Patterns of changes in the fluorescence of nadh and fad coenzymes and their relationship in skeletal muscle in the early post-mortem period (an experimental study)

Abstract

Background.The paper presents changes and mathematical models of autofluorescence of reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) and their redox ratio (RR) in rat skeletal muscle during the first 24 hours after death.Aim.To establish a pattern of change in the fluorescence intensity of NADH and FAD coenzymes, as well as their relationship in skeletal muscle during the first 24 hours after death using mathematical modeling.Material and methods.The experiment was performed on Sprague Dawley rats. The fluorescence intensity of FAD and NADH coenzymes in skeletal muscle was measured in situ in living rats under general anesthesia, 5 minutes after euthanasia, and at intervals of 1.53 h during the 24-hour postmortem period. To estimate the fluorescence intensity of NADH and FAD coenzymes and to calculate the RR, a Lasma MC-3 fluorescence measuring device with special software was used. The obtained data were analyzed by non-linear regression analysis. The summary, accuracy estimation, and significance of the regression equation coefficients were assessed using SigmaPlot 10.0 software. The significance of the regression model was tested using the Fisher F-criterion.Results.During the first 3 hours of the postmortem period, an increase in the mean values of RR and NADH fluorescence was detected; starting from 4.5 and lasting until 24 hours post mortem, their gradual decrease was observed. The relationship between NADH, RR, and time since death is characterized by the Weibull equation. The statistical significance of the NADH and RR models, based on the obtained equations, was considered high.Conclusion.The analysis of the obtained data allowed us to create mathematical models describing the relationship between RR and NADH fluorescence intensity and time after death, which confirms the non-randomness and regularity of the discovered patterns.