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ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)

Fiziologichnyi Zhurnal

is a scientific journal issued by the

Bogomoletz Institute of Physiology
National Academy of Sciences of Ukraine

Editor-in-chief: V.F. Sagach

The journal was founded in 1955 as
1955 – 1977 "Fiziolohichnyi zhurnal" (ISSN 0015 – 3311)
1978 – 1993 "Fiziologicheskii zhurnal" (ISSN 0201 – 8489)
1994 – 2016 "Fiziolohichnyi zhurnal" (ISSN 0201 – 8489)
2017 – "Fiziolohichnyi zhurnal" (ISSN 2522-9028)

Fiziol. Zh. 2016; 62(5): 27-36


Н.V. Danylovych1, Yu.V. Danylovych1, R.V. Rodik2, V.I. Kalchenko2, A.Ju. Chunikhin1

  1. Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
  2. Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;


The influence of supramolecular macrocyclic compounds сalix[4]arenes (C-97, C-99, C-107) at a concentration of 100 nM in the process of energy-dependent Ca2+-transport in isolated mitochondria of smooth muscle, as well as autofluorescence mitochondrial coenzyme NADH, FAD and hydrodynamic diameter of these organelles was investigated. Using Ca2+-sensitive fluorescent dye Fluo-4 AM it was shown that the selected calix[4]arenes can suppress energy-dependent accumulation of Ca2+ by mitochondria. Accumulation of Ca2+ (80 µM in the medium) accompanied by the growth of the fluorescent probe response from a conventional unit to a value of 1,57±0,04 (n=5). Calix[4]arenes C-97, C-99, C-107 falls fluorescent signal below the 0,88±0,08, 0,92±0,08 and 0,78±0,04 respectively. Thus, the selected calix[4]arenas lead to release of previously accumulated Ca2+ from mitochondria. Under the influence of C-97 and C-99 fluorescent signal from NADH reduced to -0,11±0,02 and -0,12±0,02, respectively, in relation to the reference value - -0,05±0,01 (n=5). Analysis of fluorescence response NADH and FAD in a suspension of isolated mitochondria suggests that the effects of test compounds on the functional activity of the electron transport chain is associated with the initial stimulation of its I-th complex and subsequent inhibition of Ca2+-dependent NADcontaining Krebs cycle dehydrogenases. Along with this, the use of photon correlation spectroscopy to assess changes in the volume of mitochondria (their hydrodynamic diameter) under the action of selected calix[4]arenes has shown that interference with the electron transport chain leads to changes in the osmotic balance between the matrix of the mitochondria and the external environment. The result is the growth of isolated organelles volume. In particular, the hydrodynamic diameter of mitochondria increased by 22±6 % and 34±8 % (n=5) in presence of C-97 or C-99. The conclusion was done about the advisability of further studies of the calyx[4]arenes effect on smooth muscle Ca2+-homeostase and mitochondrial bioenergetics in order to find effective modifiers of their functional activity.

Keywords: mitochondria; electron transport chain; Ca2+- transport; smooth muscle.


  1. Kostyuk PG, Kostyuk OP, Lukyanets EA. Intracellular calcium signaling: structures and functions. Kiev: Naukova dumka, 2010. [Ukrainian].
  3. Santo-Domingo J, Demaurex N Calcium uptake mechanisms of mitochondria. Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):907-12
  5. Pan S1, Ryu SY, Sheu SS. Distinctive characteristics and functions of multiple mitochondrial Ca2+ influx mechanisms. Sci China Life Sci. 2011 Aug;54(8):763-9
  7. Wang HW, Wei YH, Guo HW. Reduced nicotinamide adenine dinucleotide (NADH) fluorescence for detection of cell death. Anticancer Agent Med Chem. 2009; 9(9):1012-7 CrossRef  
  8. Heikal AA. Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies. Biomark Med. 2010;4(2):241-63. CrossRef PubMed PubMedCentral
  10. Gellerich FN, Gizatullina Z, Trumbeckaite S, Nguyen HP, Pallas T, Arandarcikaite O, Vielhaber S, Seppet E, Striggow F. The regulation of OXPHOS by extramitochondrial calcium. Biochim Biophys Acta. 2010; 1797(6-7):1018-27.
  12. Giuliani M, Morbioli I, Sansone F, Casnati A moulding calixarenes for biomacromolecule targeting. Chem. Commun., 2015, DOI: 10.1039/C5CC05204A. CrossRef  
  13. Danylovych HV, Danylovych IuV, Kolomiiets' OV, Kosterin SO, Rodik RV, Cherenok SO, Kal'chenko VI, Chunikhin OIu, Horchev VF, Karakhim SO. Changes in polarization of myometrial cells plasma and internal mitochondrial membranes under calixarenes action as inhibitors of plasma membrane Na+, K+-ATPase. Ukr Biokhim Zh (1999). 2012 Nov-Dec;84(6):37-48. [Ukrainian].
  15. Kosterin SA, Bratkova NF, Rursky MD. The role of sarcolemma and mitochondria in Ca2+-dependent control of myometrium relaxation. Biochemistry (Moscow). 1985; 50(8):1350-1361. [Russian].
  17. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. CrossRef  
  18. Kolomiets' OV, Danylovych IuV, Danylovych HV, Kosterin SO. Ca2+ accumulation study in isolated smooth muscle mitochondria using Fluo-4 AM. Ukr Biochem J. 2013 Jul-Aug;85(4):30-9 [Ukrainian].
  20. Bailay NTJ. Statistical methods in biology. Great Britain: Cambridge University Press; 1995.
  22. Malli R, Graier WF. Mitochondrial Ca2+ channels: Great unknowns with important functions.FEBS Lett. 2010; 584:1942-47. CrossRef PubMed PubMedCentral
  24. Wei AC, Liu T, Cortassa S, Winslow RL, O'Rourke B. Mitochondrial Ca2+ influx and efflux rates in guinea pig cardiac mitochondria: low and high affinity effects of cyclosporine A. Biochim Biophys Acta. 2011;1813:1373-81. CrossRef PubMed PubMedCentral
  26. Kolomiets' OV, Danylovych IuV, Danylovych GV. H+-Ca2+-exchanger in the myometrium mitochondria: modulation of exogenous and endogenous compounds. Fisiol. Zh. 2014;60(5):33-42. [Ukrainian].
  28. Veklich TO, Koshechkova NS, Rodik RV, Boĭko VI, Vorobets' ZD, Kosterin SO. Comparative study of calixarene effect on Na+, K+ -ATPase activity in plasma membrane of contractile and mobile cell. Ukr Biokhim Zh (1999). 2007;79(3):19-28. [Ukrainian].
  30. Agarwal B, Dash RK, Stowe DF, Bosnjak ZJ, Camara AK. Isoflurane modulates cardiac mitochondrial bioenergetics by selectively attenuating respiratory complexes. Biochim Biophys Acta. 2014; 1837(3): 354–365. CrossRef PubMed PubMedCentral

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