SDS software v.2.4. The expression levels were evaluated using the comparative cycle 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64
Statistical analysisThe data are expressed as the means ± SD. The comparisons of two groups were made using Mann-Whitney's U-test. A value of P < 0.05 was considered statistically significant.
Results
ALT, AST, and histopathology of the liverAfter the administration of APAP and MP, the plasma ALT and AST levels were significantly elevated (Fig. 1A). The histopathological analysis revealed that APAP caused hepatocellular necrosis at the pericentral regions, whereas MP caused hepatocellular necrosis at the periportal region (Fig. 1B). These two models were used for the subsequent studies.
miRNA expression at the pericentral and periportal regions of the liverThe miRNA expression at pericentral and periportal regions of the liver was determined by TaqMan microRNA array analysis. The numbers of the detected miRNAs and the miRNAs whose expression exceeded the cutoff (Ct < 32) are shown in Table 1. No large difference was observed in these numbers between the pericentral and periportal regions within a group or between the treated and control groups. Among the miRNAs with Ct values less than 32 in all of the groups, 125 miRNAs were common to all of the groups; therefore, these 125 miRNAs were used for the subsequent analyses. Fig. 2 shows the heat maps of the expression of the 125 miRNAs in each sample; note that the miRNAs are ordered based on descending expression level in the pericentral region of the control rats. The top three miRNAs that 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 7 exhibited high expression in both the pericentral and periportal regions in all of the samples are miR-709, miR-122, and miR-720. The comparison of the miRNA levels between the control rats and fasted-control rats revealed that 36 miRNAs presented higher levels and eight miRNAs exhibited lower levels at the pericentral region of the fasted-control rats (Tables 1 and 2). At the periportal region, 52 miRNAs and two miRNAs presented higher and lower levels, respectively, in the fasted-control rats compared with the control rats. Among these miRNAs, 17 miRNAs, which are shown in bold, were found in both the pericentral and periportal regions. Thus, the results demonstrate that fasting affects the expression of some miRNAs in the liver. The comparison of the miRNA levels between the pericentral and periportal regions in control rats showed that 27 miRNAs exhibited higher expression levels in the pericentral region and that 22 miRNAs presented higher expression levels in the periportal region compared with t...