Pharmaceutical composition of 1,2,4-trioxolanes with betulin in fish oil. Effect on oxidative and energy metabolism in the treatment of burns in rats

Background. Ozonation products of unsaturated fatty acids in fats and oils – 1,2,4-trioxolanes – which contribute to the normalization of oxidative and energy metabolism in various diseases, may be a promising substance for the creation of new combined anti-burn drugs.

The aim. To study the effect of 1,2,4-trioxolanes in a pharmaceutical composition with betulin, which exhibits wound-healing properties, on indicators of oxidative stress and on energy metabolism in the treatment of burns in an experiment onrats. Methods. The experiments were carried out on 50 male Wistar rats with seconddegree skin burns. We assessed the activity of glutathione reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, aldehyde dehydrogenase, superoxide dismutase and catalase, as well as the level of lipid peroxidation indicators.

Results. The properties of 1,2,4-trioxolanes were studied and a pharmaceutical composition of 1,2,4-trioxolanes with betulin from birch bark in fish oil was developed. The effect of this composition on oxidative and energy metabolism in rats during the treatment of burns was studied. The activation of NADP/H and NAD/H dependent enzymes (glutathione reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, aldehyde dehydrogenase), as well as superoxide dismutase and catalase, and the normalization of lipid peroxidation parameters under the influence of the studied composition in the treatment of burns were revealed. The effect of the composition on the energy metabolism of erythrocytes was noted, which was assessed by changes in the ratio of lactate dehydrogenase activity in the reverse and direct reactions. Morphohistological studies showed that using the composition under normal conditions and in case of a burn had no effect on the structure of the stomach wall, and also improved the condition of the burn wound on the day 10 compared to the wound condition in animals without treatment.

Conclusion. The proposed composition of 1,2,4-trioxolanes and betulin in fish oil, which allows normalizing oxidative and energy metabolism and improving the condition of a burn wound without toxic effects on the stomach wall, can be used as an element of additional therapy in the treatment of burns.

1. Porter C, Tompkins RG, Finnerty CC, Sidossis LS, Suman OE, Herndon DN. The metabolic stress response to burn trauma: Current understanding and therapies. Lancet. 2016; 388(10052): 1417-1426. doi: 10.1016/S0140-6736(16)31469-6

2. Saraf MK, Herndon DN, Porter C, Toliver-Kinsky T, Radhakrishnan R, Chao T, et al. Morphological changes in subcutaneous white adipose tissue after severe burn injury. J Burn Care Res. 2016; 37(2): 96-103. doi: 10.1097/BCR.0000000000000292

3. Vassalle C, Maltinti M, Sabatino L. Targeting oxidative stress for disease prevention and therapy: Where do we stand, and where do we go from here. Molecules. 2020; 25(11): 2653. doi: 10.3390/molecules25112653

4. Ugazio E, Tullio V, Binello A, Tagliapietra S, Dosio F. Ozonated oils as antimicrobial systems in topical applications. Their characterization, current applications, and advances in improved delivery techniques. Molecules. 2020; 25(2): 334. doi: 10.3390/molecules25020334

5. Carata E, Tenuzzo BA, Dini L. Powerful properties of ozonated extra virgin olive oil. Herbal Medicine. 2019. doi: 10.5772/intechopen.69412

6. De Almeida NR, Beatriz A, de Arruda EJ, de Lima DP, de Oliveira LCS, Micheletti AC. Ozonized vegetable oils: Production, chemical characterization and therapeutic potential. Vegetable Oil: Properties, Uses and Benefits. Nova Science Publishers, Hauppauge; 2016.

7. Menéndez S, Falcón L, Maqueira Y. Therapeutic efficacy of topical OLEOZON® in patients suffering from onychomycosis. Mycoses. 2011; 54: e272-e277. doi: 10.1111/j.14390507.2010.01898.x

8. Valacchi G, Zanardi I, Lim Y, Belmonte G, Miracco C, Sticozzi C, et al. Ozonated oils as functional dermatological matrices: Effects on the wound healing process using SKH1 mice. Int J Pharm. 2013; 458: 65-73. doi: 10.1016/j.ijpharm.2013.09.039

9. Zamora Rodríguez ZB, González Alvarez R, Guanche D, Merino N, Hernández Rosales F, Menéndez Cepero S, et al. Antioxidant mechanism is involved in the gastroprotective effects of ozonized sunflower oil in ethanol-induced ulcers in rats. Mediators Inflamm. 2007; 2007: 65873. doi: 10.1155/2007/65873

10. Zamora Z, González R, Guanche D, Merino N, Menéndez S, Hernández F, et al. Ozonized sunflower oil reduces oxidative damage induced by indomethacin in rat gastric mucosa. Inflamm Res. 2008; 57(1): 39-43. doi: 10.1007/s00011-007-7034-1

11. Kiselevich VE, Bessonova NE, Boyarinov GA, Sokolov SA, Peretyagin SP. Complex of ozonated unsaturated fatty acids and its application: Patent No. 2725980 of the Russian Federation. 2020; (19). (In Russ.).

12. Scheffler A. The wound healing properties of betulin from birch bark from bench to bedside. Planta Med. 2019; 85(07): 524-527. doi: 10.1055/a-0850-0224

13. Manzhulo IV, Tyrtyshnaia AA, Manzhulo OS, Starinets AA, Kasyanov SP, Dyuizen IV. Neuroprotective activity of docosahexaenoic acid in the central and peripheral nervous system after damage to the sciatic nerve. Neurochem J. 2020; 14(1): 101-107. doi: 10.1134/S1819712420010158

14. Ghasemi Fard S, Wang F, Sinclair AJ, Elliott G, Turchini GM. How does high DHA fish oil affect health? A systematic review of evidence. Crit Rev Food Sci Nutr. 2019; 59(11): 1684-1727. doi: 10.1080/10408398.2018.1425978

15. Sega A, Zanardi I, Chiasserini L, Gabbrielli A, Bocci V, Travagli V. Properties of sesame oil by detailed 1H and 13C NMR assignments before and after ozonation and their correlation with iodine value, peroxide value, and viscosity measurements. Chem Phys Lipids. 2010; 163(2): 148-156. doi: 10.1016/j.chemphyslip.2009.10.010

16. Zanardi I, Travagli V, Gabbrielli A, Chiasserini L, Bocci V. Physico-chemical characterization of sesame oil derivatives. Lipids. 2008; 43(9): 877-886. doi: 10.1007/s11745-008-3218-x

17. Khabriev RU. Guidelines for the experimental (preclinical) study of new pharmacological substances; 2nd edition, revised and enlarged. Moscow: Meditsina; 2005. (In Russ.).

18. Lampatov VV, Semenikhina NM, Khalimov RI. Method for simulating thermal skin damage of IIIA degree: Patent No. 2712051 of the Russian Federation. 2020; (3). (In Russ.).

19. Peretyagin PV, Solovieva AG, Luzan AS, Vorobyov EV, Didenko NV. Device for experimental modeling of thermal skin injury: Description of the utility model for the Patent No. 179126 of the Russian Federation. 2018; (12). (In Russ.).

20. Martusevich AK, Larionova KD, Peretyagin SP, Peretyagin PV, Davyduk AV. Experimental estimation of pharmacological compositions effect on microcirculation state at early postburn period. Fundam Res. 2013; 3: 332-336.

21. Dahmus JD, Bruning RS, Kenney WL, Alexander LM. Oral clopidogrel improves cutaneuos microvascular function through EDHF-dependent mechanisms in middle-aged humans. Am J Physiol Regul Integr Comp Physiol. 2013; 305: R452-R458. doi: 10.1152/ajpregu.00366.2012

22. Sies H, Berndt C, Jones DP. Oxidative stress. Annu Rev Biochem. 2017; 86: 715-748. doi: 10.1146/annurev-biochem-061516-045037

23. Seyam O, Smith NL, Reid I, Gandhi J, Jiang W, Khan SA. Clinical utility of ozone therapy for musculoskeletal disorders. Med Gas Res. 2018; 8(3): 103-110. doi: 10.4103/2045-9912.241075

24. Re L, Mawsouf MN, Menendez S, Leon OS, Sanchez GM, Hernandez F. Ozone therapy: Clinical and basic evidence of its ther-apeutic potential. Arch Med Res. 2008; 39: 17-26. doi: 10.1016/j.arcmed.2007.07.005

25. Mori TA, Burke V, Puddey IB, Watts GF, O’Neal DN, Best JD, et al. Purified eicosapentaenoic and docosahex-aenoic acids have differential effects on serum lipids and lipoproteins, LDL particle size, glucose, and insulin in mildly hy-perlipidemic men. Am J Clin Nutr. 2000; 71(5): 1085-1094. doi: 10.1093/ajcn/71.5.1085

26. Bazan NG. Omega-3 fatty acids, pro-inflammatory signaling and neuroprotection. Curr Opin Clin Nutr Metab Care. 2007; 10: 136-141. doi: 10.1097/MCO.0b013e32802b7030

27. Nakamoto K, Nishinaka T, Mankura M, Fujita-Hamabe W, Tokuyama S. Antinociceptive effects of docosahexaenoic acid against various pain stimuli in mice. Biol Pharm Bull. 2010; 33: 1070-1072. doi: 10.1248/bpb.33.1070