The role of bafilomycin as a therapeutic agent in the modulation of endo-lysosomal store of rat hepatocytes

S.V.  Bychkova; A.R. Stasyshyn; M.A.  Bychkov

doi:10.26641/2307-0404.2022.3.265768

Автор(и)

S.V. Bychkova Ivan Franko National University of Lviv, Universytetska str., 1, Lviv, 79000, Україна https://orcid.org/0000-0002-5107-3352
A.R. Stasyshyn Danylo Halytsky Lviv National Medical University, Perarska str., 69, Lviv, 70010, Україна https://orcid.org/0000-0002-6168-494X
M.A. Bychkov Danylo Halytsky Lviv National Medical University, Perarska str., 69, Lviv, 70010, Україна https://orcid.org/0000-0001-6620-1751

DOI:

https://doi.org/10.26641/2307-0404.2022.3.265768

Ключові слова:

аutophаgу, hepatocytes, bаfіlomycіn A1, ATPases, NAAD, Ca2 -store

Анотація

Endo-lysosomal system through the process of autophagy is involved in the pathogenesis of many diseases. Acidification of these organelles is carried out by V-type H+-ATPases, which is inhibited by bafilomycin A1. Endosomes and lysosomes are also important Ca²⁺-storage in a cell. Nіcotіnіc acіd adenіne dіnucleotіde phosphate (NAADP) releases Cа²⁺ from endo-lysosomes. The main purpose of the study was to found out the effect of bafilomycin A1 and NAADP on stored Ca²⁺ and on the ATPase activity of rat hepatocytes. The stored Ca²⁺ was estimated using chlorotetracycline in permeabilized hepatocytes of rats. ATPase activity was determined by level of orthophosphate spectrophotometrically. It was found that bafilomycin A1 reduces stored Ca²⁺ in permeabilized hepatocytes of rats in the micromolar range of concentration (20 and 0.04 mkM) and averted the effect of NAADP on calcium content. Lower concentrations of bafilomycin A1 (0.001 mkM) did not alter the content of stored calcium, but prevented the influence of NAADP in permeabilized hepatocytes of rats. In the subcellular fraction of rat liver bafilomycin A1 (0.001 mkM) increased Ca²⁺-ATPase and basal Mg²⁺-ATPase activities and reduced Na⁺/K⁺-ATPase activity. Preincubation of the subcellular fraction with bafilomycin A1 completely averts any changes in the activity of estimated ATPases by means of NAADP. It was concluded that the bafilomycin-sensitive store in hepatocytes of rats is NAADP-sensitive endo-lysosomal Ca²⁺-store. Using of bafilomycin A1 may be useful in treating autophagy-depended diseases.

Посилання

Bychkova S. Influence of NAADP and bafilo-mycine A1 on activity of ATPase in liver postmito-chondrial fraction. Studia Biologica. 2015;9(3-4):31-40. doi: https://doi.org/10.30970/sbi.0903.445

Bychkova S, Chorna T. NAADP-sensitive Сa2+ stores in permeabilized rat hepatocytes. Ukr. Biochem. J. 2014;86(5):65-73. doi: https://doi.org/10.15407/ubj86.05.065

Bychkova S, Hreniuh V. Activity of ATPases in postmitoсhondrial fraction of lymphoma NK/Ly cells under bafilomicine and NAADP presence mitochondria. Studia Biologica. 2015;9(2):31-38. doi: https://doi.org/10.30970/sbi.0902.413

Fay MP, Proschan MA. Wilcoxon-Mann-Whitney or t-test? On assumptions for hypothesis tests and multiple interpretations of decision rules. Stat Surv. 2010;4:1-39. doi: https://doi.org/10.1214/09-SS051

Ferents IM, Bychkova SV, Bychkov MA. Pecu-liarities of the effects of bile acids on atpase activity of the colon mucosa in patients with overweight and irritable bowel syndrome. Wiadomosci lekarskie. 2020;73(3):574-7. doi: https://doi.org/10.36740/WLek202003133

Hreniukh V, Bychkova S, Kulachkovsky O, Bab-sky A. Effect of bafilomycin and NAADP on membrane associated ATPases and respiration of isolated mitochon-dria of the murine Nemeth-Kellner lymphoma. Cell bio-chemistry and function. 2016;34(8):579-87. Epub 2016 Nov 8. doi: https://doi.org/10.1002/cbf.3231

Mauvezin C, Neufeld TP. Bafilomycin A1 dis-ruptsautophagic flux by inhibiting both V-ATPase-depen-dent acidification and Ca-P60A/SERCA-dependent auto-phagosome-lysosome fusion. Autophagy. 2015;11(8):1437-8. doi: https://doi.org/10.1080/15548627.2015.1066957

Mindell, Joseph A. Lysosomal Acidification Me-chanisms. Annual Review of Physiology. 2012;74:69-86. doi: https://doi.org/10.1146/annurev-physiol-012110-142317

Zhang B, Watt JM, Cordiglieri C, et al. Small Mo-lecule antagonists of NAADP-induced Ca2+ release in T-Lymphocytes suggest potential therapeutic agents for autoimmune disease. Nature: Scientific reports: 2018;8:16775. doi: https://www.nature.com/articles/s41598-018-34917-3