Biochemical and molecular biological characterization of rat's dermal fibroblast culture in hypoxia

Dermal fibroblast cell culture is a convenient model for studying various effects on cells, including hypoxia. Fibroblasts, in addition to collagen production, are able to synthesize biogenic amines, hormones, neuropeptides, and neurotransmitters identical to those in the central nervous and endocrine systems, which allows them to be used to study cellular disorders in various diseases. The purpose of the study was to evaluate the functional changes in the culture of dermal fibroblasts under hypoxic conditions using biochemical and molecular biological methods. In cells of fibroblast culture under normoxia (air and 5% CO2 in an incubator) and hypoxic exposure (a mixture of gases - 95% N2 and 5% CO2) at 1 and 3 hours, biochemical methods were used to determine the activity of aspartate aminotransferase (AsAT), alanine aminotransferase (AlAT), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), glucose levels, concentrations of ATP, ADP, AMP, as well as the level of mRNA expression of the hypoxia-induced factor Hif-1a and the nuclear factor Nf-κb by polymerase chain reaction. It was shown that under hypoxic exposure at 1 hour in fibroblast culture, cell viability, glucose levels and activity of LDH, AlAT, AsAT decrease, the amount of ATP decreases, after 3 hours there is a tendency to normalize all indicators. Adaptive mechanisms make it possible to normalize the functioning of cells under hypoxic conditions from 1 to 3 hours. The results obtained in the evaluation of metabolic changes at different times of hypoxic exposure (1 and 3 hours) in the culture of rat fibroblasts indicate a high adaptive capacity of connective tissue cells - fibroblasts at a lack of oxygen. The study of intracellular parameters during hypoxia, the determination of critical points depending on the time of exposure will determine the directions for further study of the mechanisms of cell adaptation, which, perhaps, will complement the tactics of compensatory effects in ischemia of tissues of various genesis. The revealed changes reflect the adaptive response of the fibroblast culture in response to hypoxic exposure.