loaʻa ka NaHS he paʻa ʻole: hiki ke ʻike ʻia ma kahi o ka hapahā o ka nui o ka sulfide mua i 24 mau hola ma hope o ka lawe ʻana i ka laʻana mai ka piko a i loko o nā ʻōmole wai ʻiole/ʻiole. Eia kekahi, ua hōʻike ʻia nā ʻiole i nā nui NaHS paʻa ʻole ma muli o ka nalowale ʻana o ka H2S i loko o ka hopena NaHS, a ʻo ka hoʻohui ʻana o NaHS i ka wai inu ʻaʻole i hoʻopilikia i ke kaumaha o ke kino, ka serum urea a me ka creatine chromium, a i ʻole ka serum sulfide holoʻokoʻa.
Ma kēia haʻawina, ʻo ka nui o ka nalowale ʻana o ka H2S mai nā hopena NaHS 30 μM i hoʻomākaukau ʻia i loko o ka wai inu ma kahi o 3% i kēlā me kēia hola. I loko o kahi hopena buffered (100 μM sodium sulfide i loko o 10 mM PBS, pH 7.4), ua hōʻike ʻia ka emi ʻana o ka sulfide ma ka 7% i ka hala ʻana o ka manawa ma luna o 8 h11. Ua pale mua mākou i ka hoʻokele intraperitoneal o NaHS ma ka hōʻike ʻana ʻo ka nui o ka nalowale sulfide mai kahi hopena NaHS 54 μM i loko o ka wai inu ma kahi o 2.3% i kēlā me kēia hola (4%/hola i nā hola 12 mua a me 1.4%/hola i nā hola 12 hope loa ma hope o ka hoʻomākaukau ʻana)8. Ua ʻike ʻia nā haʻawina mua43 i kahi nalowale mau o ka H2S mai nā hopena NaHS, ma muli o ka volatilization a me ka oxidation. ʻOiai me ka ʻole o ka hoʻohui ʻana o nā pehu, nalowale koke ka sulfide i loko o ka hopena waihona ma muli o ka volatilization H2S11. Ua hōʻike ʻia nā haʻawina i ka wā o ke kaʻina hana dilution, kahi e lawe ai ma kahi o 30-60 kekona, ma kahi o 5-10% o H2S e nalowale ana ma muli o ka mahu ʻana6. No ka pale ʻana i ka mahu ʻana o H2S mai ka hopena, ua hana nā mea noiʻi i kekahi mau hana, me ka hoʻoulu mālie ʻana o ka hopena12, ka uhi ʻana i ka hopena waihona me ka ʻili plastik6, a me ka hoʻēmi ʻana i ka hōʻike ʻana o ka hopena i ka ea, ʻoiai ʻo ka wikiwiki o ka mahu ʻana o H2S e pili ana i ka interface ea-wai.13 ʻO ka oxidation spontaneous o H2S e hana nui ʻia ma muli o nā ion metala hoʻololi, ʻoiai ka hao ferric, he mau haumia i loko o ka wai.13 ʻO ka oxidation o H2S ka hopena i ka hoʻokumu ʻia ʻana o nā polysulfides (nā ʻātoma sulfur i hoʻopili ʻia e nā pilina covalent)11. I mea e pale aku ai i kona oxidation, ua hoʻomākaukau ʻia nā hopena e loaʻa ana ka H2S i loko o nā solvents deoxygenated44,45 a laila hoʻomaʻemaʻe ʻia me ka argon a i ʻole ka nitrogen no 20-30 min e hōʻoia i ka deoxygenation.11,12,37,44,45,46 ʻO ka Diethylenetriaminepentaacetic acid (DTPA) kahi chelator metala (10-4 M) e pale ai i ka HS- autoxidation i nā hopena aerobic. I ka loaʻa ʻole o ka DTPA, ʻo ka helu autoxidation o HS- ma kahi o 50% ma luna o kahi o 3 h ma 25°C37,47. Eia kekahi, ʻoiai ua hoʻoulu ʻia ka oxidation o 1e-sulfide e ka mālamalama ultraviolet, pono e mālama ʻia ka hopena ma ka hau a pale ʻia mai ka mālamalama11.
E like me ka mea i hōʻike ʻia ma ke Kiʻi 5, hoʻokaʻawale ʻo NaHS i Na+ a me HS-6 ke hoʻoheheʻe ʻia i loko o ka wai; ua hoʻoholo ʻia kēia hoʻokaʻawale ʻana e ka pK1 o ka hopena, nona ka mahana: pK1 = 3.122 + 1132/T, kahi e piʻi ai ʻo T mai 5 a 30°C a ua ana ʻia ma nā kekelē Kelvin (K), K = °C + 273.1548. Loaʻa iā HS- kahi pK2 kiʻekiʻe (pK2 = 19), no laila ma ka pH < 96.49, ʻaʻole i hoʻokumu ʻia ʻo S2- a i ʻole i hoʻokumu ʻia i nā liʻiliʻi loa. I ka hoʻohālikelike ʻana, hana ʻo HS- ma ke ʻano he kumu a ʻae iā H+ mai kahi mole H2O, a hana ʻo H2O ma ke ʻano he waikawa a hoʻololi ʻia i H2S a me OH-.
Ka hoʻokumu ʻia ʻana o ke kinoea H2S i hoʻoheheʻe ʻia i loko o ka hopena NaHS (30 µM). aq, hopena wai; g, kinoea; l, wai. Manaʻo nā helu āpau he pH wai = 7.0 a me ka mahana o ka wai = 20 °C. Hana ʻia me BioRender.com.
ʻOiai nā hōʻike e hōʻike ana he paʻa ʻole nā ​​​​​​waihoʻo NaHS, ua hoʻohana kekahi mau haʻawina holoholona i nā waihoʻo NaHS i loko o ka wai inu ma ke ʻano he hui hāʻawi H2S15,16,17,18,19,20,21,22,23,24,25,26 me nā lōʻihi o ke komo ʻana mai 1 a 21 mau pule (Papa 2). I loko o kēia mau haʻawina, ua hoʻohou hou ʻia ka waihoʻo NaHS i kēlā me kēia 12 h, 15, 17, 18, 24, 25 h a i ʻole 24 h, 19, 20, 21, 22, 23 h. Ua hōʻike kā mākou mau hopena ua hōʻike ʻia nā ʻiole i nā ʻano lāʻau paʻa ʻole ma muli o ka nalowale ʻana o H2S mai ka waihoʻo NaHS, a ua loli nui ka nui o ka NaHS i loko o ka wai inu o nā ʻiole ma luna o 12 a i ʻole 24 h (e nānā i ke Kiʻi 2). Ua hōʻike ʻelua o kēia mau haʻawina ua noho paʻa nā pae H2S i loko o ka wai ma luna o 24 h22 a i ʻole he 2-3% wale nō ka pohō H2S i ʻike ʻia ma luna o 12 h15, akā ʻaʻole lākou i hāʻawi i ka ʻikepili kākoʻo a i ʻole nā ​​kikoʻī ana. Ua hōʻike ʻelua mau haʻawina e hiki i ke anawaena liʻiliʻi o nā ʻōmole wai ke hōʻemi i ka mahu ʻana o ka H2S15,19. Eia nō naʻe, ua hōʻike kā mākou mau hopena e hoʻopaneʻe wale kēia i ka pohō H2S mai kahi ʻōmole wai ma 2 h ma mua o 12-24 h. Hoʻomaopopo nā haʻawina ʻelua ua manaʻo mākou ʻaʻole i loli ka pae NaHS i loko o ka wai inu no ka mea ʻaʻole mākou i ʻike i kahi loli kala i loko o ka wai; no laila, ʻaʻole koʻikoʻi ka oxidation o H2S e ka ea19,20. ʻO ka mea kupanaha, loiloi kēia ʻano pilikino i ke kūpaʻa o NaHS i loko o ka wai ma mua o ke ana ʻana i ka loli o kona ʻano i ka hala ʻana o ka manawa.
ʻO ka nalowale ʻana o ka H2S i loko o ka hopena NaHS e pili ana i ka pH a me ka mahana. E like me ka mea i ʻōlelo ʻia ma kā mākou noiʻi, ʻo ka hoʻoheheʻe ʻana o NaHS i loko o ka wai e hopena i ka hoʻokumu ʻia ʻana o kahi hopena alkaline50. Ke hoʻoheheʻe ʻia ʻo NaHS i loko o ka wai, ʻo ka hoʻokumu ʻia ʻana o ke kinoea H2S i hoʻoheheʻe ʻia e pili ana i ka waiwai pH6. ʻO ka haʻahaʻa o ka pH o ka hopena, ʻo ka nui o ka hapa o NaHS i loaʻa ma ke ʻano he mau molekala kinoea H2S a ʻo ka nui o ka sulfide i nalowale mai ka hopena wai11. ʻAʻohe o kēia mau noiʻi i hōʻike i ka pH o ka wai inu i hoʻohana ʻia ma ke ʻano he mea hoʻoheheʻe no NaHS. Wahi a nā ʻōlelo aʻoaʻo a WHO, i ʻae ʻia e ka hapa nui o nā ʻāina, pono ka pH o ka wai inu ma ka pae 6.5–8.551. Ma kēia pae pH, piʻi ka wikiwiki o ka oxidation spontaneous o H2S ma kahi o ʻumi manawa13. ʻO ka hoʻoheheʻe ʻana o NaHS i loko o ka wai ma kēia pae pH e hopena i kahi ʻano kinoea H2S i hoʻoheheʻe ʻia o 1 a 22.5 μM, kahi e hoʻoikaika ai i ke koʻikoʻi o ka nānā ʻana i ka pH o ka wai ma mua o ka hoʻoheheʻe ʻana o NaHS. Eia kekahi, ʻo ka pae mahana i hōʻike ʻia ma ke aʻo ʻana ma luna (18–26 °C) e hopena i kahi loli i ka nui o ke kinoea H2S i hoʻoheheʻe ʻia i loko o ka hopena o kahi 10%, ʻoiai ʻo nā loli mahana e hoʻololi i ka pK1, a ʻo nā loli liʻiliʻi i ka pK1 hiki ke loaʻa ka hopena koʻikoʻi i ka nui o ke kinoea H2S i hoʻoheheʻe ʻia48. Eia kekahi, ʻo ka lōʻihi o kekahi mau haʻawina (5 mau mahina)22, kahi e manaʻo ʻia ai ka loli nui o ka mahana, e hoʻonui pū i kēia pilikia.
Ua hoʻohana nā haʻawina āpau koe wale nō hoʻokahi21 i ka hopena 30 μM NaHS i loko o ka wai inu. No ka wehewehe ʻana i ka nui i hoʻohana ʻia (ʻo ia hoʻi 30 μM), ua kuhikuhi kekahi mau mea kākau e hana like ana ʻo NaHS i loko o ka pae wai i ka nui like o ke kinoea H2S a ʻo ka pae physiological o H2S he 10 a 100 μM, no laila aia kēia nui i loko o ka pae physiological15,16. Ua wehewehe kekahi poʻe hiki i ka 30 μM NaHS ke mālama i ka pae plasma H2S i loko o ka pae physiological, ʻo ia hoʻi 5-300 μM19,20. Manaʻo mākou i ka nui o NaHS i loko o ka wai o 30 μM (pH = 7.0, T = 20 °C), i hoʻohana ʻia i kekahi mau haʻawina e aʻo ai i nā hopena o H2S. Hiki iā mākou ke helu i ka nui o ke kinoea H2S i hoʻoheheʻe ʻia he 14.7 μM, ʻo ia hoʻi ma kahi o 50% o ka nui NaHS mua. Ua like kēia waiwai me ka waiwai i helu ʻia e nā mea kākau ʻē aʻe ma lalo o nā kūlana like13,48.
I kā mākou noiʻi ʻana, ʻaʻole i hoʻololi ka lawelawe ʻana o NaHS i ke kaumaha o ke kino; kūlike kēia hopena me nā hopena o nā noiʻi ʻē aʻe i nā ʻiole kāne22,23 a me nā ʻiole kāne18; Eia nō naʻe, ua hōʻike ʻelua mau noiʻi ua hoʻihoʻi ʻo NaSH i ke kaumaha o ke kino i emi iho i nā ʻiole nephrectomized24,26, ʻoiai ʻaʻole i hōʻike nā noiʻi ʻē aʻe i ka hopena o ka lawelawe ʻana o NaSH i ke kaumaha o ke kino15,16,17,19,20,21,25. Eia kekahi, i kā mākou noiʻi ʻana, ʻaʻole i hoʻopilikia ka lawelawe ʻana o NaSH i ka serum urea a me nā pae creatine chromium, kahi e kūlike me nā hopena o kahi hōʻike ʻē aʻe25.
Ua ʻike ʻia ma ke aʻo ʻana ʻaʻole i hoʻopilikia ka hoʻohui ʻana o NaHS i ka wai inu no 2 mau pule i ka nui o ka serum sulfide i nā ʻiole kāne a me nā wahine. Kūlike kēia ʻike me nā hopena o Sen et al. (16): ʻaʻole i hoʻopilikia ka 8 mau pule o ka mālama ʻana me 30 μM NaHS i ka wai inu i nā pae plasma sulfide i nā ʻiole kaohi; akā naʻe, ua hōʻike lākou ua hoʻihoʻi kēia hana i ka emi ʻana o nā pae H2S i loko o ka plasma o nā ʻiole nephrectomized. Ua hōʻike pū ʻo Li et al. (22) ua hoʻonui ka mālama ʻana me 30 μM NaHS i ka wai inu no 5 mau mahina i nā pae plasma free sulfide i nā ʻiole ʻelemakule ma kahi o 26%. ʻAʻole i hōʻike nā haʻawina ʻē aʻe i nā loli i ka circulating sulfide ma hope o ka hoʻohui ʻana o NaHS i ka wai inu.
ʻEhiku mau haʻawina i hōʻike ʻia me ka hoʻohana ʻana iā Sigma NaHS15,16,19,20,21,22,23 akā ʻaʻole i hāʻawi i nā kikoʻī hou aku e pili ana i ka wai hydration, a ʻelima mau haʻawina ʻaʻole i haʻi i ke kumu o NaHS i hoʻohana ʻia i kā lākou mau ʻano hoʻomākaukau17,18,24,25,26. He molekole hydrated ʻo NaHS a hiki ke ʻokoʻa kona ʻano wai hydration, kahi e hoʻopilikia ai i ka nui o NaHS e pono ai e hoʻomākaukau i kahi hopena o kahi molarity i hāʻawi ʻia. No ka laʻana, ʻo ka ʻike NaHS i kā mākou haʻawina ʻo NaHS•1.3 H2O. No laila, ʻoi aku ka haʻahaʻa o nā ʻano NaHS maoli i kēia mau haʻawina ma mua o nā mea i hōʻike ʻia.
"Pehea lā e hiki ai i kahi hui pōkole ke loaʻa i kahi hopena lōʻihi?" Ua nīnau ʻo Pozgay et al.21 i kēia nīnau i ka wā e loiloi ai i nā hopena o NaHS ma ka colitis i nā ʻiole. Manaʻolana lākou e hiki i nā haʻawina e hiki mai ana ke pane i kēia nīnau a kuhi e loaʻa paha i nā hopena NaHS nā polysulfides paʻa hou aʻe ma waho aʻe o ka H2S a me nā disulfides e hoʻoponopono ana i ka hopena o NaHS21. ʻO kekahi mea hiki ke loaʻa i nā haʻahaʻa haʻahaʻa loa o NaHS i koe i loko o ka hopena kahi hopena maikaʻi. ʻO ka ʻoiaʻiʻo, ua hāʻawi ʻo Olson et al. i nā hōʻike ʻaʻole physiological nā pae micromolar o H2S i loko o ke koko a pono e noho i loko o ka pae nanomolar a ʻaʻole loa13. Hiki i ka H2S ke hana ma o ka protein sulfation, kahi hoʻololi post-translational reversible e hoʻopilikia ai i ka hana, ke kūpaʻa, a me ka localization o nā protein he nui52,53,54. ʻO ka ʻoiaʻiʻo, ma lalo o nā kūlana physiological, ma kahi o 10% a 25% o nā protein ate he nui i sulfylated53. Ua ʻae nā haʻawina ʻelua i ka luku wikiwiki ʻana o NaHS19,23 akā ua ʻōlelo kupanaha ʻia "ua kāohi mākou i ka nui o NaHS i loko o ka wai inu ma ka hoʻololi ʻana i kēlā me kēia lā."23 Ua ʻōlelo hewa kekahi haʻawina "ʻO NaHS kahi mea hāʻawi H2S maʻamau a hoʻohana pinepine ʻia i ka hana lapaʻau e pani iā ​​H2S ponoʻī."18
Hōʻike ka kūkākūkā ma luna nei ua nalowale ʻo NaHS mai ka hopena ma o ka volatilization, oxidation a me ka photolysis, a no laila ua hāʻawi ʻia kekahi mau manaʻo e hōʻemi i ka nalowale o H2S mai ka hopena. ʻO ka mea mua, pili ka mahu o H2S i ka interface gas-liquid13 a me ka pH o ka hopena11; no laila, e hōʻemi i ka nalowale evaporative, hiki ke hana ʻia ka ʻāʻī o ka ʻōmole wai e like me ka mea i wehewehe ʻia ma mua15,19, a hiki ke hoʻoponopono ʻia ka pH o ka wai i kahi palena kiʻekiʻe i ʻae ʻia (ʻo ia hoʻi, 6.5–8.551) e hōʻemi i ka nalowale evaporative11. ʻO ka lua, hana ʻia ka oxidation spontaneous o H2S ma muli o nā hopena o ka oxygen a me ke alo o nā ions metala hoʻololi i loko o ka wai inu13, no laila hiki i ka deoxygenation o ka wai inu me ka argon a i ʻole nitrogen44,45 a me ka hoʻohana ʻana i nā chelators metala37,47 ke hōʻemi i ka oxidation o sulfides. ʻO ke kolu, e pale i ka photodecomposition o H2S, hiki ke ʻōwili ʻia nā ʻōmole wai me ka foil alumini; Pili pū kēia hana i nā mea ʻike mālamalama e like me streptozotocin55. ʻO ka mea hope loa, hiki ke lawelawe ʻia nā paʻakai sulfide inorganic (NaHS, Na2S, a me CaS) ma o ka gavage ma mua o ka hoʻoheheʻe ʻia i loko o ka wai inu e like me ka mea i hōʻike mua ʻia56,57,58; ua hōʻike nā haʻawina ua hoʻopili maikaʻi ʻia ka sodium sulfide radioactive i hāʻawi ʻia ma o ka gavage i nā ʻiole a ua hoʻolaha ʻia i aneane nā ʻiʻo āpau59. A hiki i kēia lā, ua hāʻawi ka hapa nui o nā haʻawina i nā paʻakai sulfide inorganic intraperitoneally; akā naʻe, ʻaʻole hoʻohana pinepine ʻia kēia ala i nā hoʻonohonoho lapaʻau60. Ma ka ʻaoʻao ʻē aʻe, ʻo ke ala waha ka mea maʻamau a makemake nui ʻia o ka lawelawe ʻana i loko o nā kānaka61. No laila, ke paipai nei mākou e loiloi i nā hopena o nā mea hāʻawi H2S i nā rodents ma o ka gavage waha.
ʻO kahi palena, ua ana mākou i ka sulfide i loko o ka wai aqueous a me ke serum me ka hoʻohana ʻana i ke ʻano MB. ʻO nā ʻano hana no ke ana ʻana i ka sulfide e komo pū me ka iodine titration, spectrophotometry, electrochemical method (potentiometry, amperometry, coulometric method a me amperometric method) a me ka chromatography (gas chromatography a me ka high-performance liquid chromatography), ʻo ia hoʻi ke ʻano hana i hoʻohana pinepine ʻia ʻo ia ke ʻano spectrophotometric MB62. ʻO kahi palena o ke ʻano MB no ke ana ʻana i ka H2S i nā laʻana olaola, ʻo ia ke ana ʻana i nā hui sulfur āpau a ʻaʻole ka H2S63 manuahi no ka mea ua hana ʻia ma lalo o nā kūlana acidic, kahi e hopena ai i ka unuhi ʻana o ka sulfur mai ke kumu olaola64. Eia naʻe, e like me ka American Public Health Association, ʻo MB ke ʻano maʻamau no ke ana ʻana i ka sulfide i ka wai65. No laila, ʻaʻole kēia palena e hoʻopilikia i kā mākou mau hopena nui ma ka paʻa ʻole o nā hopena i loaʻa iā NaHS. Eia kekahi, i kā mākou noiʻi ʻana, ʻo ka hoʻihoʻi ʻana o nā ana sulfide i loko o ka wai a me nā laʻana serum i loaʻa iā NaHS he 91% a me 93%, kēlā me kēia. Kūlike kēia mau waiwai me nā pae i hōʻike mua ʻia (77–92)66, e hōʻike ana i ka pololei loiloi i ʻae ʻia42. He mea kūpono ke hoʻomaopopo ʻana ua hoʻohana mākou i nā ʻiole kāne a me nā wahine e like me nā alakaʻi o ka National Institutes of Health (NIH) e pale aku i ka hilinaʻi nui ʻana i nā haʻawina holoholona kāne wale nō i nā haʻawina preclinical67 a e hoʻokomo i nā ʻiole kāne a me nā wahine i nā manawa hiki68. Ua hoʻoikaika ʻia kēia manaʻo e kekahi poʻe ʻē aʻe69,70,71.
I ka hopena, hōʻike nā hopena o kēia noiʻi ʻaʻole hiki ke hoʻohana ʻia nā hopena NaHS i hoʻomākaukau ʻia mai ka wai inu e hana i ka H2S ma muli o ko lākou paʻa ʻole. ʻO kēia ala o ka lawelawe ʻana e hōʻike i nā holoholona i nā pae NaHS paʻa ʻole a haʻahaʻa ma mua o ka mea i manaʻo ʻia; no laila, ʻaʻole paha e pili nā ʻike i nā kānaka.
Loaʻa nā ʻikepili i hoʻohana ʻia a/a i ʻole i kālailai ʻia i ka wā o ke aʻo ʻana o kēia manawa mai ka mea kākau pili ma ke noi kūpono.
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