UDC: 611.37.018.72:615.212.7].08

Use of narcotic drugs in clinical practice for the purpose of obtaining analgesic and anti-inflammatory effects requires a comprehensive morphological study of the peculiarities of structural arrangement of organs under the conditions of exposure to opioids. The aim of our study was to establish the peculiarities of restructuring of the structural components of the pancreas under the conditions of long-time exposure to opioids in the experiment. The study included 24 adult laboratory white male rats. The test animals were divided into 2 groups, the experimental and control ones. The experimental animals were daily administered narcotic analgesic nalbuphine intramuscularly (once a day in the same interval) for four weeks, and the control animals were administered saline solution. The following research methods were used: bloodstream injection followed by translucence of sections of the pancreas and their
photographing, morphometry of the vessels of the pancreatic hemomicrocirculatory bed, histological, histochemical studies and electron microscopy of the pancreas, blood biochemistry test; statistical processing of the study results using a software package. After four weeks of opioid exposure, lesion of the pancreatic parenchyma microstructure was observed, manifested by swelling and infiltration by lymphocytes and macrophages of the pancreatic connective tissue stroma, disorganization of the exo- and endocrine parts of the parenchyma, deep destructive changes in the excretory ducts, as well as in the vessels of the hemo- and lympho-microcirculatory bed of the pancreas. At the ultrastructural level, deep dystrophic changes of exo- and endocrinocytes of the pancreas were identified, in particular, loss of regular shape, karyopyknosis and karyorrhexis of the nuclei, swelling and clearing of cytoplasm, development of microcystic degeneration of cells, loosening and disorganization of the basement membrane, which can result in impairment of exocrine function of the pancreas and complication of the process of secretory granules excretion into the lumen of the intercalated ducts. A significant decrease, compared to the control group, in the diameter of arterioles, density of exchange vessels network, as well as increase in the diameter of venules, the indicator of trophic activity of the tissue, are the evidence of destructive changes in the hemomicrocirculatory bed of the pancreas under the effects of nalbuphine. Significant changes in blood biochemistry parameters (alanine aminotransferase, aspartate aminotransferase) after a four-week administration of nalbuphine are illustrative of the process of pancreatic tissue destruction. Therefore, four-week administration of opioid leads to profound changes in the micro- and ultrastructure of the pancreas, vessels of its hemomicrocirculatory bed, and blood biochemistry parameters in experimental white rats.
Keywords: pancreas, structural changes, opioid, experiment.

Colorectal cancer is influenced by genetic mutations, lifestyle factors, and diet, particularly high fat intake, which raises bile acid levels in the intestinal lumen. This study hypothesized that bile acids contribute to tumorigenesis by disrupting ion transport and ATPase activity in the intestinal mucosa. The effects of 3-sulfo-taurolithocholic acid (TLC–S) on ATPase activity were investigated in colorectal cancer samples from 10 patients, using adjacent healthy tissue as controls, and in rodent liver function. ATPase activity was measured spectrophotometrically by determining inorganic phosphorus (P_i) in postmitochondrial fractions. Ca²⁺ dynamics were assessed in isolated mouse hepatocytes with fluorescence imaging, and rat liver mitochondria were studied using polarographic methods to evaluate respiration and oxidative phosphorylation. TLC–S increased Na⁺/K⁺ ATPase activity by 1.5 times in colorectal cancer samples compared to controls (p ≤ 0.05). In healthy mucosa, TLC–S decreased Mg²⁺ ATPase activity by 3.6 times (p ≤ 0.05), while Mg²⁺ ATPase activity in cancer tissue remained unchanged. TLC–S had no significant effect on Ca²⁺ ATPase activity in healthy colon mucosa but showed a trend toward decreased activity in cancer tissue. In rat liver, TLC–S decreased Ca²⁺ ATPase and Na⁺/K⁺ ATPase activities while increasing basal Mg²⁺ ATPase activity (p ≤ 0.05). Additionally, TLC–S induced cytosolic Ca²⁺ signals in mouse hepatocytes, partially attenuated by NED-19, an NAADP antagonist (p ≤ 0.05). TLC–S also reduced the V3 respiration rate of isolated rat liver mitochondria during α-ketoglutarate oxidation. These findings suggest that TLC–S modulates ATPase activity differently in cancerous and healthy colon tissues, playing a role in colorectal cancer development. In rat liver, TLC–S affects mitochondrial activity and ATPase function, contributing to altered cytosolic calcium levels, providing insight into the mechanistic effects of bile acids on colorectal cancer and liver function.

Bafilomycin A1 inhibits V-type H+ ATPases on the molecular level, which acidifies endolysosomes. The main objective of the study was to assess the effect of bafilomycin A1 on Ca2+content, NAADP-induced Ca2+ release, and ATPase activity in rat hepatocytes and human colon
cancer samples. Chlortetracycline (CTC) was used for a quantitative measure of stored calcium in permeabilized rat hepatocytes. ATPase activity was determined by orthophosphate content released after ATP hydrolysis in subcellular post-mitochondrial fraction obtained from rat liver as well as from patients’ samples of colon mucosa and colorectal cancer samples. In rat hepatocytes, bafilomycin A1 decreased stored Ca2+ and prevented the effect of NAADP on stored Ca2+. This effect was dependent on EGTA–Ca2+ buffers in the medium. Bafilomycin A1 significantly increased the activity of Ca2+ATPases of endoplasmic reticulum (EPR), but not plasma membrane (PM) Ca2+ ATPases in rat liver.
Bafilomycin A1 also prevented the effect of NAADP on these pumps. In addition, bafilomycin A1 reduced Na+/K+ ATPase activity and increased basal Mg2+ ATPase activity in the subcellular fraction of rat liver. Concomitant administration of bafilomycin A1 and NAADP enhanced these effects. Bafilomycin A1 increased the activity of the Ca2+ ATPase of EPR in the subcellular fraction of normal human colon mucosa and also in colon cancer tissue samples. In contrast, it decreased Ca2+ ATPase PM activity in samples of normal human colon mucosa and caused no changes in colon cancer. Bafilomycin A1 decreased Na+/K+ ATPase activity and increased basal Mg2+ ATPase activity
in normal colon mucosa samples and in human colon cancer samples. It can be concluded that bafilomycin A1 targets NAADP-sensitive acidic Ca2+ stores, effectively modulates ATPase activity, and assumes the link between acidic stores and EPR. Bafilomycin A1 may be useful for cancer therapy.
Keywords: molecular mechanisms; colon cancer; ATPase; autophagy; hepatocytes; liver; NAADP; biomarkers; bafilomycin A1; Ca2+ store 

UDC 577.352:616-006.44:542.978

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.