Integrated Autofluorescence Characterization of a Modified-Diet Liver Model with Accumulation of Lipids and Oxidative Stress
Figure 2
Curve fitting analysis: estimation of the relative contribution of the different endogenous fluorophores to the whole liver AF emission collected in vivo (a, c) and ex vivo (b, d) in control and MCD diet administered rats; NAD(P) and redox ratios calculated from in vivo (e) and ex vivo (f) data. The results are presented as mean values ± S.D. (in vivo analysis: 5 measurements per rat; ex vivo analysis: 5 measurements, per 3 liver tissue sections, per rat). Statistical analysis: preliminary inspection showed that all data within each group were normally distributed, except for NAD(P)Hbound, in vivo data. In vivo significant differences: panel a: proteins, among control and all MCD diet week groups, except for MCD diet 3, 4 versus 8/9 weeks; fatty acids, control versus MCD diet 1, 2, and 8/9 weeks, MCD diet 1, 2 versus 3, 4 weeks, and MCD diet 3, 4 versus 8/9 weeks; vitamin A, among control and all MCD diet week groups, except for MCD diet 1 versus 2 weeks, and MCD diet 4 versus 8/9 weeks; NAD(P)Hbound, MCD diet 1 versus 3, 4, and 8/9 weeks; NAD(P)Hfree, control versus MCD diet 1–4 weeks. Panel c: lipopigments control and MCD diet 1–3 versus 4, 8/9 weeks; flavins, control versus MCD diet 1, 8/9 weeks, MCD diet 1 versus 2–4 weeks, and MCD diet 2–4 versus 8/9 weeks. Panel e: NAD(P) ratio, control and MCD diet 1, 2 versus 3-4 weeks, MCD diet 3, 4 versus 8/9 weeks; redox ratio, control and MCD diet 2–4 versus 1, 8/9 weeks. Ex vivo significant differences: panel b: proteins, control versus all MCD diet week groups, MCD diet 1 versus 2–8/9 weeks, and MCD diet 3 versus 2–8/9 weeks; fatty acids, control versus all MCD diet week groups and MCD diet 3 versus 1, 2, 4, and 8/9 weeks; vitamin A, among control and MCD diet 1–3, 8/9 weeks and MCD 4 versus 1–3, 8/9 weeks; NAD(P)Hbound, control versus all MCD diet week groups; NAD(P)Hfree, control versus MCD diet 2–8/9 weeks. Panel d: lipopigments control and MCD diet 1–3 versus 4,8/9 weeks; flavins, control and MCD diet 1, 2, 4, and 8/9 versus 3 weeks. Panel f: NAD(P) ratio, control versus all MCD diet week groups and MCD diet 1 versus 2, 8/9 weeks; redox ratio, control versus MCD diet 1–3, 8/9 weeks and MCD diet 1,2,4 versus 3,8/9 weeks. Before NAD(P) and redox ratio calculation, the values of NAD(P)H in the bound form were corrected for the AF emission yield (3 : 1 NAD(P)Hbound versus NAD(P)Hfree) [12]. Redox ratios were calculated as (flavins)/(NAD(P)Htotal + flavins) [42].
