ОРИГІНАЛЬНІ ДОСЛІДЖЕННЯ: ФУНДАМЕНТАЛЬНІ НАУКИ

ORIGINAL RESEARCH: BASIC SCIENCES

DOI

UDC: (616.153.455.01:616.33-018.73-092:615.015.11]-085.37-092.9)

ATB 340 (A MODULATOR OF SULFITE OXIDASE ACTIVITY) REDUCES OXIDATIVE STRESS DURING HYPERGLYCEMIA AND IN STRESS EXPOSED GASTRIC MUCOSA IN OLD RATS

Yaroslav Pavlovskiy1, Maksym Lutsyk2, Antonina Yashchenko2, Natalia Zaichko3, John Wallace4, Oksana Zayachkivska1

1. Physiology Department of Lviv National Medical University, Lviv, Ukraine

2. Histology, Cytology and Embryology Department of Lviv National Medical University, Lviv, Ukraine

3. Biological and General Chemistry Department of National Pirogov Memorial Medical University, Vinnytsia, Ukraine

4. Physiology and Pharmacology Department of University of Calgary, Calgary, Canada pavl_jarik@ymail.com

2

Introduction. Sulfite oxidase (SO) is one of the mitochondrial enzymes involved in the synthe- sis of H2S and multiple physiological processes, including oxidative stress. The dominant sulfide oxidation products vary in a tissue-specific manner, and SO/oxidative stress which is involved in gastric mucosa (GM) injury is unclear. Our studies have shown potent cytoprotective and anti-in- flammatory effects of H2S-rich compounds, including hybrid nonsteroidal anti-inflammatory drugs (H S-NSAIDs) but their effects on age-related metabolic dysfunction in the gastric mucosa (GM) exposed to long-term postprandial hyperglycemia with high-fructose diet (HFD) is still unknown.

2

Aim. To characterize the activity of sulfite oxidase (SO) and the role of oxidative stress in the GM of older rats exposed to HFD and assess the therapeutic effect of the recently synthesized H S donor, hybrid NSAID, H S-acetylsalicylic acid (H S-ASA, ATB 340) on the GM of adult vs old rats.

2 2

Material and methods. Adult (n=24) and old (n=43) rats were divided into control groups with a standard diet (SD) and experimental groups with a 28-day high fructose level diet (HFD,

V. Kozar, 2008) with and without acute stress induction (Takagi, 1964). During 19-28 day, HFD groups consumed per os: a) placebo (1.0 ml of saline per os); b) NaHS, 100 μmol/kg/day; c) pla- cebo and stress induction; d) NaHS, 100 μmol/kg/day and stress induction; e) ASA, 10 mg/kg/ day and stress induction; f) conventional aspirin (ASA), 10 mg/kg/day and NaHS, 100 μmol/kg/ day and stress induction; g) ATB-340, 17.5 mg/kg/day and stress induction. The integrity of the GM was analyzed based on the histological severity index, which was calculated with account for morphological studies of GM samples. The indicator of oxidative stress, malondialdehyde (MDA) and the activity of SO in the GM were assessed using the standard biochemical methods.

Results. We have observed a different SO expression in the GM of adult vs old rats with SD and HFD with and without WIS. The effect of HFD resulted in an increase in the SO activity in old rats by 15%, whereas in adults this value was 14%. MDA content changes were similar. In animals that received NaHS, the changes were similar, whereas the SO activity in old rats was 23% higher. Stress induction reduced the SO activity by 35% in old animals and by 22% in adults. The difference in the MDA content amounted to 41%. The effect of exogenous NaHS in old rats under stress was the highest when combined with ASA, while the MDA content was the highest when classical aspirin was used, which also coincided with similar changes in his- tological severity index.

Оригінальні дослідження: фундаментальні науки Original research: basic sciences

Conclusion. Sulfite oxidase is the essential contributor to age-related oxidative stress in the GM under research. The use of ATB-340 (H S-aspirin) increases the H S content in the tissues, re-

2 2

duces symptoms of GM damage associated with a decrease in the SO activity and the content of

MDA, which confirms alleviation of oxidative stress and has anti-radical and anti-oxidant effects.

Disclosures. No conflicts of interest, financial or otherwise, are declared by the authors.

Author contribution. Yaroslav Pavlovskiy, Maksym Lutsyk drafted manuscript; Antonina Yash- chenko edited and revised manuscript; Natalia Zaichko, John Wallace, Oksana Zayachkivska ap- proved final version of the manuscript.

Key words: Hydrogen Sulfide, Mucosal Defense, Gastric Mucosa, Aspirin, Non-Steroidal Anti-In-

flammatory Drugs, Sulfite Oxidase, MDA, Fructose

ATB 340, МОДУЛЯТОР АКТИВНОСТІ СУЛЬФІТ ОКСИДАЗИ, ЗМЕНШУЄ ОКИСНИЙ СТРЕС У СЛИЗОВІЙ ОБОЛОНЦІ ШЛУНКА СТАРИХ ЩУРІВ ЗА УМОВ ГІПЕРГЛІКЕМІЇ ТА СТРЕСУ

Ярослав Павловський1, Максим Луцик2, Антоніна Ященко2, Наталія Заічко3, Джон Л. Уоллес4, Оксана Заячківська1

1Кафедра нормальної фізіології, Львівський національний медичний уні- верситет ім. Данила Галицького, Львів, Україна

2Кафедра гістології, цитології та ембріології, Львівський національний медичний університет ім. Данила Галицького, Львів, Україна

3Кафедра біологічної та загальної хімії, Вінницький національний медич- ний університет ім. М.І.Пирогова, Вінниця, Україна

4Кафедра фізіології та фармакології, Університет Калгарі, Калгарі, Канада

pavl_jarik@ymail.com

2

2

2

Вступ. Сульфіт-оксидаза (SO) є одним з мітохондріальних ферментів, що беруть участь у синтезі H S і численних фізіологічних процесах, включаючи окисний стрес. Продукти окис- нення сульфідів мають тканинну специфічність, однак особливості їхнього пошкоджу валь- ного впливу на слизову оболонку шлунка (GM) не відомі. В наших дослідженнях показано потужний цитопротекторний та протизапальний ефект сполук, збагачених Н S і гібридних нестероїдних протизапальних препаратів (Н S-NSAIDs), але їх вплив на вікові особливості метаболічної дисфункції у СОШ на тлі довготривалої її постпрандіальної гіперглікемії за умов гіперкалорійної високо-вуглеводної дієти фруктозою (HFD) не вивчено.

2

Мета. Охарактеризувати активність сульфітоксидази (SO) та роль окисного стресу у СОШ старих щурів при HFD та оцінити терапевтичний ефект недавно синтезованого донора H S, гібрид NSAIDS, Н S-ацетилсаліцилова кислота (H S-ASA, ATB 340) на СОШ старих тварин

2 2

порівняно із дорослими.

Матеріали і методи. Дорослих (n = 24) і старих (n = 43) щурів поділили на контрольні групи зі стандартною дієтою (SD) та експериментальні групи із дієтою з високим вмістом фруктози (HFD, V. Kozar, 2008) впродовж 28 днів, яким додатково індукували гострий стрес (WIS) (Takagi, 1964). Групи з HFD у 19-28 дні отримували per os: а) плацебо (1,0 мл фізіо- логічного розчину per os); б) NaHS, 100 мкмоль/кг/добу; в) плацебо та індукція стресу; г) NaHS, 100 мкмоль/кг/добу та індукція стресу; д) ASA, 10 мг/кг/добу та індукція стресу; е)

Оригінальні дослідження: фундаментальні науки Original research: basic sciences

ASA, 10 мг/кг/добу, NaHS, 100 мкмоль/кг/добу та індукція стресу; є) АТB-340, 17,5 мг/кг/ добу та індукція стресу). Цілісність СОШ аналізували за гістологічним індексом ураження, що обраховували враховуючи морфологічні дослідження зразків СОШ. Індикатор окисного стресу, малоновий диальдегід (MDA) та активність SO в GM оцінювали стандартними біохі- мічними методами.

Результати. Ми спостерігали різну експресію SO в GM у дорослих щурів у порівнянні зі ста- рими щурами з SD і HFD без і з WIS. Вплив HFD призводив до збільшення активності SO у старих щурів на 15%, тоді як у дорослих на 14%. Зміни вмісту MDA були схожими. У тварин, яким вводили NaHS, зміни MDA мали подібну тенденцію, тоді як активність SO у старих щу- рів була вища на 23%. Індукція стресу зменшувала активність SO у старих тварин на 35%, тоді як у дорослих – на 22%, а відмінність у вмісті MDA сягала 41%. Дія екзогенного NaHS у старих щурів за умов стресу була найвищою у разі комбінації з ASA, тоді як вміст MDA був найвищий при застосуванні класичного аспірину, що співпадало з аналогічними змінами гі- стологічного індексу ураження.

Висновок. Сульфіт оксидаза є важливим фактором вікового окисного стресу у GM, який був досліджений. Застосування ATB 340 (H S-аспірину), що збільшує вміст H S в тканинах,

2 2

зменшує прояви пошкоджень GM, пов’язаних зі зменшенням активності SO і вмісту MDA, що

свідчить про облегшення проявів окисного стресу, виявляючи анти-радикальну та анти-о- ксидантну дію.

Ключові слова: Гідроген Сульфіду, захист слизової оболонки, шлунок, аспірин, нестероїдні протизапальні засоби, сульфіт оксидаза, окисний стрес, МДА, фруктоза

Introduction

The gastrointestinal (GI) tract shows a re- markable resilience to damage induced by the beverages and foods that we ingest, which can have a wide range of osmolarity, pH and tem- perature [20]. This resistance to tissue dam- age is collectively referred to as “mucosal de- fense” and many different chemical mediators participate in GM defense, inflammation and repair [9, 18]. A new classification of human diseases will become available soon, based on consensus in expert committees that decided to include new category „stress-related dis- eases” (SRD) [7]. It is widely accepted that SRD result from complex reciprocal interaction between epigenetic and genetic factors. One of the epigenetic factors is fructose-rich nutri- tion, which results in oxidative stress, howev- er its prevention remains incompletely under- stood [3, 14]. There are age-related features in gastric mucosa (GM) defense, inflammation and repair against cytolytic agents, includ- ing prostaglandin/cyclooxygenase activities [11]. The issue of preventing and correcting the cyto-aggressive action of nonsteroidal an- ti-inflammatory drugs (NSAIDs), which widely used by elderly patients and most often caus- es GM damage to the elderly patients [1, 2,

2

6] and whatever age-related features in H S mucosal defence against oxidative stress is related to that, is open.

2

2

Important bioregulators of prostaglandin-in- dependent mechanisms of cytoprotection of the GM is sulfur compounds. Recent extensive research into their effects confirmed as potent cyto- and vasoprotective effects, including against low-grade inflammation [17]. It has been shown that Sulfhydryl (SH) groups play prominent role in gastroprotection by deplet- ing glutathione (GSH) and induction oxidative stress [11, 18]. Another important regulator is the gas transmitter H S (hydrogen sulfide), which is known for its vasodilator, antioxidant, antiapoptotic and pro-angiogenic properties, which provide cytoprotective and anti-inflam- matory effects [16, 22]. A number of stud- ies point out the unity of various ways of H S synthesis: enzymatic and non-enzymatic (the transformation of thiols and thiol-containing compounds) [5, 15, 23]. The four most im- portant enzymes involved in formation of hy- drogen sulfide are: cystathionine β-synthase

• CBS, cystathionine γ-lyase – CSE, 3-mer- captopyruvate sulfurtransferase – 3-MST, which cooperates with cysteine aminotrans-

  
  

  

  Оригінальні дослідження: фундаментальні науки Original research: basic sciences

  
  

  2

  2

  2

  2

  ferase – CAT [17]. It is known that these en- zymes have specific tissue and cellular local- ization. We have shown that for the purpose of cytoprotection of the mucosal barrier of the digestive system (esophagus, stomach and intestines) and anti-inflammatory action, the synthesis of H S is carried out using CSE, as well as the maintenance of normal microflora in the cardiovascular system under physiolog- ical conditions [16, 23]. It has been found that in activated neutrophils, H S can be oxidized to thiosulfate (under the influence of sulfite ox- idase (SO, EC 1.8.3.1) oxidizes sulfite to sul- fate), and then converted to SO2 (by enzymes thiosulfate sulfutransferase or thiosulfate re- ductase). SO is one of the enzymes involved in the utilization of H S in mitochondria too [5]. Both H S pathways enzymatic and non-enzy- matic act as crucial rheostat regulators of the redox state, affected by active forms of oxy- gen levels in cells, and have both been shown to combine together and perform key roles in cellular and subcellular survival and progres- sion under extreme factors actions. However, the full concept explaining the role of SO in GM cytoprotection in the aspect of age-relat- ed changes is still unknown. Metabolic dys- function induced by long-term fructose influ- ence is not finally clarified. Taking in account that of nonsteroidal anti-inflammatory drugs (NSAIDs), potent cytolysis’ agents for GM, are among the most commonly used medications, and their use is increasing with aging popula- tions worldwide despite of the significant risk for pro-oxidative gastrointestinal (GI) effects, bleeding and ulceration, sometimes leading to death [6, 10, 12].

  
  

  2

  Aim. To characterize the role of sulfite oxi- dase (SO) and activity of oxidative stress in GM in older rats with fructose exposed hyper- glycemia and to assess the therapeutic effect of the recently synthesized H S donor – hybrid NSAID H S-acetylsalicylic (H S-ASA, ATB 340)

  2 2

  adult vs old rat GM.

  
  

  Materials and Methods

  All experiments were carried out on adult and old rats (N=67, body weight 350±40g; n=5-

  6) in accordance with the norms of the Eu- ropean Convention for the Protection of Ver- tebrate Animals Used for Experimental and Other Scientific Purposes (1986) and in ac- cordance with the Committee on Bioethics of

  Danylo Halytsky Lviv National Medical Univer- sity (protocol № 6 from 29.03.2017). Animals were maintained under a constant 12 h light and dark cycle and an ambient temperature of 21-230C with 50±10% relative humidity. All animals were kept in raised mesh-bottom cages to prevent coprophagy. Animal from the control group were allowed free access to tap water and kept on standard diet (SD), exper- imental groups were on 28-days hypercaloric high-fructose diet (HFD by V. Kozar, 2008), with unrestricted access to 40% solution of fructose ad libitum within drinking-water [8].

  
  

  Acute stress was induced by model of Taka- gi et al. that involves short-term exposure to water-immersion stress [20]. The rats were placed in restraint cages and immersed ver- tically to the level of the xiphoid process in a water bath of 23˚ for 3.5 h. The initial and final body weights were recorded. Blood glu- cose concentrations were measured daily by glucometer (Achtung TD-4207, Germany) us- ing a blood sample from the tail vein.

  
  

  The animals were subdivided into control groups of adult rats (AR) and old rats (OR) with standard diet (SD) and vehicle (1.0 ml of physiological solution), and experimen- tal groups receiving 28-days hypercaloric high-fructose diet, HFD V. Kozar, 2008) with- out and with acute stress (Takagi, 1964). Ex- perimental groups were pre-treated 9-days per os by: 1) vehicle (adult); 2) vehicle (old);

  3) NaHS, 100 μmol/kg/day (adult); 4) NaHS, 100 μmol/kg/day (old); 5) vehicle with stress induction (adult); 6) vehicle with stress in- duction (old); 7) NaHS, 100 μmol/kg/day and induction of stress; 8) conventional aspirin (ASA), 10 mg/kg/day and induction of stress;

  9) ASA, 10 mg/kg/day and NaHS, 100 μmol/ kg/day and stress induction; 10) ATB-340

  -releasing, 17.5 mg/kg/day and induction of stress. The administration of NaHS and ATB-

  340 was performed in doses tested by J.L. Wallace, 2012-2017 [21, 22, 24].

  
  

  The rat stomach was removed immediately after euthanasia and cut with scissors in the longitudinal direction from the gastroesopha- geal junction to the pylorus. The gastric mu- cosal surface was gently washed with phos- phate-buffered (pH 7.4) saline. Samples of the GM were fixed in 10% formalin and embedded

  
  

  

  Оригінальні дослідження: фундаментальні науки Original research: basic sciences

  
  

  in parrafin. Serial sections of 5-mm thickness were stained with hematoxylin/eosin. The sec- tions were blindly evaluated by two individu- als and their scores were averaged. The GM damage was estimated by histological scor- ing (HS), including three components: a). the state of the epithelial plate on the degree of alteration (0 – no changes; 1 – stratification of the stratum corneum; 2 – focal basophilia of keratin masses; 3 – cell mass desquamation, vacuolation of basal cells, vesicular nuclei; 4 – erosion); b). Condition of subepithelial struc- tures according to stromal damage (0 – no changes; 1 – diffuse swelling of the submuco- sal basis; 2 – pronounced uneven swelling of the submucosal base and insignificant infiltra- tion; 3 – severe swelling and disorganization of the submucosal basis; 4 – perivascular or subepithelial infiltrates, perivascular hemor- rhages), c). Leukocyte infiltration of the ep- ithelial layer (0 – no leukocyte infiltration; 1

• moderate leukocyte infiltration; 2 – average leukocyte infiltration; 3 – severe leukocyte in- filtration; 4 – multiple leukocyte infiltrates).

Visualization was performed using a microscope (Swift Instruments International, Japan) and a digital video camera (Echoo-Imager 5020200 Microscope Digital Eyepiece, China).

The activity of SO was determined in GM homogenates biochemically by the rate of oxidation of sulfate anion in the presence of potassium hexacyanoferrate by standart biochemical methods. GM was homogenized at 3000 rpm (Teflon-glass) in a medium of 1.15% potassium chloride (ratio 1:3). The homogenates were centrifuged for 30 min. at 600g, the aliquots of the post-native supernatant were taken in the Eppendorf microtubules and stored at -20°C until the research was carried out [23].

2

The intensity of the oxidative component of oxidative stress was evaluated by changes in the rate of generation of an unstable free radical of oxygen – superoxide anion (O –), hydroxyl radical (OH–) and by changes in the content of terminal products of lipid peroxidation (malonic dialdehyde – MDA).

The level of MDA was determined by reaction with thiobarbituric acid (TBA) by the diagnostic set TBK-Agate (Biokont, RF). With a thiobarbituric acid the lipid peroxidation

products form a red stained complex, which is extracted with butanol, with a maximum light absorption at λ = 535 nm.

Statistical analysis of the results was carried out using programs «Excel» і «Statistica 7.0». The certainty of the changes was estimated by ANOVA Dunnett’s test.

The relative difference between the physiological and biochemical parameters against control was calculated by formula:

∆( %) = 100 • (Х - Х ) / Х ,

і n n

і

where Х – value in the main group;

Х

n

– value in the control group.

Results and discussion

Healthy adult rats on SD treated with vehicle exhibited normal gastric mucosa appearance, with histological scores of zero. The GM histological scores in old rats ranged between 1 and 2 (on a 0 to 12 scale) in HS. There was no detectable mucosal damage by HS of GM in adult rats under HFD vs old rats, however combination of WIS and HFD exposure caused sharply rise of HS (Fig. 1). The effects

2

on the GM in old rats were with irregular subepithelial exudates, submucosal vascular dilation and mild leukocyte infiltration in the gastric epithelium and submucosal clots (Fig. 2A). Influence of treatment with NaHS and ATB 340 (donors of H S synthesis) resulted in attenuated gastric injury induced by HFD, acute stress and ASA (Fig. 2B-2D).

We have observed a different expression of SO activity in GM in adult vs old rats with SD and

Оригінальні дослідження: фундаментальні науки Original research: basic sciences

Fig.2. Administration of donor of hydrogen sulfide (H2S) synthesis, NaHS (A, B, C) and ATB 340 (D) attenuates gastric injury/inflammation in HSD rats under stress induction and aspirin treatment.

HFD without and with WIS. In control groups that consumed SD, which we considered relative physiological norm, the SO activity in adult rats was 2,9 nmol/min·mg protein, in the old – 4,1 nmol/min·mg protein, the effect of HFD caused an increase in SO activity in old animals by 15%, and in adult by 14%, as compared to control (Fig. 3).

2

2

2

Changes in MDA content had the same tendency. The use of exogenous NaHS, which increases the H S content in the tissues of the body, reduced the activity of SO by 24% of adult rats, whereas in old rats it decreased by 23% compared to the groups without correction of the synthesis of H S. The obtained data allows us to interpret the effect of H S, on the background of HFD, as anti-oxidant and anti- radical. Investigation of changes of MDA levels in all experimental groups are represented on Fig. 4.

2

Regarding the importance to test age- related activity of H S enzyme SO against GM injury caused by long-term high fructose diet (HFD), we combined exposure to HFD with WIS. Stress induction showed age- related changes in the activity of SO. Older animals showed an increase in SO activity compared to adults by 18%, indicating better

Оригінальні дослідження: фундаментальні науки Original research: basic sciences

2

27%. H S-ASA shows a potent antioxidant effect, which attenuates oxidative stress. Similar results were shown in Liu L, et al study [10]. The relative difference between experimental groups and control data are represented on Tab. 1.

Conclusions. Long-term postprandial hyper- glycemia induced by HFD is ones agent that decline mucosal defense in stomach. Sulfite oxidase is the essential contributor to age-re- lated oxidative stress in GM during exposure of HFD. Supplementation of NaHS and ATB-

340 (H S-aspirin), which increases the H S

2 2

adaptive-compensatory properties of young

2

rats, while NaHS application increased SO activity by 7% and reduced MDA content by 16%. Distortion of the natural stereotype of the cytoprotective response of GM by the administration of ASA decreased the activity of SO by 20% and increased the content of MDA by 8%, while the combination of ASA and NaHS increased the activity of SO by 18% and reduced the MDA content by 7% compared to the groups without correction of H S synthesis. Our obtained results correlates with other scientific groups [2, 4].

2

2

To test effects of novel hybrid NSAIDs on changes of pro- and antioxidant balance and its impact on GI injury during aging which are still unknown, we used 7-days supplementation of conventional and H S- releasing NSAIDs during 21-28 days of HSD. Administration of the newest ATB 340, the H S-ASA hybrid drug, showed similar SO activity to the administration of ASA and NaHS, while the MDA content decreased by

content in the tissues, reduces the symptoms

of GM damage associated with a decrease in the activity of SO and the content of MDA, indicating the alleviation of oxidative stress, showing an anti-radical and anti-oxidant ef- fects.

Disclosures. No conflicts of interest, financial or otherwise, are declared by the authors.

Author contribution. Yaroslav Pavlovskiy, Maksym Lutsyk drafted manuscript; Anton- ina Yashchenko edited and revised manu- script; Natalia Zaichko, John Wallace, Oksana Zayachkivska approved final version of the manuscript.

Оригінальні дослідження: фундаментальні науки Original research: basic sciences

Reference

1. Alfonso L, Ai G, Spitale RC, Bhat GJ. Molecular targets of aspirin and cancer prevention. British Journal of Cancer. 2014;111(1):61–7.

2. Burn J, Sheth H. The role of aspirin in preventing colorectal cancer. British Medical Bulletin.

   2016;119(1):17–24.

3. Cardoso GM, Pletsch JT, Parmeggiani B, Grings M, Glanzel NM, Bobermin LD, Amaral AU, Wajner M, Leipnitz G. Bioenergetics dysfunction, mitochondrial permeability transition pore opening and lipid per- oxidation induced by hydrogen sulfide as relevant pathomechanisms underlying the neurological dys- function characteristic of ethylmalonic encephalopathy. BiochimicaetBiophysicaActa (BBA)-Molecular Basis of Disease. 2017;1;1863(9):2192-201.

4. Emilsson L, Holme Ø, Bretthauer M, Cook NR, Buring JE, Løberg M, et al. Systematic review with me- ta-analysis: the comparative effectiveness of aspirin vs. screening for colorectal cancer prevention. Alimentary Pharmacology & Therapeutics. 2016;45(2):193–204.

5. Fiorucci S, Santucci L, Distrutti E. NSAIDs, coxibs, CINOD and H2S-releasing NSAIDs: what lies beyond the horizon? Digestive and Liver Disease. 2007;1;39(12):1043-51.

6. Giustarini D, Del Soldato P, Sparatore A, Rossi R. Modulation of thiol homeostasis induced by H2S-re- leasing aspirin. Free Radical Biology and Medicine. 2010;1;48(9):1263-72.

7. Kodadek LM, Jones C. Stress Gastritis and Stress Ulcers: Prevention and Treatment. Surgical Critical Care Therapy. 2018;:231–9.

8. Kozar VV, Kudria MY, Ustenko NV, Pavlenko TO, Zhurakovska MV. The state of the humoral component of immunity under conditions of metabolic syndrome with underlying hypoestrogenia and its farmaco- logical correction. Buk Med Herald. 2009;13:141-4.

9. Laine L, Takeuchi K, Tarnawski A. Gastric mucosal defense and cytoprotection: bench to bedside. Gas- troenterology. 2008;1;135(1):41-60.

10. Liu L, Cui J, Song CJ, Bian JS, Sparatore A, Del Soldato P, Wang XY, Yan CD. H2S-releasing aspirin protects against aspirin-induced gastric injury via reducing oxidative stress. PLoS One. 2012;28;7(9):e46301.

11. Nagy L, Nagata M, Szabo S. Protein and non-protein sulfhydryls and disulfides in gastric mucosa and liver after gastrotoxic chemicals and sucralfate: possible new targets of pharmacologic agents. World Journal of Gastroenterology: WJG. 2007;14;13(14):2053-60.

12. Ralph SJ, Nozuhur S, Moreno-Sánchez R, Rodríguez-EnríquezS, Pritchard R. NSAID celecoxib: a potent mitochondrial pro-oxidant cytotoxic agent sensitizing metastatic cancers and cancer stem cells to che- motherapy. Journal of Cancer Metastasis and Treatment. 2018;4(9):49.

13. Sparatore A, Perrino E, Tazzari V, Giustarini D, Rossi R, Rossoni G, Erdman K, Schröder H, Del Sol- dato P. Pharmacological profile of a novel H2S-releasing aspirin. Free Radical Biology and Medicine. 2009;1;46(5):586-92.

14. Spinelli JB, Haigis MC. The multifaceted contributions of mitochondria to cellular metabolism. Nature cell biology. 2018 Jun 27:1.

15. Strutynska NA, Kotsiuruba AV, Budko AY, Mys LA, Sagach VF. Age-Associated Mitochondrial Dysfunction in the Heart Accompanied by Constitutive NO-Synthases Uncoupling on the Background of Oxidative and Nitrosative Stress. International Journal of Physiology and Pathophysiology. 2017;8(2):141–50.

16. Strutynska NA, Semenykhina OM, Chorna SV, Vavilova GL, Sagach VF. Hydrogen sulfide inhibits Ca2+- induced mitochondrial permeability transition pore opening in adult and old rat heart. Fiziolohichny- izhurnal. 2011; 57(6): 3-14.

17. Sulaieva OM, Uollas DL. Efekty hazopodibnykh mediatoriv: perspektyvy hastrointestynalnoi protektsii za umov vykorystannia protyzapalnykh preparativ. Suchasna hastroenterolohiia. 2016(3):114-9.

18. Szabo S, Trier JS, Frankel PW. Sulfhydryl compounds may mediate gastric cytoprotection. Science.

    1981;9;214(4517):200-2.

19. Takagi K, Okabe S. The Effects Of Drugs On The Production And Recovery Processes Of The Stress Ulcer.

    The Japanese Journal of Pharmacology. 1968;18(1):9–18.

20. Tarnawski AS. Cellular and molecular mechanisms of gastrointestinal ulcer healing. Digestive diseases and sciences. 2005;1;50(1):S24-33.

21. Wallace J, Pshyk-titko I, Muscara MN, Bula N, Pavlovsky Y, Gavriluk E, Zayachkivska O. Influence of Hydrogen Sulfide-releasing aspirin on mucosal integrity of esophageal and gastric mucosa. Pratsi nau- kovoho tovarystva im. Shevchenka. Medychni nauky. Likarskyi zbirnyk.2015(43, 27):63-74.

22. Wallace JL, Vaughan D, Dicay M, Macnaughton WK, Nucci GD. Hydrogen Sulfide-Releasing Therapeutics: Translation to the Clinic. Antioxidants & Redox Signaling. 2018;28(16):1533–40.

23. Zaichko NV, Mel’nyk AV, Iurchenko PO, Hryhor HS, Konakhovych NF. Influence of polymicroelement preparation esmin on hydrogen sulfide levels and indices of pro-and antioxidant system in the rat myo- cardium of different age. Ukrainian biochemical journal. 2014;86(3):69-76.

    
    

    

    Оригінальні дослідження: фундаментальні науки Original research: basic sciences

    
    

24. Zayachkivska O, Bula N, PavlovskyiYa, Pshyk-Titko I, Havryliuk O. Tsytoprotektorni efekty hidrohensul- fid-sporidnenoi atsetylsalitsylovoi kysloty na slyzovu obolonku stravokhodu (doklinichni doslidzhennia). Suchasna Gastroenterolohiia. 2017;1(93):15-21.

Стаття надійшла 12.11.2018

Після допрацювання 14.12.2018

Прийнята до друку 27.12.2018